D2.1 Common protocols and reference standards for

Transcription

D2.1 – Common protocols and reference standards for selected traits and species
Project no. 284181
TREES4FUTURE - Designing Trees for the Future
Start date of project: 1 November 2011
Duration of project: 4 years
Call: FP7-INFRASTRUCTURES-2011-1
Theme: INFRA-2011.1.1.13 [Research infrastructures for forestry research]
Funding Scheme: Combination of CP & CSA
D2.1 – Common protocols and reference standards for
selected traits and species
Description: Common protocols and reference standards for selected traits and species: Technical
cards are drawn up holding common protocols and reference standards at a species or at a trait level,
as appropriate, and will be made publicly available to the scientific community.
Due date of deliverable: M42
Actual submission date: M42
Organisation name of lead contractor for this deliverable: vTI (now THÜNEN)
WP2 Leader: Bart de Cuyper (VLAGEW)
Contributor(s): INRA (J-C Bastien, L Paques, A Dowkiw, A Ducousso, F Santi), ALTERRA
(SMG de Vries, J Buiteveld), CIDTG (M Touza Vazquez), CRA (F Ducci, MC Monteverdi,
R Proietti), FCBA (L Harvengt, D Micheau), IBL (J Kowalczyk, M Rzonca), ICAS (EN Chesnoiu),
INIA (R Alia, E Notivol), Metla (now LUKE) (E. Beuker), VLAGEW (B de Cuyper, M
Steenackers), vTI (now THÜNEN) (M Liesebach, V Schneck)
Dissemination level
PU Public (must be available on the website)
PP Restricted to other programme participants (including the Commission Services)
RE Restricted to a group specified by the consortium (including the Commission Services)
CO Confidential, only for members of the consortium (including the Commission Services)
X
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Table of contents
1. Summary
4
2. Context
5
3. Species oriented common protocols
3.1 Broadleaves
3.11 Beech specific protocols
3.111 Flushing
3.12 Oaks specific protocols
3.121 Flushing
3.13 Ash specific protocols
3.131 Flushing
3.14 Poplars specific protocols
3.141 Phenology
3.142 Melampsora laric-populina
3.143 Marsonia brunea
3.144 Xanthomonas
3.15 Wild cherry specific protocols
3.151 Phenology
3.152 Wood color
3.16 Broadleaves non-species specific protocols
3.161 Straightness
3.162 Forking
3.163 Branch habit
3.2 Conifers
3.21 Norway spruce protocols
3.211 Flushing
3.22 Douglas-fir specific protocols
3.221 Phenology
3.222 Frost hardiness
3.223 Needle cast
3.23 Scots pine specific protocols
3.231 Lophodermium needle cast
3.232 Pine twisting rust
3.24 Mediterranean pines
3.241 Drought
3.242 Flowering
3.243 Polycyclim
3.244 Bark thickness
3.25 Larch specific protocols
3.251 Phenology
3.252 Canker
3.253 “Drought” cracks
3.26 Conifers non-species specific protocols
3.261 Stem straightness
3.262 Basal sweep
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3.263 Forking
3.264 Branching habit
4. Traits oriented common protocols
4.1 Wood quality
4.11 General overview if available techniques and equipment
4.12 Acoustic tools
4.13 Near Infrared Spectroscopy (NIR)
4.14 Innovation and future trends
4.2 Drought resistance
4.21 Methodology and Evaluation System
4.22 Preliminary results
4.23 Conclusions
5. Reference genotypes
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1. Summary
Systematic provenance and progeny trials have been established for more than 100 years.
Many measurements and assessments were carried out within these trials. Standards were
defined for the traditional measurements like height and diameter (dbh). Numerous scoring
schemes were developed for other traits. This makes it difficult or not feasible to compare
results between institutions, but also between test series and single trials. Within the FP7
project TREES4FUTURE, this topic is addressed in “Work package 2 – Creating common
standards and protocols”.
The deliverable of “Task 2.1 – Development of common protocols and selection of reference
genotypes” presents species specific, and non-species specific common protocols for five
broadleaves (species or species groups), and five conifers (species and species groups),
respectively. The status and development of the assessment of wood characteristics and
drought are given. Furthermore, the deliverable presents a concept and a selection of
reference genotypes.
