Issue 1 - The University of Zakho

Transcription

Kurdistan Region - Iraq
Ministry of Higher Education
and Scientific Research
University of Zakho
ISSN: 2410-7549
Journal of
University of Zakho
A - Science
Volume 3
Number 1
June
2015
Journal of University of Zakho
A - Science
Volume 3
Number 1
June
2015
Journal of University of Zakho, Vol. 3(A), No.1, 2015
Contents
- THE ROLE OF K+
CHANNEL SUBTYPES IN CATECHIN INDUCED
VASORELAXATION IN RAT’S AORTIC RINGS
Omar A. M. Al-Habib and Fakhir Najim K. Sabir …………………………………………..…………1
- EFFECT OF RENAL STONE FORMATION ON SOME RENAL RELATED
PARAMETERS IN KOYA CITY
Layla Kareem Ali ……………………………………………………………………………………… 9
- TOXIC EFFECTS OF ESSENTIAL OILS OF ELATTARIA CARDAMOMUM L.
AND LAMBDA- CYHALOTHRIN ON TRIBOLIUM CONFUSUM (DUVAL)
Sofyan H. Sedo Al-Sinjari and Hani J. Al-Attar ……………………………………………………...15
- Trichodina sp. AS BIOINDICATOR FOR EVALUATION OF BIOCHEMICAL
OXYGEN DEMAND (BOD5) IN AQUACULTURE FISH FARMS (PONDS)
Karwan Sallo Najm Al-Marjan and Shamall Muhamad Amin Abdullah ..............................................27
- DIAGNOSIS OF TOXOPLASMOSIS IN SHEEP USING SEROLOGICAL (ELISA)
AND MOLECULAR TECHNIQUE IN DUHOK GOVERNORATE -KURDISTAN
REGION
Farhad Buzo Mikaeel, Jassim Abdo and Lokman Taib Omer ………………………………….……. 32
- BIOASSAY AND PATHOGENICITY OF WHEAT SEED GALL NEMATODE
ANGUINA TRITICI
Sulaiman Naif Ami, Ibrahim Esa Taher, Fathi Abdulkarem Omer…………………………………...39
- Characterization of Five Microsatellite Markers for Genetic Diversity Structure
Analysis of Walnut (Juglans regia L) in Five Village in Duhok Province
Awat Mustafa Abbas Barwary…………..…………...…………...…………...…………...…………..46
- IN VITRO MICROPROPAGATION OF VITIS VINIFERA L. IN KURDISTAN
REGION OF IRAQ
Aveen M.N. Ahmed, Diaa A. Ibrahim and Seyran Palabash Uzun…………………………………. 55
- USE OF WOOD MACERATED ELEMENTS FOR CLASSIFICATION OF SALIX L.
SPECIES IN KURDISTAN REGION- IRAQ
Saleem Esmael Shahbaz and Shamiran Salih Abdulrahman ………………………………………...67
- OCCURRENCE
OF TOMATO LEAF MINER
TUTA ABSOLUTA
MEYRICK(LEPIDOPTERA:GELECHIIDAE) IN DUHOK REGION (A)
Lazgeen Haji Assaf, Feyroz Ramadan Hassan, Halgurd Ismael Rasheed, Mohammad Abdulrahman
Sadiq, Rezgar M. Ismael, Mohammad Sh. Ahmed, Salah Adel Saeed and Farhad Mohammed Taher
Salih ………………………………………………………………………………………...…............79
Journal of University of Zakho, Vol. 3(A), No.1, 2015
- EFFECT OF FOLIAR SPRAY OF KNO3, HUMIC ACID CULTIVARS AND
THEIR INTERACTIONS ON LEAF NUTRIENTS OF OLIVE (OLEA
EUROPAEA L.) CVS. KHITHARY AND I18 TRANSPLANTS
Azad Ahmed Mayi and Gulala Mohamed Ameen Saeed ………………………………………......... 88
- EFFECT OF CULTIVARS, COMPOST, HUMIC ACID AND THEIR
INTERACTIONS ON LEAF NUTRITIONAL STATES OF SWEET CHERRY
(PRUNUS AVIUM L.)
Azad Ahmed Mayi and Nawzad Jemil Ibrahim ………………………………………..…………….. 97
- SOME TECHNOLOGICAL PROPERTIES OF SAWN BOARD Eucalyptus
camaldulensis Denh. GROWN IN ASKIKALAK
Kurdistan Hassan Yousif …………………..…………………………..………………………..… 323
HOMOGENEOUS PHOTOCATALYTIC DEGRADATION
ALIZARIN BLACK USING HYDROGEN PEROXIDE
-
OF
ACID
Mohammedamin Yasin Taha ………………………………………………………….……………. 107
- THE EFFECT OF COLOR PLASTIC MULCHES ON GROWTH, YIELD
AND QUALITY OF TWO HYBRIDS OF SUMMER SQUASH (CUCURBITA
PEPO L.)
Abduljebbar Ihsan saeid and Ghurbat Hassan Mohammed …………………….…………………... 113
- SYNTHESIS AND BIOLOGICAL EVALUATION OF SOME DIPEPTIDE
DERIVATIVES AND THEIR HETEROCYCLIC COMPOUNDS
Huda Ahmed Basheer, Sabir Ayob Mohammed and Aveen Akram Ibrahiem ………….…………..119
- ON THE ENERGY OF SOME COMPOSITE GRAPHS
Nechirvan Badal Ibrahim……………………………………………………………………………..132
CONVERGENCE OF THE BARZILAI-BORWEIN METHOD
SOLVING SLIGHTLY UNSYMMETRIC LINEAR SYSTEMS
-
FOR
Bayda Ghanim Fathi…...…………… ..…….………………………………………………………. 140
- ON GENERALIZED REGULAR LOCAL RING
Zubayda M. Ibraheem and Naeema Ali Shereef…………………………………………………...…145
- SURFACE ROUGHNESS EFFECT ON DISCHARGE COEFFICIENT OF
COMBINED CYLINDRICAL WEIR GATE STRUCTURE
Shaker A. Jalil and Safa S. Ibrahim …………………..…………………………………………...…153
Journal of University of Zakho, Vol. 3(A) , No.1, Pp 1-8, 2015
ISSN: 2410-7549
THE ROLE OF K+ CHANNEL SUBTYPES IN CATECHIN INDUCED
VASORELAXATION IN RAT’S AORTIC RINGS
Omar A. M. Al-Habib1 and Fakhir Najim K. Sabir2
Dept. of Biology, Faculty of Science, University of Zakho, Kurdistan Region - Iraq.
2
Dept. of Biology, Faculty of Science and Health, Koya University, Kurdistan Region – Iraq.
(Accepted for publication: Jan 15, 2015)
1
Abstract;
The current study aimed to investigate the role of K+ channels subtypes in relaxant effects of catechin
on rat’s aortic rings. The isometric tension of the thoracic aorta was measured using ADI PowerLab Data
Acquisition System. Catechin at concentrations (10-5 to 5x10-1M) produced more potent relaxant effects in
phenylephrine (10-6 M) precontracted aortic smooth muscle with Log IC50’s of -1.159 mg/ml as compared
to potassium chloride (60 mM) with Log IC50’s of -6.373 mg/ml . The catechin induced relaxation in aortic
rings precontracted with phenylephrine was 17.464 ± 0.068 %, where as for aortic rings precontracted
with potassium was only 5.574 ± 0.131%. In aortic rings preincubated with glibenclamide (10-5M) and
tetraethylammonium (1mM), catechin relaxant effect was enhanced with Log IC50s of -3.119 and -3.001
mg/ml, respectively. On the other hand, in aortic rings precontracted with PE and preincubated with
BaCl2 and 4-AP, catechin induced statistically non-significant relaxant effects as compared with aortic
rings of the control aortic ring which was precontracted with PE. From the results of the current study, it
can be concluded that catechin produced more potent vasorelaxant effect on aortic rings preincubated
with phenylephrine that KCl. This vasorelaxant effect of catechin is produced via the activation of both
KATP and Kca, but not Kir and Kv channels subtypes.
Keywords: K+ channel blockers, Catechin, vasorelaxation, aorta.
Introduction
C
atechins, which are also known as
flavan-3-ols, are
simple forms of
flavonoid; based on their structure, they are
classified as flavanols which include catechin,
epicatechin,
epigallocatechin,
epicatechin
gallate, and epigallocatechin gallate.
Epicatechin concentrations are high in apples,
blackberries, broad beans, cherries, black grapes,
pears, raspberries, and chocolate. The remaining
catechins such as epigallocatechin, epicatechin
gallate, and epigallocatechin gallate are found in
high concentrations in both black and green tea
(Williamson and Manach,2005 and Ghayur et
al., 2007).
Catechins has an important protective role
against many diseases such as heart disease,
stroke, atherosclerosis, hypertension, kidney
disorders, obesity, diabetes
( Crespy and
Williamson, 2004) and act as antithrombic agent
(Yang, et al, 19990). Furthermore, the
antioxidant effects of Catechins has been
recognized as they prevent the oxidation of
antioxidants such as vitamin E and even some
times elevate its level (Alessio, et al, 2003 and
Tijburg, et al, 1997). In vitro studies on
Catechins have shown that it inhibit catechol Omethyltransferase (COMT), the enzyme that
degrades norepinephrine; and thus, reflecting the
important role of the sympathetic nervous
system and its neurotransmitter norepinephrine
in the control of thermogenesis and fat oxidation
(Dulloo et al., 1999).
Catechin can induce vasorelaxant responses
via the stimulation of endothelial Nitrous oxide (
NO ) and nitrous oxide synthase ( NOS ) (Huang
et al., 1999 and Lorenz et al., 2004) or increased
the production of PGI2 (Mizugaki et al., 2000).
Catechin-evoked vasorelaxant effects appear to
be both endothelium-dependent and independent
(Huang et al., 1998). It has been indicated that
the effect of Epigallcatechin-3-gallate on rat’s
aorta may be used as an interesting model for the
subsequent development of new PDE- inhibitory
drugs for improving the pharmacological
treatment of diseases such as cardiovascular
pathology ( Alvarez, et al, 2006).
Taking into the consideration the above
informations on Catechin effects, and since no
attempt have been made to study the role of K+
channels subtypes in vasorelaxation effect of
Catechins, the current work was undertake to
shed light on the role of above channel subtypes
in vasorelaxation induced by catechin in rat’s
aortic rings.
1
Materials and Methods
Materials
Albino Rats
Adult male albino rats, Rattus rattus
norvegicus (250-350g) used in the current study
were bred in the Department of Biology, Faculty
of Science, University of Zakho. The rats were
reared in standard PVC rat cages, maintained in
the laboratory under controlled temperature (22
± 2 ℃), and photoperiod of 12-hours light/dark
cycle using automated light-switching device.
All rats were provided with standard food pellets
prepared as described by Shekha, (2010) with a
free access to water ad libitum. The animals
were acclimatized to the laboratory conditions
for 1-2 weeks before using them in the
experiments.
Methods
Isolated Aorta Preparation
The animals were injected intraperitoneally
with heparin (1500 units/ kg body weight) and
left for 30 min, to avoid blood clotting and
possible damage of endothelial layer of the aorta
(Fulton et al., 1996). Animals were then
anesthetized with Ketamine (40 mg /kg) and
Xylazine (10 mg/Kg) intraperitoneally.
The chest cavity was opened and after the
removal of excess tissue and fat, the aorta was
isolated and transferred to a beaker containing
Krebs solution aerated with carbogen [95 %
oxygen (O2) and 5 % Carbone dioxide (CO2)].
The beaker was placed in the water bath at 37 oC
and the aorta cut into small rings of about 2-4
mm long.
Measurement of Isometric Contraction in
Isolated Rat Aorta
The procedure of Shekha and Al-Habib,
(2012) was followed with some modifications to
study the vascular reactivity in the isolated aorta.
Two stainless steel wires were carefully passed
through the lumen of the aortic rings. One of
them was anchored to the base of glass organ
chamber and the other was connected to a force
transducer (Model MLT0201/RAD) coupled to
the transbridge amplifier connected to a
PowerLab Data Acquisition System and
computer running chart software (Version 7)
used for isometric tension measurement. Special
attention has been taken during the preparation
to avoid damaging the endothelium. The extents
of contraction and relaxation were expressed by
the tension recorded by the system. The
2
relaxation rate was defined as Trelaxation/ Tcontraction
expressed as a percentage.
Prior to the experiment, 10 ml of Kreb’s
solution was placed inside the glass tissue
chamber and the organ tissue bath system was
maintained at 37 oC by circulating water through
the water jacket from a circulating water bath set
at 37 oC. The aorta was continuously aerated by
carbogen (95 % O2 and 5 % CO2). The initial
tension was set at 2 g weight and left for 60
minutes. The aortic segments were initially
exposed to 60 mM K+ to test their functional
integrity. After that, the chamber medium was
changed several times until a stable resting tone
was recorded, then the experiments were started.
Statistical Analysis
The statistical analysis was performed using
two-way analysis of variance (ANOVA)
supported by Bonferroni test when carrying out a
pairwise comparison between the same dose of
different groups using Graphpad prism program,
version 6.01. Analysis of variance for repeated
measurements was applied to data consisting of
repeated observations at successive time points.
P-values less than 0.05 were considered as
statistically significant. In all figures, the
symbols (*, ** and ***) representing mean
differences are significant at the 0.05, 0.01 and
0.001 levels, respectively. The maximum
contractile responses to catechin were calculated
as a percentage of the contraction produced by
PE or KCl and expressed as the means ±
standard error of the mean (SEM). The tension
produced by vasoconstrictors (PE and KCl) was
defined as 0% tension, and the baseline tension
before adding vasoconstrictors was 100%
tension.
Results
Relaxant Effect of Catechin on Aortic Rings
Contracted by PE and KCl
Typical
traces
from
representative
experiments on the relaxing effect of different
concentrations of catechin on PE and KCl
precontracted aortic rings are shown in figure
(1). Dose-response curves for the effect of
catechin on PE- and KCl-induced contractions
are shown in figure (2). Catechin at
concentrations from 10-2 to 5X10-1 M resulted in
a highly significant relaxant effect (P<0.001) in
PE (10-6 M) as compared with KCl (60 mM)
precontracted thoracic aortic rings.
Catechin produced a more potent inhibitory
effect on PE than KCl induced contractions,
with Log IC50’s of -1.159mg/mL (Log IC50 of CI
95% between -6.708 to 4.389) and -6.373
mg/mL (LogIC50 of CI 95% between -22.430 to
9.686), respectively. Catechin produced highly
significant relaxant effects on PE-induced
contractions which was 17.464 ± 0.068% ,
whereas catechin produced a mild relaxant
effect on the aortic ring precontracted with KCl
(5.574 ± 0.131%).
Fig. 1: Typical traces showing the relaxant effects of different concentrations of catechin in rat aortic rings,
precontracted with PE (10-6M).
Fig. 2: Cumulative dose-response curve for the effects of catechin on PE (10-6M) and KCl (60mM)
precontracted aortic rings.
3
Role of Potassium Channels Subtypes in vasorelaxation Induced by Catechin
The role of K+ channels subtypes in the relaxant effect of catechin was investigated by
preincubation of aortic rings in GLIB (10-5), TEA (1mM), BaCl2 (1mM) and 4-AP (1mM) and
precontracted with PE. The Dose-response curves for Catechin effect on PE precontracted aortic rings
preincubated with K+ channel blockers are shown in figures (3, 4, 5 and 6).
In aortic rings precontracted with PE and preincubated with GLIB in the presence of catechin
showed a highly significant (P<0.001) relaxant effect at doses (3X10-3-5X10-1) and with TEA also
showed a significant (P<0.01) relaxant effect at concentrations (10-2-5X10-1) on aortic rings. The
percentages of relaxation were 31.089±0.136% and 58.392±0.110 %, and with Log IC50’s of -3.119
mg/ml (with a Log IC50 of CI 95% between -3.994 to -2.245) and -3.001mg/ml (with a Log IC50 of
CI 95% between -3.875 to -2.128), respectively. On the other hand, aortic rings precontracted with PE
and preincubated with BaCl2 and 4-AP showed limited relaxant effects which were statistically nonsignificant to catechin as compared with aortic rings precontracted with PE as control with Log IC50’s
of -1.159 mg/ml (with a Log IC50 of CI 95% between -6.708 to 4.389) and the percentages of
relaxation were 4.247 ± 0.280 and 17.464±0.068 %.
Fig. 3: Cumulative dose-response curve for the relaxant effects of catechin in aortic rings preincubated with
GLIB, precontracted with PE.
4
TEA, precontracted with PE.
BaCl2, precontracted with PE.
5
Fig. 6: Cumulative dose-response curve for the relaxant effects of catechin in aortic rings preincubated with 4AP, precontracted with PE.
Discussion
The results of the study showed that catechin
in a dose-dependent mode inhibited the
contractions induced by PE and high K+
concentration in isolated rat thoracic aorta. The
flavonoid tested in this study was found to be
more sensitive in relaxing the contraction
induced by PE. The results of one of the
previous study suggested that the reduction of
transmembrane Ca2+ influx and/or agonist
induced release of intracellular Ca2+ in these
cells may contribute to the vasorelaxant action of
catechin, although these effects do not seem to
be important for lower concentrations of the
catechin (Alvarez, et al., 2006 ).
From the results of the current study, it has
been observed that vasorelaxation-induced by
catechin was enhanced significantly by TEA and
GLIB, but not by 4-AP and BaCl2. These results
suggest that vasorelaxant responses to catechin
are mediated through increasing K+ efflux at
least, via KATP, KCa channels. There is growing
evidence that the KATP channels activity may be
modulated by NO. Kubo et al., (1993)
demonstrated that nitric oxide donors cause
activation of KATP channels in rat aorta by means
of a cyclic GMP-dependent mechanism, possibly
cyclic GMP-dependent PK.
Álvarez et al., (2006) demonstrated that
catechin-induced relaxation in rat aortic rings
was endothelium-independent and mediated by
the inhibition of phosphodiesterase (PDE) 1-5
6
isoform activity. Moreover, it was reported that
catechin has the ability to induce contractile
responses in isolated rat aorta (Sanae et al.,
2002; Shen et al., 2003 ; Álvarez-Castro et al.,
2004).
From the results of the present study it can be
concluded that the relaxant effect of catechin on
aortic smooth muscle is mediated via the
activation of KATP and Kca, but not Kir and Kv
channels.
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8
EFFECT OF RENAL STONE FORMATION ON SOME RENAL RELATED
PARAMETERS IN KOYA CITY
Layla Kareem Ali
Koya Technical Institutes, Kurdistan Region – Iraq.
(Accepted for publication: May 27, 2015)
Abstract:
The thirty stone former patients (12 men with 18 women) involved in the current study, were attended
the Shaheed Khalid Hospital in Koya city between April to November 2013. The cases diagnosed in the
hospital by clinical examination followed by kidney urinary bladder KUB x-ray and ultrasonography.
Further more thirty healthy person (15 men with 15 women) were also involved in this study as control
group.
The data of a current study revealed the dominant prevalence of the renal stone type both in men and
women was for uric acid stone type, and calcium carbonate, calcium phosphate and calcium oxalate
represent moderate types, and cysteine stone type represent the lowest type among involved patients. The
stone formation was more prevalence in age group 51-60 that represent 27% of the total included formers.
The results showed significant (p< 0.05) elevation in serum creatinine, urea and uric acid levels in both
sexes stone formers, whereas serum total protein and albumin levels were significantly lower in both men
and women stone formers as compared with the levels in normal healthy group. The hematological results
showed significantly lower numbers of RBC number and Hb content in both men and women stone formers ,
whereas, WBC count and platelets count showed higher numbers in both men and women stone formers as
compared normal healthy group .
Key words: (Renal stone formers, Hepatorenal parameters, Heamatological parameters )
Introduction
U
rolithiasis is the most common urological
disease affecting human worldwide and
stone formation contributed to genetic and
environmental factors (Harpreet Kaur et al., 2012).
Stone disease is an increasing and major public
health problem with more predominance in men
(Memon et al., 2009).
Pathogenesis of urinary stones formation is
thought to be multifactorial, the infection of
urinary tract is a major factor. Kidney stones may
contain various combinations of chemical forms,
that occur when salts in the urine precipitate and
form solid materials that resulted from a wide
variety of metabolic and environmental
disturbances (Sharda and Zia, 2006). Urinary
stones are typically classified by their chemical
composition into calcium-containing, struvite, uric
acid, or other compounds (Lieske and Segura,
2004). Stones were predominantly of mixed type,
with
calcium
oxalate
as
commonest
constitute(Qaader et al., 2006).
Creatinine, urea and uric acid are frequently
used to assess kidney function, and their elevated
serum levels indicate kidney dysfunction
(Lawrence et al., 2003). Creatinine is released
into the blood at a constant rate and freely
excreted by the kidneys. For this reason,
creatinine test is frequently used to assess kidney
function its elevation indicate kidney dysfunction
(Hamid, 2008).
Urea is one of the major nitrogenous wastes in
blood, which are toxic
end products of
catabolism and they are normally removed from
the blood and excreted by kidneys at a rate that
balances their rate of production.
Renal disease is one of the major disease
states that are associated with elevated plasma
uric acid concentration because its filtration and
secretion are impaired (Lawrence et al., 2003).
The kidneys maintain the blood volume and
regulate the mineral content in the bloodstream.
The liver converts nutrients into energy, forms
proteins and stores carbohydrates. While these
organs can be remarkably resilient in the
elimination of toxins, their other functions can be
damaged in the process.
Kidneys as the common endocrine organs are
responsible for controlling red cell production
through
erythropoiesis
by
release
of
erythropoietin (EPO) hormone, which its
deficiency leads to the decrease of RBCs and Hb
( Suresh
et al., 2012). Renal diseases are
9
associated with a variety of haemopoietic changes.
Anemia parallels the degree of renal impairment
and its most important cause is failure of renal
erythropoietin secretion. Other factors include
chronic blood loss, hemolysis and bone marrow
suppression by retained uremic factors (Suresh et
al., 2012). The current investigation aimed
measurement of the hepatorenal functions testsand
hematological parameters in stone formers.
This study included (30) patients of renal stone
former (18 men with 12 women), who attended the
Shaheed Khalid Hospital in Koya city from April
to November 2013. The diagnosis of the cases
was done in the hospital based on the clinical
examination followed by kidney-urinary bladder
KUB
x-ray,
ultrasonography,
intravenous
urography IVU and urinalysis. Furthermore this
study also included 30 healthy (15 men with 15
women) subjects at same ages free of kidney
disease signs, they were randomly selected as a
healthy group for comparison.
Five ml of venous blood samples were obtained
from both patients and healthy group by sterile
disposable syringe. After coagulation of the blood,
each blood sample was centrifuged for 3 minutes
at 4000 rpm to get a clear and cell free serum. The
serum was used for biochemical parameters assay.
The colorimetric Elitech Diagnostic kits were
used for measurement of renal function test
parameters (serum total protein, creatinine, urea,
uric acid, albumin and bilirubin. In this method, 1
ml of serum was added to flexor tube and the
concentrations of the parameters were analyzed
using automatic chemical analyzer (Benchtop
automatic biochemistry analyzer (ELITech)
(FLEXOR EL200, ELITech clinical systems),
France.
An aliquot of blood was immediately removed
and mixed with ethylene diamine tetra acetic acid
(EDTA) as an anticoagulant. The blood samples
were analyzed for blood parameters using a full
automated hematological analyzer according to the
manufacturer's protocol. The hematological
parameters (WBC, RBC, Hb, Plt, and Hct) were
analyzed on the same day a blood samples
collection using Coulter Counter.
Statistical analysis
The SPSS (statistical package for social
science) (V 20) T- test was used to analyze the
data. The P value (P<0.05) was considered to be
statistically significant.
Results
The results showed that the prevalence of the
uric acid renal stone type more dominant both in
men and women and represent 38.09% of stones.
The percentage of calcium carbonate, calcium
phosphate and calcium oxalate types were 19.04%
for each type and the remaining 4.76% was
composed of cysteine stone (Table 1).
Table 1: The prevalence of the renal stone types
Type of
stones
Cysteine
stone
Calcium
carbonate
% of
Patients
4.76
19.04
Calcium
phosphate
Calcium
oxalate
19.04
19.04
Uric acid
38.09
Results showed that renal stone formations are more prevalence in age groups 51-60 which represent
27% of the total stone formers (Table 2).
Table 2: The prevalence of the renal stone in relation to age groups
10
Age
groups
21-30
31-40
41-50
51-60
61-70
71-80
81-90
% of
patients
3.57
10.71
21.42
27
23
14.28
3.57
The results showed significant elevation in
serum creatinine level 1.137±0.143, 1.185±0.184
in both men and women stone formers respectively
as compared to normal men and women subjects
0.751±0.047, 0.630±0.057 respectively, also
serum urea level showed significant elevation in
both men and women 35.08 ± 1.415, 34.41 ±1.881
respectively, as compared to its level in normal
men and women subjects 21.21 ± 1.419, 20.5 ±
2.121 respectively. The level of the uric acid was
showed significant elevation in both men and
women stone formers 5.528 ± 0.165, 5.191 ±
0.211 respectively, as compared to its level in
normal men and women subjects 3.33 ±0.21, 3.1
±0.271 respectively. Whereas both men and
women stone former showed significant lower
serum total protein 6.725±0.172, 6.626±0.166
respectively, as compared to their levels in normal
men and women subjects 7.422±0.141,
7.283±0.148 respectively. The level of the albumin
was showed significant lower in men and women
stone
formers
4.250±0.259,
4.262±0.208
4.868±0.142 respectively. The level of bilirubin in
men and women stone formers showed nonsignificant lowering 0.725±0.138, 0.403±0.045
0.495±0.046 respectively (Tables 3 &4).
Table 3: Renal related parameters in renal stone former men compared to healthy men
Groups
Parameters
Total protein (gm/dl)
Albumin (mg/dl)
Bilirubin (mg/dl)
Creatinine (mg/dl)
Urea (mg/dl)
Uric acid (mg/dl)
Controls
M±S.E.
Patients
M±S.E.
P value
7.422±0.141
5.021±0.103
0.803±0.104
0.751±0.047
21.21 ± 1.419
3.33 ±0.21
6.725±0.172
4.250±0.259
0.725±0.138
1.137±0.143
35.08 ± 1.415*
5.528 ± 0.165*
0.005
0.012
NS
0.029
0.004
0.0011
NS= non-significant
Table 4: Renal related parameters in renal stone former women compared to healthy women
Groups
Parameters
Total protein (gm/dl)
Albumin (mg/dl)
Bilirubin (mg/dl)
Creatinine (mg/dl)
Controls
M±S.E.
7.283±0.148
4.868±0.142
0.495±0.046
0.630±0.057
Patients
M±S.E.
6.626±0.166
4.262±0.208
0.403±0.045
1.185±0.184
Urea (mg/dl)
20.5 ± 2.121
34.41 ±1.881*
0.008
0.030
NS
0.014
0.005
Uric acid (mg/dl)
3.1 ±0.271
5.191 ± 0.211*
0.002
P value
NS= non-significant
The results showed significantly lower numbers of RBC and Hb content in both men and women stone
formers, as compared with normal subjects , whereas WBC and platelets showed higher numbers in both
men and women stone formers, as compared with normal subjects (Tables 5 and 6).
Table 5: Hematological parameters in renal stone former men
Groups
Parameters
RBC x 106 cell/mm3
WBC cell/mm3
Hb (gm/dl)
Platelet x 103 cell/mm3
Controls
M±S.E.
Patients
M±S.E.
P value
5.008±0.096
6.876±0.250
14.79±0.231
227.3±11.10
4.410±0.219
8.525±0.615
13.76±0.439
273.8±14.73
0.020
0.021
0.039
0.019
11
Table 6: Hematological parameters in renal stone former women
Groups
Parameters
RBCx106 cell/mm3
WBC cell/mm3
Hb (gm/dl)
Platelet x 103 cell/mm3
Controls
M±S.E.
4.778±0.154
6.891±0.499
12.66±0.328
207.5±10.61
Discussion
The data of the current study showed that the
prevalence of the renal stone types both in men
and women are higher for uric acid stone type than
calcium carbonate calcium phosphate and calcium
oxalate types. This results are in agree with
previous results reported that approximately 80%
of stones are composed of calcium, 10% of
struvite, 9% of uric acid, and the remaining 1% are
composed of cystine (Fredric et al., 2005). In
another study reported that calcium oxalate
monohydrate is the most frequent stone
component (Alaya et al., 2011). In anther study
results indicated that calcium stone predominated
with male predominance of the most prevailing in
Egypt (Al-Ali et al., 2002).
The results showed significant elevation of
serum creatinine, urea and uric acid levels in both
men and women stone formers. Elevation of
serum creatinine in the current study confirms the
results of (Kasem (2004), who explained that this
increase in creatinine level may be due to reduced
renal function and increased acidic medium in
renal tubules and impaired metabolism as well as
kidney complications lead to higher creatinine.
Recently, another study revealed a significant
increase in serum levels of urea, creatinine, and
uric acid and a significant decrease in total
protein level in kidney stone former patients
(Kaniaw, 2013). Kidney stone cause dysfunction
of the kidney as a result of cell injuries and
increasing the accumulation of nitrogenous waste
products in the blood (Worcester et al., 2006).
Elevation of urea as a result of the damage of
some renal nephrons units leads to a defect in urea
filtration and excretion that reported in previous
study (Hayder Bawa, 2008). High serum uric acid
level might lead to stone formation and a
decreases in the solubility of calcium oxalate salts
in urine (Mehdi, 2008).
12
Patients
M±S.E.
3.508±0.190
8.446±0.433
11.08±0.310
243.4±11.75
P value
0.0001
0.032
0.003
0.035
In the current study, serum total protein and
albumin levels showed significant lowering levels
in both men and women stone formers. These
results are in agreement with another study in
Sulaimani governorate (Kaniaw, 2013), she
reported lower level of serum total protein and
albumin levels in stone former patients cause of
kidney dysfunction as a result of stone formation
and kidney tissue damage.
The results showed significantly lowering in
the numbers of RBC and Hb content in both men
and women stone formers , whereas, WBC and
platelets showed higher numbers in both men and
women stone formers. This may be caused by
impaired erythropoietin production and other
factors which suppress marrow erythropoiesis and
shortened red cell survival (Suresh et al., 2012). In
the absence of EPO, DNA cleavage is rapid and
leads to cell death (Suresh et al., 2012). RBC
survival is decreased in uremic patient’s in
proportion to the blood urea nitrogen
concentration and, it improves significantly after
intensive hemodialysis. Uremic plasma increases
the expression of phosphatidylserine on the outer
cell surface in red blood cells. This enhances the
recognition of damaged red blood cells by
macrophage, leading to their subsequent
destruction and decreased survival. (Michael et al.,
2004). Red blood cells survival is presumed to be
a toxic substance normally excreted or
metabolized by the kidneys, one such substance is
guanidine and its derivatives which appear to be a
subset of the many retained metabolites, adversely
affect erythrocyte survival (Robert and means,
2004).
In conclusion renal stone patients have lower
hematological indices, due to impaired production
of erythropoietin, and other factors like increase
haemolysis, suppression of bone marrow
erythropoiesis, hematuria and gastrointestinal
blood loss. The concentration of serum creatinine
shows negative correlation with all the
haematological parameters. And the degree of
changes depends on the severity of renal failure.
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13
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14
ISSN: 2410-7549
TOXIC EFFECTS OF ESSENTIAL OILS OF ELATTARIA CARDAMOMUM L.
AND LAMBDA- CYHALOTHRIN ON TRIBOLIUM CONFUSUM (DUVAL)
1
Sofyan H. Sedo Al-Sinjari 1 and Hani J. Al-Attar 2
Biology Department, Faculty of science, University of Zakho, Kurdistan Region – Iraq.
2
Biology Department, College of Education, University of Mousl, Iraq.
Abstract:
Toxic effect of Lambda-cyhalothrin and essential oils of cardamom seeds of Elettaria cardamomum
(L.), individually or in combination, on the larva, pupa and adult stages of confused flower beetle,
Tribolium confusum, through the topical application were studied. The percentages of mortality of larval
stage exposed to Lambda-cyhalothrin, at concentrations of 0.25, 0.35, 0.50 and 0.75 µl/ larva were 13.33,
20, 36.67 and 46.67 % respectively. While, the percentages of mortality of pupa stage at concentrations of
0.25, 0.35, 0.50 and 0.75 µl/ pupa were 13.33, 16.67, 26.67 and 43.33 respectively. For adults, exposed to
Lambda concentrations of 0.25, 0.35, 0.50 and 0.75 µl/ adult were 10.0, 13.33, 23.33 and 40.0 respectively.
The percentages of mortality of larvae exposed to cardamom oil at concentrations of 0.1, 0.3, 0.5 and 1 µl/
larva were 60, 76.67, 90 and 96.67% respectively, while the mortality of pupa exposed to concentrations of
0.1, 0.3, 0.5 and 1 µl/ pupa were 46.67, 63.33, 83.33 and 90 respectively. The percentages of mortality of the
adults exposed to concentrations 0.1, 0.3, 0.5 and 1 µl/ adult were 43.33, 60.0, 76.67 and 83.33 respectively.
The synergistic ratios for larvae, pupae and adults exposed to mixture 1 (0.03 µl / insect cardamom oil
and different concentrations of pesticide ), were 2.01, 1.70 and 1.60, respectively, where as the synergistic
ratios of larvae, pupae and adults exposed to mixture 2 ( 0.07 µl / insect cardamom oil and different
concentrations of pesticide) were 2.61, 2.34 and 3.10, respectively. This indicates that a combination of
cardamom seeds oil with different concentrations of the insecticide Lambda produced synergistic effects.
Keywords: Cardamom, Tribolium confusum, Lambda- cyhalothrin, Synergistic.
Introduction
T
he broad incorrect and uses of insecticide
had led to the development of resistance
in several species of insect to insecticide. Since
the number of insect species resistance to
insecticides was about 460 species until 2003.
The side effects of insecticides use prompted
many conservationists call to stop the use of
insecticides and prevent their production, This
call was unrealistic as evidenced by increased
production and use of insecticides throughout
the world, which of course refers to the
insecticides still is the effective means adopted
by human to control the pests, (Meister, 2010).
The most convenient alternative to reduce the
use of insecticides is the rational and correct
methods of use in order to reduce the damage
produced from their side effects. In addition
anabolic substances of insecticides increase the
effectiveness when used at low concentrations.
(Metcalf, 1972).
The cardamom, Elettaria cardamomum L.
(Maton) (Zingiberales: Zingiberaceae), called
the "Queen of spices" in India, is a tall,
perennial, reed-like herb that grows wild and
cultivated in India and Sri lanka. The essential
oils have therapeutic benefits such as
gastrointestinal, cardiovascular and neural
disorders (Lewis, 1984; Aneja and Sharma,
2010; Abbasipour et al. 2011). Some studies
showed that cardamom essential oils have toxic
effect against some stored insects (Jacobson,
1989). A study by Sivakumar et al. (2010)
revealed that cardamom essential oils showed
toxicity on cowpea weevil, Callosobruchus
maculates (F.).
The chemical composition of cardamom
varies considerably with the variety, region, and
age of the product. The content of volatile oil in
the seeds is strongly dependent on storage
conditions. Some studies showed that the main
components found in the volatile oil from E.
cardamomum were 1,8-cineol (29.7 to 36.3%)
and α-terpinyl acetate (26.1 to 31.3%) (OliveroVerbel et al. 2010 and Korikontimath et al.
1999). The later also reported that the volatile
oil of E. cardamomum contained about 11.6%
limonene, and 3% linalool,. In addition, the
basic cardamom aroma is produced by a
combination of the major components, 1,8cineole and α-terpinyl acetate (Lawrence
1979).
Lambda-cyhalothrin is a synthetic pyrethroid
insecticide, which is the analogues of pyrethrins,
and naturally occurring insecticidal compounds
produced in the flowers of chrysanthemums
(Chrysanthemum cinerariaefolium). Insecticidal
15
products containing pyrethroids have been
widely used in pests control (Amweg and
Weston 2005; Oros and Werner 2005; He et al.
2008).
The aim of the present study was to
investigate the effect of lambda and Elattaria
cardamomum essential oils applied individually
or in combination on larva, pupa and adult of T.
confusum. In addition, this paper describes for
the first time the toxicity of the essential oils of
cardamom against one important stored product
pests: the confused flour beetle, Tribolium
confusum Duval (Coleoptera: Tenebrionidae).
Insects The insect species, T. confusum, was
used during the present study. Larvae, pupae and
adults were obtained from laboratory cultures
originincubated in dark at 30± 1C° and relative
humidity 70±80% using incubators ( LAB
TECH. Korea). T. confusum was reared on
wheat flour mixed with yeast (10:1, w:w). The
5th instar larvae of T. confusum, newly formed
pupae (few hours – 24 hours) and adults ( one
week after emergence) were used in contact
toxicity experiments.
Extraction of Cardamom essential oil
Seeds of Cardamom (50g) were grinded with
electrical grinder (Tata 300 w). and the obtained
powder was extracted with 300 ml petroleum
ether ( boiling point 60-80 c°) in Soxhlet
apparatus. The extraction was condensed using
thin film Vacuum rotary evaporator to evaporate
the solvent from cardamom oils. The extracted
essential oils was transferred to glass bottles and
kept in refrigerator until needed.
Methods of Treatment (contact toxicity)
Each developmental stage (larva, pupa or an
adult confused flour beetles) was
treated
superficially (Topical application) with 5 µl of
the desired concentration of the tested material,
which was added on the upper surface of the
insect thorax (Notum) using 50 µl capacity
micro-syringe (Huang et al. 2000). The control
group was treated with appropriate solvent
(water in the case of the pesticide and petroleum
ether in extract case). After 15 minutes, the
treated groups were placed in petri dishes and
saved in incubator to avoid contamination by
16
any abnormal insect. The percentages of
mortality for larvae and adult beetles were
recorded after 24 hours, and pupa until adult
emergence.
Effect of a combination of cardamom extract
and Lambda.
The surface treatment for larvae, pupae and
adults of confused flour beetles were tested
using MSDOS system according to Probit
program which gave the lethal doses (5% (LD5)
= 0.03 µl and 15% (LD15) = 0.07 µl) of the
extract and then treated with various
concentrations of the pesticide. The mortality
rate was recorded after 24 hours of larvae and
adult, and pupa until adult emergence.
Estimating Synergistic Ratio (SR)
To calculate synergy ratios the formula of
Brattesten and Metcalf (1970) was used as
follows
Synergistic
Ratio
=
The synergistic ratio is equal to the number
of times the increase in pesticide toxicity caused
by the synergistic.
Results and Discussion
Effect results of effect of cardamom seeds
extract on of various life cycle stages of
confused flour beetle are shown in Table (1),
which indicate that the essential oil of the
cardamom seeds extracts has toxic effects on the
larvae, pupae and adult of the confused flour
beetle. The percentages mortality of the larvae
exposed to at concentrations of 0.1, 0.3, 0.5 and
1 µl/ larva were 60, 76.67, 90 and 96.67%
respectively, where as
the percentages of
mortality of pupae exposed to concentrations of
0.1, 0.3, 0.5 and 1 µl/ pupa were 46.67, 63.33,
83.33 and 90%, respectively.
The percentages of mortality of the adult
insects exposed to concentrations 0.1, 0.3, 0.5
and 1 µl/ adult were 43.33, 60.0, 76.67 and
83.33% respectively. The mortality ratio values
revealed that the essential oil of cardamom seeds
extract is more toxic to the larvae as compared
with pupae and adults (Fig. 1).
120
Mortality %
100
80
Larva
60
Pupa
40
Adult
20
0
٠.١
٠.٣
٠.٥
١
Concentrations μl / insect
Figure (1) Effect of different concentrations of cardamom seeds oil extract on larval, pupal and adult stages of
confused flour beetle.
These results are in agreement with other
studies which showed the toxicity test of
cardamom oil and different vegetable oils
against many types of insects. Ngamo et al.,
(2007) found that oils extracted from black
sesame Hyptis spicigera were toxic to adults of
various insects such as Rice weevil Sitophilus
oryzae, Maize weevil Sitophilus zeamais,
cowpea weevil Callosobruchus maculatus and
the red flour beetle Tribolium castaneum.
Furthermore, essential oils extracted from Neem
(Azadirachta indica), Tulasi leaves (Ocimum
sanctum), and seeds of pongamia (Pongamia
pinnata) have toxic effects to the wax worm
larvae (Galleria mellonella), and the their
percentages of mortality ranged between 9352% (Surendra et al., 2010).
It was found that the essential oils extracted
from leaf laurel (Laurus nobilis), myrtle (Myrtus
communis) and thyme (Thymus sipyleus) were
toxic to adult of beans weevil (Acanthoscelides
obtectus) and red flour beetle (Karaborklu et al.,
2010). Also, Padin and his coworker investigate
the toxicity effects of essential oils extracted
from nine different types of plants. They found
that all extracts were toxic to red flour beetle
adults, and the mortality rate of Ethanolic extract
from Viola arvensis was 68% (Padin et al.,
2013).
Effect of insecticide Lambda-cyhalothrin on
stages of confused flour beetle
The percentages of mortality of various
developmental stages of the flour beetle
Tribolium confusum exposed to
various
concentrations of Lambda-cyhalthrin are shown
in (Table 1). The percentages of mortality of
larval stage exposed to concentrations of 0.25,
0.35, 0.50 and 0.75 µl/ larvae were 13.33, 20,
36.67 and 46.67%, respectively; while, the
percentages of mortality of pupae exposed to
concentrations of 0.25, 0.35, 0.50 and 0.75 µl/
pupa were 13.33, 16.67, 26.67 and 43.33%,
respectively.
Adults beetles exposed to
concentrations of 0.25, 0.35, 0.50 and 0.75 µl/
adult, the percentages of mortality were 10.0,
13.33, 23.33 and 40.0%, respectively. These
results clearly indicate that the pesticide has
relatively more toxic effect against larvae as
compared with pupae and adults (Figure 2).
17
50
45
Mortality %
40
35
30
25
Larva
20
Pupa
15
Adult
10
5
0
٠.٢٥
٠.٣٥
٠.٥
٠.٧٥
Concentrations μl/ insect
Figure (2) Effect toxic effect of Lambda-cyhalothrin on various developmental stages of the flour beetle
Tribolium confusum.
The results of the current study on the toxic
effect of Lambda on flour beetles agreed with
other studies on the toxicity of lambda
insecticide and other insecticides against
different insects species. Umeda et al., (2000)
found that the insecticide lambda is extremely
toxic against cabbage looper (Trichoplusia ni)
and diamondback moth (Plutella xylostella).
Furthermore, the rates of mortality in both
insects were generally increased with the
increasing of the duration of exposure.
Additionally, the insecticide Aktara was very
toxic to 1 day aged adults of Khapra beetle
(Trogoderma granareum) and cowpea weevil
Callosobruchus maculatus and 4 weeks aged
18
adult of red flour beetle and confused flour
beetle and rice weevil (AL-Attar, 2006). The
effectiveness of the toxicity of lambda
insecticide was proven to be very toxic against
red flour beetle (Uddin and Ara, 2006). Chavan
et al. (2012) proved that the lethal dose of
insecticide lambda against teak skeletonizer
larvae (Eutectena machaeralis) after 12 hours of
treatment was 81.14 %. Naz et al. (2013)
revealed that the lambda insecticide is highly
toxic to four types of berry bug, (Halys spp.) in
comparison with Chloropyriphos insecticide and
Neem extract.
Table (1) Effect of Cardamom extract, Lambda insecticide and mixture ratios in mixtures 1 and 2 on the
percentages of mortality of Larva, pupa and adult of flour beetles
5th instar Larvae
phases
Insecticide
Conc.
μl
Mortality
%
Conc.
μl
0
0.00 z
0
0.25
0.35
Pupae
13.33
w-y
20.00 ux
0.3
16.67
x
v-
60.00
i-m
76.67 eh
90.00
cd
Conc.
μl
Mortality
%
Conc.
μl
Mortality
%
0.03
20.00
u-x
0.07
30.00
r-u
0.25
0.35
0.25
0.35
1
96.67
ab
0.75
73.33
f-i
0.75
83.33
Def
0.00
0
6.67
y
0.03
13.33
w-y
0.07
20.00
u-x
0.1
46.67 mq
0.25
0.3
63.33
0.35
0.35
0.50
0.75
0.5
1
0
0.00
z
0
0.25
10.00
xy
0.1
0.35
0.50
0.75
13.33
w-y
23.33 tw
40.00 os
0.3
0.5
1
h-l
83.33
d-f
90.00
cd
10.00
xy
43.33
n-r
60.00
i-m
76.67 fh
83.33
d-f
0.50
0.75
0.03
0.25
0.35
0.50
0.75
23.33
t-w
33.33
q-u
50.00
l-p
56.67
j-n
13.33
w-y
26.67
s-v
30.00
r-u
46.67 mq
53.33 ko
0.50
46.67
m-q
50.00
l-p
70.00
g-j
46.67 mq
13.33 wy
16.67 vx
26.67 sv
43.33 nr
0.50
33.33
q-u
43.33
n-r
63.33
h-l
0.75
z
0.5
Mortality
%
Mixture 2
36.67
0.25
p-t
0.1
Mixture 1
0.50
0
Adults
Extract
0.25
0.35
0.50
0.75
0.07
0.25
0.35
0.50
0.75
33.33
q-u
43.33
n-r
60.00
i-m
70.00
g-j
20.00
u-x
40.00
o-s
50.00
l-p
66.67
g-k
73.33
f-i
19
In the present study, the toxicity of the
insecticide varied with the developmental stages
(larvae, pupae and adult) of confused flour
beetle. The insecticide was more toxic to larvae
than pupae and adults. This difference in
sensitivity to the insecticide could be mainly
attributed to the variation of the levels of the
insecticide metabolic enzymes.
This result is in line with Saidana et al.
(2007) as it was found that the petroleum ether
extracts for several types of plants were very
effective against larvae and adults of the
confused flour beetle, but the larvae were more
sensitive than the adults. Also, the present results
agreed with Soummane et al. (2011), they
investigated the effect of the methanol extract of
aerial parts of some medicinal plants on
Mediterranean fruit fly, Ceratitis capitata, it was
found that the extract was more toxic to larvae as
compared to adults.
By comparing the effect toxic effect of
insecticide with that of cardamom oil on the
developmental stages of the flour beetle, the
results clearly indicated that the oil extract was
more effective than the insecticide lambda.
Effect of combination cardamom oil extract
and Lambda on larvae, pupae and adults.
The results of the toxic effect of a
combination cardamom seed oil and Lambda
showed the presence of seed oil enhanced the
toxicity of the insecticide lambda on larvae,
pupae and adults of the confused flour beetle,
which was increased with increasing the
synergistic ratio.
The effect on 5th instar Larvae.
As shown in Figure 1,the highest percentage
of mortality rate (73.33%) was observed in
20
larvae exposed to mixture (1), ( 0.03µl extract
and 0.75 µl insecticide). The percentage of
mortality was with increasing the concentration
the oil extract. When the of oil extract (0.07 µl)
mixed with the four insecticide concentrations
(ranging 0.25-0.75 µl), the highest percentage of
mortality
(83.33%) was observed at an
insecticide concentration of (0.75 µl).
The effect on Pupae.
The highest mortality rate (56.67%) was
obtained with the mixture (1), ( which consists
of 0.03 µl seed oil extract and 0.75 µl Lambda).
There is a proportional relationship between the
percentage of mortality oil extract concentration.
When oil extract (0.07 µl) mixed with the four
insecticide concentrations (ranging 0.25-0.75
µl), the highest percentage of mortality (70.00%)
was observed at an insecticide concentration of
(0.75 µl).
The effect on Adults.
The highest percentage of mortality (53.33%)
was observed in adult insects exposed to mixture
(1), (0.03 µl extract and 0.75 µl Lambda). The
percentage of mortality was increased with
increase the concentration of the oil extract.
When oil extract (0.07 µl) mixed with the four
insecticide concentrations (ranging 0.25-0.75
µl), the highest mortality rate (73.33%) was
observed at an insecticide concentration of (0.75
µl).
Results in Table (2) showed that the
synergistic ratios were 2.01, 1.70, and 1.60 in the
larvae, pupae and adults, respectively. Figure 3
shows the synergistic ratios of insects exposed to
mixture (1), which consist of 0.03 µl /insect of
cardamom seeds oil extract combined with
various concentrations of Lambda.
3.5
Synergistic ratio
3
2.5
2
Mixture 1
1.5
Mixture 2
1
0.5
0
Larva
Pupa
Adult
stages
Figure (3) Synergistic effect in mixtures 1 and 2 on various developmental stages of confused flourr beetle
stages
Whereas, in mixture (2) the synergistic ratios were 2.61, 2.34, 3.10 larvae, pupae and adults,
respectively, Figures 4-6 shows the addition of 0.07 µl / insect of the cardamom seeds oil extract with
various concentrations of Lambda, showed increased mortality ratios as compared to insecticide
alone and mixture (1).
90
80
Mortality %
70
60
50
Lambda
40
Mixture 1
30
Mixture 2
20
10
0
٠.٢٥
٠.٣٥
٠.٥
٠.٧٥
Concentrations μl/ Larva
Figure (4): Effect of Lambda, Mixture 1 and Mixture 2 on the percentage of mortality of larvae of confused
flour beetle.
21
80
70
Mortality %
60
50
40
Lambda
30
Mixture 1
Mixture 2
20
10
0
٠.٢٥
٠.٣٥
٠.٥
٠.٧٥
Concentration μl/ pupa
Figure (5): Effect of Lambda, Mixture 1 and Mixture 2 on the percentage of mortality of Pupae of confused
flour beetle pupae.
80
70
Mortality %
60
50
40
Lambda
30
Mixture 1
Mixture 2
20
10
0
٠.٢٥
٠.٣٥
٠.٥
٠.٧٥
Concentrations μl/ Adult
Figure (6): Effect of Lambda, Mixture 1 and Mixture 2 on the percentage of mortality of adults of confused
flour beetles.
22
The synergistic effect of cardamom seeds oil
extract on the toxic effect of lambda may be
attribute to that these materials increases the
permeability of the pesticide through the cuticle
and facilitate its incoming to the target site, this
conclusion agrees with that of Sun and Johnson
(1972); Shufeng et al. (2005) whom suggested
that synergistic effect may be due to facilitating
the entry of the pesticide through cuticle.
On the other hand, Wilkinson (1976);
Michalets (1998) and Mckinnon et al., (2008)
have suggested another mechanism for the
synergistic effect by inhibition of function
oxidative enzymes which are responsible for the
degradation (metabolized) to pesticide.
This confirms the previous conclusion that
the variation of insects sensitive to pesticides is
attributed to primarily to the contrast enzymes
levels that metabolize the pesticide, This
conclusion is in agreement with those of
Cassida et al. (1966) and Franklin (1972) that the
percentage of inhibition ratio depends on the
synergistic
concentration.
Furthermore,
Khalequzzaman and Rumu (2010) studied the
effect of mixing cardamom essential oil with
prirmiphos-methyl pesticide (1/20) and they
showed a synergistic effect by increasing
percentage of mortality against adult cowpea
weevil.
Table (2) Synergistic ratios in both mixtures 1 and 2, and their mixing ratios
Mixture 1
Phases
Extract
Conc.
μl
Insecticide
Conc.
μl
Mixture 2
SR
Extract
Conc.
μl
Adults
Pupae
5th instar Larvae
0.25
0.35
2.01
0.07
0.50
0.75
0.75
0.25
0.25
0.35
1.70
0.07
0.50
0.03
2.61
0.50
0.03
0.35
2.34
0.50
0.75
٠.٧٥
0.25
0.25
0.35
SR
0.25
0.35
0.03
Insecticide Conc.
μl
0.35
1.60
0.07
3.10
0.50
0.50
0.75
0.75
23
Refrence
Abbasipour, H., M.; Mahoudvand, F. R.; and
Hosseinpour, M. H. (2011). Fumigant toxicity
and oviposition detergency of the essential oil
from cardamom, Ellettaria cardamomum,
against three stored product insets. Journal of
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26
ISSN: 2410-7549
Trichodina sp. AS BIOINDICATOR FOR EVALUATION OF BIOCHEMICAL
OXYGEN DEMAND (BOD5) IN AQUACULTURE FISH FARMS (PONDS)
1
Karwan Sallo Najm Al-Marjan 1 and Shamall Muhamad Amin Abdullah 2
Pharmacy Dept. Medical Technical Institute. Hawler Polytechnique University, Kurdistan Region – Iraq.
**
College of Agriculture. Salahaddin University, Kurdistan Region – Iraq.
Abstract:
The present study include the using of the prevalence of the fish infestation by the protozoan
Trichodina sp. as bioindicator for evolution of the biological oxygen demand BOD5 (lowering down of the
dissolved oxygen DO) from Ainkawa fish hatchery. For this purpose, two handered and fourty (240)
finger ling fishes of Cyprinus carpio were collected from six ponds (40 samples from each pond) and fishes
were examined from December, 2012 to the end of february, 2013. Biological oxygen demands were
measured by azid modification of Winkler method (through measuring of dissolved oxygen DO) for each
pond. Results reveal that there are a direct relationship between the prevalence of fish infestation by the
Trichdina sp. and the values of BOD5. The prevalence of fish infestation in each pond increased (57.5, 40,
27.5, 45, 15, and 42.5% respectively) with the increase of the values of BOD5 (9.2 , 8.0, 5.7, 8.1, 2.9 and 8.0
mg.l-1, respectively).
Key words: BOD5, Trichodina, Water pollution, Erbil.
Introduction
he concept of aquaculture development
as a potential source of food supply is
receiving much attention in Iraq. This has
resulted in rapid growth in fish farming practice.
The intensification of such fish culturing creates
disease
problems
that
originate
from
overcrowdness or deteriorating water quality,
such as unsuitable water temperature, pH,
dissolved oxygen, BOD5 and organic mater
(Dujin, 1973 and Kugel et al., 1990).
Trichodina (Ciliophora: Protozoa) are very
common ectoparasites which in most cases are
pathogenic to both freshwater and marine fish
(Wellborn, 1967). Dogiel (1961) reported that
trichodiniasis caused by Trichodina was
stimulated by water quality and the high density
of fish in ponds. It causes irritation by feeding
on the epithelial cells covering the surface of the
gills and skin of the fish, which can result in
hyperplasia (proliferation) of the epithelial cells,
clubbing and even fusion of the gill filaments.
This affects the abilities of both gills and the
skin to maintain optimal respiratory and
excretory activities, and the ability of the skin to
maintain proper homeostatic osmoregulatory
properties. Massive infestations of these
parasites on fish can also directly result in
superficial to deep ulcerative skin lesions which
allow for secondary bacterial and fungal
T
infections to develop at the affected site (Smith
and Schwarz, 2009).
In Iraq, The first information concerning
genus Trichodina was given by Shamsaddin et
al. (1971), who recorded T. domerguei on eight
species of fishes brought from different fish
markets in Baghdad city. For the next years a
total recorded number of Trichodina reached
eighteen species (Mhaisen, 2014).