Key words: assessment, phenology, stem form, disease, wood quality, drought, reference
genotype
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2. Context
Many traits are of interest to forest tree breeders to distinguish genetic units, and to
describe genetic variation and the evaluation of genotypes along their breeding programs. A
lot of scoring systems have been developed by all tree breeders. This makes results between
institutions but also between test series and single trials difficult or unfeasible to compare.
Common protocols were used for a limited number of trial series. However, this is mostly
infrequent.
Criteria for common assessments methods are: acceptable, simple, easy to handle (fast),
low-cost, defined for a specific part in the life cycle.
The starting point for Task 2.1 was an inventory of existing national assessment schemes.
The inventory could be assembled from the results of the FP6 project TREEBREEDEX (PÂQUES
2009, DUCCI et al. 2012). The inventory was essential because the tree species beech and
oaks were not included in TREEBREEDEX. Further protocols drawn up from the literature
were added. During the workshops a selection was taken and common protocols identified.
The task involves a large number of participants, each with their own species of interest,
leading to a high number of species x traits combinations. Moreover, common protocols are
usually not transferable from one species to another. Not all species x traits combinations
can be tackled within the time span of the project, so priorities will have to be set regarding
the species as well as regarding the traits to be assessed.
A selection was made of six target species (beech, ash, wild cherry, Norway spruce, Douglas
fir, Scots pine) and four species groups (oaks, poplars, Mediterranean pines, larches) of high
interest from a European point of view (Chapter 3).
The number of common protocols varied from one each of the beech and oak species to
seven for the species group poplars. Furthermore common protocols were identified for
broadleaves and conifers. For each species/species group a core group was created and a
responsible core group leader determined.
Specific input was provided at the trait level for the assessment of wood characteristics and
drought based on the results of questionnaires (Chapter 4).
Furthermore, the concept of reference genotypes and a selection of extreme reference
genotypes with low genotype-environmental-interaction are presented (Chapter 5).
References
PÂQUES L (comp.), 2009: European survey on assessment methods of morphological traits.
Treebreedex Doc. 916: 53 pp.
DUCCI F, DE CUYPER B, PAQUES L, PROIETTI R, WOLF H (comp.), 2012: Reference protocols for assessment
of trait and reference genotypes to be used as standards in international research projects. CRA
SEL, Arezzo, Italy: 82 pp.
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3. Species oriented common protocols
3.1 Broadleaves
3.11 Beech specific protocols
Core group: Mirko Liesebach (vTI now THÜNEN),
Sven M.G. de Vries (ALTERRA ),
Marek Rzonca (IBL),
Ecaterina Nicoleta Chesnoiu (ICAS),
Ricardo Alia (INIA),
Alexis Ducousso (INRA)
3.111 Flushing (bud break)
Definition
A specific trait of beech (Fagus sylvatica L.) is flushing (bud break), which is under strong
genetic control. Results from provenance trials show large differences in leaves’ flushing
time in spring. Generally, provenances from the eastern and south-eastern part of the
distribution area, as well as provenances from high elevations, require a smaller heat sum
for flushing and thus flush earlier. Provenances from the western part of the distribution and
from low elevations where late frosts occur require a higher heat sum and flush late.
Depending on the temperature development during spring, the time differences between
the earliest and the latest individuals to flush in a set of provenances in a trial can be 4 to 6
weeks. Between the different years and over a number of sites, certain stability was found in
the ranking of flushing time in the provenances analysed.
The phenology of flushing is important for damages by late frost. In the context of climate
change that becomes more and more important. Damages by late frost can result in yield
losses and poor stem forms. Therefore, flushing is a trait which has an indirect economic
impact, apart from its adaptive value.
Description of the assessment in general
Flushing will be recorded for each tree separately. Observations on beech show that there is
no rule whether the terminal bud or other buds starts flushing.
Experience shows that in younger plants a stage is reached when approximately 3 buds show
the specific characteristics. Flushing can easily be recorded at younger plants up to a height
of 2 m (average height of a trial series)
For older and large plants it is almost impossible or at least not easy to record single buds,
i.e., when the average height of a trial site is above 2 m. In this case a stage is reached when
50 % of the crown shows the characteristics of a specific stage. This is according to the BBCH
recommendations.