In the present study, only Trichodina sp. were
selected because based on the previous study
which were done on the different fish farms in
Iraq and Kurdistan region appear that Trichodina
sp. make a large problem in all aquaculture fish
farm after Ichthyophthirius and Lernaea
cyprinacea Mama and Abdullah (2010) and AlMarjan and Abdullah (2009).
The aim of this study is to know the
relationship between fish infestation by
Trichodina sp. and the BOD5 as a step for
evaluation of the water quality of the fish farms.
Description of the Study Area
Ainkawa fish hatchery is located northwest of
Erbil city, Kurdistan region, Iraq. This project
was built in 2000 on 27 hectare, and started
working in 2005. In this region there are
eighteen ponds of different sizes, among these
six ponds were selected. Fishes of various sizes
were stocked for greater growth and artificial
reproduction (Fig. B).
27
Fig. (1): A-Map of Erbil, B-Studied stations.
This study includes two aspects, the first
deals with the isolation of the Trichodina sp.
which infect common carp Cyprinus carpio in
Ainkawa fish hatchery farms, situated northwest
of Erbil city, Kurdistan region-Iraq, for this
purpose 240 finger ling fishes were collected
from six ponds (40 samples from each pond) and
transported a live in a cool box containing tank
water (free from chloride) to the laboratory
during the period from December, 2012 to the
end of February, 2013. Each fish was examined
by taking smears from skin, fins, and buccal
cavity through slight scraping. The gills were
removed, and placed in a Petri dish containing
water then microscopically examined. Smears
were stained by Klein’s dry silver method for
observation of the adhesive disk as suggested by
Asmat (2001). All slides were examined under a
light compound microscope at 700X.
The second aspect included the measurement
of the biological oxygen demand (BOD5)
through measurement of dissolved oxygen based
on azide modification of Winkler method (Maiti,
2004). Samples of water were collected by a
special bottle (BOD bottle, 300ml capacity) from
each fish farm separately.
The prevalence of fish infestation by
Trichodina sp. in each pond were variable 57.5,
40, 27.5, 45, 15and 42.5% respectively with the
change of the BOD5 concentration in each
ponds 9.2 , 8.0 , 5.7 , 8.1 , 2.9 and 8.0 mg.l-1
respectively (Fig. 2).
The analysis of the results indicate that there
are a relation ship between the BOD5
concentration and the prevalence of fish
infestation by the Trichodina sp (Fig. 3). The
highest prevalence of the fish infestation by the
parasite (57.5%) and the highest concentration of
the BOD5 (9.2 mg.l-1) were recorded in the pond
number one. Depending on these data, we can
evaluate the quality of ponds water because high
BOD5 values is an indicator for poor quality of
water (Peter and Ludemann, 1972). On the other
hand low concentration of the BOD5 value (2.9
mg.l-1) and low prevalence of fish infestation by
Trichodina sp. (15%) was recorded in pond
number five.
Biological oxygen demand (BOD5) is
defined as the amount of oxygen required by
microorganisms in decomposing organic
materials in a sample under "aerobic condition at
20°C over a period of 5 days (Maiti, 2004).
28
High BOD5 values means presence of microorganisms (like Trichodina) in water and consumption
of small amount of oxygen, which may supported by fecal coliform results in lowering the amount of
dissolved oxygen and elevation of BOD5. Results of the present study agree with those of Othman,
(2013).
Several parasitologists and ecologists mentioned the presence of a direct relationship between the
concentration of organic materials and the prevalence of infection by the parasite especially
Trichodinid ciliates, because they can be used as potential bioindicator fish farms (Yeomans, 1997). A
strong relationship between ecological parameters and parasite infection were found from Lesser Zab
and Greater Zab rivers in north of Iraq (Abdullh, 2002). Seasonal dynamics of Trichodina sp. on
whiting (Merlangius merlangus) in relation to organic pollution on the eastern Black Sea coast of
Turkey was reported by Palm and Harry (2005).
Fig. (2): Shows percentage of infested fish by Trichodina sp. and values of biochemical oxygen demand (mg.l-1)
in the studied ponds.
Fig. (3): Shows relationship between percentages of infested fish by Trichodina sp. and values of biochemical
oxygen demand (mg.l-1) in the studied ponds.
29
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30
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31
ISSN: 2410-7549
DIAGNOSIS OF TOXOPLASMOSIS IN SHEEP USING SEROLOGICAL
(ELISA) AND MOLECULAR TECHNIQUE IN DUHOK GOVERNORATE KURDISTAN REGION
Farhad Buzo Mikaeel, Jassim Abdo and Lokman Taib Omer
Duhok Research Center, Faculty of veterinary Medicine, University of Duhok, Kurdistan Region – Iraq.
Abstract:
Toxoplasmosis is an important zoonotic diseases in human and animals. The disease caused by the
protozoan Toxoplasma gondii. It is an economically important disease of livestock, especially sheep and
goats. The present work aimed to diagnose toxoplasmosis in sheep using two methods, serological
(ELISA) and molecular tools (PCR) and comparing the serological data with the molecular results to
determine the sensitivity and the specifity of the molecular tools.
The study was carried out at Duhok Research Center, University of Duhok. Ninety sex whole blood
samples were collected from in aborted ewes in Duhok governorate during the period September 2013September 2014.The samples were collected from different localities including Aqra, Dohuk district,
Shikhan and Zakho
The serological tests showed that22(22.91 %) of the samples were positive from
examined sheep serum by ELISA, while 23(23.93%) by using PCR assay. PCR was performed on all DNA
of sheep blood samples to amplify B1 gene as a target sequence.
Good correlation between the results of PCR and ELISA were detected, there’s no statistically
significant difference, It can be concluded that ELISA combined with the PCR technique is a
recommended tool for accurate diagnosis of Toxoplasmosis but PCR is more specific for detection of T.
gondii with sensitivity of 95.45 % and specificity of 97.29%.When taking the ELISA as a reference test.
The results of PCR assay showed it’s important in the diagnosis of the carrier infected cases more than
ELISA techniques.
Keyword: Toxoplasmosis, Sheep, ELISA, PCR.
1. Introduction
Sheep are important to the economy of many
countries because they are a source of food for
humans. Sheep are commonly infected with the
protozoan parasite, Toxoplasma gondii infection
of sheep and goat poses a risk to public health,
as well as economic losses due to reproductive
failure (Dubey and Towel, 1986; Dubey and
kirkbride, 1990; Edward and Dubey, 2013). The
parasite was first described in 1954 (Buxton et
al., 2007). Ovine toxoplasmosis has a
worldwide distribution especially in temperate
sheep rearing countries where the climatic
conditions favor oocyst survival (Buxton and
Rodger, 2008). Toxoplasmosis is a major cause
of reproductive failure associated with abortion
in sheep and goat. In sheep, abortion or prenatal
mortality of lambs occur when ewes suffer a
primary infection during pregnancy (Tenter et
al., 2001).
Toxoplasmosis is diagnosed mainly by direct
smear, Immunohistochemistry, serology testing
and PCR (Mazumder et al., 1988; Ramadan et
al., 2007; Dubey, 2009, 2010)
Various ELISA methods using crude,
fractionated, or recombinant antigens have been
32
used to detect T. gondii antibodies in ovine sera.
Compared ELISA based on crude and
recombinant antigens found to have varying
degrees of specificity in naturally and
experimentally- infected sheep (CaballeroOrtega et al., 2008).
The ELISA assay for T. gondii antibodies has
been adapted for use in most domestic animals
including sheep and goat (Dubey, 2008; Dubey,
2009). There is specific ELISA assays for both
IgM and IgG subtypes. These ELISA assays are
ideally suited to screen large numbers of samples
and looking at the IgM/IgG ratio (Denmark and
Chessum, 1978). Serological analysis using
IFAT and ELISA has been widely employed in
order to detect herds infected by toxoplasma,
including swine and sheep flocks (Van der Puije
et al., 2000).
Moreover, Molecular assays such as PCR
make it possible to detect small quantities of
target DNA and potentially provide an
alternative sensitive diagnostic tool, specific
molecular diagnostics of toxoplasmosis is
generally based on the detection of a specific
DNA sequence, using different assays and
protocols, mostly from highly conserved regions
such as the B1 gene repeated 35 times in the
genome, 529 bp repetitive element with about
200-300 copies in the genome, ITS-1 (internal
transcribed spacer ) that exists in 110 copies and
18S rDNA gene sequences(Jones et al., 2000;
Habibi et al., 2012; Moazeni Jula et al., 2013;
Tavassoli et al., 2013). In general, this technique
has been proven as a useful method in diagnosis
of clinical toxoplasmosis (Dubey, 2008), it is
highly specific and sensitive and very useful
together with serological tests to differentiate the
chronic, acute or reactivated infections (Neil and
Lappin, 1991; Switaj et al 2005)
Early diagnosis of infection is of great
consequence for reducing the severity of the
disease and the risk of congenital toxoplasmosis
(Behbehani and Al-Karmi, 1980; Edward and
Dubey, 2013).
Few serological studies have been conducted
on the prevalence of T. gondii antibody in farm
animals in Kurdistan Region but no molecular
study for detection of parasite DNA have been
used. Therefore, the aims of the present study
were to estimate the prevalence of the T. gondii
antibodies in aborted ewe’s serum by using
serological tests (ELISA) and to compared with
the detection of parasite DNA using molecular
tools (PCR) in order to determine the sensitivity
and specifity of diagnostic techniques for the
diagnosis of Toxoplasmosis in sheep Duhok
Governorate,
2. Material and method:
The study was carried out in the Duhok
Research Center /Faculty of veterinary medicine,
university of Duhok. The sheep of this study
were of local breeding (Karadi), only female
aborted ewes were involved in this work.
2.1 Sampling:
Ninety sex whole blood samples were
collected from aborted ewes in Duhok
governorate during the period September 2013
to September 2014. The samples were collected
from different localities including Aqra, Duhok
district, Shikhan and Zakho.
The blood samples were collected from the
jugular veins of the sheep, each sample was
divided in to the two parts, one left few minutes
to obtain serum for serological tests, the other
part of the blood samples was collected in sterile
tubes with EDTA for PCR investigation. The
collected sera and blood samples were coded and
preserved at -20 °C until used.
2.2 Serological assay:
Sheep IgG antibodies against T. gondii were
tested using an enzyme-linked immunosorbent
assay (ELISA). The indirect Elisa (ID. VET.
Innovative diagnostics, France) was performed
using commercial kit. Optical densities (OD)
were read at 450 nm. The results were expressed
as the percentage of the mean absorbance values
of sample (S) to the mean absorbance value of
the positive (P) control sample provided with the
diagnostic kit. The resultant SP ratio was
expressed as a percentage (S/P %). According to
the manufacturer’s recommendation, sera with
S/P% ≤ 40% should be regarded as negative,
between 40 and 50% as doubtful, between 50%
≤ and < 200% as positive, and ≥ 200% as strong
positive
2.3 Polymerasechain reaction (PCR):
2.3.1 DNA extraction
Genomic DNA was extracted from blood
samples using a commercially available kit,
QIAamp DNA, blood and tissue kit (Qiagen,
Hilden, Germany). DNA extraction and
purification protocol was recommended by the
manufacturer.
Amplification was conducted in a total
volume of 25 μl. The reaction mixture contained
12.5 ul, 2X ready PCR mix (KAPA2G, PCR kit,
KAPA Biosystems; USA ) which consisted of
1.25 ul Taq-Pol, 75 mM Tris-HCL (pH 8.8),
1.5 mM MgCl2, and 0.2 mM of each dNTP. The
reaction mixture contained 12.5 ul master mix,
10 pmol of each forward and reverse primers
(Table 1), 1 ug DNA template, and 8.5 ul RNase
free water to a total volume of 25 ul.
2.3.2 Primer selection
The primer pairs used in the PCR experiment
were
TOX4
(CGCTGCAGGGAGGAAGACGAAAGTTG)
and
TOX5
(CGCTGCAGACACAGTGCATCTGGATT),
as described by ( Homan et al., 2000)
2.3.3 PCR technique
The amplification was carried out in a
thermal cycler (Ependrof, Germany) according
to the following program: an initial denaturation
step at 95°C for 7 min, followed by 35 cycles of
denaturation at 95°C for 30 sec, annealing,
primer at 55°C for 1 min, extension step at 72°C
for 45 sec. and a final extension step at 72°C for
5 min. Amplified PCR products were the PCR
products examined by electrophoresis in 1.5%
33
agarose gel, stained with ethidium bromide
solution, visualized under UV transilluminator
and photographed
Table 2. Relative sensitivity and specificity of PCR
in comparison with standard ELISA
ELISA
4. Results
In the results of the diagnosis of T.gondii
infection in sheep relied on the immunological
(ELISA) and molecular tests, as presented in
Table 1, ELISA detected anti- T. gondii
antibodies in 22 out of 96 sheep (22.91%),
whereas 23/ 96 (23.9%) of the samples were
positive by PCR. Two of the samples were
positive with PCR but they were negative with
ELISA assay and one sample was seropositive
but negative with PCR (Figure 1).
PCR
Positive
Negative
Positive
Negative
21(a)
1(c)
22
2(b)
72(d)
74
23
73
96
Sensitivity = a/ (a + c);95.45%
Specificity = d/ (b + d) 97.29
Kappa agreement *= 0.913
*
When two measurements agree by chance only,
kappa = 0. When the two measurements agree
perfectly, kappa = 1
5. Discussion:
(Fig. 1). Gel agarose electrophoresis (1.5%) for T.
gondii PCR products analysis M: 100 bp DNA size
marker Lane 1 :( control positive) T. gondii B1 gene
PCR products (529 bp) were amplified by external
primers. Lanes 2 is negative control and 3, 4,5,6,9 is
positive clinical samples for T. gondii B1 gene b, 8 is
negative clinical samples.
Table 1. Infection rate of T. gondii in examined
sheep using PCR, ELISA
ELISA
PCR
Positive
22(22.91%),
23(23.9%)
Negative
74(77%)
73(76%)
Total
96
96
Compared with ELISA The sensitivity and
specificity of PCR for detecting Toxoplasmosis
measured according to use ELISA test as
reference tools The sensitivity and specificity
were 95.45% , 97.29 respectively (Table 2).
Based upon the Kappa statistical test, a good
correlation 0.913(perfect agreement) between
the results of ELISA and PCR. Strength of
agreement based on ê was judged according to
the following guidelines: <0.2=slight; 0.2–
0.4=fair; 0.4– 0.6=moderate; 0.6–0.8=good;
>0.8=very good, and 0.8-1= perfect agreement.
34
Sheep represent an important source of meat,
milk and wool for humans in many countries,
and toxoplasmosis causes great economic losses
to sheep industry worldwide (Buxton et al.,
2007) The prevalence of T. gondii infection in
sheep and goats may be due to sheep free range
livestock associated with T. gondii infection due
to contamination of environment with oocytes.
The frequent presence of stray cats in a humid
rainy climate favor the survival of oocytes which
contributed to the Toxoplasma prevalence in
Kurdistan. The primary goal of this study was to
assess the risk of T. gondii infection to sheep in
Kurdistan and to use different tools for the
diagnosis of the parasite from aborted ewe. Out
of 96 samples 22(22.91%) were positive by
using ELISA while the 23(23.9%) samples were
positive with B1 gen using PCR two.
Two samples were positive with PCR but
negative with ELISA assay. The explanation is
that there are cases of infection with T. gondii
where serum levels of antibody are still not
enough to trigger ELISA-sensitive reactions,
ELISA is unable to detect the infection at early
stages of infection. Again, in acute phase of
infection T. gondii when death can happen in
only few days, the disease progression is too fast
to let immune system of animal to produce
ELISA-detectable levels of antibody (Nguyen et
al., 1996).One sample was seropositive but PCR
negative results can be explained by the fact that
antibodies may be present in the absent of
parasite or may be false positive reaction due to
the cross-reactivity, since several studies
suggested a closer relationship between T. gondii
sera and N. caninum (Nishikawa et al., 2002).
Detection of T. gondii DNA using PCR
minimizes the problems which the researcher
may face when using serological methods only
and facilitates the diagnosis in complex cases.
Compared to other countries, it has been found
that T. gondii seroprevalence in Duhok was
higher than that in Rahim Yar Khan (Punjab),
Pakistan (11.2%) and India (3.8%) (Ramzan et
al., 2009; Sharma et al., 2008), but lower than
the prevalence rate of toxoplasmosis in Turkey
and Iran which were (31%) for both (Taraneh et
al., 2006 and Sharif et al., 2014 respectively).
The differences could be related to differences in
ecological and geographical factors such as
temperature, rainfall or landscape differences.
The study area had overall low temperatures and
it generally thought that the prevalence and risk
of T. gondii infection decrease with decreasing
temperature because it affect the survival of
oocytes in the environment such as pastures.
In Iraq, the prevalence of the T. gondii in
sheep is higher in south than that of the north of
the country (Ali Akber et al., 2014) study
showed that the province of Babylon is the
highest in its seropositivity percentage in sheep
39.5%, followed by the province of Al-Anbar
(34.4%) and the province of Nineveh (30%) as
compared to the study of other Provinces. The
present study showed that T. gondii
seroprevalence in Duhok was lower than
Babylon, Al-Anbar and Nineveh respectively.
The differences could be due to the hot and
humid environment in south, higher prevalence
rate of toxoplasmosis in warm, moist areas
compared to those with colder and dry is
attributed to the longer viability of T. gondii
oocytes in moist or humid environments The
study area had overall low temperatures and it
generally thought that the prevalence and risk of
T. gondii infection decrease with decreasing
temperature because it affect the survival of
oocytes in the environment such as pastures.
PCR provides a simple and safe method for
accurate and early diagnosis of toxoplasmosis
(Tavassoli et al., 2009; Moazeni Jula et al.,
2013).
In General many study has been performed
using the serological and moleculare diagnosis
of T. gondii such as (Burg et al., 1989; AlSanjary and Hussein, 2012; El-Madawy and
Metawea 2013; Al-Abady et al., 2014), all those
studies agree with our conclusion that molecular
(PCR) is more sensitive and specific tools than
the serological tools for the detection of T.
gondii parasite . The sensitivity and the
specificity of PCR for detecting Toxoplasmosis
was measured according to ELISA test as a
reference test
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38
ISSN: 2410-7549
BIOASSAY AND PATHOGENICITY OF WHEAT SEED GALL NEMATODE
ANGUINA TRITICI
1
Sulaiman Naif Ami 1, Ibrahim Esa Taher 1, Fathi Abdulkarem Omer 2
Dept. Plant Protection, Faculty of Agriculture, University of Duhok, Kurdistan Region – Iraq.
2
Dept. Field crop, Faculty of Agriculture, University of Duhok, Kurdistan Region – Iraq.
Abstract :
Highest infection percentage recorded in Abo-graib and wahe CVs. (83.3%) followed by Smeto
(80.3%) while less infection percentage was in Rezgare and Al-Aez CVs. (33.3%). more galls (14 galls/
spike) formed in Sham-2 C.V. followed by Abo-graib, Smeto, Tammoz-2, Maxipak and Al-Aez
respectively. highest number of nematode (12773) occurred in large galls, while the lowest (8145) in small
galls. A. tritici population density increased dramatically with increase in galls size reached a peak (15112
second stage juveniles/gall) in big size gall. Results revealed that the vitality of second stage juveniles
reached its maximum level (8.87%) at 12 Cº with no significant differences with the validity percentage at
6 Cº while it reached its minimum level (1%) at 20 Cº.
KEYWORDS: Bioassay, pathogenicity, Wheat, Anguina tritici.
INTRODUCTION
heat (Triticum aestivum L.) is the most
economical important crops in the
world. 65% of produced wheat is consumed by
human, whereas 21% - 14% for animal feeding
and industry (Gooding and Davies, 1977).
According to USAID (2006), Iraq currently
consumes nearly 4 million tons of wheat
annually, yet only produces 500,000 tons of
qualified wheat. Thus more than 85% of
consumed wheat in Iraq imported, the current
average is 800 Kg/h. The Iraqi Kurdistan Region
produces about 50% of the wheat produced in
Iraq.
According to Bhatti,et al.,(1978) Ear- cockle
was the oldest reported disease of wheat which
was caused by wheat seed gall nematode
Anguina tritici, It is one of the major aerial
diseases and causes sustainable losses in wheat
crop of tropical and sub-tropical countries (
Kort,1972 ). It is present wherever wheat is
grown and this pest still common in Eastern
Europe and in part of Asia and Africa (Agrios,
2005). From the first record (1921) of this
nematode in Iraq (Rao, 1921), A. tritici remain
the major pest in Iraq occurred in the most wheat
growing areas by 22.9 to 45% on mexipac c.v. of
Wheat.(Al-Beldaw; et.al.1974) increased to 75%
on the same cultivar in Duhok Province in 1989
(Stephan and Antoon,1990), Ami, et.al.,( 2004)
reported that the percentage of infestation by
galls reached its maximum value (50%) in bread
wheat in Bashika – Kurdistan region of Iraq.
Ear- cockle disease reduces market price and
human consumption of wheat (Paruthi and
W
Bhatti, 1988), with significant reduction in the
protein and gluten contents of the flour product
of infested wheat with seed galls (Mustafa,
2009). This study aimed to test vitality of 2nd
stage juveniles of A. tritici under different
temperature and it’s pathogenicity toward some
common wheat cultivars in addition to its effect
on quality of wheat flour.
Materials & Methods:
Seed galls (ear-cockle) collection: Galls used in
this experiment were collected handmade from
infested bread wheat samples brought from
Faydia Silo.
Soil sterilization:
Soil was sterilized by Formalin 1% (1liter/m³) after sieving to discard stones and soil lumps,
and then placed on thick layer of polyethylene.
Formalin was mixed thoroughly with soil which
was covered by other pieces of polyethylene and
closed tightly to prevent any escape of formalin,
after 48 hours soil aerated for a week to remove
residual
of
Formalin
(Mustafee
and
Chattopadhyay, 1981).
Physical and chemical analysis of soil:
a- Soil texture: it was determined by hydrometer
method to evaluate percentage contents of clay,
silt and sand (Jaiswal, 2003).
b- Electrical conductivity (E.C) was estimated
by using Conductivity Bridge of soil extract of
1:1.( Dipak and Haldar, 2005).
C- Percentage of organic matter: was measured
in the soil by wet digestion method with
39
concentrated sulphuric acid according to (Black,
1965).
d-Soil pH by pH meter in the soil extract of 1:1
(Dipak and Haldar, 2005).
Susceptibility of wheat cultivar: nine wheat
cultivars (Abo-qraib, Aba 95, Tammoz-2,
Maxipak, Rezgare, Al-Aez, Wahe, Smeto, and
Sham-2) were sown at the rate of 5 seeds/ pots
(20 cm) in diameter containing sterilized soil
which was infested directly with five galls, in
1cm deep holes between seeds at the rate of one
gall/hole, Pots were plunged randomly in the
trenches. This experiment consisted of 18
treatments (9 wheat C.V. X 2 levels of
infestation) performed as factorial experiment in
complete Randomized design (CRD) with 3
replication for each treatment, carried out in
2014 growing season, in one fields of the
College of Agriculture, University of Duhok ,
Pots were irrigated when needed and the
following infection criteria were calculated :
1- Infection percentage according to the following equation:Number of Infected plants
Infection percentage =
× 100
Total number of plants in each pot
2-Number of galls/spike.
3-Number of seeds/spike.
4- Reduction percentage of seed number /infected spike according to the following:Seed number /spike in control treatment – seed number/spike in infested soil
×100
Seed number in control treatment
Effect of size and weight of galls on nematode
population: Galls were divided according to
their size to (Small, Moderate and large) and
weight [18.3 milligram (19.6-16.5), 11.5mg
(12.7-11.3) and 3.6mg (4, 1-3.1)]. Each galls
was opened in 6.5 Cm diameter Petri-dishes
filled with 2ml of distilled water (D.W) with the
aid of two needles then transferred to a 250 ml
beaker, volume was adjusted by adding D.W
water and number of juveniles were calculated
by the aid of counting dish and
stereomicroscope.
Effect of different temperature on vitality of
second stage juveniles: Nine Petri dishes were
divided into 3 groups and 2ml of nematode
suspension (200 second stage juveniles) was
transferred into each Petri dish and after
covering 1st and 2nd group placed in refrigerator
at 6, 12 C◦ respectively while the third group left
40
at laboratory conditions (20 c◦). Percentage of
nematode vitality was calculated according to
the following equation:Percentage of nematode vitality = (number of
moving nematodes in each replicate ÷ Total
number of nematode in the same replication) X
100. This experiment consists of 5 treatment and
run out as CRD.
Effect of different temperature on the
emergence of 2nd stage juveniles from galls:
Four different temperatures including (5, 10, 15
and 20 C◦) were selected to test their effect on
juveniles emergence from their galls which were
placed in Petri dish contained a small amount of
moist soil at the rate of one gall / petri dish.
Examining of galls was done after each 48 hrs.
for the period of 20 days to determine the
emergence of juveniles.
Results and Discussion:
1- Physical and chemical analysis of soil:
Results of soil analysis(Table,1) revealed that the texture was sandy loam which is suitable for
nematode movement as well as chemical characters involving soil salinity, organic matter and soil pH
were suitable for most plant parasitic nematode (Ismail, 1998).
Table (1): Physical and chemical analysis of soil.
Clay %
Silt %
Sand
%
17.13
23.23
59.65
Texture
Sandy loam
pH
Organic
matter
Electric
conductivity
(salinity)
8.13
1,38
0.52
Infection percentage %
2- Susceptibility of wheat cultivars:
All wheat cultivars used in this trial were infected by Anguina tritici with some significant
differences in their susceptibility. Highest infection percentage recorded in each of Abo-ghraib and
wahe CVs. (83.3%) followed by Smeto (80.3%) while less infection percentage was in Rezgare and
Al-Aez CVs. (33.3%) (Fig.1) which means there were differences in susceptibility of those cultivars
to ear-Cockle disease and also in ability of Anguina tritici to infect those cultivars.
100
80
60
40
20
0
83.33 a
83.33 a 80.33 a
76.67 a
42 b
66.67 a
41.67 b
33.33 b 33.33 b
Figure (1): Infection percentage Anguina tritici with on different wheat cultivars.
Number of galls /spike
Figure,2 illustrates that more galls (14 galls/ spike) formed in Sham-2 C.V. followed by Abograib, Smeto, Tammoz-2, Maxipak and Al-Aez respectively with significant differences between same
cvs. No galls appeared on Abae 95 and Rezgare while both showed symptom of infection, this
phenomenon indicated that A. tritici can’t completed its life cycle after climbing of seedlings, which
might be attributed that cultivar failed to supply adequate essential nutrients for nematode juveniles
(Taher, 2012).
16
14
12
10
8
6
4
2
0
14 a
10 b
7c
3d
0g
2 de
2 de
0g
0.66 ef
Figure (2): Number of galls / spike of different wheat cultivars
41
reduction percentage of
seed/spike( %)
Differences in gall number/plant are also due to host genotypes and their susceptibility to A. tritici
(Hamood and Fattah, 1989). Ear-cockle disease caused reduction in seed production in all tested
cultivars. More reduction percentage which were 93.23, 86.93 and 86.93% was recorded from
Rezgare, Abae 95 and Tammoz-2 respectively while the lowest were 36.67 and 38.87% recorded from
Smeto and Al-Aez, the same results were recorded in different wheat cultivars according to the extent
of their susceptibility (Al-Beldawi et al., 1977; Fattah, 1988; Saleh and Fattah, 1990; Stephan et al.,
2000; Mustafa 2009; Taher, 2012).
100
90
80
70
60
50
40
30
20
10
0
86.57
ab
86.93
ab
93.23 a
74.97
abc
65.1
a-d
57.2
bcd
49.97
cd
38.87 d
36.67 d
Figure (3): Reduction percentage of seeds/spike
3- Effect of gall size and weight on nematode population density:
Results indicate significant effect of gall size on the nematode population density, thus highest
number of nematode(12773) occurred in large galls, while the lowest (8145) in small galls (Table2).
This result consistent with finding of Ami, et al, (2004); Esser et al., 1991 and Taher, (2012).
Table (2): Effect of galls size on the population density of Anguine tritici.
Galls size
Nematode population density
(Nematode/ gall)
Large
12773 a ( 10114 –13881)
Moderate
11919 a ( 9766 – 12088 )
Small
8145 b ( 7255 – 9617 )
Number of juveniles/gall depend on many factors such as wheat cultivar, gall size, environment
conditions and soil biosphere characters in addition to their behavior in different geographical regions
(Taher, 2012). Weight of galls also has noticeable effect on A. tritici population density which
increasing dramatically with increase in galls size and reached a peak (15112 nematodes/gall) in big
size gall, while number decreased to lowest level (3978 nematode/gall) in small size (table, 3) which
means reduction in nematode population by 73.7%.
42
Table (3): Effect of galls weight on the population density of Anguine tritici.
Galls weight
Nematode population
density (Nematode/ gall)
18.3(19.6-16.5)
15112 a
11.5 (12.7-11.3)
9440 b
3.6 (4.1-3.1)
3978 c
Regression analysis and correlation coefficient (Fig. 4) demonstrate positive relation between galls
weight and nematode population density. From regression equation it was found that an increase of
one unit (one mg of galls) in used galls followed by increasing in nematode population density by
803.19 with high determining factor, meaning that it can rely on this equation by 90.7%. Increase in
nematode population density in big size galls and heavier ones might be due to the presence of more
space in large galls to contain more juveniles during penetration of ovary wall (Ami, et al, 2004) on
the other hand more weight and size of newly formed embryo supply more nutrient for feeding of
juvenile and increase the ability of female to laying more eggs, otherwise big and heavey galls may
belong to durum wheat Cultivars which are always bigger and have more protein contain that have
effect in female fertility as we mentioned before.
Nematode population
density / Gall
Y= 803.19X + 371.041
R2 = 90.7
20000
r = 0.952
10000
0
٣
٦
١١
Galls wieght (mg)
١٨
Figure (4): Linear relation, regression equation and correction coefficient between gall weight and nematode
population density.
4- Effect of different temperature on vitality of second stage juveniles:
The results of revealed that the vitality of second stage juveniles reached its maximum level
(8.87%) at 12 Cº with no significant differences with the validity percentage at 6 Cº while it reached
its minimum level (1%) at 20 Cº as well as the vitality percentage increased slowly by the time and
stretch 8.33% after 12 days from the time of gall opening with no significant differences with vitality
percentage after 9 days (Table 4). On the other hand the interaction between temperature degrees and
examining time showed its noticeable effect on vitality. In general maximum level of vitality was
(13%) recorded after 12 days at 12 Cº with no significant differences with vitality after 9 days at the
same temperature and with that after 9 and 12 at 6Cº while none of the juveniles moved after 3 days at
20 Cº. There are many studies on effect of planting date on infection and all are found that there are
differences in infection behaviors of juveniles ( Mustafa,2009; Radjender et al., 2009; Taher, 2012)
and most important factor in delay or early planting are temperature that influence activity of
juveniles. The sudden high temperature may not be a catalyst to re-vitality of juveniles as they were in
dormancy stage or cryptobiosis, in fact this study needs further confirmation and granting juveniles
more time to check their vitality,
43
Table 4: Effect of different temperature degrees on the percentage of vitality of second stage juveniles of A.
tritici.
Percentage of vitality (%)
1
2nd
3rd
4th
Reading
Reading
Reading
Reading
1.5 c
3c
10 a
10 a
4c
7.5 bc
11 a
13 a
0.0 c
0.5 c
1.5 c
2c
1.83 b
3.67 b
7.5 a
8.33 a
st
Temperature
6 Cº
12 Cº
20 Cº
Effect of reading
Effect of temperature
degree
8.63 a
8.87 a
1b
5- Effect of different temperature on the emergence of 2nd stage juveniles from galls: Results
illustrated that the juveniles emerged from galls after 20, 16, and 14 days at 5, 10, 15 and 20 Cº
respectively with the presence of moisture (Table 5)
Table5 : Effect of different temperature on the emergence of 2nd stage juveniles of A. tritici from the galls.
Period of time in days after which juveniles emergence ware examined
Temperature
2
4
6
8
10
12
14
16
5
10
+
15
+
20
+
+ = period after which juvenile exited from galls. - = juveniles were not exited
-which indicates that the juveniles need a
longer period of time for emergence at low
temperature that means their vitality increased
with increasing of temperature that stimulated
juveniles for getting out from galls. That means
that the temperature plays an important role in
emergence of second stage juveniles.
References
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H. S, Saharan (2009). Effect of sowing date
on the incidence of loose smut, flag smut and
seed gall ofwheat. Bangladesh J. of Agril. Res.
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of Iraq .Dept. of Agric.Iraq, Bull.7.
Saleh, H. M. and F. A, Fattah (1990). Studies on the
wheat seed gall nematode. Nemaol. Med. 18:
59-62.
Stephan Z. A.; M. S, Hassan.; A. I, Hammadi and
Antoon, B. G. (2000). New race of wheat gall
nematode Anguina tritici and susceptibility of
some cultivars to this race . Iraqi J. of Agric .5
:1-5.
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45
ISSN: 2410-7549
Characterization of Five Microsatellite Markers for Genetic Diversity Structure
Analysis of Walnut (Juglans regia L) in Five Village in Duhok Province
Awat Mustafa Abbas Barwary
Department of Biology, Faculty of Sciences, University of Zakho, Kurdistan - Region, Iraq.
Abstract:
Five microsatellites markers (WGA202, WGA009, WGA332, WGA225 and WGA069) were used to
characterize Persian walnuts (Juglans regia L.) populations in five villages (Sharanesh, Bedohe,
Kanizarke (Akre), Kashane and Kuzo) of Duhok province. The microsatellites amplified (PCR products) a
total of 186 alleles across all populations. The number of alleles per locus ranged from 4 alleles in
Sharanesh and Kashane populations to 10 alleles also in Sharanesh and Kashane populations, with an
average of 7.4. The molecular sizes of the amplified bands ranged from 158 bp to 289 bp in all
populations. The observed heterozygosity (Ho) within populations ranged from 0.59 at WGA225 to 0.65 in
the locus WGA202 with an average of 0.61. The PIC value (0.88) indicated that all markers were highly
informative and useful for genetic diversity studies in these populations. The proportion of genetic
variation presented among populations accounted for 8.4% of the total genetic diversity so it indicated a
moderate level of genetic diversity between populations. The Fis average 0.24 indicated that, there was a
regular tendency toward heterozygote deficiency and indicated the presence of inbreeding within the
populations. The phylogenetic analysis or unrooted neighbor-joining tree highlighted the genetic distance
among those five populations and separated them into two main groups placing each population according
to its genetic background. The first group consists of populations Bedohe and Sharanesh in one subgroup
and Kashane in second subgroup. The second group consists of populations Akre and Kuzo .
Keywords: genetic diversity, Duhok, microsatellites ,walnut and population.
Introduction
he genus Juglans L. is made up of 21
species of longlived deciduous trees that
produce large woody-shelled nuts. Juglans has
been divided into four taxonomic sections as
described by Manning (1978): (i) section
Cardiocaryon Dode (the east Asian heartnuts)
with three species, J. ailantifolia Carr., J.
mandshurica Maxim., and J. cathayensis Dode;
(ii) section Rhysocaryon Dode (black walnuts)
of North, Central, and South America and the
West Indies with 16 species; (iii) section
Dioscaryon Dode with one species, J. regia L.
(Persian walnut) distributed from southeastern
Europe to the Himalayan mountains; and (iv)
section Trachycaryon Dode ex Mann. with one
species, J. cinerea L. (butternut) in eastern North
America. Juglans regia L., has an exceptionally
wide natural distribution, it occurs from the
Carpathian Mountains of Eastern Europe, all
through Western Asia, the Himalayan regions of
Pakistan, India, Nepal, Bhutan and east into
China. Most of walnut populations in Kurdistan
including Duhok province have had very little
attention from scientists, therefore they are not
known if they are native trees from openpollinated seedlings or new varieties introduced
to the area by the local people for many years.
T
46
Unfortunately, the traditional methods for
characterization and assessment of genetic
variability in perennial fruit crop species, based
on
morphological,
physiological
and
biochemical studies are both time consuming
and affected by the environment. The
introduction of molecular biology techniques,
such as DNA based markers, provides an
opportunity for genetic characterization that
allows direct comparison of different genetic
material
independent
of
environmental
influences (Weising et al. 1995). Previously, in
addition to morphological identification (Zenelli
et al. 2005), various biochemical and molecular
markers have been used for genetic
characterization of walnut genotypes. These
included isozymes (Ninot and Aleta 2003; Vyas
et al. 2003; Fornari et al. 2001; Busov et al.
2002),
restriction
fragment
length
polymorphisms (RFLPs) (Fjellstrom and Parfitt
1995), randomly amplified polymorphic DNAs
(RAPDs) (Nicese et al.1998; Yan-Min et al.
2000; Li et al. 2007), intersimple sequence
repeats ISSRs) (Potter et al. 2002), simple
sequence repeats (SSRs) (Woeste et al. 2002;
Dangl et al. 2005; Foroni et al. 2005; Victory et
al. 2006; Robichaud et al 2006; Karimi et al.
2010),
amplified
fragment
length
polymorphisms (Kafkas et al. 2005; Bayazit et
al. 2007) and SNPs (Ciarmiello et al. 2011).
Simple sequence repeat (SSR), an array of
short motifs of 1– 6 bp in length, are
hypervariable and widely spread in both coding
and non-coding regions of plant and animal
genomes (Kota et al., 2003). The reproducibility,
multiallelicism,
codominance,
relatively
abundance, and good genome coverage of SSR
markers have made them one of the most useful
tools for integration of the genetic,
physiological, and sequence-based physical
maps in plant species (Powell et al. 1996 and
Kota et al. 2003).
Sample collection: Samples (fresh leaves) from
five populations of J. regia L in the Duhok
province were collected from
Bedohe
(Kanimase), kanizarke (Akre), kuzo (Zawita) ,
Kashane (Batifa) and Sharaneshe (Darkare).
Figure (1). Samples were collected according to
procedure described by Karimi et al. (2010).
Plants within 10 km from each other were
considered as belonging to the same population,
we limited our sampling to 8 trees per
population and each tree separated by a distance
of 100m from the other one. The leaves were
sampled in the early morning hours and stored at
0
–80 C until their use.
Figure (1): Names and location of five populations of J. regia L in the Duhok province. The red circle
represents location of population.
Isolation of plant genomic DNA: The DNA was isolated from the plant (leaves) according to the
method Dolye and Dolye (1987) with some modification. Three grams of fresh tissue (leaves) were
0
homogenized to powder and added to 10 ml of Extraction buffer (2x CTAB preheated at 65 C for 15
min) [2% cetyl trimethyl ammonium bromide, 50 mM 1,4-dithiothreitol, 0.3% b-mercaptoethanol,1.4
M NaCl, 100mM Tris ,20 mM ethylenediaminetetraaceticacid (EDTA), pH 8.0] and incubated at 65
0
C for40 min. The aqueous solution was extracted with 10 mL (24:1) of chloroform–isoamyl alcohol
0
and centrifuged at 5 C for 10min at 10,000 r/min, and the aqueous layer retained; 2/3 volumes of the
0
aqueous layer of 95% ethanol were added (at –20 C ) to precipitate the nucleic acids. The precipitate
was washed with 0.2 M ammonium acetate in 75% ethanol and air-dried for 5 min. The DNA was
precipitated, washed, dried, and dissolved in TE buffer.
Polymerase chain reaction amplification and electrophoresis. Samples (leaves) were obtained
from five walnut populations of Duhok province. Each population contained eight individuals walnut
tree leaves. Five primer pairs introduced by Woeste et al., 2002 were used to amplify genomic
47
DNA.(Table 1) The samples were prepared for PCR amplification in the form of single-individual
genotypes for detection genetic diversity within populations.
The SSR reaction protocol was based on the protocol of Dangle et al. (2005). Amplification
reactions were performed in a volume of 15 mL containing 1 x polymerase chain reaction (PCR)
buffer, 25 ng genomic DNA, 200 mMdNTPs, 0.2 mM of each primer, 2 mM MgCl2, and 0.5 unit of
Taq DNA polymerase For DNA amplifications, an Eppendorf thermocycler was programmed
0
0
according to the following profile: 94 C for 5min, 30 cycles at 94 C for 30 sec, the appropriate
0
0
primer annealing temperature (Table 1) for 1 min and 72 C for 40 sec; and a final elongation at 72 C
for 2 min, one cycle. After amplification, 5 mL of each sample was loaded and electrophoresed on a
2% horizontal agarose gel to control for positive amplification and to determine the approximate
amount of product. Then, 3 μl of each sample was electrophoresed on a 6% polyacrylamide gel
containing 1x TBE buffer. After electrophoresis, the gel was silver stained using the procedure of
Bassam et al. (1991). In all cases, PCR reactions were performed at least twice to ensure that allele
sizes were consistent. Allele sizing and scoring was done using a 100 bp DNA ladder (MBIFermentas, Vilnius, Lithuania) as the length reference.
Table (1) Five walnut microsatellite primers
Primers
name
Annealing
temperatures
1
WGA202
58 C
2
WGA009
50 C
3
WGA332
59 C
4
WGA225
59C
5
WGA069
45 C
0
0
0
0
0
Amplicon
size range
(bp)
Primer sequence
246-289 bp
F. CCCATCTACCGTTGCACTTT
R. GCTGGTGGTTCTATCATGGG
242-258 bp
212-230 bp
198-206 bp
158-182 bp
F.
R.
F.
R.
F.
R.
FR-
CATCAAAGCAAGCAATGGG
CCATTGCTCTGTGATTGGG
ACGTCGTTCTGCACTCCTCT
GCCACAGGAACGAGTGCT
AATCCCTCTCCTGGGCAG
TGTTCCACTGACCACTTCCA
TTAGTTAGCAAACCCACCCG
AGATGCACAGACCAACCCTC
Data analysis. The allele's data were entered in the form of single-individual genotypes. The
following parameters of genetic variation were assessed for each population: number of observed
alleles (No), observed heterozygosity (Ho), and expected heterozygosity (He) (Nei, 1987), inbreeding
coefficient (F-statistic) Fst and Fis (Nei, 1987) were calculated at each locus and over all loci using
POPGENE version 1.32 software . Polymorphic information content (PIC), availability (A), and
genetic distance (Nei, 1972) were calculated by using Power Marker version 3.25 software.Finally, in
order to visualize the relationships between provenances, and obtained tree with neighbor-joining
method based on (Nei, 1972). The phylogenetic tree was constructed by using Power Marker version
3.25 software. The tree was then viewed by using the TREEVEIW version 1.66 software.
The total number of alleles scored in five populations was 186 alleles with the sizes between 158
bp to 289 bp. The number of alleles per locus varied from 4 alleles in both Sharanesh and Kashane
populations at locus (WGA225) to 10 alleles in Sharanesh population at locus (WGA202) and
Kashane population at locus (WGA069), with an average of 7.4 alleles per locus. Table (2)
The average number of alleles per locus obtained (7.4) showed to be much higher than some other
reported studies on Juglans regia using different DNA markers. In a very recent report using SSR
markers this value were found to be 5.5 and 6.2 in (Foroni et al., 2005 and Pollegioni et al., 2011)
respectively. In other studies, however, the average of allelic variation was higher than the average
obtained in this study. For example: Victory et al (2006) using SSR marker reported the allelic
variation value as 9.13, whereas in another study using SSR marker, the average of allelic variation
was 12.9 (Pollegioni et al., 2009).
48
Table (2): A summary of the number of alleles per each population revealed by five Juglans microsatellite loci
Number of observed alleles (No) in each
population
Sharanesh
Bedohe
Akre
Kashane
Kuzo
Average number of
allele
WGA202
WGA009
WGA332
WGA225
WGA069
10
7
8
4
9
6
8
7
6
8
9
8
6
6
9
9
6
7
4
10
7
8
9
6
9
41
37
37
26
45
Sub total
38
35
38
36
39
37.2
Average
7.6
7.0
7.6
7.2
7.8
7.4
Primers
Total
186
In order to obtain reliable data analysis, the value of availability (A, number of observed alleles per
number of individuals sampled) was determined. This value was found to be high in all populations
with an average 0.91. This average indicated that the number of null allele (not amplified) was low in
all populations with an average 0.19 Table (3).
The observed heterozygosity (Ho) value per each population ranged from 0.58 in Sharanesh
population to 0.628 in Akre population with an average 0.61(Table 3). The (Ho) of a given locus
ranged from 0.59 (WGA225) to 0.65 (WGA202), with an average over all eight loci of 0.61 (Table 4).
The average expected heterozygosity (He) was 0.87 per each population and ranged from 0.84 to
0.889 in Sharanesh and Kuzo respectively (Table 3) .The value of (He) per locus also varied from 0.81
at WGA225 to 0.96 at WGA202 with an average 0.90 (Table 4) .
Table (3) Summary of Genic Variation Statistics for five Juglans population (Sharanesh, Bedohe,
Akre, Kashane and Kuzo). Availability (A), expected heterozygosity (He), observed heterozygosity
(Ho), and Polymorphic information content (PIC).
Populations
A
Ho
He
PIC
Sharanesh
Bedohe
Akre
Kashane
Kuzo
0.925
0.925
0.900
0.825
0.975
0.5893
0.6214
0.6286
0.6048
0.6179
0.8463
0.8862
0.8872
0.8722
0.8897
0.766
0.802
0.803
0.779
0.811
Average
0.910
0.6124
0.8763
0.792
The Ho in J. regia per each locus in this study was found to be higher than the average Ho (0.54)
reported in some population in the Veneto region in Northern Italy (Pollegioni et al., 2009), whereas
this value were close to the average Ho (0.69) reported in five population in Dohuk providence of
Kurdistan (Abbas and Jubrael., 2013), but lower than those reported (0.72) in the four provinces of
Iran populations (Ebrahimi et al., 2011). From this result, it may be noticed that, the observed
heterozygosity value was lower than expected, this may suggest that, there is a regular tendency
toward heterozygote deficiency, and this may also indicate the presence of inbreeding within the
populations (Coulson et al.,1998).
49
Table (4) Summary of Genic Variation Statistics for five Juglans microsatellite Loci. Availability (A), expected
heterozygosity (He), observed heterozygosity (Ho) and Polymorphic information content (PIC).
Marker
A
Ho
He
PIC
WGA202
WGA009
WGA332
WGA225
WGA069
0.95
0.85
0.95
0.925
0.875
0.6579
0.6176
0.6053
0.5946
0.6000
0.9618
0.8973
0.9098
0.8153
0.9358
0.946
0.873
0.889
0.779
0.915
Average
0.91
0.6151
0.9040
0.881
The highest polymorphic information content
(PIC) value (0.81) was observed in Kuzo
population and the lowest value (0.77) in
Sharanesh population with an average 0.79 in
each population. Tables (3) The highest PIC
value per locus was (0.94) for the primer
WGA202 and the lowest value was 0.779 for the
primer WGA225 with an average 0.88 Table (4).
This may suggest that all markers were highly
informative; because the (PIC) value was higher
than 0.50 in all loci, thus, they could be
considered as useful markers for genetic
diversity studies for J. regia populations grown
in this region (Botstein et al., 1980).
The average PIC value per locus in this study
was found to be higher than values (0.68, and
0.57) reported in (Karimi et al,. 2010 and
Pollegioni et al., 2011) respectively.
The Fst ranged from 0.05 for locus WGA069
to 0.11 in locus WGA225 with an average value
of 0.084 Tables (5). This may suggest that the
genetic differentiation in this study was a
moderate one because the Fst value was higher
than 0.05 and lower than 0.15 (Weir and
Cockerham, 1984).
Table (5) Summary of (Fst and Fis) for five Juglans
microsatellite Loci
Locus
Fst
Fis
WGA202
WGA009
WGA332
WGA225
WGA069
0.1095
0.0660
0.0788
0.1103
0.0579
0.2219
0.2560
0.2693
0.1675
0.3172
Average
0.0842
0.2492
This average value was found to be higher
than the genetic differentiation among
populations reported in other studies for J. regia
based on microsatellite markers. For example,(
50
Pollegioni et al 2011) reported the value of Fst
(0.053) in Veneto region at Northern Italy. But
this average was lower than Fst (0.122) reported
in some population in western Iran ( Karimi et
al .,2010) and Fst ( 0.106) reported by Fornari et
al (2001) who had analyzed different population
of J. regia from Europe and USA .The average
of Fst obtained here 0.084, indicated that 8.4% of
the total genetic diversity existed among
populations and 91.6% within populations.
The estimation of genetic structure in this
study was indicated that the moderate level of
genetic differentiation was observed among
these five populations according to Fst values.
This may be due to that forest trees generally
display high within-population diversity and low
differentiation among populations. Deforestation
and/or progressive selection of valuable
genotypes for nut production and removal of
vigorous trees with high wood quality leading to
the negatively affecting genetic variability
(Hamrick et al. 1992 and Müller-Starck et al.
1992).
The Fis value in this study ranged from
(0.167) for locus WGA225 to (0.317) in locus
WGA069 with an average (0.249) per each
locus. These results indicated that all five loci
have the tendency toward heterozygote
deficiency.
The average of Fis value obtained was
higher than
(0.199)
reported in some
populations in Central and Southwestern China
(Wang et al .,2008) and (0.021) reported by
Pollegioni et al (2011). The value of Fis
indicated that, there was a regular tendency
toward heterozygosity deficiency, this indicating
that the presence of inbreeding within the
populations. The genetic structure information
that obtained in this study was essential for
conservation and management of walnut tree.
Genetic distances were calculated for each
pair of populations to estimate the extent of their
divergence Table (6). The lowest genetic
distance (0.55) was found between populations
Bedohe and Sharanesh and the highest genetic
distance (1.02) was found between populations
Sharanesh and Kuzo. In the five populations, the
average genetic distance among populations
equaled 0.87.
The values of genetic distances among
populations as well as between individuals are
vary for plant breeding programs, for example
plant breeders usually selects two varieties with
a high genetic distance between them in order to
obtain the widest possible crosses. However in
other cases if the breeder wishes to introduce a
certain character which is controlled by a gene or
a group of genes for specific variety without
wide variation in the genetic material of this
variety which contains desired characters. In this
case the breeder selects close variety that has
this character within a studied variety because it
is very difficult to find genetic variation between
varieties depending on morphological characters
(Smith, 1992).
Table (6) genetic distance between five Juglans regia populations (Sharanesh, Bedohe, Akre, Kashane and
Kuzo)
Populations
Sharanesh
Beduhe
Sharanesh
00000
0.5556
Beduhe
Akre
Kashane
Akre
0.6178
0.7508
00000
Kashane
0.5970
0.8007
0.6923
00000
Kuzo
1.0234
0.7045
0.6652
0.7587
Kuzo
00000
00000
The dendrogram separated the five populations into two main groups. The first group consists of
populations Bedohe , Sharanesh and Kashane . The second group consists of populations Akre and
Kuzo . The first group divided into two subgroup, Beduhe and Sharanesh were in one subgroup and
Kashane in another subgroup. In the first cluster or group the Sharanesh and Bedohe populations had
closest genetic relationship together then the second subgroup consist of Kashane population as
detected in Figure (2). Also in second group Akre and Kuzo had close genetic relationship together.
The phylogenetic analysis highlighted the genetic distance among the populations, placing each
according to its genetic background. These results confirmed the power of microsatellites to diversify
Juglans regia in five populations.