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Depending on the scientific question and the financial resources, records will be taken
(1) once, all trees at a time (each stage) or
(2) periodically
(2.1) each stage, all trees at each assessment,
(2.2) one specific stage, only trees recently flushed at each assessment.
When the assessment should only be carried out once, the timing is important. The good
variation between the genetic units is around a mean value of the scoring schema for a site.
The assessment has to be carried out uninterrupted and should not last more than one day.
A replication should be assessed without a break.
The assessment of flushing should first be done at the beginning of the second vegetation
period in the field.
For older plants, the assessment is only to get an impression whether a genetic unit is
flushing early or late.
Scoring schema
a) For trees up to a mean height of 2 m of a trial series a 5-step-scale is recommended:
Scale values
Text
1
dormant winter bud
2
buds expanding
3
bud-burst (first green is visible)
4
leaves are flushing
5
leaves are fully expanded
Stage 1
7
Stage 2
Stage 3
Stage 4
8
Stage 5
Photos: ALTERRA (left), M. LIESEBACH (right)
b) For trees above a mean height of 2 m, a 3-step-scale is recommended:
Scale values
Text
1 (1)
dormant winter bud (late flushing)
2 (3)
bud-burst (first green is visible) (intermediate)
3 (5)
leaves are fully expanded (early flushing)
(numbers in brackets correspond to the scale mentioned under 3.1)
1
2
3
Photos: ALTERRA
Pros and cons of the described protocols
a) Pros
There is good experience in using the protocols.
Genetic units can be distinguished nicely coming at the right time for the assessment.
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Several assessment schemas were used in the past. The results of existing accessments can
only be converted backwards with losses in a few steps (see Table 3.11.1) not vice versa.
The data can be analysed as a quantitative trait by adjusting a Weibull function and can be
easily compared among sites and recording dates.
Table 3.11.1: Assessment schemas used for scoring flushing at beech trials
3-step
1= late flushing
2= intermediate
3= early flushing
5-step
1= dormant winter bud
2= buds expanding
3= bud-burst (first green is
visible)
4= leaves are flushing
5= leaves are fully expanded
6-step
2= buds swollen
3= first green is visible
4= leaves begin to
appear, individually
visible folded
5= leaves unfolded, fanshaped
6= leaves unfolded,
bright
7 step
2= buds swollen
3= first green is visible
4= leaves begin to appear
5= individually visible
folded
6= leaves unfolded,
fanshaped
7= leaves unfolded, bright
b) Cons
It is not always easy to catch the right time to have a good differentiation between the
genetic units in flushing.



Difficulties in choosing proper term of assessment,
possible subjective influence,
requires trained staff.
References
Alterra, 2005 (in Dutch): INS-CGN-FG-076 Uitloopstadia Fagus sylvatica.
LIESEBACH M, 2012 (in German): Phenotypical traits and variability of six European beech (Fagus
sylvatica L.) provenances on a test site in Schleswig-Holstein. Landbauforschung - Journal of
Applied Research in Agriculture and Forestry 62 (4): 179-192.
MADSEN SF, 1995: International beech provenance experiment 1983-1985. Analysis of the Danish
th
member of the 1983 Series. Genetics and Silviculture of Beech, Proceedings from the 5 Beech
Symposium of the IUFRO Project Group P1. 10-100, 19-24 September, 1994, Mogenstrup,
Denmark, 35-44 + 288
MALAISSE F, 1964: Contribution à l’étude des hêtraies d’Europe occidendentals. Note 4: Quelques
observations phénologiques de h1etraies en 1963. Bull. Soc. r. Bot. Belg. 97: 85-97.
MATRAS J, et al., 2005 (in Polish): Growth at the young age and genetic variability of European beech
(Fagus sylvatica L.) populations in Poland. Scientific report for years 2001-2005 with
recommendations for practise, Warsaw, 105 p.
ROBSON TM, ALIA R, BOZIC G, CLARK J, FORSTEUTER M, GÖMÖRY D, LIESEBACH M, MERTENS P, RASZTOVITS E,
ZITOVÁ M, VON WÜHLISCH G, 2011: The timing of leaf flush in European beech (Fagus sylvatica L.)
saplings. Monografias INIA: Serie forestal, vol. 22: 61-80.