The unrooted tree determined in this study observed that the populations separated one from
another according to their genetic differentiation that present among them and not according to the
geographical distribution. This may be attributed to the fact that all populations belong to Duhok
province and the area or geographical distance between populations is not very wide to produce high
amount of genetic differentiation
among populations. In addition , the habitat of
the
populations are approximately similar including climate, temperature, rainfall , landform whereas
suitable for growth of walnut tree in this region. (Abbas and Jubrael., 2013)
Figure (2): Unrooted neighbor-joining tree representing the genetic relationships between five Juglans regia
populations (Sharanesh, Bedohe, Akre, Kashane and Kuzo
51
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@@
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54
ISSN: 2410-7549
IN VITRO MICROPROPAGATION OF VITIS VINIFERA L. IN KURDISTAN
REGION OF IRAQ
Aveen M.N. Ahmed1, Diaa A. Ibrahim2 and Seyran Palabash Uzun3
Biology Dept. , Faculty of science, University of Zakho , Kurdistan Region - Iraq.
2
Scientific Research Center, Faculty of science, University of Duhok , Kurdistan Region - Iraq.
3
Department of Bioengineering and science, Kahramanmaraş Sütcü imam University, Turkey.
1
Abstract
This investigation was carried out at plant biotechnology laboratories of Scientific Research Center in
Faculty of Science, University of Duhok, Kurdistan Region of Iraq, during the period from April 2013 to
January 2014. The objectives of the study were to investigate the influence of different plant growth
regulators in micropropagation on two local cultivars of grapes Vitis vinifera L. The most effective explant
disinfestations were obtained by using NaOCl in a ratio of 1:1 water for 10 min which was very effective
in reducing contamination in the established cultures resulting in a higher percentage of contaminations
free cultures. At initiation stage, the highest initiation rate was achieved by using specific concentrations
of BA on basal MS media. At multiplication stage, BA alone and BA combined with GA3 which was
showed different responses in the measured parameters. At rooting formation stage two auxins were
tested including IAA and NAA to estimate their ability for root formation. The in vitro propagated of
grape plantlets were gradually transferred in successful way from lab. To field conditions, which didn’t
showed any morphological abnormalities.
Keywords: In vitro, Micropropagation, Vitis vinifera L.
Introduction
rapevine "Vitis vinifera L." belongs
to the family Vitaceae. Vitaceae
comprise woody climbers, vines, trees, shrubs
and succulent’s trees (Timmons et al., 2007;
Chen and Wen, 2007) which are important food
sources. Vitaceae family is valuable raw material
for the production of wine, medicine and
perfumery (Gashkova, 2009). Different varieties
produce different types of dried fruit; the dried
fruits are the raisins (Kishmish), Currants
(Zabib, and the small seedless variety of grape
also known as Kishmish). The fruit juice can be
concentrated and used as a sweetener. Leaves are
cooked. Vitis vinifera is not found wild in
Kurdistan Region of Iraq although it is very near
to the region where it is native to it, i.e. the area
between Taurus range and the southern shores of
Caspian Sea. But it is extensively cultivated
throughout Kurdistan, especially in the
mountains where grape yards are usually not
irrigated, often on mountainsides of gentle slope,
open oak forest, and small moist plains.
Micropropagation of grape through tissue
culture provides a valuable tool for a more
efficient and rapid multiplication of plant. Plant
cell and tissue culture offers means not only for
rapid mass multiplication of existing stocks but
also for the conservation of important, elite and
rare plants. Micropropagation of selected vitis
G
genotypes can be carried out, among others, by
the culture of intact or fragmented shoot apical
meristems, axillary-bud, microcuttings or
through adventitious bud formation (Heloir et al.,
1997). However, most efficient protocols have
been reported for muscadine or other than V.
vinifera grapes (This and Graves, 1992; Qiu et
al., 2004), while studies with cultivars of V.
vinifera L. have met with less success (Chee and
pool,1983; Zatiko and Molnar, 1985).
The use of apices and axillary buds for the in
vitro propagation of various species and cultivars
of Vitis is documented (Gray and Fischer, 1985)
.Working on tendril explants of cultivars of
grape, reported somatic embryogenesis and low
frequency conversion of embryos to plants.
Many authors have found that the use of MS
medium supplemented with BA at (1-3 mg l⁻¹).
Apical meristem explants were successfully
taken from cultures grown on MS medium with
5 µM BA. Cytokinins at different concentrations
were tested: 5, 10 and 20 µM BA; 10, 20 and 40
µM kin; 0.5, 1 and 5µM TDZ (Gray and Benton,
1991). It has been shown that MS medium was
the most suitable medium for multiplication stage
and BA was the best cytokinin to obtain higher
proliferation rate. The range of BA that was used
in MS medium ranged between 0.5-2 mg1-1
showed a better response. Generally ,
micropropagated plants are difficult to transplant
55
for two primary reasonds: a heterotrophic mode
of nutrition and poor control of water loss
(Trigiano and Gray, 1996). This investigation
was carried out to test the effects of different
plant growth regulators on shoot multiplication
to establish a micropropagation protocol of
grapes cultivars via (shoot tips and axillary buds
segments).
Several experiments were carried out during
the period from April 2013 to January 2014, in
plant biotechnology laboratories of Scientific
Research Center, Faculty of Science, Duhok
University, Kurdistan Region of Iraq to study the
effects of growth regulators on in vitro
propagation of Vitis vinifera L.
All culture glasswares were carefully washed
with detergent, and distilled water. Before
culturing and using the laminar air flow cabinet,
the UV light of the laminar was switched on for
24 hours for sterilizing before culturing.
Murashige and Skooge (1962) medium was used
as basal medium. Stock solutions, for
macronutrients, micronutrients, vitamins and
growth regulators at (100 ppm) were prepared
and used for the preparation of the media. Also,
the media was supplemented with 100 mg1⁻¹
myo-Inositol, 30 g sucrose and 7 g Agar. The
medium was dispensed in equal sizes (25 ml)
into culture vessels. Finally, they were
autoclaved at 121˚C and 1.04 kg/ m² and then
left at room temperature for solidifying to be
ready for culture after 24 h (Karim, 2008).
Two kinds of explants (apical shoot tips and
lateral buds) of 1.5-2 cm long were taken from
Vitis vinifera L. field plants, the explants were
rinsed under running tap water for 30 mins and
drops at dishwashing liquid detergent for 5 min
then the explants were transferred to the laminar
air flow cabinet to complete the following
combinations:
A. 5% NaOCl and distilled water (2:1) + drops
of tween-20 for 5 min.
B. 5% NaOCl and distilled water (1:1) + drops
C. 5% NaOCl and distilled water (1:2) + drops
The explants were then rinsed with
sterilized distilled water three times, for 5 min,
followed by removing the ends of explants that
exposed to sterilization. The explants were
cultured in jars containing 25 ml of MS medium
56
alone and supplemented with (1 mg1⁻¹) BA as
an initiation media according to Deniel, (2009).
To determine the suitable combination of
plant growth regulators for explants (lateral
buds) establishments, the sterilized explants
were cultured on MS medium supplemented
with BA (0.0 and 1.0 mg1⁻¹). Because of the
limited numbers of grape vine apical shoot tips,
the in vitro growing shoot segments with two
nodes (1-3 cm) were taken as lateral buds
explants. Measurements were recorded after 4
weeks including shoot number / explants and
mean length of shoot. After 4 weeks of culture,
the shoots were cut into segments of shoot tips
and two nodes (≥ 1 cm long) and were cultured
on multiplication MS medium. Multiplication
stage was divided into two experiments, at the
first experiment the explants were cultured on
MS medium supplemented with BA alone at
different concentrations (0.0, 0.5, 1.0, 1.5 and
2.0 mg l⁻¹) and same BA protocols combined
with GA3 (0.25 mg l⁻¹). For the second
experiment to determine the effect of these
combinations on the studied parameters.
After 4 weeks of incubation in multiplication
medium, the plantlet microshoots were
transferred to rooting media. To determine the
most suitable auxins concentrations and salt
strength of MS media for rooting, microshoots
were cultured on:
A. MS medium supplemented with different
concentrations of NAA (0.0, 0.5, and 1.0 mgl⁻¹).
B. MS medium supplemented with different
concentrations of IBA (0.0, 0.5, and 1.0 mgl⁻¹).
After 4 weeks, data were recorded
including rooting percentage, number of
root/explant and mean length of roots. For all the
treatments, the cultures were incubated in the
growth room at 24±1˚C under 16 h photoperiod
by white fluorescent tubes under light intensity
of 1000 lux. Three replicates were cultured in
jars containing 25 ml of MS medium.
The rooted plantlets were taken from rooting
media and washed thoroughly with water to
remove adhering media, immersed in a beaker
containing 1 gml⁻¹ Benlet fungicide for 10 mins.
and washed with distilled water then the rooted
plantlet were transferred to pots containing
autoclaved peatmoss, loam and Styrofoam
(1:1:0.5) (v:v:v). The pots were placed in
sterilized box and covered by polyethylene cover
during the first week. The post were enclosed in
polyethylene bags, which were closed and
placed in a shaded area of temperature –
controlled greenhouse set at 23-25c. The plants
were irrigated with a nutrient solution containing
1/4 strength of MS salts. After 8 to 10 days, the
bags were opened and after another 8 to 10 days,
the bags were removed and plants were grown
under regular greenhouse condition (Toma,
2009).
The experiments were arranged according to
Complete Randomized Design (C.R.D) using
three replicates for each treatment. Data scored
in percentage were subjected to arcsine
transformation before statistical analysis and
then converted back to percentages for
presentation. Data were analyzed and means
were compared with each other using Duncanʼs
multiple range test to evaluate the significant
differences between the means.
The results showed that the combination (B)
which have been used in surface sterilization of
both types of
explants gave the highest
percentage of healthy - uncontaminatedexplants which was achieved 70 % in Be-dandk
cultivar ,while was reached 66 % in apical shoot
tips of Des-alaneez cultivar. On the other hand,
healthy lateral bud explants showed a higher
initiation response (95 %) in Be-dandk cultivar,
followed by (90 %) in Des-alaneez lateral buds
cultivar. Therefore, the second treatment was
selected for disinfesting the explants for later
experiments.
As it is known that the minimum
concentration and duration of sterling treatment
is more preferable to insure not damaging the
treated explant (Razdan, 2003). Sodium
hypochlorite proved to be the best sterilant ever
being effective not only as decontaminant but
also easy to remove from explants in a minimal
damage to the explant tissue. And the
sterilization was efficient in eliminating various
pathogens such as bacteria and fungi, as well as
prevention of browning of explant in grapevine
tissue culture (Dalal et al., 1991). The higher
initiation response in lateral buds it might be due
to the higher content of endogenous hormones.
These results are seems to be similar to those
obtained by Razdan (2003) and Amin (2007).
At
multiplication
stage,
microshoots
produced at initiation stage were used as
explants sources for shoot multiplication stage.
The ability of different concentrations of BA
(0.0, 0.5, 1.0, 1.5, and 2.0 mg l⁻¹) alone and
same levels of BA supplemented with (0.25 mg
l-¹) GA3 were used to induce Vitis vinifera L.
shoot multiplication.
Table (1): Effect of different BA concentrations and BA+ GA3 combination on highest shoot length (cm) of
Grapevine plants after four weeks in culture .
Hormone concentration mgl-1
Cultivars
Be- dandek
Des –alaneez
Effect of
cultivar
×
concentration
Effect of
hormone
× Conc.
Concentration
effect
Effect of
cultivar ×
hormone
2.05a
1.15c
1.60b
1.32c
Hormones
0
0.5
1.0
1.5
2.0
BA
BA + GA3
BA
BA + GA3
1.43c
0.70c
1.15b
1.37a
2.27ab
1.18 ac
1.57ab
1.67a
2.20ab
1.48a
1.93a
1.23a
2.70a
1.42ab
2.07a
1.15a
1.63bc
0.97bc
1.27b
1.20a
1.07e
1.73bc
1.84ba
2.06a
1.30ed
1.26ed
1.62bc
1.58dc
1.61bc
1.23e
Hormone
Effects
1.29c
1.92b
2.07b
2.38a
1.45c
1.82a
1.03c
1.43c
1.36c
1.28c
1.08c
1.24b
1.16b
1.67a
1.71a
1.83a
1.27b
Bedandek
Des
alaneez
BA
BA+GA3
Effect of
cultivars
1.60a
1.46a
Different letters represent significant differences according to Duncan’s multiple range tests at
5% levels.
57
Table (1) shows the effects of BA levels and BA enriched with (0.25 mg l-1) GA3 after four weeks
in culture on highest shoot length of grapevine plant.
The effect of the hormones clarify that the media supplemented with different concentration of BA
increased significantly and reaches a highest length (1.82 cm) when compared with the BA + GA3
(1.24 cm).
Table (2) Generally, the data clarify that there is no significant differences between the two
cultivars used in this investigation, whereas the effect of cytokinins induce significant increase in this
parameter by using the combination of BA with GA3 (1.09 cm) when compared with the medium
supplemented with BA alone (1.01 cm). In the same table, the result elucidate that the shoot length
average reaches (1.17 cm) in Be-dandek cultivar because of the interaction between cultivars and plant
growth regulators, which was reduced significantly (0.88 cm) by adding BA + GA3.
Contrastive response has been shown in Des-alaneez cultivar whereas the average shoot length
increased significantly in the medium enriched with BA + GA3 (1.31 cm) while achieve (0.85 cm) in
same cultivar cultured in different levels of BA alone.
Table (2): Effects of different BA concentrations and BA+GA3 combination on shoot length average (cm) of
Grapevine plants after four weeks in culture.
Cultivars
Be- dandek
Des –alaneez
Effect of
cultivar
×
concentration
Effect of
hormone
× Conc.
Concentration
effect
Effect of
cultivar ×
hormone
1.17a
0.88b
0.85b
1.31a
Hormones
0
0.5
1.0
1.5
2.0
BA
BA + GA3
BA
BA + GA3
1.10bc
0.41b
0.63bc
1.13a
1.40ab
0.88ab
0.97ab
1.30a
1.07bc
1.02ab
0.97ab
1.50a
1.60a
1.13a
1.13a
1.28a
0.70c
0.93ab
0.57c
1.35a
0.76c
1.14bac
1.04bac
1.37a
0.82bc
0.88bc
1.13bac
1.23ba
1.21ba
0.96bac
Hormone
Effects
0.87bdc
1.18bac
1.02bdc
1.37a
0.63d
1.01b
0.77dc
1.09bac
1.26ba
1.21ba
1.14bac
1.09a
0.82c
1.14ba
1.14ba
1.29a
0.89bc
Bedandek
Des
alaneez
BA
BA+GA3
Effect of
cultivars
1.02a
1.08a
5% levels.
Table (3) declares that the addition of BA levels in general to MS multiplication media was
valuable in case of number of shoots per explants after four weeks in culture. Since the highest number
of shoots per explants (2.9) in the medium enriched with different levels of BA which decreased
significantly (1.78 shoot / explant) in the same concentration of BA supplemented with 0.25 mgl-1 GA3
(Figure 1). On the same manner, both cultivars (Be-dandek and Des alaneez) recorded the highest
significant shoot numbers per explants (2.99 , 2.81) respectively when cultured in the media
supplemented with BA whereas, achieved (1.8 , 1.77) in the medium containing different
concentrations of BA + GA3. Although that, the data didn’t shown any significant differences between
the two cultivars.
58
Tableb(3): Effects of different BA concentrations and BA+GA3 combination on number of shoots/ explant of
grapevine plants after four weeks in culture .
Cultivars
Be- dandek
Des –alaneez
Effect of
cultivar
×
concentration
Effect of
hormone
× Conc.
Concentration
effect
Effect of
cultivar ×
hormone
2.99a
1.80b
2.81a
1.77b
Hormones
0
0.5
1.0
1.5
2.0
BA
BA + GA3
BA
BA + GA3
1.50b
1.83a
1.10c
1.53a
3.03cb
1.67a
2.60bcd
1.83a
3.17ba
2.00a
2.73bcd
1.90a
3.20ba
2.00a
3.50ab
2.00a
4.07a
1.50a
4.10a
1.56a
1.67bc
2.35ba
2.58a
2.60a
2.78a
1.32c
2.22ba
2.32ba
2.75a
2.83a
1.30c
2.82b
2.95b
3.35b
4.08a
Hormone
Effects
2.90a
1.68c
1.75c
1.95c
2.00c
1.53c
1.78b
1.49c
2.28b
2.45ba
2.68ba
2.81a
Bedandek
Des
alaneez
BA
BA+GA3
Effect of
cultivars
2.40a
2.29a
5% levels.
Table (4). It is clear that using BA plant growth regulator in multiplication medium
cause highly significant increase in number of leaves / explant (8.19) when compared with
BA+GA3 combination which reduce the number of leaves (4.35) (Figure 1). Although there
was no significant differences between the two investigated cultivars in this parameter; The
results indicate that the interaction between the cultivars and hormone concentration
increase the number of leaves per explant (8.21 and 8.17) in Be-dandek and Des alaneez,
respectively in the media containing BA only which was decreased significantly in the same
interaction by adding GA3 (4.82 and 3.89mgl⁻¹) for both cultivars.
Table (4): Effects of different BA concentrations and BA+GA3 combination on number of leaves / explant of
Cultivars
Be- dandek
Des –alaneez
Effect of
cultivar
×
concentration
Effect of
hormone
× Conc.
Concentration
effect
Hormones
BA
BA + GA3
BA
BA + GA3
Bedandek
Des
alaneez
BA
0
4.17cb
3.33a
2.47c
3.43b
0.5
1.0
1.5
11.53a
9.00ab
8.97ab
4.77a
4.83a
6.00a
10.17a
10.33a
11.60a
3.50b
3.77b
5.43b
2.0
7.40c
5.17ab
6.30bc
3.30b
Effect of
cultivar ×
hormone
8.21a
4.82b
8.17a
3.89b
3.75c
8.15a
6.92ba
7.48a
6.28ba
2.95c
6.83ba
7.05a
8.52a
4.80bc
3.32c
10.85a
9.67a
10.28a
6.85b
Hormone
Effects
8.19a
3.38c
4.13c
4.30c
5.72cb
4.23c
4.35b
3.35c
7.49a
6.98ba
8.00a
5.54b
Effect of
cultivars
6.52a
6.03a
BA+GA3
5% levels.
59
Shoot multiplication traits were influenced by addition of BA more than mixing it with GA3 and
this could due to the number of double bonds on its molecule structure (Mohammed, 1985) which is
provided with three bonds in its side chain and consequently increase the activity. In addition to
presence of benzene ring on BA structure improve its efficiency and become the most effective
cytokinin (Wasfy, 1995). Positive role have been seen on multiplication stage (shoot numbers, length
and leaves numbers), may be due to two causes:
First, is that the cytokinin increase the enzymes and proteins and RNA synthesis in the cells which
promote bud growth (Al-Rifae and Al- Shobaki, 2002). The second, releasing lateral buds from the
dominance of terminal buds without the need of removing the apical bud by promoting formation of
xylem tissues of buds which facilitate the transporting of water and nutrients. (Mohmmed and Younis,
1991).
One of the major physiological effects of the auxins is the stimulating of adventitious roots
formation in both in vitro and in vivo cutting (Hartmann et al., 2002). At rooting stage, in vitro shoots
derived from multiplication stage were used and two kind of auxins (IBA and NAA) as followed: MS
medium supplemented with different concentration of NAA (0.0, 0.5, and 1.0 mg⁻¹). MS medium
supplemented with different concentration of IBA (0.0, 0.5, and 1.0 mg⁻¹). Roots were initiated at
rooting formation stage, following 4 weeks of incubation on rooting media of above specific
concentrations, thereafter the roots were adventitiously initiated at cut margins of the shoots.
The percentage of root formation was significantly affected by the different treatments tested on
Vitis vinifera L.. Table (5). It is clear that increase the concentration of investigated auxins from 0.5 to
1.0 mgl-1 caused significant increase (82.0% and 86.33%) in rooting percentage of grape plants
compared with free media of auxins.
Concerning the differences in rooting abilities of grape, regarding the combination between types
of hormones and concentration of auxins, 0.5 and 1.0 mg1⁻¹ NAA was nearly similar in both
concentration which produced the highest significant rooting percentage (87.83% and 87.33%),
respectively when compared with control treatment (68.61%).Whereas 1.0 mg1⁻¹ concentration of
IBA produced the highest rooting percentage (85.33%) and decreased significantly in control
treatment (58.33%). On the other hand, no significant difference was seen between cultivars.
Table (5): Effects of different Auxin (NAA and IBA) concentrations on rooting percentage (%) of grapevine
plants after four weeks in culture.
Auxin concentration mgl-1
Cultivars
Be- dandek
Des –alaneez
Effect of cultivar
× concentration
Hormones
NAA
IBA
NAA
IBA
Bedandek
Des alaneez
Effect of hormone
×
Conc.
Concentration effect
NAA
IBA
Effect of
cultivar ×
hormone
84.18a
74.22a
78.33a
72.33a
0
68.89bc
53.33c
68.33bc
63.33bc
0.5
95.67a
82.00bc
80.00bc
70.33a-c
1.0
88.00bc
87.33bc
86.67bc
83.33bc
61.11b
88.83a
87.67a
65.83b
75.17ab
85.00a
Hormone
Effects
68.61bc
87.83a
87.33a
81.26a
58.33c
76.17ab
85.33a
73.28a
63.47b
82.00a
86.33a
Effect of
cultivars
79.20a
75.33a
5% levels.
60
Data in Table (6) clarify that there is a significant difference in the number of roots formed on
explants of Vitis vinifera L. as a result of tested treatments. The highest number of roots/explant
(14.44) was recorded in Bae-dandk cultivar compared with Des-alaneez (7.039 roots/explant).
Regarding the effects of the auxins on this parameter it is obvious that the effect of NAA was more
valuable of IBA which increase the roots significantly number to (13.33 roots / explant) compared
with IBA (8.15 root/explant)(Fig 1).
Increasing the concentration of auxin from 0.5 to 1.0 mgl-1 increase the roots number (14.13
root/explant) significantly according to control treatment (4.33 roots / explants).
Table (6): Effects of different Auxin (NAA and IBA) concentrations on root numbers/ explants of grapevine
plants after four weeks in culture.
Cultivars
Be- dandek
Des –alaneez
Effect of cultivar
× concentration
Hormones
NAA
IBA
NAA
IBA
Bedandek
Des alaneez
Effect of hormone
×
Conc.
Concentration
effect
NAA
IBA
Effect of
cultivar ×
hormone
19.56a
9.33b
7.11b
6.97b
0
2.83f
2.83f
4.00ef
7.67c-e
0.5
29.67a
11.50bc
8.50cd
5.40d-f
1.0
26.17a
13.67b
8.83cd
7.83c-e
2.83c
20.58a
19.92a
5.83b
6.95b
8.33b
Hormone
Effects
3.42c
19.08a
17.50a
13.33a
5.25c
8.45b
10.75b
8.15b
4.33b
13.77a
14.13a
Effect of
cultivars
14.44 a
7.039 b
5% levels.
Valuable inhibition in the number of roots formation was shown in Des alaneez cultivar. It is clear
that both auxins (NAA, IBA) weren't too much effective to increase the roots numbers although of
simple increase when compared with control treatments. Combined cultivars with type of auxin
significantly increased the number of roots (19.50 roots/explants) which were formed on shootlets of
Bae-dandek cultivar when cultured on MS supplemented with NAA compared with Des-alaneez when
cultured on MS supplemented with NAA and IBA (7.11, 6.97 roots/explants) respectively.
Root length average was affected significantly as a result of adding investigated auxins to the
medium. Table (7) clarify that there was significant increase in root length in Bae dandek cultivar (4.6
cm) compared with Des alaneez (2.27 cm). Hormone effect declares that the IBA induce more
significant increase in root length average (5.17 cm) than NAA (1.69 cm).
61
Table (7): Effects of different Auxin (NAA and IBA) concentrations on root lengths average (cm) / explants of
Cultivars
Be- dandek
Des –alaneez
Effect of cultivar
× concentration
Hormones
NAA
IBA
NAA
IBA
Bedandek
Des alaneez
Effect of hormone
×
Conc.
Concentration
effect
NAA
IBA
Effect of
cultivar ×
hormone
2.59b
6.61a
0.80c
3.73b
0
3.50bc
4.00b
0.93c-e
3.33b-d
0.5
2.27b-e
7.90a
0.67e
4.37b
1.0
2.00b-e
7.92a
0.80de
3.50bc
3.75ab
5.08a
4.96a
2.13b
2.52b
2.15b
Hormone
Effects
2.22bc
1.47c
1.40c
1.69b
3.67b
6.13a
5.71a
5.17a
2.94a
3.80a
3.55a
Effect of
cultivars
4.60a
2.27b
5% levels.
Finally the result of root formation stage, indicate that the auxins have an important role in
promotins adventitious root establishment on shoot end cuttings (Abdul, 1978; Saleh, 1991).
Adventitious root formation is a complex process that is affected by multiple endogenous factors
including phytohormones and environmental factors. Both auxins NAA and IBA varied in their
effects on grapevine rooting whereas NAA was more effective in the case of rooting percentage and
root number per explants while IBA was preferable in its effect in the stimulation of root length, these
results was similar to finding of those of Danial et al., (2009). And confirm that the presence of these
auxins had positive influence on rhizogenesis under in vitro conditions.
62
A
B
C
D
E
63
F
G
H
I
Figure (1):
A. Initiation shoots from lateral buds on MS medium enriched with 1.0 mgl-1 of BA after 4 weeks in culture.
(Bae dandek , Des alaneez Cultivars respectively in all picture)
B. Multiplication stage: Effects of different BA concentrations and BA+ GA3 combination on highest Shoot
Length (cm) of grapevine plant
C. Multiplication stage: Effects of different BA concentrations and BA+ GA3 combination on Shoot Length
Average (cm).
D. Multiplication stage: Effects of different BA concentrations and BA+ GA3 combination on number of Shoots
/ explant.
E. Multiplication stage: Effects of different BA concentrations and BA+ GA3 combination on number of leaves
/ explant.
F. Rooting stage. Effect of different IBA and NAA on rooting percentage (%) of Grapevine plants.
G. Rooting stage: Effects of different IBA and NAA on number of roots of Grapevine plants.
H. Rooting stage: Effect of different IBA and NAA on roots length average of Grapevine plants.
I.Acclimatization stag
64
CONCLUSIONS
It can be concluded that treating the explants
with Sodium hypochlorite + distilled water (1:1)
was more effective in reducing explants
contamination. At initiation stage, BA showed
better results for establishing aseptic Vitis
vinifera culture from lateral buds while in
initiation stage, lateral bud explants were more
effective in producing shoots than apical bud
explants. Hence, shoot multiplication traits were
influenced by addition of BA more than mixing
it with GAȝ
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66
ISSN: 2410-7549
USE OF WOOD MACERATED ELEMENTS FOR CLASSIFICATION OF
SALIX L. SPECIES IN KURDISTAN REGION- IRAQ
1
Saleem Esmael Shahbaz 1 and Shamiran Salih Abdulrahman 2
Faculty of Agriculture, University of Duhok, Kurdistan Region – Iraq.
2
Faculty of Science, University of Zakho, Kurdistan Region – Iraq.
Summary:
Characters of wood macerated elements were investigated for taxonomic application. Wood samples
from 5 stems of each Salix acmophylla, S. alba, S. babylonica, S. aegyptiaca, and S. Purpurea were collected
from different geographical regions of kurdistan to clasp most possible variation. Wood of most species
contains vessel elements described of primitiveness together with those descriptive of more or less
advanced. Vessels of different species are similar in possessing the simple perforation plate. The
differences are huge in vessel length and vessel width and hence in length/width ratio. Differences are also
huge in vessel type or the end wall characters of vessel elements described of primitiveness and
advancement, occurrence or absence of spiral thickening, cell walls pitting intensity and distribution
within and between species. All these features are found to be important for providing taxonomic
application. Libriform fibers are the only fibers present in the wood of this genus, no fiber trachieds
occur. All species have fibers possessing dentition, almost exclusively at the distal parts, very rarely
situated throughout the fiber length. Dentition morphological structure, distribution, size and orientation
form important diagnostic characters.
Key words: Vessel elements, Libriform fibers, Salix acmophylla, Salix Alba, Salix babylonica, Salix aegyptiaca,
Salix purpurea.
Introduction
T
here is about 526 species of Salix in the
world (Fang-Zhen, 1987), most of which
are distributed in the Northern hemisphere with
only a few species in the Southern hemisphere.
In Asia, according to the same researcher, there
are about 375 species, making up 71.29 percent
of the total Salix species in the world with the
center of origin and development in the region
between 20-40 º N in East Asia. In Iraq, 6
species are described, they are S. alba, S.
acophylla Boiss., S. babylonica L., S. purpurea
L., S. aegyptiaca L., S. triandra L. All of Iraqi
representative species occur in Kurdistan region
with the exception of S. triandra which has a
very limited area of distribution at Tigris river
banks near Rashidia north of Mosul city. Salix
species are moisture demanding plants,
commonly found in the lower and middle forest
zones, in the latitudinal range 439-1840m
(Abdulrahman, 2011), at river banks, moist
mountain valleys, in riparian thickets, occasional
in the moist steppe region. Willow species S.
alba and S. aegyptiaca and S. babylonica need
high soil moisture content and lower mean
annual temperature compared to similar
requirements for S. acmophylla and S. purpurea.
In Kurdistan region, the species S. acmophylla is
of especially high drought tolerance (Townsend
and Guest, 1980). All willow species with the
exception of S. Babylonia is found in the
wilderness. Both S. alba and S. acmophylla are
abundant and common in the region, while S.
purpurea and S. Aegyptiaca are rare and
occasional, especially in the western and central
mountain sectors, they may be absent from the
eastern sector. Only male plants of S. purpurea
have been observed in Iraq.
According to Stace (1981) anatomical
features are of particular value to scientists who
need to identify small scraps of plant material.
Some
recent
scientific
works
relate
characteristics of the secondary xylem of Salix
species to environmental conditions and
pollution in Canada and Eastern Europe (Cooper
and Cass 2001; Chavchavadze et al. 2002;
Sizonenko and Chavchavadze 2002). On the
other hand, Wagner (2009) studied the wood
anatomy of young and adult samples of Salix ×
rubens, aiming to improve the knowledge about
the most used species for basketry in South
Brazil by relating wood anatomy characters of
xylem quantity and ratio of pith/xylem to wood
flexibility.
The vessel elements that originated in a
precursor to the angiosperms may have been
subsequently lost in some lineages like
Amborellaceae,
Tetracentraceae,
Trochodendraceae, and Winteraceae. These
67
basal lineages were described by Cronquist
(1981) as primitively vesselless. Moreover
Cronquist considered the vessels of Gnetum to
be converging with those of angiosperms. When
data of vessel element characters of density,
vessel lumen diameter, vessel length, and vessel
clustering, in four endemic Salix taxa from the
Lake Athabasca-Canada were compared by
Cooper and Cass (2001) with the putative sister
species for each endemic, it revealed similar
vessel element densities to their associated sister
species. In addition to that, the vessel element
lumen diameter and length were significantly
different in some of the species pairs. The
authors indicated that the structural differences
for these endemic willows appear to be related to
their open sand habitat. The fiber dimorphism
noticed in Dubautia and many other genera in
which there are bands of thinner-walled fibers at
intervals within a background of thick-walled
fibers are probably a division of labor between
fibers serving for photosynthate storage and
fibers serving for mechanical strength (Sherwin,
1988). According to Sherwin (2014), most
authors contrast wider, thinner-walled, shorter
fibers with narrower, thicker-walled fibers that
have narrower Lumina. For the same author, the
evolutionary significance of fiber dimorphism in
the form of few small changes in fiber structure
can result in the achievement of different
functions. Moreover, Sherwin (1984) indicated
that the simple slit-like pits of libriform fibers
cause minimal loss in mechanical strength
compared to bordered pits.
Many researchers consider Salix as one of the
most difficult genus for identification; there is
still disagreement regarding the identity, number
and distribution of species (Heywood et al.
2007; Mabberly 2008). The easy process of
hybridization between species in the wild,
together with their phenetic plasticity and
different time of flowers and leaves
development, made it difficult to observe all of
these relevant characters on a single plant or
specimen (Salih, T. et al. 2014). In many
instances morphological characters are not
sufficient to delimit closely related species of
Salix. Therefore, in addition to morphological
features, anatomical characters may provide
valuable information for the classification of
Salix (Arihan and Güvenç 2011).
The taxonomic relationships among the Salix
species are not well understood and are still
under debate. It is always felt for this genus that
wood macerated elements should be viewed
68
carefully in an attempt to provide useful
taxonomic traits to identify taxa and demonstrate
taxonomic relationships between species
representative of Kurdistan-Iraq.
Material and Methods
Wood samples from Salix acmophylla, S.
alba, S. babylonica, S. aegyptiaca, and S.
Purpurea were collected from different
geographical regions to clasp most possible
variation. Samples of 2-3mm thick and 2-4cm
long were prepared from growth increments 4-5,
starting from the pith of the 5stems of each
species, to avoid juvenile wood of it. The
herbarium vouchers of them were deposited in
the
Biology
department/Faculty
of
Science/University of Zakho.
Wood samples were macerated using
Franklin solution (Franklin 1946). A sample of
fibers was removed from the maceration and
spread over a glass slide. 25 measurements of
each of the following parameters were surveyed:
1. Libriform fiber length in mm.
2. Libriform fiber diameter (at the midpoint) in
µm.
3. Libriform fiber wall thickness in µm.
4. Vessel element length in mm.
5. Vessel element diameter (at the midpoint) in
µm.
Characteristics of the Genus:
Vessel element and libriform fiber
dimensions are given in table (1). The overall
range of vessel length and vessel diameter is
found to be 0.2-0.55mm and 95.2-209µm
respectively. These values together with STDs
indicate high variance in vessel dimensions.
The vessel elements have simple perforation
plates, inter-vessel pits are bordered pits, wood
of most species contain vessel elements
described of primitiveness together with those
descriptive of more or less advanced.
Libriform fibers are the only fibers present in
the wood of this genus, no fiber trachieds are to
be observed. Fibers range 0.55- 1.20mm in
length, 13.2-30.8µm in diameter at the midpoint,
1.5-7.0µm in cell wall thickness; statistics given
in table (1) indicate low variability in measured
parameters of the fiber. All species have fibers
possessing dentitions, almost exclusively at the
distal parts, vary rarely situated throughout the
fiber length.
S. alba
Fibers of S. alba seems rather homogenous,
mostly straight, thin-walled with small slit-like
pits. Only distal dentitions occur on one side of
some fibers, in most cases the dentitions of one
end differ from dentitions of the other end, few
fibers are with very smooth walls, containing no
dentitions. Actually the overall fiber characters
refer to a considerable modification and
evolutionary process figure (1).
Vessels of S. alba exhibit high variability in
shape and size. The wood tissue contains
numerous vessel types at different level of
advancement. According to Ghose (1984) and
Klotz (1978) the vessel elements with simple
perforation plate, horizontal end wall and least
value of vessel length/vessel width are advanced,
while those of scalariform perforation plate,
sloping end wall and high vessel length/vessel
width value are primitive. Considering these
criteria, it is apparent that vessel type of S. Alba
lay between the two extremes, none of them is
more advanced or more primitive.
Number of lateral inter-vessel pitting areas
differs from vessel to vessel, regardless of the
level of advancement or primitiveness; it is a
characteristic
feature
of
this
species.
Table (1): Mean, Standard Deviation (STD) and Range for Libriform Fibers and Vessel Elements.
Characters
S . alba
S.
S.
S.
S.
acmophylla
babylonica
aegyptiaca
purpurea
Mean
1.01
0.93
1.10
0.88
0.90
Libriform fiber
STD
± 0.11
± 0.14
± 0.12
± 0.1
± 0.13
length (mm)
Range
0.77-1.19
0.66-1.15
0.8-1.2
0.69-1.04
0.55-1.1
Mean
20.93
19.70
23.22
20.8
20.2
STD
±2.96
± 2.90
±3.11
± 2.35
± 2.93
Range
15.4-25.3
13.2-24.2
19.5-30.8
16.5-24.0
15.2-26.0
Mean
3.4
4.2
4.1
3.1
4.3
STD
± 0.93
± 1.08
± 1.35
± 0.80
± 1.45
Range
2.2-4.4
1.5-6.0
2.0-5.3
2.1-4.1
1.8-7.0
Mean
0.32
0.38
0.40
0.37
0.31
STD
± 0.09
± 0.07
± 0.06
± 0.11
± 0.07
Range
0.22-0.50
0.26-0.52
0.27-0.49
0.26-0.57
0.20-0.55
Mean
135.9
143.6
140.4
142.9
153.7
STD
± 16.27
± 21.85
± 18.5
± 31.35
± 26.4
110-180.5
99-165
95.2-220
108-209
Libriform fiber
diameter
(μm)
Libriform fiber
wall
thickness
(μm)
Vessel
element
length (mm)
Vessel
element
diameter
(μm)
Range
115.5187
69
Each value represents 25 replicate
S. acmophylla
The fibers of this species express a high
range of variation in fiber length and fiber cell
wall thickness (table 1). Wide differences occur
in fiber overall shape, some of them are quite
straight; others are winding, especially at their
distal parts (figure 2). Differences also occur in
dental number, shape and distribution; mostly
concentrated at the two fiber ends while some
fiber possesses dentition distributed along the
fiber length, forming a significant diagnostic
character. Like S. alba, dentitions occur only on
one side of some fibers, moreover dentitions of
one end often differ from the dentition of the
other end. Fiber wall is relatively thin, but
thicker than that of S. alba, with small slit-like
pits.
As figure (2) shows, vessel members of S.
acmophylla are highly variable. End wall slopes
of vessel elements show a wide range of
variations such as, both end walls transverse,
both end walls oblique and mixed type, i.e., one
end wall transverse and the other oblique, the
length/width ratio differences are also high
indicating differential evolutionary process. The
same figure refers to vessel shapes of short and
transverse end wall possessing apomorphous
character states in common together with
members exhibiting pliesiomorphous character
states of both end walls oblique. Moreover,
mixed vessels of one end wall transverse and the
other oblique are also found in the mixture of the
macerated elements.
S. babylonica
The fiber length and fiber diameter
measurements of this species are found higher
with moderately thick cell wall compared to
other Salix species (table 1). Fibers are ranging
between straight forms to winding or slightly
curved or zigzag with highly tapering distal
ends. Dentitions differ in shape, number and
size. Only distal fiber ends have dentitions,
middle portions possess none of them. Number
of dentitions are usually few, often one of them
is conspicuous, either oriented upward to the
fiber tip or perpendicular to the surface (figure
3), it is worth mentioning that the presence of
such distinct dentition provide a significant
taxonomic application. Pits, like other species,
are small slit-like.
Vessel members macerated from the wood of
this species show a great resemblance to one
70
another, only two types of vessels are distinct;
one possessing transverse and oblique end wall,
while the other has oblique ends similar to those
of very primitive vessel members, but not to the
foraminate which contains several round
perforations. These vessel members have lateral
inter-vessel pits, mostly located around the
perforation plate.
S. aegyptiaca
Fiber length and the fiber cell wall thickness
are inferior to the corresponding measurements
of other species. Fibers are rather irregular in
shape; few seem to be straight with no or with
very small dentitions. One of the distinct
diagnostic character is the sudden reduction in
the fiber diameter at both ends (figure 4) leaving
three portions, two of them are strongly tapering
distal portions and one constituting the median
portion, the two tapering distal part may or may
not equal the mid portion.
Statistics given in table (1) refer to high
variability in length and diameter of vessel
members and the occurrence of the high rate of
large sized vessel in the macerated mixture.
Vessel shapes displayed in (figure 4) shows
different evolutionary levels (Ghose and Das,
2001), ranging from most developed or
advanced to the most primitive, but with higher
ratios for the longest primitive members in the
wood mixture. All vessel types of (i) both the
end walls transverse (often typically quadrate)
(ii) both the end walls oblique and (iii) mixed
type are present. The occurrence of minute intervessel pitting and spiral thickening is a useful
tool for identifying this species.
S. purpurea
The average fiber cell walls of this species
are significantly thicker than cell wall of other
species. Few fibers are found to exhibit straight
forms; most are zigzag, curved or winding.
Dentitions are restricted to the distal ends,
ranging in number from 1to 6, on one side of the
fiber. The two ends are often dissimilar in shape,
number and size of dentitions. One of the
diagnostic characters is the presence of course
irregular of up to 6 dentitions on only one side of
some fibers (figure 5).
The average length of vessel elements of S.
purpurea is significantly lower than that of other
species, but their average vessel diameter is
significantly higher than other species (table 1).
The advanced vessel type of both end walls
transverse is common, one end wall transverse
and the other oblique is also common, while
both the end walls with very slightly oblique is
less common. The lower length/width ratio and
the increased number of both end walls
transverse are evidence of advancement of the
wood tissue of this species more than any other
Salix species.
From comparing different species, it reveals
great differences in intensity of inter-vessel
pitting; some members are even without lateral
pitting. The former type of vessels which appear
to be quadratic in cross section are very
abundant, while the latter is rather rare and
appear circular in cross section, a structure
probably indicating the trend of wood tissue
evolution. For the first type of the one end
truncate and the other oblique, the length/ width
ratio is highly variable probably referring to the
different environmental adaptation. According to
Haarer (1952) the result may be influenced by
parental gene tree and other environment
condition effect.
Figure (1): S. alba. 1. Vessel element, both ends slightly oblique, inter-vessel near adjoining vessels, 2. Vessels
of variable ends and areas of pitting, 3. libriform fibers, 4. Medium part of a libriform fiber showing cell wall
thickness and cell lumen, 5. Distal part of a libriform fiber showing dentitions on one side of the fiber.
71
Figure (2): S. acmophylla: 1. Two ends by end connected vessel elements, both with two end
transverse, 2. Two ends by end connected vessel elements, both with two ends oblique (140x), 3.
Typically, straight-shaped libriform fiber, 4. Distal part of a fiber showing minute irregular dentitions,
5. Fiber cell wall and lumen with a fiber tip, 6. An irregular-shaped libriform fiber (below), Vessel
member and numerous variously shaped libriform fibers (above).
72
Figure (3): S. babylonica. 1. Vessel element of mixed end walls with intensive lateral pitting, 2. A
vessel element of mixed end walls with no lateral pitting, 3. High length/width vessel element with
libriform fibers, 4. A primitive type of a vessel element with both ends steeply oblique, 5. Two ends
by end connected vessel elements, both with two end transverse, 6. Libriforms of different length and
shapes, 7. Fiber cell wall thickness and lumen, 8 and 9. Distal fiber portions showing variously
oriented dentitions.
73
Figure (4): S. aegyptiaca. 1. Vessel member, very short with both ends transverse, 2 and 3. Vessel
members of more or less transverse end walls, 4. Vessel member with one end wall transverse and the
other oblique, 5. Vessel member with both ends oblique, 6. A very primitive vessel member with the
both end walls steeply sloped, 7. Different types of libriform fibers, 8. Distal fiber portions showing
different types of dentitions.
74
Figure (5): S. purpurea. 1. Vessel member with one end oblique and the other transverse, 2. Vessel
member both end walls transverse, 3. Two vessel members, one (above) withmixed end walls, the
other (below) with both end walls slightly oblique, 4. Libriform fibers, 5. Fiber cell wall thickness and
cell lumen, 6 and 7. Two types of distal portions showing variable kinds of dentitions.
75
Important taxonomic applications:
A. Fiber dentition:
1. Dentitions of most fibers occur only on the
distal ends: S. alba, S. babylonica, S. aegyptiaca,
S. purpurea.
2. Dentitions of some fibers occur along the
length of the fiber: S. acmophylla.
B. Dentition size and orientation:
1. One of dentitions on the distal portion is large
and conspicuous, either oriented upward to the
fiber tip or perpendicular to the surface: S.
babylonica.
2. None of the dentitions are so large and
conspicuous, little differences occur between
them: S. alba, S. acmophylla, S. aegyptiaca, S.
purpurea
C. Fiber structure:
1. Sudden reduction in the fiber diameter at both
ends, dividing the fiber into three portions, two
at both ends, one median: S. aegyptiaca.
2. Gradual reduction in the fiber diameter
towards ends: S. alba, S. acmophylla, S.
babylonica, S. purpurea.
D. Vessel length:
1. Vessel element never exceeds 0.55mm in
length: S. alba, S. acmophylla, S. babylonica, S.
purpurea.
2. Vessel element exceeds 0.55mm and reaching
0.57mm in length: S. aegyptiaca.
E. Vessel morphology:
1. Vessels with minute inter-vessel pitting and
spiral thickening: S. aegyptiaca.
2. Vessels with larger-sized inter-vessel pitting,
with no spiral thickening; S. alba, S. acmophylla,
S. babylonica, S. purpurea.
F. Vessel type:
1. Wood macerated mixture containing vessel
type with both the end walls steeply oblique: S.
babylonica, S. aegyptiaca.
2. Wood macerated mixture containing no vessel
type of both end walls steeply oblique: S. alba,
S. acmophylla, S. purpurea.
References
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ISBN: 978-3-662-21716-0 (Print) 978-3-66221714-6 (Online). Springer Series in Wood
Science.
Sherwin Carlquist (1984). Vessel grouping in
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Aliso, 10(4), 1984, pp. 505-525.
Sizonenko, O.Y. and Chavchavadze, E. S. (2002).
Wood anatomy in species of the genus salix
(salicaceae) from northern regions of Russa.
Botansheskii zhurnal 87: 67-77.
Stace, C. (1980). Plant taxonomy and biosystematics.
Edward Arnold publisher Ltd, London.
Townsent, C. C. and Guest, E. (1980). Flora of Iraq 4:
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78
ISSN: 2410-7549
OCCURRENCE OF TOMATO LEAF MINER TUTA ABSOLUTA
MEYRICK(LEPIDOPTERA:GELECHIIDAE) IN DUHOK REGION (A)
Lazgeen Haji Assaf 1, Feyroz Ramadan Hassan 2, Halgurd Ismael Rasheed 2, Mohammad Abdulrahman Sadiq 1, Rezgar M.
Ismael 1, Mohammad Sh. Ahmed 1, Salah Adel Saeed 1 and Farhad Mohammed Taher Salih 1
1
2
General Directorate of Agriculture-Duhok, Kurdistan Region – Iraq.
Dept. of Plant Protection, School of Plant Production, Faculty of Agriculture and Forestry, University of Duhok, Kurdistan
Region – Iraq.
(Accepted for publication: Feb 10, 2015)
Abstract:
The aim of this work is to investigate the population density and infestation percentage of Tuta
absoluta on tomato crop Lycopersicum esculentumMill under field conditions in three locations at Duhok
province/ Kurdistan region/ northern of Iraq in 2012. The average number of mines/ leaflet, larvae/ leaf
and larvae/ fruit during the study season were 2.33, 0.34 and 0.61 respectively. The results showed a
significant differences in infestation percentage and number of male per trap among the three locations.
The highest percentage of infestation was recorded in September as 74.00, 72.00 and 60.00 for Summel,
Shekhan and Zawita respectively. The maximum number of males/ trap/ week was 1205.40 recorded on
26/8/2012 in Summel. Concerning the use of pheromone trap for pest monitoring, linear regression
analysis results was significant between trap catches and the number of mines per leaf and the infestation
rate of leaves.
Keywords: Tuta absoluta, population dynamics, Pheromone traps, Tomato.
1- Introduction
omato (Lycopersicon esculentum) is one
of the economic important crops of Iraq.
It is consumed as fresh table tomato and as an
essential raw material for a variety of food
processing industries. According to Food and
Agriculture Organization 2010, tomato was
planted in highland in the world reached to 1.2
million hectare and in Iraq the planted area
reached to 842.296 hectare. In Duhok province,
more than 12000 hectare of land was planted
with tomato in 2012. Tomato production faces
many problems from several causes such as
seasonal weather, temperature, humidity,
diseases and insect pests as thrips, whitefly,
tomato fruitworm, leaf miner, leafhopper, aphid
and mites.
The tomato borer,Tuta absoluta (Meyrik)
(Lepidoptera:Gelechiidae ) is one of the most
destructive pest of tomato crops in South
America (Epp,2010).After its first detection in
eastern Spain in 2006,its rapidly invaded various
other European countries and spread through the
Mediterranean basin (Desneux et al.2010).In
Iraq,tomato borer T. absolutawas first detected
in Autumn 2010 from sex pheromonetraps
installed in Rabia region, Nineveh province
(AbdulRazzak et. al.2010). Since that time,this
pest spread quickly in all tomato growing
areas,destroying entire open and protected
T
fields.Ramireze et. al.(2010) reported that the
damage by T.absoluta can reach to 100% in
unprotect crops andwas considered in its region
distribution area as asignificant tomato insect
pest(Leite et.al. 2010),due not only to the
intensity of its attack but also to its occurrence
during all crop cycle (Oliveira et. al. 2008).It
can develop on other Solanaceous plants, like
potato (Pereyra and Sanchez,2006),eggplant and
wild species (Garacia and Espul, 1982). Abbes
and Chermiti, 2011 reported that the trap catches
can be correlated with larval damage. In this
way, the minimal amount of spraying required to
control the pest population.
The aim of this work is to study the
occurrence and infestation level of T. absoluta
on tomato crops Lycopersicum esculentum Mill
underopen field conditionsin three sites as
Summel, Zawita and Shekhan at Duhok
governorate during 2011- 2012 crop season .
79
2- Material and methods
2-1: Study area
For general observations , initial survey was
done before starting the project in order to put
the hand on the predominant areas in which the
tomato planted and settled in Duhok
provinces.To carry out
this work,
threecommercial fieldslocated in three sites
within Duhok governorate ( 36° 54' N, 43° 8' E
) were choose as Shekhan (483 m a.s.l and 34
km far form Duhok centre ), Zawita (855m a.s.l.
and 16 km far from Duhok centre)and Summel (
473 m a.s.l and 16km far from Duhok centre).
The average degree of temperature and relative
humidity was obtained from Metrological
Station of Duhok Governorate .
2-2: Experiment procedure
To determine the tomato infestation by the
tomato leaf miner, two methods were followed
as:
1- leaves sampling for recording larvae and
mines of T. absolutae :
To calculate the larvae and mines number of
T.absoluta, fields were divided to five subarea
approximately
0.2 ha. Each subarea with
asimilar number of plants to ensure that all the
area represented in the samplings. Weekly
sampling from May to September were carryout
and in each field 25plants were randomly
selectedbeing five in each subarea. At each
selected plants five leaflets were collected and
singly packed in labelled plastic boxesthen
transported to the laboratory.
At the laboratory,with aid of Binocular, each
leaflet was examined and the number of mines
and larvae per leaflet was registered (Leite et.al.
2001). Also the number of larvae / fruit was
recorded during this study.
2- Trapping methods for adult males
harvesting :
This experiment was conducted in two fields,
Summel and Shekhan site. Theadults of
T.absoluta
were
monitored
by
using
pheromonelure TUA-500 and the TUTASAN®
trap (TUTASAN® is a water trap specifically
conceived to early detect and catch the tomato
80
leaf miner males, each trap contains of one plate
and one pheromone container (green basket with
a lid)).The pheromone lure was put in the green
basket, closed up with the lid and fixed at the top
of the plate. The plate was filled with water till
reaching the maximum level. The male insects
are attracted by the sex pheromone lure from the
top of the trap, and fall down into water. In each
field six traps were well positioned and installed
0.5 m above ground level. Four traps at the tops
of tomato plants (one in each direction) and two
within the field centre. Sex pheromone lure were
renewed every four weeks and the number of
captured individuals was recorded every week
during study period from May to September
2011-2012.
Note: Pest management strategies were
applied by farmers and based largely on
chemical applications (Insecticide) which