10
SCHÜLER S, LIESEBACH M, 2015: Latitudinal population transfer reduces temperature sum requirements
for bud burst of European beech. Plant Ecol 216: 111-122.
SCHÜLER S; LIESEBACH M, WÜHLISCH G VON, 2012 (in German): Variation in flushing time of European
beech: effects of provenance, year and environment. Landbauforschung - Journal of Applied
Research in Agriculture and Forestry 62 (4): 211-219.
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3.12 Oak (Quercus spp.) specific protocols
Core group:
Marek Rzońca (IBL),
Sven M.G. de Vries (ALTERRA),
Alexis Ducousso (INRA),
Mirko Liesebach (vTI now THÜNEN)
3.121 Flushing (bud break)
Definition
Tree phenology changes seasonally due to both climate and endogenous rhythms. A specific
trait for oak species is flushing, (bud break). Bud break is the first visible step in the annual
cycle of trees growth. In oaks species, early wood, including large vessels and a part of the
annual radial stem growth is achieved before leaf expansion in spring. Oaks have two types
of flushing, early and late. Long term observations show about 2-3 week differences
between these types. In the southern range (northern Italy) flushing starts on about 20 April,
in the northern range (southern Scandinavia) on 20 May (for Quercus petraea, Q. robur).
Spring flushing dynamics and terms of dormancy are largely genetically determined. The
variability of flushing is important for damages by late frost, causes in yield losses and poor
stem forms. Consequently, flushing is a specific visible trait, which affects the oaks’ adaptive
potential and economic value.
The assessment of oak flushing is similar to beech.
Bud break should be assessed separately for each tree in the field.
Due to size of observed material (height of tree) flushing can be more easily recorded for
younger tree, especially those up to 2 m of height. Taller trees should be assessed with
another scale. In this case is not possible to observe single buds. The BBCH recommendation
states that when 50 % of the crown achieves the specific characteristics, the stage is
reached.
The issue of the date of assessment, it should be carried out in several ways:
 One time (all trees in one date)
 Periodically
- In each stage (all trees at severals date and each assessment)
- In only one specific stage (only trees recently flushed at each assessment)
When the assessment is carried out, the most important factors is to check for a good time
for observation. If the assessment starts too early, buds are not variably developed and
cannot be observed at all stages of the scale. The same problem appears when assessment
starts too late. The good variation among trees is about a mean value of scoring schema for
an each field trial.
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Scoring scheme
a) For trees up to a mean height of 2 m, a 5-step-scale is recommended:
Scale values
Text
1
dormant winter bud
2
buds expanding
3
bud-burst (first green is visible)
4
leaves are flushing
5
1
2
3
4
5
Photos: MLUV
b) For trees above a mean height of 2 m. a 3-steps-scale is recommended:
Scale values
Text
1 (1)
dormant winter bud (late flushing)
2 (3)
bud-burst (first green is visible) (intermediate)
3 (5)
leaves are fully expanded (early flushing)
numbers in brackets correspond to the scale mentioned under a)
1
2
3
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1
2
3
Photos: E. N. CHESNOIU
a) Pros



There is a lot of good experience in using this protocol, the scale is in practice. The
results of existing accessments can only be converted backwards with losses in a few
steps (see Table 3.12.1) not vice versa.
it is possible to aggregate some steps for comparing assessments (Table 3.12.1),
this scheme has been discussed among a broad range of foresters (e.g., Treebreedex).