targeted to controlling T. absoluta and other
lepidopteran pests (Noctuidae: Heliothinae).
Data were statistically analysed as Factorial
experiment using Randomize Complete Block
Design (RCBD) with five replications and SAS
program was used. The means were compared
using Duncan's multiple range test (DMRT)at P
≤ 0.05. Also the relation between trap catches,
the infestation rate and the number of mines per
leaf was performed by a linear regression
analysis.
3-Results and Discussion
3-1: Leaves sampling for recording larvae
and mines of T. absoluta
The data represented in table (1) showed that
the general mean number of T. absoluta
mines/leaflet was 3.37, 2.46 and 1.2 for
Shekhan, Summel and Zawita, respectively.
T. absoluta mines resorted to concentrate on
leaves on the middle and upper levels.
According to fig. (1), the number of leaf mines
was initially low in May and June and increasing
during August and September to reach a
maximum of24.33 on 26/8/2012.Allache and
Demnati (2012) mentioned that in Biskra
(Algeria), during the first phonologic stages of
the crop, tomato plants are free from attack of T.
absoluta .
Table1: Mean mines, larvae number and the infestation percentage of leaves during study season.
Sample
sites
Average
No. of
mines/
leaflet
Average
No. of
larvae/lea
flet
Average
No. of
larvae
/fruit
Leaves Infestation Percentage %
May
June
July
August
September
Shekhan
3.37 a
0.36 b
0.54 c
0
10.67
B
67.20
a
69.00
A
72.00
a
Summel
2.46 b
0.46 a
0.68 a
0
34.00
A
68.00
a
73.33
A
74.00
a
Zawita
1.20 c
0.20 c
0.61 b
0
14.00
b
29.60
b
58.00
B
Average
2.33
0.34
0.61
0
19.56
54.93
66.78
60.00
b
65.33
Means followed by a common letter within the same Column are not significantly different at the 5%
level by Duncan's Multiple Range Test (DMRT).
The maximum number of leaf mines was closely related to the density of T. absoluta larvae, that
the damage caused by larvae on tomato leaves in three locations led to high density of mines rising up
to an average of 7.20 mine/ leaf starting from the beginning of July (fig.1).
Harizanova et.al.(2009) pointed that the leaves were the most heavily damaged plant parts with an
average of 9.42 and 8.75 mines per leaflet on the middle and upper layer of the canopy respectively,
followed by the fruits. Increase of the larvae number on leaves caused a high infestation rate reached
to 73.33%, 69% and 58% in August for Summel, Shekhan and Zawita.The maximum number of
mines/ leaf recorded in September which was 31.4, 28.6 and 14.6 (fig. 2) in Shekhan, Summel and
Zawita, respectively.
No. of mines/ leaf
25
20
15
10
5
0
May
June
July
August
September
sampling date
Fig.1: Average number of T. absolutamines / leaf on tomato leaves in three regions in 2012.
81
Statistical analysis showed a significantdifferences in leaf mines caused by T. absoluta larvae
amongthe three locations. It could be concluded that in Zawita the mean number of mines (fig. 2), the
infestation rate and the number of larvae per mine(tab. 1) was the lowest because it was located in
mountainous location which was characterized by (855 m) above sea level and the low temperature
degree.
Mines number/ leaf
35
30
25
20
15
10
5
0
Shekhan
Summel
Zawita
May
June
July
August September
sampling date
Fig.2: Evolution of the number ofT. absolutamines/ month/location on tomatoleaves inthree sites in 2012.
Lacordaire and Feuvrier (2010) reported that the number ofT. absoluta was influenced perhaps by
abiotic factors like temperature, insecticide application and by biotic factors i.e absence of natural
enemies.
The average number of larvae/ leaflet (tab. 1) was 0.46, 0.36 and 0.20 for Summel, Shekhan and
Zawita , respectively. It was frequent according to Lacordaire and Feuvrier (2010) that the larvae of T.
absoluta left their galleries and reinstalled in another leaflet or leaf as suggested by Torres et.al.
(2001),which also added that this low number might be due to insecticide applications which limited
the development of the larvae.
3-2 Trapping methods for adult males harvesting :
According to fig. 3, the average number of adults male captured by pheromone traps was variable
during study period in the two sites. T. absoluta was present from May until September in Summel,
while in Shekhan it was noted from June to September (fig. 4). For the two sites, the adults captured
increased towards the end of plant cycle. It was relatively low in June with an average of 10.9 adults/
trap/week and their number became relatively high, as their attack became intense towards the end of
crop cycle that reached to a maximum of 1055.40 adults/trap/week on 3/9/2012 (fig.3). An increase in
temperature was detected at this time in the year. These results agreed with Nannini et.al. (2010), who
mentioned that in Southern Sardinia (Italy), the highest number of moths caught in traps were in
September-October. These results also matched with those found by Miranda et.al. (1998) and
Lacordaire and Feuvrier (2010), who underlined the occurrence and increase in leaf miner captures
during the crop season. During this gradual increase, each renewal of the capsules was often followed
by an increase in the number of trapped adults.
82
1200
Males / trap
1000
800
600
400
200
0
May
June
July
August
September
sampling date
Fig.3: Average number of T. absolutatrapped males of two regions
The average number of captured males/week was respectively about 378.73 and 337.23 in Summel
and Shekhan. Although the number of adults captured in Summel was more abundant than that in
Shekhan and there was a significant differences between these two population, the figure of the
evaluation of mass trapping in Summel field shows a similar shape of that in Shekhan (fig. 4). Indeed
after 16.80 males caught on 19/6/2012, the number of trapped males continued to increase speedily to
exceed 1205.40 0n 26/8/2012.
1200
Males/ Trap
1000
800
600
400
200
0
Summel
Shekhan
May
June
July
August
September
Sampling Date
Fig.4: Average number of trapped males/ month/ site of T. absoluta for two regions
83
To investigate that the sex pheromone trap reflects the real level of damage in tomato crops and the
infestation rate of plants depending on the number of trapped males of T. absoluta, the relation
between trap catches, the infestation rate and the number of mines per leaf was performed by a linear
regression analysis .
Analysis showed a significant relationship between the number of mines /leaf and the number
of trapped males in both Summel and Shekhan(fig. 5 A, B). However, linear regression between
infestation rate and the number of catches in sex phermone traps was also significant in two regions
(fig. 6 A, B). These results agreed with Abbes and Chermiti (2011) who reported that the number of
leaves and leaflets were differentially affected by the densities of T. absolutainfestation (p≤0.05)
A
B
Fig. 5: Linear regression between the number of caught males of Tuta absoluta and the number of mines per leaf
in Summel (A) (R2= 0.88) and Shekhan (B) (R2= 0.53) in 2012.
84
A
B
Fig. 6: Linear regression between the number of caught males of Tuta absoluta and the infestation rate of leaves
in Summel (A) (R2= 0.37) and Shekhan (B) (R2= 0.31) in 2012.
Results of linear regression betweenthe
number of trapped males and theinfestation level
suggest the utilization ofthe sex pheromone traps
as good indicators of the infestation rate of
thecrop. Some authors recommend 30trapped
males per trap per week tostart chemical curative
measures.Monitoring is very important in
IPMprograms; it allows the detection of theearly
infestations. Using of chemicalsbased on data
from pheromones traps areusually more effective
in controlling firstattacks and avoiding the
rapidcolonization
of
the
crop.
This
cansignificantly reduce the number ofsprayed
insecticides, and preserve naturalenemies of the
pest.
References
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Marks
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First record of tomato borer (tomato moth)
Tuta absoluta (Meyrick) (Lepidoptera:
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Arab and Near East Plant Protection
Newsletter no. 51, p 31.
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-Allache F andDemnati F 2012. Population Changes
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Gelechiidae). J. Appl.Entomol. 122: 43-47.
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used in the EPPO region. http://
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-Garcia, M.F. and Espul, J.C. 1982. Bioecologia de la
polilladel tomae (Scrobipalpulaabsoluta) en
Mendoza,Republica Argentina. Rev. Invest.
Agropecuarias INTA, 17,135-146.
Harizanova V., Stoeva A. and Mohamedova M.
2009. Tomato leaf miner, Tuta absoluta
(Povolny) (Lepidoptera: gelechiidae) – first
record inBulgaria. Agricultural science and
technology 1(3) 95 - 98, 2009.
-Lacordaire A.I. and Feuvrier, E.2010. Tomate,
traquer Tuta absoluta. Phytoma, 632, 40-44.
-Leite, G.L.D., Picanço, M., Guedes, R.N.C. and
Zanuncio, J.C.2001. Role of plant age in the
resistance of Lycopersicumhirsutum f.
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Hort., 89, 103-113.
-Miranda, M.M.M., Picanço, M.C., Zanuncio, J.C and
Guedes,R.N.C. 1998. Ecological life table of
Tuta
absoluta(Meyrick)
(Lepidoptera:
Gelichiidae). Biocont. Sci. andTech., 8, 597606.
- Nannini M., Atzori F., Foddi F., Pisci R. andSanna
F. 2010. A survey of Tuta absoluta (Meyrick)
(Lepidoptera: Gelichiidae) outbreaks in tomato
greenhouses in southern Sardinia (Italy).
International Symposium on Plant Protection
-Oliveira, A.C.R.D., Veloso, V.D.R.S., Barros,
R.G.,Fernandes, P.M. and Souza, E.R.B.D.
2008. Captura deTuta absoluta (Meyrick)
(Lepidoptera: Gelichiidae) comarmadilha
luminosa
na
cultura
do
tomateiro
tutrado.Pesqui. Agropecu. Trop., 38(3), 153157.
-Pereyra, P.C. and Sanchez, N. (2006). Effect of two
Solanaceous plants on developmental and
population parameters of the tomato leafminer,
Tuta
absoluta(Meyrick)
(Lepidoptera:
Gelechiidae). Neotropical Entomology 35:
671-676.
-Ramirez, L, Ramirez, N., Fuentes, L.S., Jiminez, J.
andHernandez-Fernandez,
J.
2010.
Estandarización de unbioensayo y evaluación
preliminar de tres formulacionescomerciales
de Bacillus thuringiensis sobre Tuta
absoluta(Meyrick) (Lepidoptera: Gelechiidae).
Rev. Colomb.Biotecnol., 12(1), 12-21.
-Torres, J.B., C.A. Faria, W.S. Evangelista & D.
Pratissoli. 2001.Within-plant distribution of
the leaf miner Tuta absoluta(Meyrick)
immatures in processing tomatoes, with notes
onplant phenology. Int. J. Pest Manag. 47:
173-178.
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87
ISSN: 2410-7549
EFFECT OF FOLIAR SPRAY OF KNO3, HUMIC ACID CULTIVARS AND
THEIR INTERACTIONS ON LEAF NUTRIENTS OF OLIVE (OLEA
EUROPAEA L.) CVS. KHITHARY AND I18 TRANSPLANTS
Azad Ahmed Mayi 1 and Gulala Mohamed Ameen Saeed 2
Department of Horticulture, School of Plant Production, Faculty of Agriculture and Forestry, University of Duhok,
Kurdistan Region – Iraq.
2
Department of Horticulture, College of Polytechnic Agriculture , University of Helebje, Kurdistan Region – Iraq.
1
Abstract:
This study was carried out during the growing season (2012) in Bakrajo Nursery Station/ Sulaimani,
Kurdistan Region-Iraq. Uniform and healthy olive (Olea europaea L.) cvs. Khithary and I 18 transplants
of (2) years old were used. Filled with river loamy soil to investigate the effect of three levels KNO3 (0, 100
and 200 mg.L-1), three humic acid concentrations (0,150 and 300 mg.L-1) and their interactions on leaf
nutrients of Olive cvs. Khithary and I 18 transplants. The results are summarizing as follows: Khithary
significantly dominated over cv. I 18 in total leaf chlorophyll, P, K, Zn. However cv. I 18 significantly
increased N and Fe. The interactions between cv. Khithary with KNO3 significantly increased P, K, Fe.
While, cv. I 18 significantly increased N and Fe. The interactions between cv. I 18 +300ppm of humic acid
significantly increased Fe. While Khithary with humic acid caused the highest values of N, P, K, Zn. The
interactions between KNO3, humic acid and cv. Khithery affected significantly on most of the leaf
nutrients characteristics. While, 200 ppm KNO3+0 humic acid with cv. Khithary increased all parameters
except leaf Zn.
* Part of M.Sc thesis of the second author
Key words: KNO3, Humic acid, Olive transplant
INTRODUCTION
O
live belongs to the botanical order,
Ligustrales, family (Oleaceae), this family
includes (30) genus including (Olea) which has
(600) species. Olive is botanically called (Olea
europaea L.). Commercial olives belong to the
(Europaea) species, this species has two
subspecies: oleaster and sativa (Bartolucci, and
Dhakal 1999). World olive production performs
an important role in the economy of many
countries such as Spain, Italy, Greece, Turkey
and Tunisia. Olive is an important perennial crop
in many agricultural regions of the
Mediterranean countries, as it is the most
important olive growing region. The olive tree
yield has two main products: oil and table olives
(Sibbett, et al, 2005). In Iraq, olive trees
growing in some areas of central North Iraq and
Kurdistan Region, Nineveh is the governorate
leading olive producer, its cultivation in Nineveh
spreading in an area including villages of
(Baashiqa, Bahzany, Fadiliya, Sheikh Uday,
Dhecan, Sinjar), Diyala, Kirkuk, Baghdad, Erbil,
Duhok, Aqrah, Bamarni followed by Babylon
(Shaima, 2012).
The importance of olive fruit is due to heavy
loading and dietic value, as the fruit is a good
source of vitamins (A, B, C, D, E) and minerals
88
like (K, Ca, Mg and P) Ibrahim, (2005). In
addition, olive oil is filled with monounsaturated fatty acids and has many antioxidative properties as phenolic acid (Shaima,
2012). Potassium takes part in many important
processes, regulating the opening and closing of
stomata, the transport of organic and inorganic
ions within the plant; promoting the maturity,
yield, size and quality of the fruit. Sufficiency
level of (K %) in olive leaves were (0.8-1.3),
which sampled from the mid-length of current
year's young shoots that do not bear fruit
(Ashraf, et al,. 2004). Organic fertilizers are
natural materials and good medium for the
interaction of micro-organisms and provide plant
with nutrients as well as having an indirect role
in nutrition by the activity of micro organisms.
So using organic and bio-fertilizers instead of
the chemical forms could be the way to produce
the natural healthy fruits. In this respect, the
organic fertilization improved vegetative growth
and nutritional status (Farag, 2006).
This investigation aimed to:
Study the effect of KNO3 and humic acid on
leaf nutrients parameters of (Khithary and I 18)
transplants in the climate at Kurdistan Region.
Find out a fertilization program can replace the
mineral which will be beneficial for organic
production of olives. Save human health and
environment.
Impact of both olive cultivars which newly
entered to the region on the vegetative growth of
olive transplants. In addition to study the
possibility of the production transplanting with
proper size, in a short period of time.
Materials and Methods:
The study was carried out during (2012) in
Bakrajo Nursery station/ Sulaimani, Kurdistan
Region-Iraq, located on 15km southwestern of
Sulaimani city. Uniform and healthy olive (cvs.
Khithary and I 18) transplants of (2) years old
were chosen, (Khithary is originated in Syria and
I 18 is originated in Spain). The experiments
were started in (March 15th 2012), as transplants
were grown in pots each of (5 kg) weight. Three
KNO3 concentrations (0, 100 and 200 mg.L-1),
KNO3 compound of 44% K2O, 13% N and 37%
K. (Restrep-Diaz et al,. 2009). Humic acid (HA)
concentrations (0,150 and 300 mg.L-1), abo
najmeh20, compound of %20 humic acid, %20
organic potassium and %10 organic carbon,
Naser company for agrochemical-Syria), were
sprayed at 15th April and repeated at same
concentrations in 15th May.
Experimental design and statistical analysis:
The experiment was arranged in factorial
experiment. The completely Randomized Block
Design (R.C.B.D) was used, the experiment
comprised of (18) treatments with three
replicates, each replicate was presented by five
pots each pot contained one transplant (Al-Rawi,
and Khalafalla1980).
The obtained data were tabulated and
statistically analyzed by computer using SAS
system (1996). The differences among various
treatment means were tested with Duncun
multiple range test at (5%) level. SAS Institute
(1996).
Parameters:
The following measurements were recorded
on November 25th
1-Nitrogen concentration was determined with
Microkjeldahl.
2-Phosphorus concentration was determined
with
colorimetric
methods
using
Spectrophotometer Pharmacia LKB method.
3- Potassium concentration was determined with
using flamephotometer.
4- Iron and Zing concentration (ppm) was
determined
by
Atomic
Absorption
Spectrophotometer.
RESULTS AND DISCUSSION
1- Leaf nitrogen%
Table (1) shows that the transplant when
treated with 200 mg KNO3.L-1 gave the highest
value of nitrogen content (1.56%) and the lowest
value (1.44%) recorded from untreated
transplants. Nitrogen content% in the leaves of
transplants untreated with humic acid
significantly increased leaf nitrogen contents
compared with other treatments. Leaves nitrogen
content% differed between the two cultivars,
leaves of 'I 18' cultivar contained significantly
higher nitrogen content (1.52%) compared with
'Khithary' leaves (1.46%). The interactions
between KNO3 and humic acid significantly
influenced nitrogen content in transplants leaves
that treated with 0 mg humic acid.L-1 and 200 mg
KNO3.L-1 which gave the highest value (1.76%)
and the lowest value (1.26%) obtained with 200
mg KNO3.L-1 plus 300 mg humic acid.L-1. KNO3
and cultivar interactions displayed cv. 'I 18'
transplants treated with 200 mg KNO3.L-1 gave
the highest percentage of nitrogen (1.58%).The
interactions between humic acid and cultivar
significantly increased nitrogen content%
untreated 'Khithary' transplant which gave the
highest value (1.75%) and the lowest value
(1.23%) was recorded from in 'Khithary'
89
Table (1): Effect of KNO3, humic acid and their interactions on leaf nitrogen (%) content of olive transplants
cvs. 'Khithary' and'I 18'.
Mean
KNO3 ( mg.L-1)
HA
Var*HA
effect of
Var.
( mg.L-1)
0
100
200
Var.
1.57 c-e
1.72 bc
1.95 a
1.75 a
150
1.32 g
1.32 g
1.54 d-f
1.39 d
300
1.39e-g
1.16 h
1.14 h
1.23 e
0
1.45e-g
1.48d-g
1.57c-e
1.50 c
150
1.53d-f
1.64b-d
1.79 b
1.65 b
300
1.35 g
1.53d-f
1.37 fg
1.42 cd
Mean effect of KNO3
1.44 b
1.47 b
1.56 a
Khithary
1.43 b
1.40 b
1.54 a
I 18
1.45 b
1.55 a
1.58 a
0
1.51 cd
1.60 bc
1.76 a
1.62a
150
1.43 d-f
1.48 de
1.66 ab
1.52b
300
1.37e-g
1.35 fg
1.26 g
1.32c
I 18
Khithary
0
1.46 b
1.52 a
Mean effect of HA
Var* KNO3
HA* KNO3
Means within a column, row and their interactions followed with the same letters are not
significantly different from each other’s according to Duncans multiple ranges test at 5% level.
transplants when treated with 300 mg humic
acid.L-1. Results of KNO3, humic acid and
cultivars interactions indicated that spraying
'Khithary' olive cultivar with 200 mg KNO3.L-1
plus 0 mg humic acid.L-1 was the most potent
treatment which gave (1.95%) nitrogen% while
the lowest nitrogen coincided with untreated
'Khithary' olive cultivar(1.14%)
2- Leaf phosphorus%.
Table (2) reveals that the transplant treated
with 200 mg KNO3.L-1 gave the highest value of
phosphorus content (0.47%) and the lowest
value (0.39%) recorded in untreated transplants.
Humic acid concentration decreased leaf
phosphorus
content%,
while
untreated
transplants gave the highest value of leaf p%
compared with other treatments. Khithary
cultivar leaves contained higher phosphorus
content (0.45%) compared with leaves of 'I 18'
(0.41%). The interactions between KNO3 and
humic acid significantly influenced phosphorus
90
leaf content when treated 200 mg KNO3.L-1
which gave the highest value (0.66%) and the
lowest value (0.37%) was recorded from 200 mg
KNO3.L-1 plus 300 mg humic acid.L-1. In the
case of KNO3 and cultivar interaction, it was
found that 'Khithary' leaves treated with 200 mg
KNO3.L-1 gave the highest percentage of
phosphorus (0.53%) compared with other
interactions. The interactions between humic
acid and cultivar showed that phosphorus
content % of untreated 'Khithary' transplants
gave the highest value (0.56%) and the lowest
value (0.39%) was recorded from 'I 18'
transplants when treated by 150 mg humic
acid.L-1. Results of KNO3, humic acid and
cultivars interactions indicated that spraying
'Khithary' olive cultivar with 200 mg KNO3.L-1
plus 0 mg humic acid.L-1 was the most potent
treatment which gave (0.85%) phosphorus, while
the lowest phosphorus content (0.33%) in
Khithary transplant when treated with 0 mg
KNO3. L-1 plus 300 mg humic acid.L-1.
Table (2): Effect of KNO3, humic acid and their interactions on leaf hosphorus (%) content of olive transplants
cvs. 'Khithary' and 'I 18'.
I 18
Khithary
Var.
KNO3 ( mg.L-1)
HA
Mean
Var*HA
( mg.L-1)
0
100
200
0
0.35 ef
0.48 bc
0.85 a
0.56 a
150
0.41 b-f
0.43b-e
0.38d-f
0.41 b
300
0.33 f
0.49 b
0.37d-f
0.40 b
0
0.46 bd
0.36 ef
0.46 bd
0.43 b
150
0.39c-f
0.39c-f
0.38c-f
0.39 b
300
0.42b-f
0.41b-f
0.38d-f
0.40 b
0.39 b
0.43 b
0.47 a
Mean effect of KNO3
Var.
Var*
Khithary
0.36 d
0.47 b
0.53 a
KNO3
I 18
0.43 bc
0.38 cd
0.41 cd
0
0.41 bc
0.42 bc
0.66 a
0.49a
150
0.40 bc
0.41 bc
0.38 c
0.40b
300
0.38 c
0.45 b
0.37 c
0.40b
HA*
KNO3
effect of
0.45 a
0.41 b
Mean effect of HA
Means within a column, row and their interactions followed with the same letters are not
significantly different from each other’s according to Duncans multiple ranges test at 5% level.
3- Leaf potassium (%)
Table (3) shows that spraying olive transplant with KNO3 or humic acid at both levels not affect
the leaf potassium content%. Leaves of 'Khithary' cultivar contained significantly higher potassium
content (0.99%) when compared with leaves of 'I 18' (0.77%). The interactions between KNO3 and
humic acid significantly influenced potassium content in the leaves of transplants when untreated
transplant gave the highest value (0.98%). KNO3 and cultivar interactions showed that the leaves of
'Khithary' transplants treated with 100 mg.L-1 KNO3 contained the highest percentage of potassium
(1.08%) compared with other interaction between KNO3 and cultivar. The interactions between humic
acid and cultivar non affecter on potassium content% of the 'Khithary' transplants when treated with
300 mg humic acid.L-1 which gave the highest value (1.00%). Results of KNO3, humic acid and
cultivars interactions indicated that spraying 'Khithary' olive cultivar with 100 mg KNO3.L-1 plus 300
mg humic acid.L-1 was the most potent treatment which gave the highest value (1.14%) potassium,
while the lowest potassium content (0.62%) in 'I 18' transplant was found when treated with 100 mg
KNO3.L-1 plus 0 mg humic acid.L-1.
91
Table (3): Effect of KNO3, humic acid and their interactions on leaf of potassium (%) content of olive
transplants cvs. 'Khithary' and 'I 18'.
KNO3 ( mg.L-1)
HA
Mean effect
Var.
Var*HA
-1
( mg.L )
of Var.
0
100
200
1.04 ab
1.00 ab
0.92 a-d
0.98 a
150
0.92 a-d
1.12 a
0.91 a-e
0.98 a
300
0.93 ab
1.14 a
0.92 a-d
1.00 a
0
0.92 a-c
0.62 e
0.92 a-d
0.82 b
150
0.89 a-e
0.77b-e
0.78 b-e
0.81 b
300
0.80 b-e
0.63 de
0.63 c-e
0.69 b
0.92 a
0.88 a
0.85 a
Khithary
0.96 ab
1.08 a
0.91 bc
I 18
0.87 bc
0.67 d
0.78 cd
0
0.98 a
0.81 ab
0.92 ab
0.90a
150
0.90 ab
0.94 ab
0.84 ab
0.90a
300
0.87 ab
0.88 ab
0.77 b
0.84a
I 18
Khithary
0
Mean effect of KNO3
Var* KNO3
HA* KNO3
0.99 a
0.77 b
Mean effect
of HA
Means within a column, row and their interactions followed with the same letters are not significantly different
from each others according to Duncans multiple ranges test at 5% level.
4- Leaf iron (ppm)
In table (4), iron content (ppm) in leaves differed significantly between the two cultivars, 'I 18'
cultivar contained significantly higher iron content (82.70 ppm) when compared with 'Khithary' (78.69
ppm).
I 18
Khithary
Table (4): Effect of KNO3, humic acid and their interactions on leaf iron (ppm) concentrations of olive
transplants cvs.'Khithary 'and 'I 18'.
KNO3 ( mg.L-1)
Mean
HA
Var.
Var*HA
effect of
( mg.L-1)
0
100
200
Var.
0
34.68 i
60.28 h
101.40 a
65.46 f
150
68.91 f
95.69 b
99.79 a
88.13 b
300
86.61 c
75.23 e
85.63 c
82.49 c
0
63.36 g
93.97 b
77.79 d
78.37 d
150
95.11 b
66.43 f
68.01 f
76.52 e
300
84.96 c
95.08 b
99.59 a
93.21 a
72.27c
81.11 b
88.70 a
Mean effect of KNO3
78.69 b
82.70 a
Var*
Khithary
63.40 e
77.07 d
95.61 a
KNO3
I 18
81.15 c
85.16 b
81.80 c
0
49.02 g
77.13 f
89.59 b
71.91c
150
82.01 e
81.06 e
83.90 d
82.33b
300
85.79 c
85.16cd
92.61 a
87.85a
HA*
KNO3
Mean effect of HA
Means within a column, row and their interactions followed with the same letters are not significantly different
from each others according to Duncans multiple ranges test at 5% level.
92
It was clear that treated transplants, with 200
mg KNO3.L-1 gave the highest value of iron
content (88.70 ppm. Results of KNO3, humic
acid and cultivars interactions indicated that
spraying 'Khithary' olive cultivar with 200 mg
KNO3.L-1 plus 0 mg humic acid.L-1 was the
most potent treatment which gave (101.40 ppm)
iron,. The interactions between KNO3 and humic
acid significantly influenced iron content in leaf,
when treated with 200 mg KNO3.L-1 plus 300
mg humic acid.L-1 gave the highest value (92.61
ppm) and the lowest value (49.02 ppm) was
recorded in untreated transplants. Regarding
KNO3 and cultivar interactions, leaf of 'Khithary'
transplants treated with 200 mg KNO3.L-1
contained the highest percentage of potassium
(95.61ppm) compared with other interaction
between KNO3 and cultivar.The interactions
between humic acid and cultivar had
significantly increased in iron content (ppm) in
cv. 'I 18' transplant when treated with 300 mg
humic acid.L-1 giving the highest value (93.21
ppm) and the lowest value (65.46 ppm) was
recorded in untreated transplants of cv.
'Khithary'.
5- Leaf zinc (ppm).
In table (5), it was notice that the transplant
when treated to 200 mg KNO3.L-1 gave the
highest value of zinc content (17.02ppm). Zinc
content in leaf of transplants treated with 300 mg
humic acid.L-1 increased significantly. Zinc
content in leaves differed significantly between
the two cultivars, 'Khithary' leaves cultivar
contained higher zinc content (13.48 ppm) when
compared with 'I 18' (12.68 ppm). The
interactions between KNO3 and humic acid
significantly influenced zinc content in the
leaves when treated with 200 mg KNO3.L-1 plus
0 mg humic acid.L-1 by giving the highest value
(20.75ppm).
Table (5): Effect of KNO3, humic acid and their interactions on leaf Zinc (ppm) concentrations of olive
transplants cvs. 'Khithary ' and ' I 18'.
I 18
Khithary
Var.
HA KNO3
Mean
Var*HA
effect of
( mg.L-1)
0
100
200
0
6.54 g
13.84 d
14.81 cd
11.73 cd
150
7.84 f
14.84 cd
14.25 cd
12.31 c
300
14.87cd
15.24 c
19.12 b
16.41 a
0
6.53 g
11.85 e
26.68 a
15.02 b
150
15.11 c
7.42 fg
12.55 e
11.69 cd
300
7.85 f
11.43 e
14.67 cd
11.32 d
9.79 c
12.44 b
17.02 a
Khithary
9.75 d
14.64 c
16.06 b
I 18
9.83 d
10.23 d
17.97 a
0
6.53 e
12.85 c
20.75 a
13.38
b
150
11.47 d
11.13 d
13.40 c
12.00
c
300
11.36 d
13.34 c
16.90 b
13.86
a
Mean effect of KNO3
Var* KNO3
KNO3 ( mg.L-1)
HA
Var.
13.48 a
12.68 b
Mean effect of HA
Means within a column, row and their interactions followed with the same letters are not significantly
different from each others according to Duncans multiple ranges test at 5% level.
93
Regarding KNO3 and cultivar interactions, it
was found that the leaves of 'I 18' transplants
treated with 200 mg.L-1 KNO3 contained the
highest percentage of zinc (17.97 ppm)
compared with other interactions between KNO3
and cultivar. The interactions between humic
acid and cultivar had significantly increased
zinc contents in' Khithary' transplants when
treated with 300 mg. humic acid.L-1 giving the
highest value (16.41ppm) and the lowest value
(11.32 ppm) was recorded from the untreated
transplants of cv. 'I 18'. Results of KNO3, humic
acid and cultivars interactions indicated that
spraying 'I 18' olive cultivar with 200 mg
KNO3.L-1 plus 0 mg humic acid.L-1 was the most
potent treatment which gave (26.68ppm) zinc,
while the lowest zinc (6.53ppm) was recorded in
untreated 'I 18' transplant.
Discussions:
1- KNO3: It is clear from studied parameters that
the effect of KNO3 on nutrient composion
characteristics significantly affected and
improved all parameters, the results may be due
to role of K and N in plants such as
photosynthesis
reactions,
nucleic
acid
metabolism,
protein
and
carbohydrate
biosynthesis due to increased leaf mineral
content. (Hafez, and El-Metwally 2007).
Potassium takes part in many important
processes, regulating the opening and closing of
stomata, the transport of organic and inorganic
ions within the plant, (Ibrahim, 2005) and
(Elloumi, et al,. 2009).
2- Humic acid: For the effect of humic acid, the
same tables that show studied parameters
indicates that leaf nutrient status gave the
highest value. The reason for the positive effect
of humic acid may be due to the role of (HA) to
stimulate plant growth by acting on mechanisms
involved in: cell respiration, photosynthesis,
protein synthesis, water and nutrient uptake and
enzyme activities (Nardi et al,.1996, Chen et al,.
2004, and Ali, et al,.2007). Whereas direct
effects are various biochemical actions exerted at
the cell wall, membrane or cytoplasm and
mainly of hormonal nature (Varanini and Pinton
2001 and Chen et al,. 2004). The hormone like
activities of HAs is well documented in various
papers, in particular auxin, cytokinin and
gibberellins like effects (Piccolo et al,. 1992 and
Pizzeghello, et al,. 2002).
3- Cultivars: It's clear from most tables that the
vegetative growth characteristics significantly
94
differed between the two cultivars. The
differences between the cultivars in leaves
nutrient such as (N, P, K, Fe and Zinc, may be
ascribed to the differences in genotype
characteristics (Jordao, et al,. 1999). In addition,
the genetic integrity of the plant species might
influence particular nutrient uptake efficiency
(Popovic et al,. 1999). Then, these differences in
nutrient uptake efficiency between cultivars may
cause differences in vegetation growth
characteristics. Also, the differences in growth
vigor between the two cultivars may be
attributed to the response of different cultivars to
the local environmental conditions according to
the genetic variation between the cultivars
(Gaafar and Saker 2006 and Khalifa 2007).
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95
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96
ISSN: 2410-7549
EFFECT OF CULTIVARS, COMPOST, HUMIC ACID AND THEIR
INTERACTIONS ON LEAF NUTRITIONAL STATES OF SWEET CHERRY
(PRUNUS AVIUM L.)
1
Azad Ahmed Mayi 1 and Nawzad Jemil Ibrahim 2
Department of Horticulture, College of Agriculture, University of Duhok, Kurdistan Region – Iraq.
2
Department of Horticulture, Ministry of Agriculture and irrigation, Kurdistan Region – Iraq.
Abstract:
The present study was carried out on sweet cherry (Prunus avium L.), during the growing season of
(2012-2013) on private sweet cherry orchard, located in the Bibad village near Amadi town / Duhok
governorate- Kurdistan region-Iraq. The field experiment was done in the orchard that contained two
sweet cherry cultivars (“Nefertity” and “Berlit”). The application of compost was done in December 25th
2012, at (0, 2, 4 and 6 kg /tree), foliar spray of humic acid done at (0, 100, 200 and 300 ppm) and repeated
after two weeks. The results are summarized as follows: Nefertity cultivar significantly dominated over
Berlit cultivar in leaf N, P content, whereas Berlit cultivar dominated leaf (N, K, Ca). Compost specially at
4kg/tree has significantly improved leaf P, K,whereas, compost at 6 kg/tree leaf nutrient N. Humic acid
specially at 300 ppm has significantly increased the leaf nutrients N, P, while, 200 ppm increased the leaf
K, while, 100 ppm increased the leaf N, K. The interactions between cultivars, compost and humic acid
has affected significantly most of the studied parameters. The more effective treatment interactions was
Nefertity + 6 Kg /tree of compost plus 200 ppm of humic acid which increased significantly P,K, but
Nefertity + 6 Kg /tree of compost plus 300ppm of humic acid treatment increased leaf N content.
*Part of M.Sc thesis of the second author
Key word: Cherry , Compost, Humic acid
INTRODUCTION:
he sweet cherry (Prunus avium. L.)
belongs to the Rosaceae family, subfamily Prunidaecea (Rodrigues and Antunes,
2008). It is believed to be originated from the
regions between the Black and Caspian Sea of
Asia Minor. Seed spreading by birds carried it to
Europe, where the earliest cultivation of sweet
cherry was reported. Further spread to North
America via English colonists occurred in the
seventeenth century. (Webster and Looney,
1996). Sweet cherry is one of the most popular
temperate fruits. According to (FAO, 2010),
cherries produced worldwide, and an important
horticultural crop - approximately 2.2 million
tons of cherries were produced worldwide in
2009 (Anonymous, 2010). Compost is a resource
of converting organic waste material, such as
food waste, yard waste and manure, into a matter
called humus, a nutrient-rich soil amendment.
Humus is an essential element in maintaining
healthy soil and plant, making composting a
useful tactic for nurturing productive agricultural
fields, ornamental plants and grasses (Chiumenti,
2005).The compost has an important role in the
agriculture sector because it contains a high
amount of elements necessary for plant growth
and soil improvement, the use of compost as a
T
fertilizer for plant in Iraq and Kurdistan has a
large space and this is backward in the field of
agriculture when we compare with the
developed countries. Thus, encouraging to use
the compost as a first studies in Kurdistan and
Iraq in order to encourage our farmer to use the
compost as a plant fertilizer. The risks and
problems posed by heavy metals in fertilizers
and other soil inputs have increasingly drawn the
attention
of
farmers,
environmental
organizations, consumers, and public policy
makers.
This study evaluates a wide spectrum of soil
amendments and fertilizers used in organic
agriculture, including biosolids, major nutrient
fertilizers, industrial wastes, composts, liming
materials and micronutrient sources with a focus
on inputs used in organic agricultural production
in Iraq. Humic substances are no one single
chemical recognized as humic acid, since the
chemical makeup has never been completely
defined. These materials are composed of
complicated organic mixtures which are
associated together in a random manner,
resulting in extraordinarily complex materials. It
has been suggested that no two molecules of
humus are exactly the same (Mikkelsen, 2005).
The cherry fruit is desirable in Kurdistan in
97
relations as far as consumption and also
desirable by farmers to cultivate but the
production is very little so far. Therfor, this
study is as the first one on the nutrition of cherry
fruits in order to improve the quality and
increase their production.
This investigation aimed to study the
response of two cultivars to local environmental
condition, and their response to fertilized by
organic matter (compost and humic acid).
However, it also hopes to confirm the risk of
heavy metal concentration in compost to find a
fertilization program that can replace the
minerals which will be beneficial for organic
production of sweet cherry, since there are little
or no studies in Kurdistan about the role organic
fertilization in yield and quality of sweet cherry
according the aims were to evaluate the
interaction effect of cultivars, compost, humic
acid on leaf nutrient statutes and some heavy
metal concentration of two cultivars of sweet
cherry.
MATERIALS AND METHODS
This study was carried out in the Bibad
village near Amadi town/ Duhok governorate/
Kurdistan region of Iraq. The orchard is situated
at latitude: 37.05◦N, and longitude 43.29◦ E and
at an altitude of 1202 m above the sea level. The
experiment included two cultivar of sweet cherry
"Nefertity" and "Berlit". Application of compost
at different levels (0, 2, 4 and 6 kg /tree), and
foliar spray of humic acid at concentrations (0,
100, 200 and 300 ppm). The compost were
98
carried out is consisting from resides waste of
Dohuk city, is produced in Kawsha manufacture
of compost fertilizer. The orchard experiment of
compost application was done in December 25th
2012, by working hole around the tree under
brunch projection and mixed with the soil in
order to investigate the effect of soil application
of four levels of compost, (0, 2, 4 and 6 kg /tree).
The humic acid is a liquid content analysis w/w,
organic matter 5%, (K2 O) 1%, total humic +
fulvic acid 15%. The foliar spray of humic acid
was done in April 15th 2013, after full bloom at
four concentrations (0, 100, 200 and 300 ppm)
and replicated the same concentrations after two
weeks after the first spray.
Statistical analysis
All the obtained data were tabulated and
statistically analyzed with computer using SAS
system (SAS, 1996). The experiment followed
Randomized Complete Block Design in factorial
Experiment; the experiment comprised of 32
treatments with three replicates, each replicate
was presented by one tree of each cultivar. The
differences between various treatment means
were tested with Duncun Multiple Range Test at
5% level, (Al- Rawi and Khalaf-Alla, 2000).
Measurements:
Leaf nutrients states
1- Total nitrogen (N %).
2- Total phosphorus (P%).
3- Total potassium (K %).
4-Total calcium (Ca%).
RESULTS
1- Leaf nitrogen content (%)
2.5
a
2
a
b
a
c
b
ab ab a
1.5
1
Berlit
0.5
0
6
Nefertity
Leaf N content (%)
d
2
200
4
100
0
Cultivar
300
0
Compost (Kg)
Humic acid (ppm)
(columns with the same letters are not significantly different from each other according to Duncan's multiple rang
Figure (1) Effect of cultivar, compost and humic acid on leaf N content (%) of sweet cherry.
Nefertity
Berlit
Humic acid
(ppm)
Cultivar
Table (1): interactions effect of cultivar, compost and humic acid on leaf N content (%) of sweet cherry.
Compost (Kg/tree)
Cultivar ×
Humic
0
2
4
6
0
1.13 h
1.37 gh
1.82 a-f
1.92 a-c
1.56 c
100
1.62 b-g
1.47 fg
2.03 a
1.81 a-f
1.73 b
200
1.86 a-e
1.51 e-g
1.92 a-c
2.02 a
1.83 ab
300
1.98 ab
1.93 a-c
1.78 a-f
1.99 a
1.92 a
0
1.54 d-g
1.74 a-f
2.04 a
2.01 a
1.84 ab
100
1.61 c-g
1.83 a-f
1.88 a-d
1.88 a-d
1.80 ab
200
1.31 gh
1.86 a-e
1.58 d-g
2.09 a
1.71 bc
300
1.30 gh
1.90 a-d
1.57 c-g
2.10 a
1.72 bc
Cultivar × Compost
Cultivar
Berlit
1.65 de
1.57 ef
1.89 a-c
1.93 ab
Nefertity
1.44 f
1.83 bc
1.77 cd
2.02 a
Humic acid × Compost
Humic
acid
(ppm)
0
1.34 e
1.56 d
1.93 ab
1.97 ab
100
1.62 cd
1.65 cd
1.96 ab
1.84 a-c
200
1.59 d
1.68 cd
1.75 b-d
2.05 a
300
1.64 cd
1.92 ab
1.68 cd
2.05 a
Means of each factor and their interaction followed by the same letter are not significantly different from each
others according to duncans multiple ranges test at 5% level.
99
Figure (1) clearly shows that there are no significant differences between two cultivar on leaf
nitrogen content. Soil application of compost at a level (6 kg/tree) had significantly effect which
recorded (1.98 %), and the lowest value was recorded in control (1.55 %). Foliar spray of humic acid
at a concentration (300 ppm) were significantly effective, which registered (1.82 %), while the lowest
values were recorded in control (1.70 %). The interaction between cultivars and humic acid on leaf N
indicate that cultivar Berlit with humic acid at a (300 ppm) was the best treatments when compared
with other treatment. The results of interaction between application (6 kg/tree) of compost plus (200
ppm) of humic acid, produced the better treatment (2.059 %), while the lowest concentration was
observed in control (1.464 %).
2. Leaf phosphorus content (%):
Figure (2) shows the phosphors concentration in cultivar Nefertity increased significantly
compared with cultivar Berlit. Obviously soil application of compost at a (4 kg/tree) was better
treatment when compared with other treatments which recorded (0.949%), and the lowest value was
recorded in control (0.437%). The same figure illustrates that the humic acid at a concentrations (300
ppm) was suggestively better treatment which registered (0.823 %), but the lowest values were
recorded in control (0.468 %).
However, the interaction between Nerfertity plus (6 kg/tree) of compost gave the highest value
(1.306 %) when compared with control. The interaction between cultivar Nefertity and (100 ppm) of
humic acid gave the highest value compared with other treatment.
1
a
a
0.9
a
a
ab
0.8
b
Leaf content (%)
0.7
0.6
0.5
b
b
c
b
0.4
0.1
00
Berlit
0.2
Cultivar
Nefertity
0.3
6
2
200
4
0
Compost (Kg)
300
100
0
Humic acid (ppm)
(columns with the same letters are not significantly different from each other according to Duncan's multiple rang test at 5% leve).
Figure (2) Effect of cultivar, compost and humic acid on leaf P content (%) of sweet cherry.
Manifests the interaction between (4 kg/tree) of compost with (100 ppm) of humic acid, produced
the better treatment which registered (0.917) when compared with other treatments.
The same tableillustrates that the interaction effect of cultivar Nefertity + (4 kg/tree) compost +
(100 ppm) humic acid provided the best treatment which was (1.26 %) when compared with another
treatments.
100
Humic acid
(ppm)
Nefertity
Berlit
Cultivar
Table (2) : Interactions effect of cultivar, compost and humic acid on leaf P content (%) of sweet cherry.
0
2
4
6
0
0.17 e
0.21 c-e
0.27 c-e
0.30 b-e
0.24 d
100
0.27 c-e
0.33 b-e
0.56 b-e
0.66 bc
0.46 c
200
0.60 b-d
0.50 b-e
0.61 b-d
0.67 bc
0.59 bc
300
0.63 bc
0.67 bc
0.68 bc
0.72 b
0.67 b
0
0.22de
0.54 de
0.66 bc
1.20 a
0.65 b
100
0.40 de
0.68 bc
1.26 a
1.33 a
0.92 a
200
0.53 b-e
0.61 b-d
1.28 a
1.36 a
0.94 a
300
0.65 bc
0.60 b-d
1.29 a
1.32 a
0.96 a
Compost (Kg/tree)
Cultivar ×
Humic
Cultivar × Compost
Cultivar
Berlit
0.42 c
0.43 c
0.53 c
0.59 c
Nefertity
0.45 c
0.61 c
1.12 b
1.30 a
Humic
acid
(ppm)
0
0.19 e
0.37 c-e
0.468 cd
0.75 ab
100
0.33 de
0.51 b-d
0.91 a
1.00 a
200
0.56 bc
0.561 b-d
0.94 a
1.01 a
300
0.64 bc
0.63 bc
0.98 a
1.02 a
Means of each factor and their interaction followed by the same letters are not significantly
The interaction between cultivar Berlit + (100 ppm) was the best treatment when compared with
other treatments. The interaction between (6 kg/tree) of compost + (200 ppm) of humic acid produced
the best treatment.
3. Leaf potassium content (%):
Figure (3) shows that the leaf potassium in cultivar Berlit had significant effect compared to
cultivar Nefertity. Obviously application of compost at a level (4 kg/tree) was the best treatments
which noted (1.701 %), when compared with other treatment. The foliar spray of humic acid at a (100
ppm) was a better treatment among the humic concentration which registered (1.746 %), value (1.926
%), when compared with other treatments. In the same table the interaction between cultivar Nefertity
+ (6 kg/tree) of compost + (200 ppm) of humic acid provided the maximum value (2.011 %), when
compared with other treatments. The interaction of cultivars Berlit plus (2 kg/tree) of compost has the
best treatment to increase the leaf potassium, (1.608 %) when compared with other treatments.
101
2
a
b
1.6
b
a
a
a ab
ab
a
b
1.4
1.2
1
0.8
0.6
0.4
0.2
0
6
Berlit
Nefertity
Leaf potassium content (%)
1.8
2
200
4
100
0
0
Cultivar
300
Compost (Kg)
Humic acid (ppm)
(columns with the same letters are not significantly different from each other according to Duncan's multiple rang test at 5% leve).
Figure (3): Effect of cultivar, compost and humic acid on leaf K content (%) of sweet cherry.
Humic acid
(ppm)
Nefertity
Berlit
Cultivar
Table (3): Interactions effect of cultivar, compost and humic acid on leaf K content (%) of sweet cherry.
0
2
4
6
0
1.21 jk
1.39 g-j
1.41 f-j
1.48 d-j
1.37 d
100
1.84 a-c
1.87 a-c
1.93 ab
1.66 b-i
1.82 a
200
1.77 a-e
1.77 a-e
1.85 a-c
1.84 a-c
1.81 ab
300
1.689a-h
1.75 a-f
1.84 a-c
1.91 a-c
1.80 ab
0
1.04 k
1.32 i-k
1.42 f-j
1.36 h-i
1.29 d
100
1.61 b-i
1.64 b-i
1.81 a-d
1.57 c-i
1.66 bc
200
1.63 b-i
1.74 a-f
1.70 a-g
2.01 a
1.77 ab
300
1.68 a-h
1.70 a-g
1.63 b-i
1.47 e-j
1.62 c
Compost (Kg/tree)
Cultivar ×
Humic
Cultivar × Compost
Cultivar
Berlit
1.63 ab
1.70 a
1.76 a
1.72 a
Nefertity
1.49 b
1.60 ab
1.64 ab
1.607ab
Humic
acid
(ppm)
0
1.13 f
1.36 e
1.42 de
1.42 de
100
1.73 a-c
1.76 a-c
1.87 ab
1.61 cd
200
1.70 a-c
1.76 a-c
1.78 a-c
1.92 a
300
1.68 bc
1.73 a-c
1.73 a-c
1.69 a-c
Means of each factor and their interaction followed by the same letters are not significantly
102
4. Leaf calcium content (%)
2.5
a
ab
ab
b
ab
a
1.5
1
0.5
00
200
6
Berlit
Nefertity
Leaf Ca content (%)
ab
b
b
2
a
2
4
100
0
0
Cultivar
300
Compost (Kg)
Humic acid (ppm)
(columns with the same letters are not significantly different from each other according to Duncan's multiple rang test at 5% leve)
Figure (4) effect of cultivar, compost and humic acid on leaf calcium content (%) of sweet cherry.
Humic acid
(ppm)
Nefertity
Berlit
Cultivar
Table (4) interactions effect of cultivar, compost and humic acid on leaf Ca content (%) of sweet cherry.
0
2
4
6
0
2.05 b-d
2.08 a-d
2.08 a-d
1.88 cd
2.02 b
100
2.14 a-d
2.08 a-d
2.18 a-d
2.04 b-d
2.11 b
200
2.20 a-d
2.15 a-d
2.27 a-d
2.13 a-d
2.18 b
300
2.30 a-c
2.31 a-c
2.54 ab
2.673 a
2.46 a
0
1.66 d
1.74 cd
1.98 b-d
2.27 a-d
1.91 b
100
1.78 cd
1.85 cd
2.03 b-d
2.27 a-d
1.98 b
200
1.84 cd
2.06 a-d
2.04 b-d
2.19 a-d
2.03 b
300
1.92 b-d
1.89 cd
2.00 b-d
2.12 a-d
1.98 b
Compost (Kg/tree)
Cultivar ×
Humic
Cultivar × Compost
Cultivar
Berlit
2.17 a
2.15 a
2.27 a
2.18 a
Nefertity
1.80 b
1.88 b
2.01 ab
2.21 a
Humic
acid
(ppm)
0
1.86 b
1.91 b
2.03 ab
2.07 ab
100
1.96 b
1.96 b
2.10 ab
2.15 ab
200
2.02 ab
2.10 ab
2.15 ab
2.16ab
300
2.11 ab
2.10 ab
2.27 ab
2.40 a
Means of each their interaction followed by the same letters are not significantly different from
each others according to Duncans multiple ranges test at 5% level.
103
Figure (4) shows leaf Ca content in cultivar
Berlit had a significant effect when compared
with cultivar Nefertity. The results clearly show
the soil application of compost at a level (6
kg/tree) was significantly increased the leaf Ca
content, (2.201 %), when compared with other
treatments.
Foliar spray of humic acid at (300 ppm) was
better treatment which registered (2.224 ppm),
when compared with other treatments. Table (4),
the interaction between cultivar Berlit plus (2
kg/tree) of compost was the best treatment when
compared with other treatments. The results of
combining between cultivar Berlit plus humic
acid at a (300 %) indicated the highest
significant effect when compared with other
treatments. Also, the interaction between (6
kg/tree) compost plus plus (300 ppm) humic acid
produced the best treatment (2.400 ppm), when
compared with control. The interaction between
cultivar Berlit + (6 kg/tree) compost + (300
ppm) humic acid which provided the best
treatment (2.673 %) when compared with other
treatments.
DISCUSSION
1. The effect of cultivar on leaf nutritional state,
may be ascribed to the differences in genotype
characteristics for root growth, nutrient
absorption efficiency and photosynthesis process
efficiency (Jorda, et al, 1999). Also, the response
of different cultivars to the local environmental
condition according to the genetic variation
between the cultivars (Gaafar and Saker, 2006 ;
Khalifa, 2007).
2. The effect of compost may be due to the
improvement of soil physical, biological
properties and chemical properties resulting
more release of nutrient elements available
which absorbed by plant root and its effect on
the physiological process, in addition to water
use efficiency, also adequate nutrient quantities
of nitrogen, phosphorus, and potassium, which
increase both rate of leaf expansion as well as
cell division which subsequently leads to larger
individual leaves and higher photosynthesis
activities (Abd El-Wahab, 2011). May be
attributed to a higher nutritional uptake mainly
by greater expansion of root system due to
increased supply of photosynthetic productions
in the leaves, attributed to presence of plant
growth regulators, which are produced by
increased activity of microbes such as fungi,
104
bacteria, yeasts, actinomycetes and algae
(Arancon et al., 2004 ).
3.The effect of humic acid may be acting on
mechanisms involved in: cell respiration,
photosynthesis, protein synthesis, water and
nutrient uptake, enzyme activities. (Ali et. al.,
2007). The hormone like activities of (HA) is
well documented in various papers, in particular
auxin-, cytokinin and gibberellins like effects
(Pizzeghello et al., 2002). Also, the effect of
humic acid may be due to the role of (HA) to
increase of cation exchange capacity which
affects the retention and availability of nutrients,
or due to a hormonal effect, or a combination of
both. (Chun hua et al., 1998).
Conclusions:
1. The Nefertity cultivar was superior on the
Berlit cultivar in increasing vegetative growth,
and most leaf nutrients.
2. Soil application of compost at a level 4 kg/tree
was more than other levels and control in
increasing most vegetative growth parameters
and most leaf nutrients.
3. Foliar spraying of humic acid at concentration
300 ppm had more effects .
4. The interaction of Nefertity cultivar + 4 Kg of
compost + 300 ppm of humic acid had more
effect on increasing in most studied parameter.
2. Recommendations:
The flowing points of view can be
recommended:
1. Testing other cultivars of sweet cherry.
2. Testing the different level and time of
application of compost as well testing other
organic fertilizer such as animal manure chicken
manure and municipal.
3. Testing the effect of spraying with humic acid
on the other sweet cherry cultivar.
4. Farmers are recomended to use the compost in
their orchards like other organic fertilizer, such
as animal manure without any fear of toxic
materials, soil pollution and harmful effect of
heavy metals.
References:
Abd El-Wahab, M. A. (2011). Reducing the Amount
of Mineral Nitrogen Fertilizers for Red Globe
Grapevines by Using Different Sources of
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Ali V. K.; H. Çelik; M. A. Turan and B. B. Aºýk.
(2007). Effects of Soil and Foliar Applications
of Humic Substances on Dry Weight and
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agricultural faculty, department of soil science
and plant nutrition. aust. J. basic & Appl. Sci.,
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Al-Rawi, A. A. and A. Khalaf- Alla (2000). Analysis
of Experimental Agriculture Design. Dar AlKutub for Printing and Publishing. Mosul
University. (In Arabic).
Anonymous, (2010). FAOSTAT, FAO Statistical
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Nations),
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Arancon N. Q.; C. A. Edwards; P. Bierman; J.D.
Metzger, S. Lee and C. Welch (2004). Effects
of Vermicomposts on Growth and Marketable
Fruits of Field-Grown Tomatoes, Peppers and
Strawberries. Pedobiol. 47: 731-735.
Chiumenti, A. (2005) Modern Composting
CompostingTechnologies. Biocycle, JG Press
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Chun hua, L.; R.J. Cooper and D.C. Bowman, (1998).
Humic
acid
Application
Affects
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FAO (2011), Joint FAO/WHO Food Standards
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on
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Gaafar, R. M. and M. M. Saker (2006). Monitoring of
Cultivars Identity and Genetic Stability In
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Ibrahim, A. M. and G. A. Abd El-Samad (2009).
Effect of different irrigation regimes and
partial substitution of N-miniral by organic
manures on water use, growth and
reproductively of pomegranate trees. European
J. Sci. Res. 38(2): 199-218.
Jorda O; L. Durarte; F. Calouro and A. Silva. (1999).
Relationship Between Fruit and Foliar Mineral
Levels In Twenty Olive Cultivars Grown In
Portugal. Acta Hort. (ISHI) 286: 267-270.
Khalifa, G. H. F.H. (2007). Effect of Planting Date
and on Growth and Yield Characteristics of
Two Variety of Strawberry (Fragaria x
ananassa Duch), M.Sc. Thesis, Agriculture
and Forest college, Mosul University, Ministry
of Higher Education and Scientifics. Research.
Iraq.
Mikkelsen R.L. (2005) Humic Materials for
Agriculture, Better Crops/Vol. 89 (No. 3).
Pizzeghello D.; G. Nicolini and S. Nardi, (2002).
Hormone-Like Activities of Humic Substances
in Different Forest Ecosystems. New Phyto,l
155, 393-402.
Rodrigues and D.Antunes (2008). Influence of
Nitrogen and Potassium on Yield, Fruit
Quality and Mineral Composition of Kiwifruit.
International j. of energy and Environment.
Issue 1, volume 2.
SAS Institute, Inc (1996). The SAS system. Relase
6.12. Cary, NC.
Webster A.D. and N.E. Looney (1996). The
taxonomic classification of sweet and sour
cherries and a brief history of their cultivation,
crop physiology, production and uses, CAB
international: wallingford, oxon, UK pp 3–24.
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106
ISSN: 2410-7549
SOME TECHNOLOGICAL PROPERTIES OF SAWN BOARD Eucalyptus
camaldulensis Denh. GROWN IN ASKIKALAK
Mohammedamin Yasin Taha
School of Forestry Dept. of Forestry, Faculty of Agriculture, University of Duhok, Kurdistan Region-Iraq.
ABSTRACT
This study deals with wood density and static bending boards property of Eucalyptus camaldulensis
Den. It was conducted using standardized, defect-free test specimens. Boards of air drying showed lowest
value (749.117 kg/cm²) of static bending when compared with kiln (812.267kg/cm²) and solar
(815.267kg/cm²) drying respectively, whereas, the quarter sawing boards recorded the lowest rates
(586.633kg/cm²) when compared with flat sawn boards (1008.467kg/cm²), Also 2cm thickness level
achieved the lowest rate (747.367kg/cm²) when compared with other two levels 4cm (812.650 kg/cm²) and
6cm (837.633kg/cm²) of thickness. But, there were no any significant effects of wood density on the studied
factors. E. camaldulensis in Kurdistan Region of Iraq has potential for traditional uses and by itself it can
be harvested by applying suitable techniques at plantation areas, in saw mills and drying for utilization.
KEYWORDS: Wood Density, Static bending, E. camaldulensis, Sawn Board
Introduction
K
urdistan Region of Iraq is located in
Northern Iraq. According to its fertile
soil and appropriate environmental conditions
have made this region known for its natural
forests. Most prominently, the flowing of a
number of big rivers through this region such as
Tigris and some others lead to the prospect of
establish artificial stands depending on river for
irrigation.
About 30-40 species of the genus were
introduced to Iraq during the last century
Shahbaz, (2010).
Literature mentioned that the two main of
them being E.camaldulensis and E. microtiheca,
were considerably succeeded in middle and
southern of country Rooitzsch and Reader,
(1969). E. camaldulensis Den. has been used for
pulp, chipboard, fire wood, shelter belts and
others. Appropriate temperature and humidity
rates as well as precipitation in Kurdistan
provided well situations for Eucalypt throughout
the year. The shortage of manufacturing wood
products in Iraq, so including in Kurdistan
Region makes it difficult to have a clear
overview of the prospects of eucalypt
exploitation (Taha, 2013).
Eucalypts
species
are
recognized
simultaneously of the fastest wood producing
trees. It has the capability to produce
approximately100m³/capita. Some investigations
in their studies (Myburg, et al., 2006,Acosta,et
al.,2008 and Iglesias, et al., 2008) reported that
there are about 18 Million hectares of Eucalypt
in the world, and they expected the total areas of
Eucalyptus will increase to reach up to 20
Million hectares in 2010 cited by (Taha,2013). It
is
distinguished
that
environmental
circumstances might have their important
belongings on the wood properties, and for the
reason of the lack of researches on eucalypts
wood in Iraq principally those associated to its
drying, this study was designed to investigate the
wood density and static bending (MOE)
properties of dried sawn board of trees grown in
Kurdistan Region.
Ten trees of Eucalyptus camaldulensis with
DBH 35-45cm were selected, felled, and logged
in a stand at Khabat district (Askikalak) in Erbil
Governorate, Kurdistan Region of Iraq where it
lies at N 36° 15'; E 43° 38’ and located at 252m,
It is37km far from Erbil city. All logs were
bucked (2m length) then they were sawed by
sawing table with band saw (SIPA 100 Saw)
preliminarily; the logs then were converted to
planks. After that, the resulted planks were
sawed into two types of board; quarter sawn
board and flat sawn board at three different
thickness 2,4, and 6cm. Logs were used for
producing lumber for air drying, kiln drying, and
solar drying methods respectively.
Method of randomizing
The following two characteristics were
examined:
Wood density and static bending, before
applying the test, make sure that the samples be
107
free from defects and splits. The tests concerned
load the experiment specimens at Mid-length at
prepared sawed boards.
With the aim of collect material to be used
for present study determined by using ASTM D143-94
and
ASTM
D-2395-93
procedures(Anonymous 1996) get 10 store dried
(Air, Solar, and kiln) boards of (Eucalyptus
camaldulesis) for each properties randomly. To
evaluate wood properties of sawed boards, in
situation (temperature 20-25°C, 35-40% relative
humidity and wood moisture content 12%)
defect free specimens (5×2×2cm) for wood
density (kg/ cm²). And (30×2×2cm) for static
bending (MOE) were tested by using the
universal Strength Testing Machine .
The collected data so collected were analyzed
statistically using experiment; it was included of
three factors:
Factor A: Drying method; with 3 levels: (AD;
Air Drying, KD; Kiln Drying and SD: Solar
Drying)
Factor B: Thickness of board; with 3 levels:
(Th1; 2cm, Th2; 4cm, and Th3; 6cm)
Factor C: Board kind; with 2 levels: (FS; Flat
Sawn board and QS; Quarter Sawn board).
Numbers of treatment combinations were 18.
Ten boards were chosen randomly to represents
replication of each treatment combination. The
experiment was statistically analyzed as factorial
RCBD by using SAS program version 0.9 SAS,
(2002). Statistical differences between treatment
combination means were tested by Duncan
Multiple Range test at 5% level Duncan, (1955).
The influence of interaction between drying
methods, board thickness, and sawing methods
on the studied parameters:
Effect of drying method, board thickness, and
sawing method on wood density and static
bending of dried board as shown in Table (1)
Table (1): Analysis of variance of wood density and static bending as affected by drying method (DM), board
thickness (Th), and board kind (BK).
Wood
Density
F Value
Wood
Density
Pr > F
Static
Bending
F Value
Static
Bending
Pr > F
2
2.09
0.1276
4.45
0.0123
2
2.21
0.1135
13.72
0.0001
1
3.08
0.0815
750.65
0.0001
DM×Th
4
1.06
0.3780
3.83
0.0054
DM× BK
2
0.73
0.4814
3.77
0.0252
Th× BK
2
0.96
0.3846
2.66
0.0731
DM×Th×
BK
4
1.06
0.3778
3.77
0.0060
Variable
s
Drying
Methods
(DM)
Thicknes
s
(Th)
Board
Kind
(BK)
108
d.f
Table (1) refers that drying method (DM), thickness (TH), and board kind(BK) and their interaction
could not affect significantly on wood density. Mean values of (Tab. 2) indicate that moderate values
of wood density have been obtained in almost all treatments.
Accordingly, differences should be so small that they would not be enough to give statistical
significances. The results, also agreed with what has been found by Lima et al., (2008) who mentioned
that density varies from a minimum of 0.319 g/cm³ to a maximum of 0.731 g/cm³. In general, wood
structures formed in early stages of tree growing that have low density.
Table (2): Mean values of wood density as affected by drying method, board thickness, and board kind.
--------------------------------------------------------------------------------------------------------------------------Drying Methods Board Thickness Board Kind (BK) MD TH Mean of (DM)
(DM)
(TH) cm
Flat
Quarter
--------------------------------------------------------------------------------------------------------Air Drying(A)
2
4
6
0.570
0.601
0.608
0.609
0.680
0.789
0.590
0.641
0.699
0.643
(a)
--------------------------------------------------------------------------------------------------Kiln Drying(K) )
2
0.570
0.609
0.609
4
0.601
0.680
0.685
0.670
6
0.608
0.789
0.715
(a)
--------------------------------------------------------------------------------------------------Solar Drying(S)
2
0.570
0.609 0.606
4
0.601
0.680 1.065
0.824
6
0.608
0.789 0.800
(a)
---------------------------------------------------------------------------------------------------Mean of (BK)
0.644
0.780
0.712
(a)
(a)
--------------------------------------------------------------------------------------------------TH BK
T1
0.578
0.616
0.601(a)
T2
0.655
0.939
0.797(a)
T3
0.689
0.786
0.738(a)
--------------------------------------------------------------------------------------------------DMBK
A
0.593
0.693
K
0.648
0.692
S
0.690
0.957
However, the diminishing of wood density alongside tangential direction (flat sawn board) can be
associated to differentiations between chemical compositions in wood structures and existing of
heartwood near to the pith compare to sapwood Akhtari, et al., (2012).The results were achieved from
the study confirmed that the density of dried boards is located within the middy category (0.56 -0.75
g/cm³) according to the classification of the IAWA, (1989).
The studied factors were affected differently on static bending (MOE) (Tab.1). While board
thickness showed moderate significant effects, drying method affected at lower level, and board
kind(BK) high levels of confidence (p<0.05). Drying method showed high statistical influences on
static bending. Boards dried by (AD) possessed the lowest values (Tab.3) because the density values
were obtained in this study show the difficulty of different drying situations clearly.
109
Table (3): Mean values of MOE as affected by drying method, board thickness, and board kinds.
Drying Methods Board Thickness Board Kind (BK) MDTH Mean of (DM)
(DM)
(TH) cm
Flat
Quarter
Air Drying(A)
4
6
2
746.000
553.400 649.700
1036.200 584.500 810.350
749.117
1056.200 614.400 835.300
(b)
--------------------------------------------------------------------------------------------------Kiln Drying(K) )
2
1018.000 556.400 787.200
4
1038.200 586.400 812.300
812.267
6
1058.200 616.400 837.300
(a)
--------------------------------------------------------------------------------------------------Solar Drying(S)
2
1021. 000 559.400 790.200
4
1041.200 589.400 815.300
815.267
6
1061.200 619.400 840.300
(a)
Mean of (BK)
1008.467
586.633
797.550
(a) (b)
--------------------------------------------------------------------------------------------------TH BK
T1
928.333
556.400
742.367(b)
T2
1038.533 586.767
812.650(a)
T3
1058.533 616.733
837.633(a)
DMBK
A
946.133
584.100
K
1038.133 586.400
S
1041.133 589.400
*Means of each factor and their interactions
followed by the same letters are not significantly
different from each other according to Duncan's
multiple ranges test at 5% level.
The air dried boards (765.117 kg/cm²), when
compared with kiln and solar drying
respectively. While board thickness increased,
static bending values increased too. Thin boards
(2 cm thick.) bended only by (742.367 kg/cm²),
while thickest ones (6cm thick.) deflected from
straight line by (837.633 kg/cm²)as a mean may
be caused by existence of different drying
situation. The scholarships Bolza and Kloot
(1963), Bryce (1967) and Louppe et al., (2008)
were attributed by reason of low age and fast
growth level.
Ogunsanwo,
(2000),
Oluwafemi
and
Adegbenga, (2007) through their studies on
Eucalyptus camaldulensis were clarified that the
values of density and mechanical properties
achieved, They could be of use its wood in
various aspects as building, construction,
flooring, cabinetry, and furniture.
110
References:
Acosta, M.S.; Marco, M.; Piter,J.C.; Sossazitto,
M.A.; Villalba, D.I. andCarpinetti,L.
(2008).Physical and mechanical properties
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- Duncan, D. B. (1955). Multiple Range and
multiple F. tests. Biometrics, 11: 1-42.
-IAWA Committee. 1989. IAWA list of
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221–332.).
-Iglesias, T.G. and Wilstermann,D.(2008).
Eucalyptus universals. Globalcultivated
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Forestry
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ologics.Retrieved from http://git –
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(2008).Aproveitamento
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N.C.;Nogueira
TecnologiasAplicadasaoSetorMadeireiro
III.JerônimoMonteiro, ES, I Simcatem,
III,Chapter 10, p. 255-290.
-Louppe D, AA Oteng-Amoako, M Brink (eds).
2008. Plant Resources of Tropical Africa
7(1).Timbers 1. PROTA Foundation
Wageningen, Netherlands/ CTA, 704pp.
-Myburg; A.A., Potts; B, Marques; CM, Kirst;
M, Gion; JM and GrattapagliaD, Grima –
Pettenati, J. (2006).Eucalyptus. In: Kole C
(ed) Genome Mapping &Molecular
Breeding in Plants Volume 7: Forest
Trees. Springer, pp in press.
-Ogunsanwo,O.Y.(2000).Characteristic of wood
properties
of
Plantation
grown
Triplochiton sclroxyloninOmo forest
reserve, Ogun State. Nigeria. PhD Thesis.
University of Ibadan.
-Oluwafemi, O.A. andS.O. Adegbenga,(2007).
Preliminary report on utilization potential
of Gliricidiasepium(Jacq.) stead for
timber. Res.J.Forestry,1:80.85.
-Roitzsch-Reader J.E., (1969). Forest Trees in
Iraq. University of Mosul, Iraq.
-SAS Institute, Inc. (2002). Statistical Analysis
System Ver. 9.0.SASinstitute Inc.,Cary,
NC.,USA.
-Shahbaz, S. Esmael.( 2010) Trees and shrubs. A
field guide to the trees and shrubs of
Kurdistan
region of Iraq. University of Duhok.
University of Duhok Publication.
-Taha, MA.Y.(2013).Characteristics of Sawn
board
Produced
from
Eucalyptus
Camaldulensis Dehn. Grown in Askikelek
/ Kurdistan Region of Iraq .PhD Thesis,
School of Forestry, Faculty of Agriculture
&Forestry
the
University
of
Dohuk.Kurdistan RegionofIraq.
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112
ISSN: 2410-7549
THE EFFECT OF COLOR PLASTIC MULCHES ON GROWTH, YIELD AND
QUALITY OF TWO HYBRIDS OF SUMMER SQUASH (CUCURBITA PEPO
L.)
Abduljebbar Ihsan Saeid and Ghurbat Hassan Mohammed
Department Of Horticulture, Faculty of Agriculture, University of Duhok, Kurdistan Region – Iraq.
Abstract
This experiment was carried out at the vegetable research farm of Horticulture Department/ Faculty
of Agriculture /Duhok University on two hybrids of summer squash during spring in the season of 2014,
to study the effect of five color plastic mulches (without cover, transparent, black, red and blue) on two
hybrids of summer squash (Amjed and Alexandra F1). The results showed that covers plastic mulches led
to positive significant differences in leaf area (cm2), leaves chlorophyll content%, , fruit number/plant,
early yield and total yield as well as fruit weight (g ), fruit length (cm) and fruit diameter (mm) as
compared to without cover. There were no significant differences between the two hybrids in all detected
traits. The interaction between covers plastic mulches and two hybrids was significantly enhanced all
detected traits, since summer squash (Alexandra hybrid) covered with transparent plastic mulch were
characterized by the highest values of leaf area (cm2), leaves chlorophyll content% and fruit diameter
(mm), and summer squash (Amjed hybrid) covered with blue plastic mulch gave positive significant of
fruit number/plant and total yields ton/ hectare.
Keywords: mulches, summer squash, hybrids
Introduction
S
ummer squash (Cucurbita pepo L.) is one
of the most important vegetable crops in
Iraq especially during spring. In addition to
cultivation in fall season in some areas it
consists of annual crops including cucurbitaceae
and Summer squashes are planted for its fruits,
which are the edible parts of the plant after
cooking. It has a medium food value due to
some nutritional elements (P, Ca and Fe), and
some vitamins (Matlob et al., 1989). Yield of
summer squash per unit area is remained too low
which is about (12.82 and 13.09 t/ha.) in 1999
and 2000 respectively (Annual Statistic Book,
2000), as compared to the world production.
To increase vegetable production, many
applications such as coverage are used.
Coverage vegetables can promote early yield
and reduce fruit defects. However, coverage can
reduce evaporation from the soil surface, prevent
weed growth, raise soil temperature, reduce
costs, reduce insect number and increase yield
(Ekinci and Dursun, 2006). Polyethylene
mulches benefit to adjust the soils microclimate
in order to prolong the growing season and
increase plant growth (Tarara, 2000). Black
plastic
mulch
has
intense
shortwave
transmittances and high shortwave absorption,
which causes quickly increased soil temperatures
(Heibner et al., 2005) white plastic mulch is
prefers during the summer season in warm
regions because white plastic maintain soil
moisture and providing cooler temperature.
Kasperhauer (1992) mentioned that red plastic
increased yield in some crops, believed that it is
generate a positive phytochrome response, found
that improved yield quality due to used colored
plastic (Brown and Channel-Butcher, 2001) This
study aimed to determine the effect of different
coverage Plastic color on growth, some quality
properties and yield in two hybrids of summer
squash.
The experiment was conducted at the
vegetable research farm, Faculty of Agriculture,
University of Duhok, on summer squash during
spring season of 2014. Seedlings were growing
in first of April 2014 at a distance 40 cm
between plants and 1.5 m between the rows.
The experiment comprised the effect of two
hybrids namely (Amjed and Alexandra), five
coverage (without coverage, transparent, Black,
Red and Blue). Each treatment was replicated
three times. A replicate contained ten plants per
one and was implicated in a completely
randomized block design (RCBD). The soil was
well softened, and then it was divided into rows
and in this study all plants received the regular
agricultural practices that usually carried out in
the vegetable crops. Coverage was done before
planting the seedling. Data were analyzed by
using SAS program (SAS, 2001).
113
Experimental measurements
Three plants were selected randomly from
each experimental unit to measure:
Table (1) shows that transparent cover plastic
mulch caused significant increase in leaf area
(cm2) as compared to without coverage and
insignificant increase in Chlorophyll content %.
As for there was no significant differences
between its two hybrids on leaf area (cm2) and
Chlorophyll content %.
Concerning the effect of interaction between
covers plastic mulches and hybrids observed that
interaction between transparent cover and
Alexandra hybrids was significant in its effect
in leaf area (cm2) and chlorophyll content% by
the highest values of (317.20) cm2 and (54.90)
respectively. The increase in growth was
attributed to sufficient soil moisture at the root
zone and minimized the evaporation loss due to
covers. The extended retention of moisture and
availability of moisture also lead to higher
uptake of nutrient for proper growth and
development of plant. Similar findings have also
been obtained by Dean Ban et al. (2004), Ansary
and Roy (2005) in watermelon, Angrej-Ali and
Gaur (2007) in strawberry, Aruna et al. (2007) in
tomato.
1-Vegetative growth characteristic
a-Leaf area (cm2)
b-Leaf chlorophyll content%
2-Yield characteristic
a-Early yield: The first three harvests from
each treatment were weighted to considered as
an early yield.
b-Total yield: the total yield was measured
by harvested all fruit from each treatment along
the harvesting period were weighted to calculate
the total yield Kg per plant and ton per hectare.
c- Fruit number per plant:
Number of
fruits per plant along the harvesting period was
counted from each experimental unit, starting
from the commence of harvesting and lasted to
the end of the growing season and calculated.
3-Fruit quality:
five fruits from each treatments were
randomly taken for determining average fruit
character as follows:
a- Fruit fresh weight (g )
b- Fruit length (cm)
c- Fruit diameter (mm)
d- Fruit dry weight (gm).
Table (1) Effect of covers plastic mulches on leaf area (cm2) and Chlorophyll content % of two hybrids of
summer squash
covers
AM
without
coverage
White
Black
Red
Blue
hybrids effect
leaf area (cm2)
hybrids
AL
Covers
effect
Chlorophyll content %
hybrids
Covers
effect
AM
AL
209.40bc
184.30c
196.8ob
50.93b
53.ooab
51.97a
302.60ab
317.20a
309.90a
53.23ab
54.90a
53.62a
220.20a-c
252.80a-c
236.50b
54.00ab
52.67ab
53.80a
234.70a-c
276.90a-c
255.80ab
52.63ab
53.33ab
52.98a
269.20a-c
237.30a-c
253.20ab
52.40ab
52.33ab
52.37a
247.30a
253.70a
52.83a
53.07a
Means within a column, row and their interaction following with the same latter are not
significantly different according to Duncan multiple range test at the probability of 0.05 level
114
Data presented in Table (2 and 3) are clearly shown that covers caused significant increases in all
yield characteristics as compared with without coverage. In case of cultivars there was no significant
increase in all yield characteristics.
The interaction between covers and hybrids was significant in its effect. since Amjed hybrid and
blue cover were confined by highest value in fruit number/plant (29.00), total yield (4.50 kg/plant) and
total yield (65.68 t/ ha) as compared with the lowest values of these traits for without covers which
gave (17.33, 2.28kg/ plant and 33.36 t/ha) respectively and the interaction between red polyethylene
and Alexandra hybrid gave by the highest value in early yield kg/plant as compared with without
cover. Plant under polyethylene produced larger fruit and have higher yield per plant because of better
plant growth due to favorable hydro-thermal regime and complete weed free environmental.
Dickerson et al. (2003) reported earlier yield under plastic mulch. The above results were in
agreement with those of Dean Ben et al. (2004), Ansary and Roy (2005), Cenobio et al. (2007), and
Arancibia and Motsenbocker (2008) in watermelon.
Table (2) effect of covers plastic mulches on fruit number and early yield(kg) of two hybrids of summer squash
covers
AM
without
coverage
Transparent
Black
Red
Blue
hybrids effect
Fruit number/plant
Hybrids
AL
Early yield(kg)
hybrids
AM
AL
Covers
effect
Covers
effect
17.33c
19.00c
18.17c
0.10de
0.08e
0.09c
24.89ab
25.11ab
25.00b
0.47ab
0.48ab
0.48a
26.89ab
24.22b
25.56b
0.32bc
0.27dc
0.29b
25.11ab
26.00ab
25.56b
0.46a-c
0.52a
0.49a
29.00a
28.78a
28.89a
0.41a-c
0.34a-c
0.38ab
24.64a
24.62a
0.35a
0.34a
Table (3) Effect of covers plastic mulches on Total yield kg/plant and Total yield t/ha of two hybrids of summer
squash.
Covers
AM
without
coverage
Transparent
Black
Red
Blue
hybrids effect
Total yield Kg /plant
Hybrids
Covers
effect
AL
AM
Total yield t/ha
hybrids
Covers effect
AL
2.28bc
2.71c
2.50b
33.36bc
39.60c
36.48b
4.03a
4.39a
4.21a
58.80a
64.12a
61.48a
4.08a
3.73ab
3.90a
59.52a
54. 56ab
57.04a
4.16a
4.34a
4.25a
60.84a
63.36a
62.08
4.50a
4.42a
4.46a
65.68a
64.60a
64.32a
3.81a
3.92a
55.64a
57.24a
115
Data in Table (4 and 5) show that transparent cover plastic mulch had positive effect on fruit length
(cm) and fruit diameter (mm) and no positive effect on fruit weight (g) and fruit dray weight (g). For
the effect of hybrids there was no significant effect between two hybrids on fruit weight (g), fruit
length (cm) and fruit dray weight (g).
Also the interaction between transparent covers plastic mulches and Amjed hybrid was high
significant effect on fruit length (16.27cm) and fruit diameter (40.51mm) and the interaction between
black cover and amjed hybrid give high significant effect on fruit weight (188.59g). Among all
mulching treatment, maximum fruit weight was recorded in black cover. It appears that black
polyethylene mulch have induced favorable conditions conducive to a attainment of fruits of higher
weight. The highest fruit length was due to congenial soil moisture results higher uptake of nutrition
for better growth of fruit, the reduction in evaporation losses of soil moisture caused by mulches
covered the soil surface in row of summer squash. The above results were in agreement with those of
Ansary and Roy (2004), and Arancibia and Motsenbocker (2008) in watermelon, Aruna et al. (2007)
in tomato.
Table (4) Effect of covers plastic mulches on fruit weight (g ) and fruit length (cm) of two hybrids of summer
squash.
covers
AM
without
coverage
Transparent
Black
Red
Blue
hybrids effect
Fruit weight(g )
hybrids
AL
Cover
effect
Fruit length(cm)
hybrids
AM
AL
Cover
effect
130.72b
142.51ab
136.61a
13.17b
12.86b
13.01c
161.29ab
174.65ab
167.97a
16.27a
15.48ab
15.88a
188.59a
153.11ab
170.85a
14.83ab
13.57b
14.20a-c
165.63ab
166.16ab
165.89a
15.32ab
14.87ab
15.09ab
154.77ab
153.63ab
154.20a
14.04ab
13.15b
13.60bc
160.20a
158.01a
14.73a
13.99a
Table (5) Effect of covers plastic mulches on fruit diameter (mm) and fruit dray weight (g ) of two hybrids of
summer squash.
Covers
AM
without
coverage
Transparent
Black
Red
Blue
hybrids effect
Fruit diameter(mm)
hybrids
Covers
effect
AL
Fruit dry weight(g )
hybrids
Covers
effect
AM
AL
34.55a-c
29.86c
32.21c
3.68a
3.71a
3.70a
40.51a
39.58a
40.05a
3.50a
3.94a
3.72a
37.87ab
34.53a-c
36.20a-c
3.93a
4.28a
4.11a
39.60a
36.32a-c
37.96ab
4.00a
3.86a
3.93a
35.56a-c
31.73bc
33.64bc
4.03a
4.24a
4.14a
37.62a
34.40b
3.83a
4.01a
116
References
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117
@@
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118
ISSN: 2410-7549
SYNTHESIS AND BIOLOGICAL EVALUATION OF SOME DIPEPTIDE
DERIVATIVES AND THEIR HETEROCYCLIC COMPOUNDS
Huda Ahmed Basheer, Sabir Ayob Mohammed and Aveen Akram Ibrahiem
Chemistry Department, Faculty of Science, University of Zakho, Kurdistan Region – Iraq.
Abstract:
The protected dipeptide esters (3a-e, 4a-l) is prepared by the reaction of compounds (1a-e, 2a-d) with
dicyclohexylcarbodiamide (as coupling reagent) and amino acid esters .Thereafter hydrazides (5a-e, 6a-j)
are obtained by the reaction of corresponding esters with hydrazine hydrate. Hydrazones (7a-e, 8a-e) are
synthesized by the reaction of the above hydrazides with p-nitro benzaldehyde , which was cyclized to 2,5disubstituted 1,3,4- oxadiazole (9a-e, 10a-e) through lead oxide and to phthalazines (13a-b) through
hydrochloric acid. 1,3,4- oxadiazole -2- thione (11a-b, 12a-e) were prepared by the reaction of the
corresponding hydrazides with carbon disulfide in alcoholic potassium hydroxide. Hydrazides were
reacted with ammonium thiocyanate to afford thiosemicarbazide (14a-c) which were cyclized to 1,2,4triazole -3- thione (15a-b) in sodium hydroxide medium. The structures of the synthesized compounds
were confirmed by physical and spectral methods. The antibacterial activity of the prepared compounds
(5d, 7a, 9e, 10d, 11a, 12e, 13a, 14b) against the gram +ve and –gram –ve Bactria (Staphylococcus aureus,
Bacillus subtilis, Escherichia coli and Proteus mirabilis) were studied and discussed.
Introduction
Oxadiazole derivatives displayed wide
spectrum of activities such as antibacterial(
Salimon et al 2011; Jumat et al 2010),
antimicrobial (Karthikeyan et al 2008; Gaonkar
and Rai 2006), anti-fungal (Kanthiah et al 2011;
Srivastava et al 2010) analgesic (Amir and
Kumar 2007), as anticonvulsant (Zargahi et al
2005; Rastogi et al 2006), antitumor (Bezerra et
al 2005), anti-tubercular (Dhoel et al 2005),
antihypoglycemic (Goankar et al 2006) and antihepatitis B viral activities (Tan et al 2006).
Compounds bearing 1,3,4- Oxadiazole nucleus
are known to exhibit unique anti-edema and antiinflammatory activity (Husain and Ajmal 2009;
Franski 2005; Amir and Kumar 2007). Some of
2,5-disubstituted-1,3,4-oxadiazole
derivatives
used against 60 tumor cell lines derived from
nine cancer cell type, anti-tumor activity against
leukemia, colon and breast cancer (Wagle et al
2008; Aboraia et al 2006) While compounds
having 1,2,4-triazole nucleus are very important
in the field of medicinal chemistry, as fungicidal
(Royr et al 2005), antibacterial (Singh and Singh
2009), antimicrobial (Hussain et al 2008),
antimycotic (Zamani et al 2004), antidepressive
(Clerici et al 2001), cardiotonic (Onkol et al
2004) and anticonvulsant (Parmar et al 1974)
activites. Triazole ring derivatives are known to
possess anti-inflammatory (Moise et al 2009),
analgesic (Shenone et al 2001) antihaemostatic
activity (Kamble and Sudha 2006) and for antiHIV-1 activity by examination of their inhibition
of HIV-1-induced cytopathogenicity in MT-4
cells and by determination of their inhibitory
effect on HIV-1 reverse transcriptase (Wu et al
2007) On the other hand, heterocycles
containing the phthalazine moiety are of interest
due to their pharmacological and biological
activities (Jain and Vederas 2004). Some of the
phthalazinone derivatives have found application
in clinical medicine due to interesting
vasodialator
(Demirayak
et
al
2004),
anticonvulsant (Zhang et al 2009), antidiabatic
(Boland et al 1993), antiallergic (Hamamoto et
al 1993), and antiasthmatic (Yamaguchi et al
1993). Phthalazinone nucleus has been proved to
be a versatile system in medicinal chemistry
such as, aldose reductase inhibitor activities
(Kashima et al 1998). According to the above
and due to the significance of peptides in
antibiotic activity; we made a combination of
peptide and heterocycles nucleus in a goal to
increase the biological effects.
119
120
Experimental
Uncorrected melting points were determined
by using bibby scientific limited stone,
Staffordshire, ST 15 OSA, UK., IR spectra were
recorded as KBr disc in the (400-4000 cm-1)
range by using (spectrum one B FT-IR
spectrometer), 1H NMR spectra in δ units (ppm)
relative to an internal standard of tetramethyl
silane on 1H NMR (NMR: BRUKER400 MHz
UltrashieldTM) in DMSO-d6, In Department of
chemistry, faculty of Science, Diçle University,
in Turkey. Amino acids esters and benzoyl
amino acids were prepared according to
literature procedure (Basheer 2000), p-toluene
sulfonyl amino acids were prepared following
literature reported procedure (Qreenstien and
winitz 1961).
Synthesis of protected dipeptide ester (3a-e,
4a-l)
To a solution of 0.01 mole protected amino
acid and 0.01mole of amino acid ester in 50 ml.
of dichloromethane is added 0.01 mole N,N'dicyclohexylcarbodiimide the mixture is allowed
stirring over night at room temperature ,The
precipitated dicyclohexylurea is removed by
filtration and the filtrate washed with water ,
diluted hydrochloric acid , water , half saturated
sodium bicarbonate solution and water, and
finally dried over anhydrous sodium sulfate.
Evaporation of the solution gives residual
mixture of crystals and oil. This is treated with a
small amount of diethylether and filtrate;
although the material is quite soluble in
diethylether and hence is lost in appreciable
amount when this solvent is employed, the white
precipitate is recrystallized from acetone –
petroleum ether.
Synthesis of protected dipeptide hydrazide
(5a-e, 6a-j)
A mixture of protected dipeptide ester
(0.01mole) and hydrazine hydrate (0.2mole) in
absolute ethanol (70 ml) was refluxed for (3)
hours. The solvent was evaporated under
reduced pressure and the residue was
crystallized from methanol.
Synthesis of 4-nitrobenzaldehyde protected
dipeptide hydrazone (7a-e,8a-e)
A
mixture
of
p-Nitrobenzaldehyde
(0.01mole) and (0.01mole) of protected
dipeptide hydrazide (5a-e,6a-e) in (20 ml)
ethanol was refluxed for 2 hrs. The solvent was
concentrated and the precipitate was filtered and
recrystallized from benzene.
Synthesis of 2-protected dipeptide residue, 5(4´-nitrophenyl) -1, 3, 4-oxadia-zole (9a-e,
10a-e)
To a homogenous solution of hydrazone (7ae,8a-e) (0.01mole) in 20 ml glacial acetic acid,
lead oxide (pbO2) (0.01mole) was added, the
mixture was stirred at 25 C◦ for 1 hr. The
reaction mixture was diluted with ice- water and
left to stand for 24 hrs. The precipitate was
filtered off and recrystallized from benzene.
Synthesis of 5-protected dipeptide residue 1,3,4-oxadiazole -2-thione(11a-b, 12a-e)
(0.05mole) of the protected dipeptide
hydrazide (5a-b,6a-e) was dissolved separately
in (70 ml) 0.5% ethanolic potassium hydroxide.
(0.1 mole, 6 ml) of carbon disulfide was added
gradually and the resulted mixture was refluxed
for 16 hours until the evolution of hydrogen
sulfide was ceased (checked by filter paper
moister with lead acetate). The solvent was
evaporated under reduced pressure and the
residue was poured on crushed ice, diluted with
ice-water, acidifies with diluted HCl, filtered and
dried recrystallized from chloroform.
Synthesis of 1-(benzoyl dipeptide residue)-7nitrophthalazine (13a-b)
(0.002mole) of the hydrazones (8a-b) in (10
ml) of amyl alcohol (saturated with HCl gas)
was heated on steam bath for (1.5 hrs.) and then
reflexed for (1hr.). The reaction mixture was
cooled, washed with (10 ml) of (20%) sodium
hydroxide and then with water until neutralized.
Evaporation of the solvent and recrystallization
from ethanol afforded the product (13a-b).
Synthesis Substituted thiosemicarbazide (14ac)
A mixture of (0.003mole) benzoyl dipeptide
hydrazide (6a-c), (0.009mole) of ammonium
thiocyanate, (4 ml) hydrochloric acid in (25 ml)
absolute ethanol, was reflexed for 22 hrs. The
solvent was evaporated and the residue poured
on crushed ice. The precipitated product was
filtered, dried and recrystallized from ethanol.
Synthesis 5-substituted-1,2,4-triazole-3-thione
(15a-b)
A mixture of substituted thiosemicarbazide
(14a-b) (0.0012mole) and (7.5 ml) 1% aqueous
sodium hydroxide solution was refluxed for 3
hrs. , the mixture was treated with charcoal and
121
the charcoal then removed by hot filtration. The
solution was acidified by 10% hydrochloric acid
with cooling; the precipitate was filtered, and
recrystallized from methanol.
Antibacterial assay
Discs of filter paper (6mm diameter) were
sterilized at 140 Cº for 1hr. and impregnated
with 1ml. of stock solution (10mg. /ml, 1mg. /
ml, 0.1mg. /ml, and 0.01mg. /ml) of each
compound and then dried-DMSO (dimethyl
sulfoxide) was used as a solvent for compounds
(5d, 7a, 9e, 10d, 11a, 12e, 13a, 14d). The same
solvent was used for antibiotics. Blank paper of
DMSO was used as control. The inoculated
plates were incubated at 37 Cº for 24 hrs. And
the inhibition zones (mm) were measured. In all
experiments, the mean of each triplicate was
measured (Garrod et al 1981).
Results & Discussion
Protected dipeptide esters were prepared from
the reaction of p-toluene sulfonyl amino acid
with amino acid ester by using N,Ndicyclohexylcarbodiimide (DCC) as coupling
group. The synthesized compounds (3a-e,4a-l)
were characterized by their IR and 1H NMR
spectra, the IR spectra data provide evidence in
support of structures (3a-e,4a-l) for these series
of compounds in which characteristic bands at
3373 -3268cm-1 for N-H stretching, 1651-1610
cm-1 for C=O (phCONH) stretching, 16601626cm-1 for C=O amide stretching, 17511721cm-1 for C=O ester stretching, as illustrated
in Table (1). The 1H NMR spectra (Table 4) of
the compounds (3a,e 4c,j) indicated the presence
of the ethyl group resonating triplet signals at the
region of 1.3 -0.7ppm for -CH3 group and
quartet for -CH2- group at the region ٤-3.1 ppm,
aromatic protons appeared in the expected range
δ 7.9-7.0 ppm, finally the two amide protons
occurred at the relatively downfield positions of
8.9- 8.1. Protected dipeptide hydrazides were
prepared by the reaction of their corresponding
ester with hydrazine hydrate in absolute ethanol.
The structures of hydrazide compounds (5a-e,
6a-j) are confirmed on the basis of the following
evidence. The IR showed the characteristic
absorption bands as follow 3253-3325 cm-1
(NH), 1629-1603cm-1 C=O (phCO-) and 16561605 cm-1 (C=O amide). In addition of absence
of band for C=O of ester, as illustrated in Table
(2).The 1HNMR spectrum of the compounds
(6e,6j) showed a signal at 4.5ppm indicating the
122
presence of NHNH2 protons, absence the protons
of the ethyl group and other signals were
observed at appropriate places, as illustrated in
Table (4).
Hydrazone compounds (7a-e, 8a-e) were
prepared by the condensation reaction of
protected dipeptide hydrazide with 4nitrobenzaldehyde. Hydrazone compounds (7ae,8a-e) were confirmed by the IR spectroscopy
which showed the absorption bands of (C=N,
C=C), amide, C=O (phCO-) and amine groups
appeared stretching vibration at (1600-1530cm1
), (1690-1629cm-1) (1625-1600cm-1), and
(3467-3251cm-1), respectively. While at
stretching
vibration
(1538-1513cm-1)
Asymmetric stretching of aromatic NO2, and
(1345 - 1326 cm-1) Symmetric stretching of
aromatic NO2, as shown in Table (2). The 1H
NMR spectrum of the compounds (7d,e, 8c,e )
showed a signal at ( δ 7.3- 7.1ppm) (parikh
1974) indicating the presence of CH=N proton
and other signals were observed at appropriate
places, Table (4).
The hydrazone were cyclized to 2,5disubstituted- 1,3,4- Oxadiazole (9a-e,10a-e) by
their reaction with lead oxide. The IR
characterization absorption bands of the
oxadiazoles (9a-e,10a-e) were given in Table
(3).The main absorption bands for imine and
amide groups appeared at (1600-1561cm-1)
,(1656 -1600 cm-1) and (1686-1633cm-1)
for(C=N, C=C),C=O (phCONH)and C=O, while
at (1525-1519 cm-1) (1346-1340cm-1) represent
NO2
asymmetric,
symmetric
stretching,
respectively. The N-H groups of these
compounds appeared at (3435- 3258cm-1) as
broad bands.
1
H NMR spectral analyses of compounds
(9a,c-d,10b-d) exhibited aromatic protons in the
expected range (8.4-7.1ppm), hiding peak due to
(CH=N) protons, and other signals were
observed at appropriate places, as illustrated in
Table(5).
Oxadiazole -2-thione (11a-b,12a-e) synthesis
was performed by the reaction of hydrazides and
carbon disulfide in alkaline medium. The
mechanism of the reaction is accomplished by
nucleophilic attack of the enol hydrazide form at
the carbon atom of carbon disulfide. The formed
xanthat salts underwent intra nucleophilic attack
followed by hydrogen sulfide elimination. The
IR spectra of the compounds (11a-b,12a-e)
showed absorption bands at (1266-1154cm-1)
corresponds to the thione stretching vibration,
and others gave the following vibrational
absorption bands (1597-1529cm-1) , (16671632cm-1) and (3430- 3246cm-1) which were
assigned to (C=C, C=N), (C=O amide), and (NH) respectively as illustrated in Table (3). The
1
H NMR spectrum of compounds (11a,12b,e)
revealed the resonance peaks that appeared at
7.1-8.2 ppm could be assigned to the
contribution of aromatic protons, and the
remaining signals has appeared in the
expectation of places, as illustrated in Table (5).
Phthalazine compounds (13a-b) were
prepared through ring closure of hydrazone by
using hydrochloric acid in amyl alcohol. The
main absorption bands of substituted Phthalazine
compounds which includes stretching vibrations
of C=N, C=C, C=O amide at (1605-1585cm-1),
and (1630, 1629 cm-1) respectively. While the
absorption bands at (3435, 3327 cm-1) were
assigned to (N-H) stretching vibrations, as
shown in Table (3).
Hydrazides
were
converted
to
thiosemicarbazide (14a-c) by its reaction with
ammonium thiocyanate / hydrochloric acidic.
The structure of compounds (14a-c) was
confirmed by IR in which strong bands for C=S,
C=O, C=O (phCO) and N-H stretching were
observed at (1225-1085), (1631–1630), (16151610) and (3152–3114) cm–1, respectively Table
(3).
Cyclocondensation
of
the
thiosemicarbarbazide with aqueous sodium
hydroxide afford 5-substituted-1,2,4-triazole-3thione. The IR spectra of the synthesized
compound (15a-b) showed the presence of N-H
stretching bond at(3326-3127cm-1) and detection
of C=N stretching bond at (1582 -1577 cm-1) for
evidence of ring closure of triazole ring Table
(3).
The antibacterial activity of the compounds
(5d, 7a, 9e, 10d, 11a, 12e, 13a, 14b) were
evaluated using various species of bacteria,
Staphylococcus aureus,
Bacillus subtilis,
Escherichia coli and Proteus mirabilis. The
result showed that the compounds (7a-12e) were
active in inhibiting the growth of nearly all
organisms used as indicated from the diameter of
inhibition zone Table (6). Blank discs DMSO
did not show any activity. According to the data
(Table 6), it was evident that the activity of
tested compounds decreased considerably at
lower concentration (0.01mg/ml).
However, compounds (7a, 9e, 11a and 12e)
showed the higher antibacterial effect on
Bacillus subtilis, Proteus mirabilis then
Staphylococcus aureus and Escherichia coli, as
indicated from the diameter of inhibition zone.
123
Table(1): Physical properties and spectral data of compounds (3a-e, 4a-l)
IR (KBr) υ cm-1
Comp
No.
R,R´
M. p.
˚C
Yiel
d
%
C=C
C=O amide
C=O
ester
NH
(OH)
UV
DMS
O
max
(nm)
3a
H,H
78-79
65
1597m
1655s
1730s
3373b
264
3b
-CH(CH3)2 , H
113-115
80
1535m
1632s
1741s
3277b
265
3c
-CH3,-CH2CH(CH3)2
98-100
63
1598m
1655s
1716s
3268b
264
3d
-CH2CH(CH3)2 , H
127-128
64
1539m
1647s
1736s
3303b
265
3e
-CH(CH3)CH2CH3,
H
98-100
77
1555S
1647S
1744s
3340b
265
4a
H , -CH2C6H5
102-103
64
1545b
1650w,1642s
1739s
3295b
263
4b
H,H
140-141
43
1573m
1626s
1744b
3327s
266
1744b
(3389
)
266
3339s
4c
H ,-CH2C6H4OH
120-122
55
1555s
1647s,1632
m
4d
-CH(CH3)2 ,H
82-83
45
1580w,1546m
1660w,1651s
1751s
3299b
267
135-137
44
1578w,1536s
1635s,1620w
1742s
3323b
264
CH(CH3)CH2CH3,
4e
-CH(CH3)2
4f
-CH(CH3)2,CH(CH3)2
139-140
57
1578w,1537s
1633s
1742s
3276b
262
4g
-CH(CH3)2,CH2C6H5
155-156
76
1578w,1548m
1631b
1723s
3303b
261
4h
-CH3,-CH2CH(CH3)2
113-115
50
1536b
1655w,1633s
1743s
3279b
265
4i
-CH3 , H
133-134
55
1577w,1534m
1640m,1631
s
1721s
3325b
262
4j
-CH3, -CH2C6H5
97-98
72
1577w,1535s
1656m,1632
s
1736s
3302b
268
4k
-CH3, -CH2C6H4OH
80-81
60
1532b
1640s,1610w
1739s
3324b
(3400
)
263
127-128
69
1578w,1530m
1660m,1631
s
1740s
3284s
264
-CH(CH3)CHCH3,
4l
- CH2C6H5
124
Table(2):Physical properties and spectral data of compounds (5a-e,6a-j, 7a-e, 8a-e)
-
NH
(OH)
3307b
3309b
3283b
3336b
3253b
UV
DMS
O
max
nm
264
264
270
264
264
1645s,1603w
-
3284b
265
1636s,1610w
-
1640s,1629m
-
3307b
3410b
3287b
1530w
1629b
-
3287b
262
94
1532m
1630b
-
3283b
263
102-104
90
1578w
1532m
1628b
-
3323b
263
102-103
96
1541m
1628s
1610w
-
3325b
263
163-164
97
1538m
1633s
-
3267b
267
126-127
98
1540m
1640s
1628w
-
3284b
266
201-202
94
1631b
-
3281b
267
1688m
1644s
1519s
1343s
1522s
1345s
1522s
1345s
1520s
1344s
1520s
1345s
1522s
1345s
1533m
1342m
1538s
1328m
1513m
1326s
1525s
1345s
M. p.
˚C
Yie
ld
%
H,H
-CH(CH3)2 , H
-CH3, CH2CH(CH3)2
CH2CH(CH3)2 , H
CH(CH3)CH2CH3,H
116-117
133-135
112-113
102-103
188-189
91
67
85
66
60
6a
H , -CH2C6H5
180-181
68
6b
H , CH2C6H4OH
116-117
94
6c
-CH(CH3)2 ,H
-CH(CH3)2 ,
-CH(CH3)2
-CH(CH3)2 ,
-CH2C6H5
-CH3 ,
CH2CH(CH3)2
140-142
Comp
No.
R,R´
5a
5b
5c
5d
5e
6d
6e
6f
6g
6h
6i
6j
-CH3 , H
CH3 ,
-CH2C6H5
-CH3 ,
- CH2C6H4OH
CH(CH3)CHCH3
CH2C6H5
IR (KBr) υ cm-1
C=C
C=O amide
NO2
1636s
1639s
1639s
1651s,1605m
1634s,1656w
94
1542m
1545m
1533m
1533m
1555m
1576m1
546w
1577w
1542m
1578w
152-153
96
204-205
1578w
1533m
1597m
1540m
1599m
1545w
1600m
1535w
1595m
1586w
7a
H,H
167-169
78
7b
-CH(CH3)2 , H
205-206
52
7c
-CH3, CH2CH(CH3)2
161-163
81
7d
CH2CH(CH3)2 , H
183-185
92
7e
CH(CH3)CH2CH3,H
180-182
93
1597m
8a
-H,CH2C6H5
170-171
74
1587m
1560w
1686m
1649s
1686s
1656s
1690m
1646m
1656b
1610w
8b
-CH(CH3)2,
-CH2C6H5
180-181
82
1590m
1630b
8c
-CH3, -CH2C6H5
149-150
84
1578m
1632b
8d
-CH3, -CH2C6H4OH
117-119
79
8e
-CH(CH3)CH2CH3, CH2C6H5
181-183
92
1578m
1530w
1599m
1578w
1635s
1625m
1629s
1600m
1656b
3353b
3433b
3467b
3394b
3251b
3279b
3288b
3271s
3324b
3424b
3282b
264
262
341
262
337
260
337
260
335
264
336
264
335
262
371
262
371
262
371
261
345
262
125
Table (3): Physical properties and spectral data of compounds (9a-e, 10a-e, 11a-b, 12a-e, 12a-c, 13a-b, 14a-c,
15a-b)
-1
IR (KBr) υ cm
Com
No.
R R'
M. p.
Cº
Yiel
d
%
C-O-C
9a
-H,H
201-203
70
1153m
248-249
81
1162m
197-198
94
1162s
194-195
92
1162b
195-196
79
1160m
179-181
66
1106w
268-271
43
1106w
230-231
61
1162w
205-207
45
1153m
228-230
64
1125w
-CH(CH3)2
-H
-CH3
CH2CH(CH3)2
9b
9c
9d
9e
10a
10b
10c
10d
10e
CH2CH(CH3)2 -H
-CHCH2CH3
CH3
-H
-H ,
-CH2C6H5
-CH(CH3)2,
CH2C6H5
-CH3 ,
-CH2C6H5
-CH3 ,
CH2C6H4OH
-CHCH2CH3
CH3
CH2C6H5
NO2
C=C
C=N
1519s
1344s
1520s
1345m
1519s
1344s
1520m,
1340s
1521s
1343s
1522s
1345s
1525s
1346s
1520s
1342s
1525s
1346s
1596m
1561w
1597s
C=O
amide
NH
C=S
1648s
3364b
-
1656m
3433b
-
UV
DMS
O
max
(nm)
341
264
335
261
338
263
339
263
335
259
338
261
337
263
334
260
340
265
1593s
1570w
1592m
1586m
1600w
1596m
1649s
3258b
-
1648m
3435b
-
1643s
3298b
-
1587s
1647s
1600w
3421b
-
1597m
1633b
3434b
-
1595w
1586w
1597m
1570w
1686s
1656s
1681s
1629m
3394b
-
3430b
-
1524s
1345s
1595m
1580w
1633b
1605w
3277b
-
335
262
11a
CH(CH3)2, H
205
64
1093m
-
1575m
1548m
1656s
3323s
3246s
1161s
344
274
11b
CH2CH(CH3)2,H
161-162
71
1092m
-
1595m
1542m
1667s
3363s
3255s
1163s
327
268
12a
-H , -CH2C6H5
75-76
79
1065m
-
1643s
3411b
1154m
12b
-CH(CH3)2, CH2C6H5
82-83
68
1058m
-
1637s
3273b
1157m
12c
-CH3 , -H
163-164
45
1188m
-
1632s
3326b
1266m
12d
-CH3 , -CH2C6H5
150-151
60
1059w
-
1635s
3430b
1158m
1632s
3287b
1155m
1628m
3435b
1630m
3327b
12e
13a
13b
14a
14b
14c
1590w
1531m
1597m
1540w
1577m
1535m
1585m
1529w
1577w
1529s
1605s
1595s
1596s
1585s
CH(CH3)CH2CH3
-CH2C6H5
-CH3 ,CH2C6H4OH
CH(CH3)CH2CH3
-CH2C6H5
93-95
85
1059w
-
292-293
48
-
-
>300
43
-
-
-CH(CH3)2 ,-H
212-213
65
-
-
1596m
122-123
73
-
-
1600m
212-214
68
-
-
1610m
-CH(CH3)2,
-CH2C6H5
CH(CH3)CH2CH3CH2C6H5
15a
-CH(CH3)2 ,-H
214-215
85
-
-
1582m
15b
-CH(CH3)2,
-CH2C6H5
152-153
65
-
-
1577m
126
1630s
1610m
1631s
1615m
1631s
1612m
1625m16
05m
1629b
1519s
1343s
1519s
1345s
350
275
335
270
345
271
348
274
350
270
334
260
340
265
3152b
1085m
261
3138b
1225m
276
3114b
1096m
275
3127b
1094s
3326b
1216s
335
263
338
260
Table (4): 1H NMR spectral data of compounds (3a,e, 4c,j, 6e,j, 7d,e, 8c,e)
Com
. No.
R , R'
-H , -H
3a
3e
4c
-CHCH2CH3
CH3
-H
-H
-CH2C6H4OH
-CH3,-CH2C6H5
4j
6e
-CH(CH3)2
CH2C6H5
6j
-CHCH2CH3
CH3
-CH2C6H5
7d
CH2CH(CH3)2 H
7e
-CHCH2CH3
CH3
-H
8c
-CH3
-CH2C6H5
8e
-CHCH2CH3
CH3
CH2C6H5
I
H NMR (400 MHz, DMSO-d6, δ, ppm)
1.2-1.3 (t, 3H) CH3CH2; 2.5 (s, 3H) CH3ph; 3.3, 3.5 (s, 4H) 2CH2CO; 3.63.7(q, 2H) -CH2CH3 7.4-7.8(m, 4H) ArH; 8.9(s, 1H) NH; 8.1(s, 1H) NH.
0.7-0.9 (m, 6H) 2CH3; 1.2-1.3 (t, 3H) CH3CH2; 2.5(s, 3H) CH3ph; 1.0-1.1 (m,
2H) CHCH2CH3; 3.5-3.6 (m, 2H) CH2CO; 4.0-4.1 (quartet, 2H) COCH2CH3 ;
1.5-1.7(m, 2H) 2CH; 7.3-7.8(m, 4H) ArH; 8.2-8.3(s, s, 2H) 2NH.
1.0(t, 3H) CH3CH2; 3.1(q, 2H) COCH2CH3; 3.4(d, 2H) phCH2CH, 3.9(s, 2H)
CH2CO; 4.2(d, 1H) COCH CH2ph; 7.0-7.9(m, 9H) 2ArH; 8.4(s, 1H) NH; 8.7
(s, H) NH; 8.9(s, 1H) OH.
1.1(t, 3H) CH2CH3; 1.3(d, 3H) CHCH3; 2.9-3.3 (m, 2H) 2CHCO; 3.9 (q, 2H)
COCH2CH3; 4.5 (d, 2H) phCH2CH; 7.2-7.9(m, 10H) 2ArH; 8.3 (d, 1H) NH;
8.5(d, 1H) NH.
0.7(d,6H)(CH3)2CH; 1.0(m,1H)CH(CH3)2; 2.1(d,2H) phCH2CH;
2.8(m,2H)2CHCO; 4.5(b, 2H)NH2; 7.2-8.2 (m, 10H) 2ArH; 8.5(d, H) NH; 9.3
(s, 2H)2NH.
0.7(m, 6H) 2CH3; 1.2(m, 2H)CH2CH3; 1.7(m, 1H) CH2CHCH3;
1.9(d,2H)phCH2CH, 2.9(m,2H)2CHCO; 4.5 (d,2H) NH2;7.1-8.0(m, 10H)
2ArH; 8.2(s, 1H) NH; 8.9(s, 1H)NH; 9.1(s, 1H) NH.
0.8(m,6H)2CH3; 1.2(m, 1H) CH(CH3)2; 1.4(m, 2H) CH2CH; 2.5(s, 3H)
CH3ph; 4.2(t, 1H) COCHCH2; 5.1(q,2H)CH2CO; 7.2(s,1H) CH=N; 7.1,7.8
(dd,4H) ArH; 7.4(s,1H)NH; 7.5,8.2(d,d, H)ArH; 8.8(s,1H) NH; 8.9(s,1H) NH.
0.9(m, 6H)2CH3; 1.2(m, 1H)CH; 1.5(m, 2H) CH2CH3;2.3(s, 3H) CH3ph;
3.6(m, 1H)CHCO; 3.9(d, 2H) CH2CO; 7.3(s, 1H) CH=N; 7.4(s, 1H) NH; 7.7,
8.1(dd, 4H) ArH; 7.8(s,1H)NH; 7.9,8.1 (dd, 4H)ArH 8.4(s,1H)NH; 9.0(s, 1H)
NH.
1.1(d, 3H) CH3CH; 2.4(d, 2H) CH2CHph; 2.8(m, 1H) COCHCH3; 3.0(t, 1H)
COCHCH2; 7.1(s, H) CH=N; 7.3-8.3 (m, 14H) 3ArH; 4.2, 4.5(b, 2H)2NH;
9.1(s, H) NH.
0.8(m, 6H) 2CH3; 1.3(m, 2H) CH2CH; 1.9(m, H) CHCH3; 2.8 (d, 2H)
phCH2CH; 4.3(m, 2H) 2COCH; 7.1(s, 1H)CH=N; 7.4-8.1(m,14H) 3ArH;
4.5(s, 1H) NH 8.4(s, 1H) NH; 9.1(s, 1H)NH.
127
Table (5): 1H NMR spectral data of compounds (9a,c-d, 10b-d, 11a, 12b,e)
Com
. No.
R , R'
-H,H
9a
9c
9d
10b
10c
10d
11a
12b
12e
-CH3,
CH2CH(CH3)2
-CH2CH(CH3)2 ,
-H
-CH(CH3)2,
CH2C6H5
-CH3 ,
CH2C6H5
-CH3 ,
CH2C6H4OH
CH(CH3)2, H
CH(CH3)2,
CH2C6H5
-CHCH2CH3
CH3
CH2C6H5
128
I
H NMR (400 MHz, DMSO-d6, δ, ppm)
2.4(s, 3H)CH3ph; 3.9(d, 2H)NHCH2; 4.3(s, 2H) CH2CO; 7.4,7.7(dd,4H)ArH;
7.9,8.2 (dd,4H) ArH; 8.0(s,2H)2NH.
0.9(d,6H)(CH3)2CH; 1.2(m,H)CH(CH3)2; 1.5(d, 3H) CH3CH; 1.7(m, 2H)
CH2(CH)2; 2.5(s, 3H) CH3ph;2.8(t,H)NHCHCH2; 7.1-7.9(m, 8H) 2ArH; 8.9(s,
1H)NH;8.5(s, 1H) NH.
0.8(d,6H)(CH3)2CH; 1.3(d,2H)CH2CH;1.7 (m,1H) CH(CH3)2; 2.4(s,3H)
CH3ph; 3.7 (t,2H) CH2NH; 4.1(m,1H) COCHCH2; 7.3,7.9(dd,4H) ArH; 7.68.3 (dd,4H) ArH; 8.5(s, 2H)2NH.
0.9(d, 6H)(CH3)2CH; 1.8(d, 2H)phCH2CH; 1.4(m, 1H)CH(CH3)2; 2.1(t,
1H)COCHCH2ph; 2.8 (d,1H) COCHCH; 4.5(s,1H)NH; 5.6(s,1H)NH; 7.18.2(m, 14H)3ArH.
1.1(d, 3H) CH3CH; 1.6(d, 2H) phCH2CH; 2.0(m,1H) COCHCH3; 2.3(t,1H)
CHCH2ph; 4.5(s,1H)NH; 5.7 (s,1H)NH; 7.2-8.2(m, 14H)3ArH.
0.8(d, 3H) CH3CH; 1.0(d, 2H) phCH2CH; 1.4 (t, 1H) CHCH2ph; 2.1(q, H)
COCHCH3; 4.5(s, 1H) NH; 5.6(s,1H)NH;7.4-8(m,5H)ArH;8.1,8.4
(dd,8H)2ArH; 8.9(s, 1H)OH.
0.9(d, 6H) (CH3)2CH; 1.3(m, 1H) CH(CH3)2; 2.5(s, 3H) CH3ph; 2.7 (d, 2H)
CH2NH;4.1(d,1H)COCHCH;5.6(b,1H)NH; 7.2,7.5(dd,4H)
ArH;7.4(s,H)NH;8.6(s, 1H)NH.
0.7(d, 6H)(CH3)2CH; 1.3(m, H)CH(CH3)2; 1.8 (t,1H) NHCHCH2;2.3
(d,2H)CH2CH; 3.6(d,1H) CHCO; 4.2 (b,1H) NH, 7.2(d, 1H)NH; 8.4 (s,1H)
NH; 7.6-8.2(m, 10H)2ArH.
0.9 (m, 6H) 2CH3; 1.2(m, 2H) CH2CH3; 1.6 (m,2H) 2CH; 2.1(d,2H) CH2Ph;
4.3(d,1H) CHCO; 5.2(b, 1H) NH; 7.1-7.8(m, 10H) 2ArH; 8.2 (s, 1H)NH,
8.8(s, H) NH.
Table (6): the antibacterial activity of compounds (5d, 7a, 9e, 10d, 11a, 12e, 13a, 14d); and diameter of
inhibition zone(cm).
Comp.
Mg/m
No.
l
DMSO
5d
7a
9e
10d
11a
12e
13a
14b
10
-
1.2
1.8
-
1.8
0.9
-
-
-
Staph.
1
-
-
1.3
-
1.0
-
-
-
-
Aureus
0.1
-
-
0.9
-
-
-
-
-
-
0.01
-
-
0.6
-
-
-
-
-
-
10
1.5
1.6
2.1
2.0
1.6
2.2
1.4
-
-
Bacillus
1
0.7
1.2
1.4
1.2
1.2
1.4
1.0
-
-
subtilis
0.1
-
0.7
1.0
0.6
0.7
0.7
0.7
-
-
0.01
-
-
0.6
-
0.5
-
-
-
-
10
-
0.6
1.2
-
-
0.8
0.6
-
-
Escherichia
1
-
-
0.8
-
-
-
-
-
-
coli
0.1
-
-
0.5
-
-
-
-
-
-
0.01
-
-
-
-
-
-
-
-
-
10
1.2
0.8
1.4
-
1.2
1.4
-
-
-
Proteus
1
0.7
-
1.0
-
0.8
0.8
-
-
-
mirabilis
0.1
-
-
0.8
-
-
-
-
-
-
0.01
-
-
0.5
-
-
-
-
-
-
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H@5d,7a,9e,10d,11a,12e,13a,14bI
@
@
131
ISSN: 2410-7549
ON THE ENERGY OF SOME COMPOSITE GRAPHS
Nechirvan Badal Ibrahim
Dept. of Mathematics, Faculty of Science University of Duhok, Kurdistan Region-Iraq.
ABSTRACT:
Eigenvalues of a graph are the eigenvalues of its adjacency matrix. The energy of a graph is the sum of
the absolute values of its eigenvalues, was studied by (Gutman 1978 ). This paper divided in to three
parts, in part one spectra and nullity of graphs are defined ( Brouwer and Haemers, 2012) and (Harary,
1969). In the second part graph products an their spectra is studied (Shibata and Kikuchi 2000) and
(Balakrishnan and Ranganathan , 2012). In the last part, we proves the energy of some graph products
including Cartesian, tensor, strong, skew and inverse skew which are applied of some graphs.
Keywords: Graph product, Spectra, Energy.
1. INTRODUCTION
L
et G be a graph of p vertices with adjacency matrix A, then A is a real symmetric matrix and so
the eigenvalues of A are real and hence can be ordered. The eigenvalues of A, λ1 ≥ λ2 ≥ ··· ≥
λp are called the eigenvalues of G and form the spectrum of G. The energy E(G) of a graph G is
defined as the sum of absolute values of its eigenvalues. That is E(G)=∑ |λ |.The study of
properties of E(G) was initiated by ( Gutman, 1978 ). All graphs considered in this paper are finite,
simple and undirected.
In this part, we look at the properties of graphs from their eigenvalues. The set of eigenvalues of a
graph G with its multiplicities is known as the spectrum of G and denoted by Sp(G).
Definition 1.1: The adjacency matrix A(G) or A=[aij] of a labeled graph G with vertex set V(G),
V(G)=v1, v2, …., vp is a pp matrix in which aij =1 if vi and vj are adjacent, and 0 if they are not.
Adjacency matrices define graphs up to isomorphism. Moreover, the adjacency matrix of a
graph G is a symmetric 0, 1 matrix having zero entries along the main diagonal, and in which the sum
of the entries in any row or column is equal to the degree of the corresponding vertex. Because of this
correspondence between graphs and matrices, any graph theoretic concept is reflected in the adjacency
matrix.
Definition 1.2: The characteristic polynomial of the adjacency matrix A(G) of a graph G with p
vertices is called the characteristic polynomial of G, denoted by (G; x) with the convention that the
coefficient of the highest order term is positive:
(G;x)=Det(xIp-A(G))=(-1)pDet(A(G)-xIp).
Therefore, the characteristic polynomial of a graph G of order p is a polynomial of degree p:
(G; x) =a0xp+a1xp-1+…+ap-1x+ap.
It has two practical forms, explicitly as a polynomial in the variable x, or as product of linear
factors. Thus,
(G; x)=∑ a x =∏ (x −  ).
Definition 1.3: The eigenvalues of a graph G of order p are defined to be the eigenvalues of the
adjacency matrix associated with the graph G. That is, if G has adjacency matrix A(G), then the
eigenvalues of G are those p (not necessarily distinct) numbers  which satisfy the determinant
equation Det(A(G)-Ip)=0, viz each  is a root of the polynomial equation A(G)-Ip = 0.
Equivalently, a number  is an eigenvalue of G if there exists a non-zero p1 vector X (called an
eigenvector of  ) such that A(G)X = X.
Since A(G) is a real symmetric ( 0, 1) matrix, its eigenvalues 1, 2, …, p must be real by the
next theorem and can be ordered as 1  2  …  p. See (Balakrishnan, 2004)
132
Theorem 1.4: The eigenvalues of a real symmetric matrix are real. ■
Theorem 1.5: The sum of the eigenvalues of any simple graph is zero. ■
Definition 1.6: Let G be a graph of order p and H is a subgraph of G of order m, m  p, then H is
called a partial cover of G if every component of H is isomorphic with K2 or a cycle graph. If m = p,
then H is called a spanning cover of G.
Theorem 1.7: (Sachs’ Theorem) The coefficients ai’s of (G; x) are given by
ai=∑ (−1) ( ) 2 ( ) ,
Where the summation extends over all partial covers H on i vertices of G, and where k(H) and c(H)
denote respectively, the number of components and of cycles in H.
Definition 1.8: The spectrum Sp(G) of a graph G is defined as the eigenvalues of its adjacency
matrix, that is, another matrix of two rows, the first row consists of the eigenvalues of the graph G
and the second row consists of the multiplicities of the corresponding eigenvalues. That is if the
distinct eigenvalues of G are 1, 2, …, k and their multiplicities are m1, m2, …, mk, respectively,
then we write

 …  Sp(G)= m
m
… m
Or just as  ,  , …,  .
If G is a disconnected graph with components G1, G2, …, Gk, then the spectrum of G is the “union”
of the eigenvalues of the components of G in some manner because of the fact that
(G; x)=∏ (G ; x).
Spectra of graphs can be obtained using the fact that the coefficients of the characteristic
polynomial are integers. It follows that the sum of k-th powers of eigenvalues are integers too. Since
the coefficient of the highest power term xp of the characteristic polynomial (G; x) is equal to 1,
hence any eigenvalue of G which is rational must be an integer, and for any square matrix with real
entries, the sum of its eigenvalues is equal to its trace.
To specify the spectrum of a graph G with order p, the coefficients aisof the characteristic
polynomial (G; x)=∑ a x
are of important use. Thus, we seek the coefficients ais first.
Certainly a0 =1, and using Theorem 1.7, we can easily verify that a1=0, -a2 = q and -a3 is twice the
number of triangles in G. And to find the remaining coefficients apply Theorem 1.7.
Definition 1.9: A graph G is said to be a singular graph provided that its adjacency matrix A(G) is a
singular matrix. The algebraic multiplicity of the number zero in the spectrum of the graph G is called
its nullity (degree of singularity), and is denoted by (G).
Lemma 1.10:
i) The eigenvalues of the cycle graph Cp are of the form2cos
, i = 0, … , p − 1, and
2,ifp ≡ 0(mod4),
0,otherwise.
ii) The eigenvalues of the path graph Pp are of the form2cos
, i = 1, … , p, and
η(Cp)= η(Pp)=
1, ifpisodd,
0, ifpiseven.
iii) The spectrum of the complete graph Kp, Sp(Kp)=
iv)
The
Sp(Kp1,p2)=
p−1
−1
1, ifp = 1,
,and η(Kp)=
1
p−1
0, ifp > 1.
complete
bipartite
graph
Kp1,p2,
spectrum
of
the
p p
0
− p p
, and η(Kp1,p2)=p1+p2-2, for all p1, p2.
1
p +p − 2
1
2. Graph Products
133
In this part, we study some graph products and determine the spectra of some of them.
Let G1 =(V1, E1) and G2=(V2, E2) be vertex disjoint non-trivial graphs.
Definition 2.1: The Cartesian product G1G2 of the two graphs G1 and G2 is the graph with vertex
set V(G1G2)=V1V2 and two vertices (u1, v1) and (u2, v2) are adjacent in G1G2 if, and only if, [u1 ≡ u2
and v1v2E(G2)] or [u1u2E(G1) and v1 ≡ v2].
It is clear that: p(G1G2)=p(G1)p(G2) and q(G1G2)=p(G1)q(G2)+q(G1) p(G2)
Definition 2.2: The tensor product G1G2 of the two graphs G1 and G2 is the graph with vertex set
V(G1G2)=V1V2 and two vertices (u1, v1) and (u2, v2) are adjacent in G1 G2 if, and only if, [u1 u2
E(G1) and v1v2E(G2)].
It is clear that: p(G1G2)=p(G1)p(G2) and q(G1G2)=2q(G1) q(G2).
Definition 2.3: The strong product G1⊠G2 of the two graphs G1 and G2 is the graph with vertex set
V(G1⊠G2)=V1V2 and two vertices (u1, v1) and (u2, v2) are adjacent in G1⊠G2 if, and only if, [u1 ≡ u2
and v1v2E(G2)] or [u1u2E(G1) and v1 ≡ v2] or [u1 u2  E(G1) and v1v2E(G2)].
It is clear that: p(G1⊠G2)=p(G1)p(G2) and q(G1⊠G2)=p(G1)q(G2)+q(G1) p(G2)+2q(G1) q(G2).
Definition 2.4: The skew product G1G2 of the two graphs G1 and G2 is the graph with vertex set
V(G1G2) = V1V2 and where (u1, v1) and (u2, v2) are adjacent in G1G2 if, and only if, [u1 ≡ u2 and
v1v2E(G2)] or [u1u2E(G1) and v1v2E(G2)].
It is clear that: p(G1G2) = p(G1)p(G2) and q(G1G2) = p(G1)q(G2)+2q(G1)q(G2).
Definition 2.5: Let G1 = (V1, E1) and G2 = (V1, E1) be vertex disjoint non-trivial graphs. The inverse
skew product G1 G2 of the two graphs G1 and G2 is the graph with vertex set V(G1 G2) = V1V2 and
where (u1, v1) and (u2, v2) are adjacent in G1G2 if, and only if, [u1u2 E(G1)and v1 ≡ v2] or
[u1u2E(G1) and v1v2E(G2)].
It is clear that: p(G1 G2) = p(G1)p(G2) and q(G) = q(G1)p(G2)+2q(G1)q(G2).
Lemma 2.6: (Shibata and Kikuchi, 2000) Let G1 and G2 be two graphs with orders p1 and p2,
respectively. Then
i) A(G1G2) = (Ip1 A2)+(A1A2).
ii) A(G1G2) = (A1Ip2)+(A1A2). ■
Corollary 2.7: Let Sp(G1)={ λ1, ..., λp1} and Sp(G2)={µ1,..., µp2}, and let A1 and A2 be the adjacency
matrices of G1 and G2; respectively. Then
(i) A(G1G2)=( Ip1 A2)+(A1Ip2) and Sp(G1G2)= {λi+µj: 1≤ i ≤ p1; 1≤ j ≤ p2}.
(ii) A(G1G2)=(A1A2) and Sp(G1G2)= {λiµj : 1≤ i ≤ p1; 1≤ j ≤ p2}.
(iii) A(G1 ⊠ G2) =(A1A2)+(Ip1A2)+(A1Ip2) and Sp(G1 ⊠ G2)= { λiµj + λi+µj : 1≤ i ≤ p1; 1≤ j ≤ p2}.■
3. Energy of a Product Graphs
In this part, we discuss another application of eigenvalues of graphs. The energy E(G) of a graph G
is defined as follows:
Definition 3.1: Let G be a graph on p vertices, and let its ordinary spectrum (i. e., the spectrum of its
adjacency matrix) consist of the numbers λ1, λ2, ..., λp. Then, The energy E(G) of a graph G is defined
as follows:
E(G) = ∑ |λ |.
In the next proposition, the energy of some special graphs is studied by (Balakrishnan, 2004) and
(Li, Shi, and Gutman, 2012).
Proposition 3.2: The energy of some special graphs is defined as follows:
1) E(Kp)= 2p-2.
134
2) E(Kp1,p2)=2 p p .
3) E(Pp)=
( (
)
( (
( (
( )
( )
(
)
)
( )
4) E(Cp)=
)
)
− 2, ifp = 0(mod2),
)
− 2, ifp = 1(mod2).
, ifp = 0(mod4),
, ifp = 2(mod4),
)
, ifp = 1(mod2).
In the following, we determine the spectra of skew and inverse skew products.
Let A1 and A2 be the p1×p1 and p2×p2 adjacency matrices of G1 and G2 have eigenvalues λi,1≤ i ≤ p1
and µj, 1≤ j ≤ p2, respectively.
Theorem 3.3: The p1p2 eigenvalues of the skew product G1G2 are µj+λiµj, ∀ i, j; 1 ≤ i ≥ p ;1 ≤ j ≥
p .
Moreover, if X1, ..., Xp1 are the eigenvectors of A1 corresponding to λ1, ..., λp1, and Y1, ...,Yp2 are
the eigenvectors of A2 corresponding toµ1,...,µp2, then XiYj are the eigenvectors of G1G2
corresponding to µj +λiµj, 1≤ i ≤ p1; 1≤ j ≤ p2.
Proof: By Lemma 2.6, we have:
A(G1G2)=[(Ip1A2)+(A1A2)]
Assume that X(Y) is an eigenvector of G1(G2) corresponding to the eigenvalue λ(µ). Then
A(G1G2) (XY)=[(Ip1A2)+(A1A2)] (XY)
=(XA2Y)+( A1XA2Y)
=(XµY)+( λXµY)
=( µ+ λµ)(XY). ■
Theorem 3.4: The p1p2 eigenvalues of the inverse skew product G1 G2 are λ i+λiµj, for all 1≤ i ≤ p1;
1≤ j ≤ p2.
Moreover, if X1, ..., Xp1 are the eigenvectors of A1 corresponding to λ1, ..., λp1, and Y1, ...,Yp2 are
the eigenvectors of A2 corresponding to µ1, ..., µp2, then XiYj are the eigenvectors of G1  G2
corresponding to λi+λiµj, 1≤ i ≤ p1; 1≤ j ≤ p2.
Proof: By Lemma 2.6, we have:
A(G1 G2)=[( A1 Ip2)+(A1A2)]
Also, assume that X(Y) is an eigen vector of G1(G2) corresponding to the eigen value λ(µ). Then
A(G1 G2) (XY)=[( A1 Ip2)+(A1A2)] (XY)
=(A1XY)+( A1XA2Y)
=(λXY)+(λXµY)
=(λ+ λµ)(XY). ■
Thus, as a result for the spectra of the above two products we have:
Corollary 3.5: Let Sp(G1)={ λ1, ..., λp1} and Sp(G2)={µ1, ..., µp2}. Then
(i) Sp(G1G2)= {µj+λiµj : 1≤ i ≤ p1; 1≤ j ≤ p2}.
(ii) Sp(G1 G2)= {λi+ λiµj: 1≤ i ≤ p1; 1≤ j ≤ p2}.■
Proposition 3.6: The energy of the Cartesian product of Kp1 and Kp2 where p1, p2 >1, is given by
E(Kp1Kp2 )= 4(p1p2-p1-p2+1).
Proof: By Corollary 2.7, we have:
135
Sp(Kp1Kp2)
p −1
−1
p −1
=
+
1
p −1
1
−1
p +p −2
=
p −1
1
p −2
p −1
p −2
p −1
−2
p p −p −p +1
Thus, by Definition 3.1, we get:
E(Kp1Kp2 ) =p1+p2-2+p1p2-p1-2p2+2+p1p2-p2-2p1+2+2p1p2-2p1-2p2+2
= 4(p1p2-p1-p2+1).
Therefore, Kp1Kp2 is singular, if and only if p1=p2=1, this gives the case K1K1=K1. ■
Proposition 3.7: The energy of the Cartesian product of Kp and Kp1,p2 is
E(KpKp1,p2 )= 2(p-1)( p1+p2+ p p -1).
Proof: By Lemma 1.10 and Corollary 2.7 it follows that the spectrum of KpKp1,p2 is
p−1
1
−1
+
p−1
p+ p p −1
1
p p
0
− p p
=
1
p +p −2
1
p−1
p− p p −1
p p −1
p +p −2
1
p−1
−1
(p − 1)(p + p − 2)
−( p p + 1)
p−1
Thus, by Definition 3.1, we have:
E(KpKp1,p2)=(p + p p − 1) + (pp + pp − 2p − p − p + 2) + (p − p p − 1) +
( p p p − p p − p + 1) + (pp + pp − 2p − p − p + 2) + ( p p p − p p + p −
1)=2pp1+2pp2+2 p p p - 2p -2p -2p +2-2 p p
=2p(p1+p2+ p p -1) -2(p1+p2+ p p -1)= 2 (p-1)( p1+p2+ p p -1). ■
Moreover, if p≠ p p +1≤ 0 , so
i) If p1=p2=1, then KpK1,1 is (p -1) singular.
ii) If either p1 or p2 is not 1 while p= p p + 1 , then KpKp1,p2 is 1- singular.
Lemma 3.8: The energy of the Cartesian product of Kp1 and Kp2,p2 is
E(Kp1Kp2,p2)=(p − 1)(6p − 2).
Proof: Put p1=p2 in Proposition 3.7, we get the result. ■
Proposition 3.9: The energy of the Cartesian product of Kp1,p2 and Kp1,p2 is
E(Kp1,p2Kp1,p2 )=4 p p (p + p − 1).
Proof: By Corollary 2.7, we have
p p
p p
0
− p p
0
− p p
+
1
p +p −2
1
1
p +p −2
1
2 p p
p p
0
p p
0
− p p
=
1
p + p − 2 1 p + p − 2 2p + 2p − 4 p + p − 2
2 p p
p p
0
− p p
−2 p p
=
1
2p + 2p − 4 2p + 2p − 2 2p + 2p − 4
1
0
1
− p p
p +p −2
−2 p p
1
Thus, by Definition 3.1, we have:
= 2 p p + 2p p p + 2p p p − 4 p p + 2p p p + 2p p p − 4 p p + 2 p p
E(Kp1,p2Kp1,p2 )= 4p p p + 4p p p − 4 p p = 4 p p (p + p − 1). ■
Moreover, the nullity of Kp1,p2Kp1,p2 is 2p1+2p2-2.
136
Lemma 3.10: The energy of the Cartesian product of Kp1,p1 and Kp2,p2 is
E(Kp1,p1Kp2,p2 )=2(4p p − p − p ).
p 0
−p p
0
−p
+
=
1 2p − 2
1
1 2p − 2
1
p +p
p p −p
p
0
1
2p − 2
1
2p − 2 (2p − 2)(2p − 2)
−p
2p − 2
p −p
1
−p −(p + p )
2p − 2
1
E(Kp1,p1Kp2,p2 ) = 8p p − 2p − 2p = 2(4p p − p − p ). ■
Moreover, the nullity of Kp1,p1Kp2,p2 is (2p1-2)(2p2-2), provided that neither p1 nor p2 is 1. If
p1=p2=1, then the nullity of K1,1K1, 1 is 2.
Proposition 3.11: Let G1and G2 be two graphs on p1 and p2 vertices, respectively. Then E(G1 G2 ) ≤
p2 E(G1 )+p1E(G2 ).
Proof: Let λ1, λ2, …, λp1 and μ , μ , … , μ be the eigenvalues of G1 and G2; respectively. Then
E(G1G2)= ∑ ∑ [ λ + μ ] ≤ ∑ ∑ |λ | + |μ |≤ ∑ ∑ |λ | + ∑ ∑ |μ |
≤ p2 E(G1 )+p1E(G2 ). ■
Proposition 3.12: The energy of the tensor product Kp1Kp2 is given by:
E(Kp1Kp2 ) = 4(p1-1) ( p2-1).
Proof: By Lemma 1.10 and Corollary 2.7, we have:
p −1
−1
p −1
−1
1
p −1
1
p −1
(p − 1)(p − 1) −p + 1 −p + 1
1
=
1
p −1
p − 1 (p − 1)(p − 1)
E(Kp1Kp2 ) = p1p2-p1-p2+1+p1p2-p2-p1+1+ p1p2-p2-p1+1+p1p2-p1-p2+1
= 4(p1p2-p1-p2+1) = 4(p1-1) (p2-1). ■
Proposition 3.13: The energy of the tensor product Kp1,p2Kp1,p2 is given by:
E(Kp1,p2  Kp1,p2 )=4p p .
p p
p p
0
− p p
0
− p p
=
1
p +p − 2
1
1
p +p − 2
1
p p
0
−p p
0
0
0
1
p +p − 2
1
p + p − 2 (p + p − 2) p + p − 2
p p
0
−p p
Sp((Kp1,p2  Kp1,p2 )=
.
2
4(p + p − 2) + (p + p − 2)
2
Then, by Definition 3.1, we get:
E(Kp1,p2  Kp1,p2 )=4p p .
And the nullity of Kp1,p2 Kp1,p2 is4(p + p − 2) + (p + p − 2) . ■
−p p
1
0
p +p −2
Proposition 4.14: (Balakrishnan 2004) Let G1and G2 be two graphs on p1 and p2 vertices, respectively.
Then E(G1G2 )= E(G1) E(G2).
Proof: By Definition 3.1, we have:
E(G1  G2 )= ∑ ∑ |λ μ | = ∑
∑
|λ ||μ |= ∑
|λ |∑
|μ |= E(G1 ) E(G2 ). ■
137
p p
1
Proposition 3.15: The energy of the strong product of two graphs is the sum of energy of their
Cartesian and tensor products. ■
Proposition 3.16: The energy of the skew product of two graphs G1 and G2 is related as:
E(G1  G2 ) ≤ p1 E(G2 )+ E(G1 ) E(G2 ).
Proof: Let the spectra of G1and G2 be {λ1, ..., λp1} and {µ1, ..., µp2}; respectively.
Then
E(G1G2)=∑ ∑ [|μ + λ μ |] ≤ ∑ ∑
μ + λ μ ≤∑
∑ ∑ |λ μ | ≤ p1 E(G2 )+ E(G1 ) E(G2 ). ■
∑
|μ | +
∑
|λ | +
Example 3.17: Let G1=K2 and G2=P3, then the skew product of K2 and P3 is given by:
1 −1
Sp(K2)=
and Sp(P3)= √2 0 −√2
1 1
1 1
1
For j=1 and i=1, 2, we have: μ + λ μ = 2√2, μ + λ μ = 0.
For j=2 and i=1, 2, we have: μ + λ μ = 0, μ + λ μ = 0.
For j=3 and i=1, 2, we have: μ + λ μ = −2√2, μ + λ μ = 0.
Then, E(K2P3)=4√2.
And by Proposition 3.16, we have E(K2P3)≤ 2 ∗ 2√2 + 2 ∗ 2√2 = 8√2.
Moreover, equality does not hold for any pair of non-empty simple graphs.
Proposition 3.18: The energy of the inverse skew product is given by
E(G1  G2 ) ≤ p2 E(G1 )+ E(G1 ) E(G2 ).
Proof: Let the spectra of G1 and G2 be {λ1, ..., λp1} and {µ1, ..., µp2}, respectively.
Then
E(G1

G2)=∑ ∑
λ + λ μ ≤ ∑ ∑ [ μ + λ μ ]≤∑
∑ ∑ |λ μ | ≤ p2 E(G1 )+ E(G1 ) E(G2 ). ■
Example 3.19: Let G1=K2 and G2=P3, then the inverse skew product of K2 and P3 is given by
1 −1
Sp(K2)=
and Sp(P3)= √2 0 −√2
1 1
1 1
1
For i=1 and j=1, 2, 3, we have: λ + λ μ = 1 + √2, λ + λ μ = 1, λ + λ μ = 1 − √2.
For i=2 and j=1, 2, 3, we have: λ + λ μ = −1 − √2, λ + λ μ = −1, λ + λ μ = −1 + √2.
Then, E(K2 P3)= 2+4√2.
And by Proposition 3.18, we have E(K2  P3)≤ 3 ∗ 2 + 2 ∗ 2√2 = 6 + 4√2.
REFERENCES
- A.E. Brouwer and W.H. Haemers; (2012), Spectra of Graphs, Springer, NewYork.
- F. Harary, ;( 1969), Graph Theory, Addison-Wesley, Reading, MA. Fundamentals, Vol. E83-A, No. 3, pp.459464.
- I.Gutman; (1978), The Energy of a Graph, Ber. Math. Statist. Sekt. For schungszenturm Graz. 1031–22.
- R. Balakrishnan, and K. Ranganathan; (2012), A Textbook of Graph Theory, Springer, New York.
- R. Balakrishnan; (2004), The energy of a graph, Linear Algebra and its Applications 387, pp.287–295.
X.
Li,
Y.
Shi,
and
I.
Gutman;
(2012),
Graph
Energy,
Springer.
http://books.google.iq/books?id=90U4suPlWS4C&printsec=frontcover&hl=ar#v=onepage&q&f=false.
- Y. Shibata, and Y. Kikuchi,; (2000), Graph products based on the distance in graphs, IEICE Trans.
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139
ISSN: 2410-7549
CONVERGENCE OF THE BARZILAI-BORWEIN METHOD FOR SOLVING
SLIGHTLY UNSYMMETRIC LINEAR SYSTEMS
Bayda Ghanim Fathi
Dept. of Mathematics, Faculty of Science, University of Zakho, Kurdistan Region-Iraq.
Abstract:
Due to its simplicity and numerical efficiency, the Barzilai and Borwein (BB) gradient method has
received numerous attentions in different scientific fields. In this paper, the sufficient condition for
convergence of the BB method when the coefficient matrix of linear algebraic equations is slightly
unsymmetric with positive definite symmetric part is presented.
Keywords: Unsymmetric linear algebraic equations, Barzilai and Borwein gradient method, symmetric and
skew-symmetric matrices, eigenvalues, condition number.
1. Introduction
A significant development that has completely changed our perspectives on the effectiveness of
gradient methods is due to Barzilai and Borwein ( Barzilai andBorwein, 1988). They proposed two
choices of step size the gradient method. The computational cost and typical behaviour of the
algorithm for both choices of step size are quite similar. Their method aimed to accelerate the
convergence of the steepest descent (SD) method.
The main idea of Barzilai and Borwein’s approach is to use the information in the previous
iteration to decide the step size in the current iteration. The Barzilai-Borwein (BB) method requires
few storage locations and inexpensive computations. Therefore, several authors have paid attention to
the BB method. Their method is strongly related to Quasi-Newton (QN) algorithms (Dennis and
Schnabel, 1983) and (Fletcher, 2000).
It is known that the (BB) method ( Barzilai andBorwein, 1988) converge when this method is
applied to solve linear systems of the form
Ax =b,
where A is symmetric and positive definite. For some finite difference discretizations of elliptic
problems, one gets positive definite matrices that are almost symmetric. Practically, the BB method
works for these matrices. However, the convergence of this method is not guaranteed theoretically.
For quadratics that the BB method has been shown to converge (Raydan. ,1993) and its
convergence is R-linear (Dai and Liao, 2002). For the case of = 2, the method is R-superlinearly
convergent ( Barzilai andBorwein, 1988). For more details on the Barzilai and Borwein method see
(Raydan, 1991) and (Dai and R. Fletcher, 2005). The BB algorithm for quadratic case is summarized
in Algorithm 1.
Algorithm 1 Barzilai and Borwein (BB)
1. Given ∈ ℝ , choose arbitrary > 0, for instance,
2. For k=0, 1, 2, …(until convergence) do
3.
=
−
4.
=
−
5.
=
−
(
,
)
(
,
6.
=
,( or
=
(
,
)
(
,
)
)
=
(
(
,
,
)
,(or )
=
(
(
,
,
)
)
)
)
7. End for
This paper presents the proof of convergence of the BB method when the Euclidian norm ( ) of
the unsymmetric part of a positive definite matrix is less than some value related to the smallest and
the largest eigenvalues of the symmetric part of the given matrix. This means that the restriction of
be symmetric is removed, and required only that its symmetric part ( + )/2 be positive definite.
140
Let us begin with some notations:
1- The Euclidean inner product of any two vectors , ∈
is defined by
( , )=
,
and the induced Euclidean norm (or 2-norm) of is
║ ║ = ( , ).
2- Let be an matrix and the associated matrix norm is given by
A ≔ Sup‖ ‖ ║Au║.
It is known that can be represented as
= 0+ ,
where
and the symmetric and skew-symmetric of a matrix , i.e.
= ( + )/2,
= ( − )/2.
4- Let be positive definite, then the symmetric part
is also positive definite. Hence
and
are invertible and the eigenvalues of
are all positive real numbers. If
,…,
are the eigenvalues of
such that
0<
≤… ≤
then
, …,
are the eigenvalues of
.
5- The condition number of is defined to be
:=║ ║║
║.
Since
is symmetric,
=
≥ 1.
6- The Rayleigh-quotient of any vector
∈
with respect to is defined by
( , )
( )=
.
( , )
The lemmas that will be used frequently in the proof of the main theorem are follows:
Lemma1. For any vector ,
║ ║ ≤ ( , )≤
‖ ║ .
Proof. For a symmetric matrix positive definite matrix , the Rayleigh quotients are bounded by the
smallest and the largest eigenvalues of the matrix. Thus, the following relation holds for any u
( ,
)
≤
≤
.
║ ║
Hence the proof is complete.
Lemma 2. For any vector ,
)≤
║ ║ ≤( ,
║ ║ .
Proof. Similar to proof Lemma 1.
Lemma 3. For any vector ,
( , )=( ,
) and
Proof. We have
Since
is a real number,
= ( ) =
Therefore ( ,
) = 0 and ( ,
( ,
( ,
)=0
)=( ,
=−
)=( ,
.
)+( ,
)
).
Lemma 4 (Cauchy-Schwarz inequality for a positive definite matrix).
If
is a positive definite matrix, then for any u andv
|( ,
)| ≤ ( ,
)( ,
).
141
2. Convergence of the BB Method
In this section, the proof of convergence of the BB method applied to the slightly unsymmetric
linear system
= with the positive definite symmetric partis given.
Theorem 1. If
‖ ‖<
(−1 + 1 +
then the BB method, defined by
=
−
(2.1)
where
(
,
)
=
,
( or
=
(
,
)
),
(
,
(
)
,
)
),
(2.2)
converges.
Proof. Using (2.1), (2.2), Lemma (3), Lemma (1) and since
is the Rayleigh quotient of the
symmetric positive definite matrix (i.e. 0 < ≤
≤
for all ),we have
( , )=(
−
−
)
, )−2 (
)+ (
=(
)
, , ,
)−2 (
=(
,
,
)+(
,
)
+ (
,
)+(
,
)
)− (
=(
,
,
)−2 (
,
)
+ (
,
)
= ( ,
) − ( , ) − 2 (
,
)
)+(
+ ( ,
,
)
(2.3)
(
)
=
− (
)−2 (
,
)
,
,
+ (
,
).
(2.4)
) for any , using Lemma 2 and since
Assuming ≔ ‖ ‖ and
≔(
,
≤
≤
, we obtain
(
,
)≥
=
.
(2.5)
By using Lemma 2 twice, we have
‖
‖
‖
(
,
) ≤
‖ ≤
≤
(
,
)≤
.
(2.6)
Using Lemma 4, we get
,
≤
(
,
From (2.3) to (2.7) and utilizing Lemma 3,
≤
−
+2
=
(1 −
+2
+
).
For the purpose of convergence, we need
1−
+2
+
< 1,
2
+
−
< 0,
multiplying by
, we have
+2
−
< 0.
This satisfies when
<−
+
+
=
)
=
.
(2.7)
+
−1 + 1 +
.
This completes the proof.
3. Numerical Experiments
In this section, two numerical experiments are presented to show the rate of convergence of the
Barzilai-Borwein (BB) algorithm for solving the linear system of equations
= where is slightly
unsymmetric positive definite matrix. They demonstrate that if the sufficient condition
142
‖ ‖<
(−1 + 1 +
)
Satisfies, then the BB algorithm convergence. Simulations were run in MatLab 27.
Example 1. For the first experiment, a matrix
1.0000
−0.0007
0.0013
−0.0004
0.0000
−0.0017
−0.0021
−0.0009
1.6667
0.0015
−0.0014
0.0024
0.0011
0.0021
−0.0001
−0.0000
2.3333
−0.0023
−0.0014
−0.0020
0.0028
with size
can be taken as
−0.0013 −0.0007 0.0020
−0.0022 −0.0013 0.0012
−0.0007 −0.0029 0.0018
3.0000 −0.0007 −0.0019
0.0026
3.6667 −0.0024
0.0007 −0.0033 4.3333
−0.0027 0.0023
0.0020
0.0021
0.0008
0.0003
0.0027
0.0026
0.001
5.0000
where
= 7, =
, = 1,2, … , ; = 1,2, … , such that
are equally spaced real
numbers between 1 and 5when = , and
is a random number between -1 and 1 when
. The
symmetric part of ( ) is symmetric positive definite matrix and its condition number = 5 where
= 1 and
= 5. ‖ ‖ = 0.0041. The value of
−1 + 1 +
= 0.0427.
=
Figure 1 shows the rate of convergence
(
)
,
(
,
)
of the BB algorithm as a function of
number of iteration =300
Figure 1: Rate
of convergence as a function of for Example 1.
Example 2. For the second experiment, a matrix
11.0000
0.0000
0.0000
0.0000
0.0000
0.7621
11.0000
0.0000
0.0000
0.0000
with size
can be taken as
0.7621 0.4057 0.0579
0.7919 0.9355 0.3529
11.0000 0.9169 0.8132
0.0000 11.0000 0.0099
0.0000 0.0000 11.0000
where = 5, =
, = 1,2, … , ; = 1,2, … , such that
= 11 when = , and
=
( ) when
.
is symmetric positive definite matrix and its condition number = 1.1801
where
= 10.3449 and
= 12.2082. ‖ ‖ = 1.0117and the value of
−1 +
1+
= 3.4204.
Figure 2 shows the rate of convergence
of iteration
= 100.
=
(
)
,
(
,
)
of the BB algorithm as a function of number
143
Figure 2: Rate
of convergence as a function of
for Example 2.
References
Barzilai, J. and Borwein, J. M.: Two-point step size gradient methods. IMA Journal of Numerical
Analysis, 8, pp. 141–148, 1988.
Dai, Y. H. and Fletcher, R.: On the asymptotic behaviour of some new gradient methods.
Mathematical Programming, 103(3), pp541–559, 2005.
Dai, Y. H. and Liao, L. Z.: R-linear convergence of the Barzilai and Borwein method for
unconstrained optimization. IMA Journal of Numerical Analysis, 1, pp. 1–10, 2002.
Dennis, J. E. and Schnabel, R. B: Numerical Methods for Unconstrained Optimization and Nonlinear
Equations. Prentice-Hall, Englewood Cliffs, NJ, 1983.
Fletcher, R.: Practical Methods of Optimization. Wiley, 2nd edition, 2000.
Raydan, M. M.: On the Barzilai and Borwein choice of steplength for the gradient method. IMA
Journal of Numerical Analysis, 13, pp. 321–326, 1993.
Raydan, M. M.: Convergence propreties of the Barzilai and Borwein gradient method. PhD thesis,
Rice University, 1991.
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144
ISSN: 2410-7549
ON GENERALIZED REGULAR LOCAL RING
Zubayda M. Ibraheem and Naeema Ali Shereef
Dept. Of Mathematics ,College of Computer and mathematical Sciences, University of Mosul, Iraq.
Abstract:
A ring R is called a generalized Von Neumann regular local ring (GVNL-ring) if for any a∈R, either a
or (1-a) is π-regular element. In this paper, we give some characterization and properties of generalized
regular local rings. And we studied the relation between generalized regular local rings, Von Neumann
regular rings, Von Neumann regular local rings (VNL-rings) and exchange rings.
Key words: local, π-regular , exchange rings .
1- Introduction:
T
hroughout this paper, R will be an associative ring with identity. For a∈R, r(a), ℓ (a) denote the
right (left) annihilator of a. We write Y(R), (R) for the right singular ideal and the Jacobson
radical of R, respectively. An ideal of a ring R is said to be essential if and only if has a non-zero
intersection with every non-zero ideal of an R.A ring R is reduced if R contains no non-zero nilpotent
element. A ring R is said to be Von Neumann regular (or just regular) if and only if for each a in R
,there exists bin R such that a=aba [7]. In[3] Contessa first introduced and characterized a VNL-ring,
and gave many properties, a ring R is called Von Neumann regular local rings (VNL-rings), if for any
a∈R, either a or (1-a) is Von Neumann regular element. A ring R is called local ring, if it has exactly
one maximal ideal [1]. A ring R is called π-regular ring if and only if for each a in R, there exists b in
R and a positive integer n such that n= n b n [4]. clearly that every π-regular ring is GVNL-ring. A
ring R is said to be strongly commuting regular if for each
,
in R
)2[6].
thereexista
∈ ℎ ℎ (
)=(
)2a(
A ring R is called Exchange ring if for any a∈R, there exists an idempotent element e ∈R such that
e∈aR and (1-e) ∈(1-a)R [5].
2- GVNL-Rings
This section is devoted to give the definition of generalized Von Neumann regular local ring(
GVNL-rings) with some of its characterization and basic properties .
Definition 2.1: [2]
A ring R is called a generalized Von Neumann regular local ring(GVNL-ring) if a or (1-a) is πregular for every a∈R .
Examples:
1-) Let (Z2 ,+, .) be a ring and let G= :
is cyclic group, then
Z2 G= 0, 1, , 1 + is not regular, but π-regular ring.
2-) Let R be the set of all matrix in Z2 which defined as:
R=
0
: , ,
∈
It is easy to find the elements of R
, , , , , , ,
0 0
1 0
0 1
0 0
A =
=
,A =
,A =
,
0 0
0 0
0 0
0 1
0 1
1 1
1 0
1 1
= ,
=
,
, =
0 1
0 1
0 1
0 0
Thus R is not regular but R is π-regular.
3-) Let : R=Z4⊕ Z4, then it is easy to check that ,
(3, 2)and(1, 1) − (3, 2)arenotregular. So R is not a VNL-ring, but R is π-regular. Thus R
is a GVNL-ring.
Proposition: 2.2:
A ring R is GVNL-ring iff an R is generated by an idempotent element for every a∈R and some
positive integer n.
R=
145
Proof:
Suppose that R is a GVNL-ring. Then either a or (1-a) is π-regular. Let a∈R. Then there exists an
element b∈R and appositive integer n such that n= n b n , let e= n b. Clearly e is an idempotent
element in R. We shall show that n R=eR. Let c ∈ nR, then, c=anr for some r∈R. But, n= n b n,
so
C= n b nr = e . nr ∈eR, n R ∈ R. Therefore nR ⊆ eR …(1)
On the other hand if d∈eR, then d=e s for some s∈R, but, e= n b, so, d=anbs ∈ nR
Therefore eR⊆ nR …(2). Thus n=eR
Now, if (1-a) is π-regular, then there exists z∈R and n∈Z+such that
(1- )n=(1- )nz (1- )n,
let e=(1- )nz, we shall show that (1- )nR=eR.
Let w∈(1- )n R, then w=(1- )n s , for some s ∈R. But (1- )n=(1- )n z(1- )n .
so ,w=(1- )n
n
n
n
n
n
z(1- ) s=(1- ) z(1- ) s=e(1- ) s ∈eR, therefore (1- ) R ⊆ eR.
Now if d∈eR, then d=es for some s∈R. But e=(1- )nz, so, d=(1- )n zs ∈ n R
Thus eR⊆(1- )n R. Therefor (1- )n R=eR.
Conversely, let ∈R, e is an idempotent in R such that n R=eR for some positive integer n. Then
e= n b for some b∈R and n=e c for some c∈R, so e n= n b n= , so e n= n b n= e e c=e c= n .
Hence an = e n = n b n. Therefore is π- regular.
Now, for (1- )∈R, let e be an idempotent element in R such that
(1- )n R=eR, for some positive integer n .Then e= (1- )nb for some b∈R
and (1- )n =e c ,for some c∈ R . So, e(1- )n =(1- )n b (1- )n, and e(1-a)n=e e c, hence e(1- )n
= e c=(1-a)n , then (1- )n=e(1- )n=(1- )n b (1- )n
Thus(1- ) is π-regular and therefore R is GVNL- ring.
#
Proposition 2.3:
Let R be a commutative ring and let P be a primary ideal of R. Then P is a maximal ideal if R/P is
a GVNL-ring .
Proof:
Let a∈ , Then + ∈ / .Since R/P is GVNL-ring, then either ( +p) or
((1- )+p) is πregular element in R/P . Let (a+P) be π-regular in R/P. Then let (b+P)∈ R/P and n∈ Z+ such that
n
+P =( +P)n=( +P)n (b +P) ( +P)n
=( n +P)(b +P) ( n+ P)= n b n +P= n b n +P
That is ( n – n b n) ∈P, then n (1- b n) ∈ P. Assume that n ∉p .
Since P is primary, then (1-b n) m ∈P, m ∈ Z+ such that ,
!
(1 - b n)m=1 -[∑
C (-1)k-1 b a ( ) ] n ∈p where =
.
)!
!(
(
)
Now, let z=∑ (−1)
.
n
n
Then (1-z ) ∈P, Thus 1+p=(z+P) ( +P)=(z n-1 +P)(a+P) .
Therefore (a+p) has inverse in R/P. Now, if ((1-a)+P) is π-regular element in R/P, then there exists
n∈Z+and (ℓ+P)∈R/P such that
((1-a)n+P)=((1-a)+P)n=((1-a)+P)n(ℓ+P)((1-a)+P)n=((1-a)n+P)(ℓ+P)((1-a)n+P))
=(1-a)n ℓ (1-a)n+P
Then (1-a)n - (1-a)n ℓ(1-a)n ∈P, that is (1-a)n(1-ℓ(1-a)n)∈P
Let (1-a)n ∉P, since Pis primary ideal, Then (1-ℓ(1-a)n)m ∈ P, m∈Z+such that
(−1) ℓ (1 − ) ( ) ](1 − ) ∈ [1- ℓ(1-a)n]m=1-[∑
(
)
(
)
(−1)( ) ( )( ) ] (1-a)n ∈p
=1- [∑ C )(− 1)( ) ∑
=1-[∑
Where
∑
(
=
)
(
)
(− 1)
a
(
)(
)
] (1-a)n ∈p
!
!(
)!
(
)
(
)
∑
Now, let w=∑
(− 1)
( )( )
n
Then 1-w(1-a) ∈P which implies that 1+p=(w+p)(1-a)n+p = (w(1-a)n-1+P)(a+P) .
Hence ((1-a)+P) has inverse in R/P, thus R/P is a division ring. Therefore P is a maximal ideal in R
. #
146
Theorem 2.4: Le t be a regular ideal of a ring R. Then R is GVNL-ring if and only if R/ is GVNLring .
Proof:
Let R be GVNL-ring. Then either a or (1-a) is π- regular element in R, for all a∈R.
Now, if a is π-regular, then there exists b∈R and n∈Z+ such that n= n b n
Hence (a+I) n= n+ = n b n +
=( n+ ) (b+ ) ( n + )
= ( + )n (b+ )( + )n
Thus ( + ) is π-regular element in R/
Now, if (1- ) is π-regular in R, then there exists d∈ R and n∈ Z+ such that
(1- )n=(1- )n d (1- )n
Hence ((1- )+ )n=((1- )+ ) n (d+ ) ((1- )+ )n
=((1- )n+ ) (d+ ) ((1- )n+ I)
=(1- )n d (1- )n+
= (1- ) n+I
Therefore ((1-a+ ) is π-regular in R/
Conversely, let R/ be GVNL and ∈R. Then either ( + ) or ((1- )+ ) is π-regular element in R/ .
Then there exists (b+ )∈ R/ and a positive integer n such that
( n+ ) = (a+I)n (b+ )( + ) n = ( n+ )(b+ )( n+ ) = an b an+
Hence n+ = n b n+ , so n - n b n∈
Since is regular, there exists c∈ such that
n
- n b n = (n - n b n) c( n - n b n)
n
= n b n+ n c n - n b n c n - n c n b n+ n b n c n ban
= n ( b+c - b n c-c n b+b n c n b) n
= n w n, where w=(b+c-b n c-c n b+b n c n b)
Thus a is π-regular in R.
Now, if (1-a)+ is π-regular, then there exists q+ and n∈Z+ such that,
(1-a)n + = ((1-a)n + )(q+ )( (1-a)n + )
So (1-a)n-(1-a)n q(1-a) n ∈ , since is regular then
(1- )n-((1- )n q (1- )n) =[(1- )n-(1- )n q (1- )n ] w [(1- )n-(1- )nq(1- )n]
=[(1- )n w (1- )n - (1- )n w (1- )n q (1- )n]
-[(1- )n q (1- )n w (1- )n]+[ (1- )n q (1- )n w (1- )n q (1- )n]
= (1- )n [w-w (1- )n q-q (1- )n w +q(1- )n w (1- )n q] (1- )n
n
n
n
(1- ) =(1- ) q(1- ) +(1- )n[w-w (1- )n q-q (1- )n w+ q(1- )n w (1- )n q] (1- )n
= (1- )n[q-w-w(1- )n q-q(1- )n w +q(1- )n w(1- )n q] (1- )n
= (1- )n Z (1- )n
Thus (1- ) is π- regular in R. Therefore R is a GVNL –ring.
#
Proposition2.5:
Let R be a GVNL-ring. Then (R) is nil ideal.
Proof:
Let 0≠ ∈ (R) Since R is GVNL-ring, then either a or (1-a) is a π- regular element of R, if a is πregular, then there exists an element b∈R and n∈Z+ such that n = n b n, then n - n b n = 0. Hence
n
(1- n b) n=0 Since ∈ (R), therefore
∈ (R), and n b ∈ (R). Thus (1- n b) is invertible,
n
so there exists u∈ R such that
u (1b)=1 , it follows that u ( n - n b n) = n, thus a is
nilpotent element. Now, if (1-a) is π-regular, since a ∈ (R), then (1-a) is invertible, and then (1-a)n
invert able, thus (1-a) is nilpotent element and Therefore (R) is a nilideal.
Corollary: 2.6:
Let R be a reduced GVNL – ring Then (R)=0
Proof
Suppose that (R) ≠ 0, then there exists a ∈ (R) and by ( prop .2.5) a is a nilpotent element in R.
But R is reduced, then a=0.therefore (R) =0. #
147
Corollary 2.7:
Let R be a reduced GVNL- ring, Then R is VNL- ring .
Proof:
R is exchange ring [ 2,Theorem 2.2].
Now, R is reduced exchange ring for any a∈R .
n
= n n for some positive integer n and ∈R. Clearly e = n
.If a is π- regular in R, then
where
e
is
an
idempotent
element
in
R
,
so
((1-e) a) n =(1-e) n=(1-e) e n=0, and hence (1-e)a=0 Therefore = e a= ( n-1 ) , is regular.
Now, if (1- ) is π-regular, then (1- ) n = (1- )n z(1- )n for some positive integer n and z∈R.
Clearly e = (1- )n z, so (1-e) (1- )n = (1-e) e (1- )n = 0 .
Hence (1-e) (1- ) = o .
Thus (1-a) = e(1-a) = (1-a)n z(1-a) = (1-a)(1-a)n-1z (1-a) = (1-a)w(1-a)
That is (1- ) is regular. Therefore R is a VNL-ring.
#
Proposition 2.8:
Let R be a GVNL-ring with r (a) ⊆ r (1-a)n for a∈R . Then Y(R) is a nilideal.
Proof:
Let a ∉ 0, 1 be an element in Y(R). Then Y(R) is an essential right ideal of R. Now, since
r(a)⊆r(an), then r(an) is also an essential right ideal of R.
Since R is a GVNL-ring, then either a or (1-a) is π-regular element in R. If a is π-regular, then
there exists b∈R and n∈Z+, such that an=an b an .
Now, consider r ( n) ∩b n R, and let ∈ r( n)∩b nR, Then n =0 and =b n r for some r∈R. So,
n
b n r=0, which implies n r=0, yielding =0.
Therefore r ( n) ∩ b n R=0, since r( n) is a non-zero essential right ideal of R, then b n=0, and
hence n=0 .
Now, if (1- ) is π-regular, then there exists c∈R and n∈Z+ such that
(1-a)n = (1- )n
n
c (1- ) .
Now, since r(a) ⊆ r( n)⊆ r (1-a)n, then r((1- )n) is also an essential ideal of R. Consider r((1- )n)∩
c(1- )n R. Let ∈ r((1- )n)∩c((1- )n) R, then (1-a)n =0, and =c(1- )nr for some r∈R. So, (1-a)n c (1)n r=0, Which implies (1-a)n r=0, yielding =0. Therefore r((1- )n) ∩ c(1- )n R=0 .
Since r((1- )n) is a non-zero essential right ideal of R, then c(1- )n=0 .
And hence (1-a)n=0, and then 1= , a contradiction .
Thus n=0 and therefore Y(R) is nilideal.
#
Theorem 2.9:
Let R be a ring with (r(
Then R is GVNL-ring
n+1
)) ⊆ r( n) and r( (1- )n)⊆ r((1- )n) for a∈R, if R/r( ) is GVNL-ring.
Proof:
Assume that R/r( ) be a GVNL-ring, then either +r( ) ∈R/r( ) or
(1- )+r( ) is π-regular element. Now, if +r( ) is π-regular element for ∈R then there exists
b+r( ) ∈R/r( ) and n∈Z+, such that
n
+ r( )=( +r( ))n
= ( +r( ))n (b+ r( ))( +r( ))n
= ( an +r( ))(b+ r( ))( n +r( ))
= n b n+ r( )
Then n - n b n ∈ r( ), that is
( n- n b n ) =0, hence n+1(1-b n) =0.Then (1-b n ) ∈ r( n+1)⊆ r( n), which implies that n(1-b
n
) =0, then we get n = n b n
Thus is π-regular element in R .
Now, if (1- )+r( ) ∈R/ r( ) is π-regular for ∈R, then, there exists c+r( ) ∈R/ r( ) and n∈Z+ such
that (1- ) n +r( )=((1- ) + r( ))n
148
=((1- ) + r( ))n (c+ r( )) ((1- )+ r( ))n= ((1- )n +r( ))( c+ r( ))((1- )n+ r( ))
=(1- )n c (1- )n+ r( ), then (1- )n - (1- )n c (1- )n ∈r( ), that is
((1- )n - (1- )n c (1- )n )=0, so (1- )n ( 1- c(1- )n)=0 then
(1-c (1- )n) ∈ r( (1- )n) ⊆ r((1- )n, hence (1- )n (1-c(1- )n)=0
Thus (1- )n - (1- )n c (1- )n=0. Hence (1- ) is π- regular element in R
Therefore R is GVNL –ring. #
Theorem 2.10:
If R is a reduced ring and every maximal ideal of R is a right annihilator.
Then R is GVNL-ring.
Proof:
Let a∈R, we shall prove that n R+r( )=R, if not there exists a right maximal ideal M containing n
R+ r( n). If M=r(b) for some 0≠b∈R, we have
b∈ℓ ( n R+r( n))⊆ ℓ( n)=r( n), which implies that b∈M=r(b), then b2=0,
and b=0 a contradiction. Therefore nR+ r( n)=R
In particular n c+d=1, c∈R, d∈r( n), then n c n= n which proves
is π-regular element. Now, if (1-a)∈ R, we shall prove that.
((1- )n +r((1- )n)=R , if not there exists a maximal right ideal M containing
(1- )n R+ r(1- )n , if M=r(c) for some 0≠c∈R, we have
c∈ℓ(1- )n R+r(1- )n ⊆ℓ (1- )n=r(1- )n. Which implies c∈M=r(r), then c2=0.
And hence c=0 a ctraduction. Therefore (1- )n R+ r(1- )n=R
Inparticular
(1- )n
+
=1
Where
∈R
and
∈r((1- )n)
Then,
n
n
)
(1 −
(1- ) +0 = (1- ) .
Thus (1- ) is π-regular element in R. Therefore R is GVNL-ring.
#
Now, we have the following result to obtain the relation between GVNL-ring and exchange ring .
Proposition 2.11:
If R is GVNL-ring, then eRe is also GVNL-ring for every idempotent element e in R .
Proof:
For ∈eRe, or (1-a) is π-regular in R. If is π-regular, then there exists b in R and n∈Z+ such
that n = n b n, so, n = ( n e) b (e n) = n (ebe) n. Thus is π-regular in eRe.
If (1- ) is π-regular, then there exists c∈R and n∈Z+ such that
(1a)n=(1- )nc(1- )n
.
Now ,(e-a)n = (e-ea)n = (e(1-a))n = e(1-a)ne .
Hence e-a is π-regular in eRe.
Therefore, eRe is a GVNL-ring . #
Now, we give the following proposition which is due to YING Zhi- ling in [8]
Proposition 2.12:
If R a GVNL-ring, then for every idempotent element e in R, either eRe or
regular ring .
Proof:
Let R be a GVNL-ring and e ∈R be an idempotent element in R. Then
(1 − )
R≅
(1 − )
(1 − ) (1 − )
If ∈eRe and ∈(1-e)R(1-e) are two non-π-regular elements. Then both
0
1−
0
a=
and 1- =
0 1−
0
are also non-π-regular, a contradiction.
From proposition 2.12 , clearly every GVNL-ring is an exchange ring .
(1-e) R(1-e) is a π-
#
149
The following corollary is given in [8]
Corollary 2.13:
For an abelian ring R, R is GVNL-ring if R is an exchange ring
Proof:
For ∈R, let 1= and 2=1- . Then
1 R+ 2R=R, then by [5,proposition 1-11] ,there exists an orthogonal idempotent
e1 and e2 such that e1∈ e2 R and e1+e2=1 .
Now, e1= 1 b1 and e2= 2b2 for b1, b2 ∈R, so e1 a1=(e1 a1) (e1 b1)(e1 a1)
and e2 a2=(e2 a2) (e2 b2)(e2 a2) is regular.
Since R is abelian, e1 a1 and e2 a2 is also strongly regular.
Thus we can suppose that
e1a1 b1=e1 b1 a1 and e2 a2 b2=e2 b2 a2, implying that, e1 a1=(e1 a1)2 (e1 b1) and e2a2=(e2a2)2 (e2 b2) . By
(proposition2.12) either e1 R=e1 R e1 and e2 R =e2 R e2 or
(1-e1)R(1-e1)and(1-e2)R(1-e2)areπregularife1Re1and e2 Re2 are π-regular, then R being a finite direct product of e1 Re1 and e2 Re2 is πregular. If (1-ej)R(1-ej) is π-regular for j=1, 2, then there exists m> 0 and ∈R for j=1, 2 such that (
(1-ej) aj )mCj((1-ej) aj)m = ((1-ej)aj)m = (1-ej) ajm),
ej a j= (ejaj)2 (ej bj), we have
[ejaj+(1-ej) aj]m [(ej bj)m +(1-ej) ] [ejaj+(1-ej)aj]m
= [ej aj + (1- ej) aj]m = ajm
That is a1, a2 is also π-regular. Therefore R is GVNL-ring.
#
Now, to give the relation between GVNL-ring and strongly commuting regular rings.
Theorem 2.14: [6]
Suppose that R is a strongly commuting regular ring. Then R is π-regular .
Proof:
By strongly commuting regularity of R, for each ∈R, there exists an element c in R such that
2
= 4 c 4 . With suppose c = 2 c 2. Then we have
2
= 2 ̅ 2 .Which implies that R is π –regular . #
We observe the following corollary .
Corollary 2.15:
I f R is a strongly commuting regular. Then R is GVNL-ring .
3- The GVNL – Ring without zero Divisor element .
In this section, we give some results about GVNL-ring without zero –divisors and some relation
with other rings like division ring, local ring, π-regular ring, simple ring, VNL-ring .
Proposition 3.1:
Let R be a GVNL-ring without zero divisors. Then every element a∉ 0, 1 in R is invert able .
Proof:
Let Rbe a GVNL-ring and 0≠ ∈ . ℎ ℎ (1 − ) −
. If
is π-regular element in R, then there exists b∈R and n∈Z+ Such that
n
= n b n, and then we have n - n b n=0
.
That is, n - n b n= n ( 1- b n)= ( n-1 (1-b n))=0 . Since R is without zero divisors then ,
n-1
( 1-b n)=0, thus ( n-2 (1-b n))=0 and hence … (1-b n)=0, then 1-b n=0 Thus 1=b n ,
which implies that 1=(b n-1) , that has left inverse.
Now, since 1=( b n-1) , then, = ( b n-1) and thus ( 1- b n-1) =0.
Hence (1- b n-1) ∈ ℓ ( ) = 0, which implies that 1= (b n-1),
Thus has right inverse and therefore is invertable element in R .
Now, if (1- ) is π-regular element in R, then there exists c∈R and n∈Z+ such that ,
(1- )n - (1- )n c (1- )n =0,that is (1- )n[1-c(1- )n]= (1- )n (1-a)n-1 [1-c(1-a)n]=0 .
150
Since R is without zero divisor and ∉ 0, 1 .
Thus (1- ) ((1- )n-2(1- c(1- )n)=0, and hence … (1- ) ( 1-c(1- )n)=0.
Then ( 1-c (1- )n)=0, which implies that, 1=c(1- )n
That is 1=( c (1- )n-1) (1- ). Hence (1-a) has left inverse .
Now, since 1=( c (1- )n-1) (1- ), then,
(1- )=(1- ) ( c(1- )n-1) (1- ), and then (1-(1- ) c (1- )n-1) (1- )=0 thus,
(1- (1- )c (1- )n-1)∈ ℓ (1- ) =0 which implies that 1=(1- ) (c (1- )n-1) .
And hence(1- )has right inverse.Therefore (1- )is invertible element in R. #
Corollary 3.2:
Let R be a GVNL-ring without zero-divisors. Then R is a division ring .
Corollary 3.3:
Let R be a GVNL-ring without zero divisors. Then R is;
1- Local ring. 2- VN-regular ring and reduced ring.
2- VN-regular ring and reduced ring .
3- π-regular ring .
Finally we give the following result .
Proposition 3.4:
If is a proper ideal of GVNL-ring R. Then every element of
every GVNL-ring has non-zero divisors is simple.
is zero divisors in R. Especially
Proof:
Let
0≠
∈
,
since
R
is
GVNL-ring,
then
either
or
(1- )
is
π-regular element in R. If is π-regular element, then there exists an element b∈ R and a positive
n
(1-b n)=0 gives (1-b
integer n such that n= n b n , if. a is not zero divisors then
n
)=0, and hence 1∈ , a contradiction.
Hence, is zero divisors . Now, if (1- ) is π-regular element, then there exists c∈R and a positive
(1- )n (1- c (1- )n)
integer n such that (1- )n =(1- )n c (1- )n . If (1- ) is not zero divisor, then
n
=0 gives, 1-c(1-a) =0 thus
1=c ( 1- )n ∈ a contradiction.
If R has nonzero divisors, there is not a proper ideal of R. Hence R is simple. #
REFERECES
1-Burton, D.m.(1970), " A first Course in Rings and Ideals, " Addison Wesley Pulishing Company.
2-CHEN W.X., TONG W.T. (2006), " On non-commutative VNL-rings and GVNL-rings
"(J).Glasgow Math J, 48:11-17 .
3-CONTESSA M. (1984), " On certain classes of pm-rings"(J).Comm. Algebra, , 12:1447-1469.
4-Mc Coy, N.H.(1939)," Ggeneralized regular rings", Bull.Amer.Math.Soc.Vol.45, pp.175-178.
5- NICHOLSON.W.K. (1977), " Llifting idempotents and exchange rings". Transacati -ons of the
American Math. Society, vol.229, pp.269-278.
Amer. Math.Soc., 229:269-278 .
6-Sh.A.Safarisabet and Marzieh. Farmani , ( 2013), " Strongly commuting regular rings " J.Basic
.Appl.Sci. Res.3(1) vol 3(1), pp.707-713 .
7-Von Neumman, J.(1936), " On regular rings". Proc. Nat. Scad. Sciences U.S.A, Vol, 22, pp.707-713.
8-YING Zhi-ling, (2011), " Some Charactrizations of GVNL_Rings", Journal of Nanjing University of
Posts and Telecommunications (Natural Science), Vol.31 No.5.
151
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152
ISSN: 2410-7549
SURFACE ROUGHNESS EFFECT ON DISCHARGE COEFFICIENT OF
COMBINED CYLINDRICAL WEIR GATE STRUCTURE
1
Shaker A. Jalil1 and Safa S. Ibrahim2
Assistant Professor, Water Recourses Engineering Department, Faculty of Engineering, University of Duhok, Iraq
2
Assistant Lecture, Petroleum Engineering Department, Faculty of Engineering, University of Zakho, Iraq
ABSTRACT:
The aim of this research is to investigate the effect of surface roughness on the performance of weir
and gate. An experimental study in a laboratory flume is carried out to study flow over and under
cylindrical weir gate in combined structure as flow measurement device. Four models having different
diameters were tested in a laboratory flume. In each model, the surface was roughed four times. The
results of the test show logical negative effect of the increase of surface roughness on the performance. The
performance of the combined structure improved with decrease ratio of roughness to the upstream head
(Ks/H) and with the increase of the total head to the diameter of the weir (H/d). Empirical relations were
obtained to estimate the variation of discharge coefficient (Cd) in terms of some dimensionless parameters.
Within the limitations of the present experimental work an equation to predict the discharge is proposed
with R2 of 0.936. Finely the contribution of the gate increases relative to the weir when the surface
roughness increases.
KEYWORDS: Combined discharge measuring structure, cylindrical weir, gate, surface roughness, discharge
coefficient.
1. INTRODUCTION
T
he most common and important water
measurement structures that used for
controlling, adjusting the flow in irrigation
channel and diverting the flow from a main
channel to a secondary channel are weirs and
gates. One of the weirs demerits is they need to
be cleaned of sediment and trash periodically.
Sluice gates are used extensively for flow
control and water measurement for long time.
One disadvantage of the gates is they retained
the floating materials. In order to maximize their
advantages, weirs and gates can be combined
together in one device, so that water could pass
over the weir and below the gate simultaneously.
One of the combined weir-gate structures is
cylindrical weir- gate structure. Regarding the
form of the combined cylindrical weir-gate
structures, it has some advantages including easy
design by using commercial pipes, sediments
and floating materials flow, high flow discharge
coefficient than other replaceable structures and
its being economic and combination of those
structures in one device can minimize the
disadvantages of separate use of each device.
The performance of the combination has been
studied by many investigations, Negm et al.
(2002) presented the effects of hydraulic and
geometrical parameters, viscosity and surface
tension in the combined flow over rectangular
weirs and rectangular gates with sharp crested.
They found that the trend of variation in sloping
bed cases was similar to that horizontal bed.
Hayawi et al. (2008) studied the characteristics
of free flow through the combined triangular
weir and rectangular gate. They found that the
values of theoretical discharge was inversely
proportional to the geometrical dimensionless
parameters and directly to distance between the
bottom of the weir and the upper edge of the
gate. Hayawi et al. (2009) investigated the
coefficient of discharge (Cd) for a combined
rectangular weir with semi-circular gate. They
found that the average value of (Cd) was equal to
0.695. Also, obtained that the value of (Cd)
increase with the increases of the distance
between the weir and gate opening with average
value of (Cd) 0.74. Jalil and Sarhan (2013)
studied the flow over a sharp crested weir and
under gate in combined oblique structure as flow
measurement structure. They found that the
value of
range from 0.403 to 0.623 with
different effect parameters. Masoudian et al.
(2013) experimentally studied the effects of
canal size on discharge coefficients of
cylindrical weir-gate by used two flumes. They
found that the value of ( ) in large canals were
(0.75 to 1.05) more than in small canals (
0.55 to 0.9) which can be as a result of
surpassing effects of walls, canal size, surface
tension and viscosity on discharge coefficient in
153
small canals since the ratio of boundary layer
thickness to the canal width in small canals was
considerably more than the ratio in large canals
and whereas, the amount of flow velocity in
boundary layer was low, led to reduce discharge
coefficient. Severi et al. (2013) performed the
effect of vertical movement of cylindrical weirgate on free flow hydraulic. Experimentally
resulted that the gate opening changes has
inverse effect related to discharge coefficient, so
that the maximum and minimum discharge
coefficient were visible in cylindrical weir and
cylindrical gate, respectively. Furthermore, in a
constant diameter and constant discharge,
discharge coefficient changes had an increasing
process by the decreases of the gate opening
height. Besides, in a constant discharge and gate
opening height, discharge coefficient decreases
with increase in the structure diameter. Khassaf
et al. (2013) investigated the coefficient of
discharge and characteristic of free flow for a
combined weir (have two rectangular notches
with trapezoidal notch between them) and
semicircular sluice gate. The results show that
the values of (Cd) range from (0.358 to 0.426)
with an average value of (0.392). On the other
side the effect of surface roughness are studied
by numerous research on different hydraulic
structure, such as, Othman et al. (2011) and
Ghobadian et al (2013) studied the effect of
size and surface roughness of cylindrical weir.
Mohammed et al. (2011) studied the effect of
bed roughness of free overfall in a rectangular
channel with different bed slope. Jalil et al.
(2014) investigate the effects of surface
roughness sizes on the discharge coefficient for
broad crested weirs.
Previous studied sources the discharge
coefficient of combined weir-gate structure
usually expressed for a smooth case. Therefore,
the main objective of this investigation is to
study the effects of different diameters and
surface
roughness
on
the
hydraulic
characteristics of the combined cylindrical weir
and gate structure.
2. THEORETICAL BACKGROUND
The conservation principles of the energy and
continuity can lead to the theoretical base of the
free flow over weir and under the gate as two
parts of the hydraulic flow measurements
structure. Theoretically if no energy lost, a wellknown equations for evaluating the discharge
over the weir and under the gate can be
presented as in Equations (1 and 2) respectively,
the overall discharge of the structure is the
addition of the two equations which presented in
Equation 3. Figure 1 shows the definition sketch
for the flow with the geometric parameters.
Figure 1. Combined weir gate structure
Where: Qth = discharge passing over the weir and under gate (L3 /T),
154
Qw= discharge passing over the weir (L3 /T),
Qg= discharge passing under gate (L3 /T),
H = upstream head (L),
d= diameter of the pipe (L),
h= head depth of water over the weir (L),
a= gate opening (L),
B = weir and gate length (L), and
g = acceleration due to gravity (L/T2).
The proportion of the actual to the theoretical values of discharge is well known as coefficient of
discharge (Cd), then the actual discharge of total structure can be written as in Equation 4.
Where: Cd = coefficient of discharge.
The actual value of discharge is affected by all the factors imposed in the flow phenomena. This
coefficient simulates the effect of all factors entering the physical process of flow. Based on Equation
4 and used the dimensionless parameters introduced by earlier studies, the following wide function
relationship can be written as in Equation 5.
Where: Ks= roughness size (L),
Re = Reynolds number, and
We = Weber number.
The values of Reynolds number and Weber number are not affected due to turbulent flow and
neglecting surface tension. Equation 5 can be written as
3. EXPERIMENTAL WORK
The models of study were made from plastic pipes have four different diameters (d = 4, 6.3, 9, and
11 cm). Each model was roughed four, three models were roughed by uniform aggregates of size 2.36,
3.35, and 4.75 mm, and the fourth model left as it is a smooth plastic, see Figure 2. The experimental
investigation was carried out in a horizontal flume of working length 2.4 m, having a rectangular cross
section of 0.25m height and 0.075m width. The depth of flow was measured, using point gauge with
venier scale reading to 0.1 mm, at center line of flume. The flow rate was measured by a volumetric
tank. Details of the experimental program are shown in Table 1.
Figure 2. Cylindrical weir-gate structure model
155
Table 1. Details of the cylindrical weir-gate structure models tested during the experimental program
Models
Diameterd (m)
1
0.04
2
0.063
3
0.09
4
0.11
State of Surface Ks (mm)
Smooth
2.36
3.35
4.75
Smooth
2.36
3.35
4.75
Smooth
2.36
3.35
4.74
Smooth
2.36
3.35
4.75
4. RESULTS AND DISCUSSIONN
The data collected from the tests of the four structural diameter sand two roughness are presented
in Figure 3, which shows the comparison between the smooth surface of the cylindrical combined
structure and the rough surface of Ks=4.35 mm. It is clear that the total discharge flowing over weir
and under the gate is increase with the total head. The effect of surface roughness is also clear, it can
be notice that for a certain total head and pipe diameter the total discharge is more for the smooth, so
the head increase with increase of surface roughness (Ks) for a certain value of discharge. This
increase in head caused by the head loss due to increase of friction force.
0.0025
Smooth
Kr=4.35
0.0020
d=0.040 m
d=0.040 m
)s
/ 0.0015
(m
cta
0.0010
Q
d=0.063 m
d=0.063 m
d=0.090 m
d=0.090 m
d=0.110 m
d=0.110 m
3
0.0005
0.0000
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
H (m)
Figure 3. Variation of the discharge with total head for different pipe diameter and roughness
The experimental data is presented in Figure 4, which shows the effect of structure diameter on the
discharge for all surface roughness heights (Ks). It can be pointed that there is a slight decrease in
discharge value with the increase in surface roughness for large pipe diameter, while the effect of
surface roughness seen to be more effective for the small pipe diameter.
156
0.0025
d=0.04 m
0.0020
)s
/ 0.0015
3
(m
tc
a 0.0010
Q
d= 0.11 m
Smooth
smooth
Ks=2.35 mm
Ks=2.35 mm
Ks=3.35 mm
Ks=3.35 mm
Ks=4.75 mm
Ks=4.75 mm
0.0005
0.0000
0
0.02
0.04
0.06
0.08
0.1
0.12
0.14
0.16
0.18
H (m)
Figure 4. Variation of the discharge with total head for different roughness
The calculated value of discharge coefficient (Cd) in Equation 4 was studied with the dimensionless
parameters in Equation 5. It is clear that the value of (Cd) decreases with the increase of roughness
(Ks) for a fixed value of (h/H) and increases with the increase of the diameter of the cylindrical
combined structure as shown in Figure 5. It can be also noticed that for a fixed value of surface
roughness the rate of changes in value of (Cd ) depends on the value of the diameter.
0.80
0.90
0.70
0.80
0.70
0.60
0.60
0.50
d=0.09 m
d 0.40
C
d
C
Smooth
0.30
0.20
0.10
Smooth
0.40
Ke=2.36 mm
0.30
Ks=3.35 mm
0.20
Ks=4.75 mm
0.10
0.00
d=0.11 m
0.50
Ks=2.36 mm
Ks=3.35 mm
Ks=4.75 mm
0.00
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0
0.05
0.1
0.15
h/H
0.2
0.25
0.3
h/H
Figure 5. Variation of the discharge coefficient and h/H for different Ks
In addition, the value of the coefficient of discharge increases with the increase of (H/d), the rate of
change in value is also depends on the diameter, the negative effect of the surface roughness on the
performance of the structure by reducing the value of (Cd) is also can noted in Figure 6.
0.90
0.90
0.80
0.80
0.70
0.70
0.60
d
C
0.60
0.50
d=0.04 m
0.40
Smooth
0.30
d
C
0.50
d=0.063 m
0.40
Smooth
Ks=2.36 mm
0.30
Ks=2.36 mm
0.20
Ks=3.35 mm
0.20
Ks=3.35 mm
0.10
Ks=4.75 mm
0.10
Ks=4.75 mm
0.00
0.00
1
1.2
1.4
1.6
1.8
2
2.2
2.4
2.6
1
H/d
1.2
1.4
1.6
1.8
2
H/d
Figure 6. Variation of the discharge coefficient and H/d for different Ks
157
The collected experimental data of sixteen models and the geometric parameters are used to
calculate the dimensionless parameters in Equation 6. The calculated discharge coefficient (Cd) from
Equation 4 varies in the range from 0.4616 to 0.7876, with Standard Error 0.0067091 and Standard
Deviation 0.0867006,as shown in Table 2 with other parameters which they have highest significant
correlation with the independent variable (Cd). The highest negetive Pearson Correlationis Factor
0.913 between the (Cd) and (Ks/a) at the 0.01 level (2-tailed).
Table 2. Descriptive analysis of the calculated coefficient of discharge
The facilities built in the features of SPSS 17(statistical software package) are employed to get
statistical analysis and mathematical relations between the variables. The regression has been used to
find correlation models between the dependent (Cd) variable and the independent parameters in
Equation 6. The models studied include 18 mathematical relations of three type’s models, linear,
square and power. The best and simplest forms of equations were the linear models; they have the
highest coefficient of determination reaches to 0.940. Nonlinear Regression shows that the square and
power models haven’t very high value of R2 as the linear models, the highest power equation has R2 is
equal to 0.731. To show some of the models, 9 models have been chosen Table 3
Table 3. The regression models analysis
No.
1
2
3
4
Ks
h
h
H
h
d
Ks
Ks
 0.269  2.144  0.499  1.210  0.031  2.676
 5.173
a
a
B
d
H
h
d
H
Ks
h
H
Ks
d
h
Ks
Ks
Cd  0.228  0.957
 0.983  0.414  2.405
 0.026  0.094  4.709
 0.263
a
H
d
H
h
B
B
h
Ks
h
H
Ks
d
h
Cd  0.216  0.297
 1.073  0.428  2.522
 0.021  0.150
a
H
d
H
h
B
Ks
h
H
Ks
Cd  0.396  0.149
 1.069  0.385  3.661
a
H
d
H
0. 604
 Ks 
Cd  0.074

 a 
6
 Ks 
Cd  0.076

 a 
 Ks 
Cd  0.520

 a 
7
 Ks 


 B 
0. 551
0 .042
0 .967
 Ks 


 B 
 Ks 


H 
0 .959
0.087
h
 
B
 0.095
 Ks 


 d 
 Ks 


 d 
0.393
H 
 
a 
2
2
2
2
0.104
2
0 .40
H 
 
d 
 Ks 


H 
0. 162
H 
 
a
 Ks H Ks d 
Cd  0.524
 
 
 a d d a
8
158
0.940
Cd  0.134  1.075
5
9
R2
Equation
d 
 
a
0 .153
0.935
0.928
0.912
0 .349
0.731
0. 448
d 
 
a
0.710
0 .040
0.686
0.029
0.671
2
2
2
 Ks 
 Ks 
h
H 
 Ks 
 h
a
d
d 
Cd  -3.560 
  123 .06
  0.081   0.229    61 .693
  1.346   0.05   0 .003   0.1 
 a 
 B 
B
d 
H 
H
h
h
a
2
0.418
The first four equations in the table have R2 more than 0.9 and have the simplest form. These
equations have been found by stepwise linear regression of the dependent variable (Cd) with all twelve
independent dimensionless variables in Equation 5. The first linear equation in the table can be
sugested to calculate the discharge coeffecient for rough combined hydralulic structures, which based
on stepwise method of including independent variables in model to determine statistical significance
predictors of the higher correlation in the matrix at confidence level of 95%. After including seven
independent variables the Adjusted R square is equal to 0.937 and standard Error of estimate equal to
0.0364, this relation is presented in Equation 7.
Cd  0.134  1 .075
Ks
h
h
H
h
d
Ks
Ks
 0.269  2.144  0 .499 1 .210  0. 031  2.676
 5 .173
a
a
B
d
H
h
d
H
(7)
The statistical analysis output details for the proposed Equation 7 are shown in Table (4 to 6).
Table 4. Stepwise regression analysis
Model Summaryk
Change Statistics
Model
10
R
.970
j
Adjusted R
R Square
Square
.940
.937
Std. Error of the
Estimate
.02383332376
R Square
Change
F Change
.002
df1
6.098
Sig. F
Change
df2
1
158
.015
DurbinWatson
1.014
j. Predictors: (Constant), ks/a, h/a, h/B, H/d, h/H, d/h, ks/d, ks/H
k. Dependent Variable: cd
Table 5. Stepwise regression analysis
ANOVA a
Sum of
Squares
Model
10
Regression
Residual
Total
Mean
Square
df
1.417
8
.177
.090
158
.001
1.507
166
F
Sig.
311.835
.000
j
j. Predicto rs: (Constant), ks/a, h/a, h/B, H/d, h/H, d/h, ks/d, ks/H
k. Depende nt Variable: cd
Table 6. Stepwise regression coefficients
Coefficientsa
Stand ardized
Coeffi cients
Unstandardized
Coefficients
Model
10
B
(Constant)
Std. Error
.134
.057
ks/a
-1.075
.159
h/a
-.269
.047
h/B
2.144
.358
H/d
.499
.032
h/H
-1.210
.086
d/h
.031
.005
Beta
t
2.345
Sig.
.020
-1.625
-6.767
-2.142
-5.671
2.592
95.0% Confi dence
Interval for B
Lo wer
Bound
Upper Bound
.021
.246
.000
-1.389
-.761
.000
-.363
-.175
5.994
.000
1.438
2.851
1.419
15.672
.000
.436
.562
-1.105
-14.065
.000
-1.379
-1.040
.409
6.792
.000
.022
.041
ks/d
-2.676
.793
-.944
-3.373
.001
-4.242
-1.109
ks/H
5.173
2.095
.939
2.469
.015
1.035
9.310
a. Dependent Variable: cd
The sugested coefficient of discharge in the formulations above based on the assumtion that the
same discharge coefficient for the weir and the gate, but the flow in this combined structure is
composed of two parts, each of them performs on its phenomena of flow. Within the limetation of this
experimental work , the contribution percent of weir and gate in the tatal discharge can be found by
linear regression analysis. The model of the relation is pesented in Equation 8.
159
Qact  F (C g  Qth .
Where:
gate
 C w  QTh . weir )
(8 )
= percent of theoretical flow under gate,
= percent of theoretical flow over weir and
F = interaction factor which is a function of structure geometry. The model summary and ANOVA of
regression analysis for the experimatal measrements are shown in Table 7, while the coefficients in
Table 8.
Table 7. The regression models analysis for Equation 8
Model Summaryd,e
R
Model
1
R Square
.976
2
Adjusted R
Square
b
Std. Error of the
Estimate
a
.953
.953
.22416
c
.978
.977
.15492
.989
a. Predictors: H/d
b. For regression through the ori gin (the no-intercept model), R
Square
measures
c. Predictors:
H/d, the
ks/Hproportion of the variability in the dep endent
d. Dependent Variable: F
e. Linear Regression through the Origin
Model Summaryd,e
R
Model
1
R Square
.976
2
Adjusted R
Square
b
Std. Error of the
Estimate
a
.953
.953
.22416
c
.978
.977
.15492
.989
a. Predicto rs: H/d
b. For regression through the origin (the no-intercept model), R
Square
measures
c. Predictors:
H/d, the
ks/Hproportion o f the variability in the dep endent
d. Dependent Variable: F
e. Linear Reg ression through the Ori gin
ANOVAa ,b
Sum of
Squares
Model
1
Regression
Residual
Total
2
Regression
Residual
Total
Mean
Square
df
169.616
1
169.616
.050
8.341
166
177.957
173.997
d
167
2
86.999
3.960
165
.024
d
167
177.957
F
Sig .
c
3375.575
.000
3624.884
.000
a. Dependent Variable: F
b. Linear Reg ression through the Ori gin
c. Predictors: H/d
d. This total sum of squares is not corrected for the constant because the constant is zero
e. Predicto rs: H/d, ks/H
160
e
Table 8. Coefficients for models in Equation 8
Coefficientsa,b
Unstandardized
Coefficients
B
Std. Error
Standa rdized
Coefficients
Beta
95.0% Confidence
Inte rval for B
Lower
Upper
Bound
Bound
Model
1
H/d
.608
.010
.976
t
58.100
Sig.
.000
.587
.62 8
2
H/d
.754
.013
1.211
58.011
.000
.728
.77 9
-10.004
.740
-.282
-13.511
.000
-11.465
-8.54 2
ks/H
a. Dependent Variable: F
b. Linear Regression through the Ori gin
The obove analysis shows that contribution percent of the theoratical flow is different. The relation
which give an idea on the contribution percent is presented in Equation 9 and 10. It can be also notice
that contribution of the gate increase with the increase of roughnessas shown in Figure 7.
Qact  F ( 0.677Qth. gate  0.498QTh. weir )
F  0.754
H
Ks
 10.004
d
H
(9)
(10)
Figure 7. Relation between weir and gate discharge for different roughness’s
161
5. CONCLUSION
The effect of surface roughness on the
performance of flow over rounded weir and
under gate in combiened structure was studied
experimantaly. From the statistical analysis of
experimental data the following conclusions may
fixed with in the experimental limetation:
1. The performance of the combined structure
affected negativity with increases of surface
roughness, Cd decreases with increase of Ks/H
2. The value of Cd increases with the increase of
H/d for constant (Ks) and decreases with the
increase of h/H
3. The value of Cd range from 0.4616 to 0.7876.
4. Within the limitations of the present
experimental work a discharge prediction
Equation 7 is developed with mean percent error
of 0.036 %.
5. The contribution of the gate increases with
increase of roughness, percent contribution is
49.8% for the weir while for the gate 67.7%
from its theoretical discharges.
6. REFERENCES
Ghobadian, R., Chaghabagi, F., Farmanifard, M., and
Ahmadi, A. (2013) .Effect of Crest Roughness
on Flow Characteristics over Circular Weirs.
Civil Engineering Infrastructures Journal,
Vol. (46), No. (2).
Hayawi, H. A. M., Hayawi, G. A. M., and Alniami,
A. A. G. (2009). Coefficient of Discharge for a
Combined Hydraulic Measuring Device. AlRafidain Engineering, Vol. (17), No. (6).
Hayawi, H. A., Yahia, A.A., and Hayawi, Gh. A.
(2008). Free Combined Flow over a
Triangular Weir and under Rectangular Gate.
Damascus University Journal, Vol. (24), No.
(1).
162
Jalil, Sh. A., and Sarhan, S. A. (2013). Experimental
Study of Combined Oblique Weir and Gate
Structure. ARPN Journal of Engineering and
Applied Sciences, Vol. (8), No. (4).
Jalil, Sh. A., Ibrahim, S. S. and Jafer, R. A. (2014).
Surface Roughness Effects on Discharge
Coefficient of Broad Crested Weir. Research
Journal of Applied Sciences, Engineering and
Technology, Vol. (7), No. (24), Pp. 52275233.
Khassaf, S.I., Yost, S. A., and Abbas, H. A. (2013).
Coefficient of Discharge for a Compound
Weir Combined with Semicircular Gate.
International Journal of Advance Research,
Vol. (1), No. (9).
Masoudian, M., Fendreski, R., and Gharahgezlou, M.
(2013). The Effects of laboratory Canal Size
and Cylindrical Weir-Gate Diameter on
Discharge
Coefficient.
Tech
Journal
Engineering and Applied Sciences, Vol. (3),
No. (15), Pp. 1630-1634.
Mohammed, M. Y., Taee, A. Y., and Al-Talib, A. N.,
(2011). Gravel Roughness and Channel Slope
Effects on Rectangular Free Overfall.
Damascus University Journal, Vol. (17), N0.
(1).
Negm, A. M., AL-Brahim, A. M., and Alhamid, A.
A. (2002). Combined-Free Flow over Weirs
and below Gates. Journal of Hydraulic
Research, Vol. (40), No. (3).
Othman, k. I., Chimeran, T. A., and Al-Hafith, I. A.
I., (2011). Effect of Size and Surface
Roughness of Cylindrical Weirs on over Flow
Characteristics. Al-Rafidain Engineering, Vol.
(19), No. (2).
Severi, A., Masoudian, M., Kordi, E., Roettcher, K.
(2013). Experimental Investigation on
Discharge Coefficient Changes of Moveable
Cylindrical Weir-Gate. Khavaran Institute of
Higher Education.

.   
              


              
            (H/d)   (Ks/H

    0.936R 2 

:‫اﻟﺨﻼﺻﺔ‬
‫اﻟﮭﺪف ﻣﻦ ھﺬا اﻟﺒﺤﺚ ھﻮ دراﺳﺔ ﺗﺄﺛﯿﺮ ﺧﺸﻮﻧﺔ اﻟﺴﻄﺢ ﻋﻠﻰ أداء اﻟﻤﻨﺸﺄت اﻟﻤﺘﻜﻮﻧﺔ ﻣﻦ اﻟﮭﺪار‬
‫ إﺟﺮﯾﺖ دراﺳﺔ ﺗﺠﺮﯾﺒﯿﺔ ﻓﻲ ﻗﻨﺎة اﻟﻤﺨﺘﺒﺮﯾﺔ ﻋﻠﻰ ﺟﺮﯾﺎن ﻓﻮق ھﺪار داﺋﺮي وﺗﺤﺘﮭﺎ ﻓﺘﺤﺔ‬. ‫واﻟﺒﻮاﺑﺔ‬
‫ ﺗﻢ اﺧﺘﺒﺎر أرﺑﻌﺔ ﻧﻤﺎذج ﻣﺨﺘﻠﻔﺔ‬.‫ﺑﺒﻮاﺑﺔ داﺋﺮﯾﺔ اﻟﺸﻜﻞ ﻓﻲ ﻣﻨﺸﺄ ﻣﺸﺘﺮك ﯾﻌﻤﻞ ﻛﺠﮭﺎز ﻟﻘﯿﺎس اﻟﺘﺼﺮﯾﻒ‬
‫ﺑﯿﻨﺖ اﻟﻨﺘﺎﺋﺞ اﺧﺘﺒﺎر ﺗﺄﺛﯿﺮ‬. ‫ وﻟﻜﻞ ﻧﻤﻮذج ﺗﻤﺖ ﺗﺨﺸﯿﻦ اﻟﺴﻄﺤﺔ أرﺑﻊ ﻣﺮات‬،‫أﻗﻄﺎر ﻓﻲ ﻗﻨﺎة ﻣﺨﺘﺒﺮﯾﺔ‬
‫اﺛﺒﺖ ان أداء اﻟﻤﻨﺸﺄ ﯾﺘﺤﺴﻦ ﺑﺎﻧﺨﻔﺎض ﻗﯿﻤﺔ اﻟﻨﺴﺒﺔ‬.‫ﺳﻠﺒﻲ ﻣﻨﻄﻘﻲ ﻟﺰﯾﺎدة ﺧﺸﻮﻧﺔ اﻟﺴﻄﺢ ﻋﻠﻰ اﻷداء‬
‫( ﻣﻊ زﯾﺎدة ﻗﯿﻤﺔ اﻟﻨﺴﺒﺔ‬Ks/H) ‫اﻟﺘﻲ ﯾﻤﺜﻠﮭﺎ ﺧﺸﻮﻧﺔ اﻟﺴﻄﺢ اﻟﻰ اﻟﺤﻤﻮﻟﺔ اﻟﻜﻠﯿﺔ ﻓﻲ ﻣﻘﺪﻣﺔ اﻟﻤﻨﺸﺄ‬
‫ﺗﻢ اﻟﺤﺼﻮل ﻋﻠﻰ ﻋﻼﻗﺎت ﺗﺠﺮﯾﺒﯿﺔ ﻟﺘﻘﺪﯾﺮ‬. (H/d) ‫ﺑﯿﻦ اﻟﺤﻤﻮﻟﺔ اﻟﻜﻠﯿﺔ ﻣﻘﺪم اﻟﻤﻨﺸﺄ اﻟﻰ اﻟﻘﻄﺮ اﻟﻤﻨﺸﺄ‬
‫ ﻓﻲ ﺣﺪود اﻟﻌﻤﻞ اﻟﺘﺠﺮﯾﺒﻲ اﻟﺤﺎﻟﻲ‬.‫ ﻣﻊ ﻣﻌﺎﯾﯿﺮ ﻋﺪﯾﻤﺔ اﻟﻮاﺣﺪات‬Cd ‫ﺗﺒﺎﯾﻦ ﻓﻲ ﻗﯿﻤﺔ ﻣﻌﺎﻣﻞ اﻟﺘﺼﺮﯾﻒ‬
‫ ﻛﻤﺎ ﺗﻢ ﺗﺸﺨﯿﺺ‬،0.936 = R2 ‫ﺗﻢ اﻗﺘﺮاح ﻣﻌﺎدﻟﺔ ﻟﻠﺘﻨﺒﺆ ﻋﻦ ﻗﯿﻤﺔ ﻣﻌﺎﻣﻞ اﻟﺘﺼﺮﯾﻒ ذا ﻣﻌﺎﻣﻞ اﻟﺘﺤﺪﯾﺪ‬
.‫زﯾﺎدة ﻣﺴﺎھﻤﺔ ﺑﻮاﺑﺔ ﻓﻲ اﻟﺘﺼﺮﯾﻒ ﻣﻘﺎرﻧﺔ ﺑﺎﻟﮭﺪار ﻋﻨﺪ زﯾﺎدة ﺧﺸﻮﻧﺔ اﻟﺴﻄﺢ‬
163
‫طوظارا زانكؤيــا زاخــؤ‬
‫أ ‪ -‬زانست‬
‫ثةربةندا (اجمللد) ‪3‬‬
‫هذمارا (العدد) ‪1‬‬
‫خزيران‬
‫حزيران‬
‫‪2015‬‬
‫َ‬
‫عرياقي‬
‫َ‬
‫هةريا كوردستانا‬
‫وةزارةتا خويندنا باال‬
‫وتو َيذينةوةي زانسيت‬
‫زانكــؤيــا زاخــــؤ‬
‫‪ISSN: 2410-7549‬‬
‫اقليم كوردستان العراق‬
‫وزارة التعليم العالي‬
‫والبحث العلمي‬
‫جامعــة زاخـــــو‬
‫طــــوظـــــارا‬
‫زانكؤيــا زاخــؤ‬
‫أ ‪ -‬زانست‬
‫مـجــلـــة‬
‫جامعة زاخــو‬
‫أ ‪ -‬علوم‬
‫ثةربةندا (اجمللد) ‪3‬‬
‫هذمارا (العدد) ‪1‬‬
‫خزيران‬
‫حزيران‬
‫‪2015‬‬

Issue 1 - The University of Zakho

Transcription

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