Table 3.12.1: Assessment schemas used for scoring flushing at oak field trials
3-step
1= late
flushing
5-step
1= dormant
winter bud
2= buds
expanding
2=
intermediate
3= bud-burst
(first green is
visible)
4= leaves are
flushing
3= early
flushing
5= leaves are
fully expanded
5-step BBCH
1=00= buds are closed
and covered in brown
scales
2=01= the majority of
leaf buds have started
to expand
3=03= the majority of
leaf buds have finished
expanding and are
ready to open
4=07= start of budburst
and leaf growth
6-step
1= dormant
winter bud
8-step
0=dormant winter
buds
2= buds
swollen
1=buds swollen
3= buds begin
to open
4= individual
small leaves
are visible
5=09= the tips of the
first leaves are clearly
visible
5= leaves are
big, but still
folded
2=buds swollen and
elongated
3=buds burst, first
green is visible
4=buds begin to
open, leaves
gathered into a
bunch
5=individual small
leaves are visible
6=leaves are big, but
still folded, main
shoot starts to grow
7=leaves unfolded
and smooth, main
shoot is growing
6= leaves
unfolded and
smooth
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b) Cons




It is not always easy to catch the right time to have a good differentiation between the
genetic units in flushing.
Difficulties in choosing proper term of assessment,
possible subjective influence,
requires trained staff.
References
BREDA N, GRANIER A, BARATAUD F, MOYNE C, 1995: Soil water dynamics in an oak stand. Part I: soil
moisture, water potentials and water uptake by roots. Plant and Soil 172: 17–27.
MARKIEWICZ P et al., 2006 (in Polish): Variability of the growth and quality of pedunculate oak
(Quercus robur L.) populations and families in Poland. Scientific report for years 2002-2006 with
recommendations for practise, Sekocin Stary, 45 p.
SECONDAT N, 2008: Study of the phenology of the sessile oak (Quercus petraea). Masters project,
University of Bordeaux, 13 p.
MLUV [Ministerium für Ländlicher Entwicklung, Umwelt und Verbraucherschutz des Landes
Brandenburg, Landeskompetenzzentrum Forst Eberswalde] (ed.) 2009 (in German): In the foodsteps of climate change: Phenology of forest trees. Leaflet.
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3.13 Ash specific protocols
Core group: Joukje Buiteveld (ALTERRA),
Arnaud Dowkiw (INRA),
Bart de Cuyper (VLAGEW)
3.131 Flushing (bud burst)
Bud burst is the period of the initiation of growth from a bud till elongated shoots with
expanded leaves. The trait is under strong genetic control (high heritability) in ash and is
influenced by environmental factors and tree health conditions. The most important
environmental factors influencing the timing of flushing are spring temperature and winter
chilling. Data from provenance trials indicate large differences in flushing time between
provenances. In general, northern and western provenances flush later than southern and
eastern ones. Also, provenances from high elevations flush later than the ones from low
elevations. There is a strong positive correlation between flushing dates between the years.
Flushing is an important adaptive trait. Early flushing provenances/trees are prone to
damage from late spring frost. This damage may reduce the growth of the tree and affect
the tree’s fitness and stem form. There is evidence from more recent studies that flushing is
correlated with susceptibility to ash dieback, and that late-flushing trees are more
susceptible to infection by fungus.
Evaluation of bud burst dynamics is based on observations over a period of time with a
scoring system describing the different stages for the development of the bud and unfolding
of the leaves. It will be assessed on single trees. In principle the terminal bud is scored in
young trees up to a height of 5m (average height of a trial series). For older trees (height
above 5m) it is difficult to score the single terminal buds. Here a stage is reached when 50%
of the crown meets the described criteria of a specific developmental stage (conform BBCH
recommendations).
Depending on the research objectives/available resources different ways of assessment are
possible:
• Observation only once in spring. The time is chosen so that a maximum variation of the
stages occurs in the population of trees to be studied.
• (Julian) day when a particular developmental stage is reached.
• Observation of every tree at steady intervals (of 3 days or a week) during the whole
flushing period (gives better insight in the dynamical aspects of bud burst behaviour
during the whole observation period).
Each tree is preferably scored on a single day.
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Scoring scheme
For young trees of a trial site up to 5 m, a 5-point scale is recommended:
Scale values
Text
1
dormant bud
2
swelling of bud, slight greening of bud scales
3
buds begin to burst, first green visible
4
bud burst, petioles of leaves visible, no lengthening of twig
5
bud burst, petioles of leaves visible, twig has started lengthening,
1
2
3
4
5
Photos: INRA
For older trees (above 5 m) a 3-point scale is recommended:
Scale values Description
1 (1)
dormant bud
2 (3)
buds begin to burst, first green visible
3 (5)
bud burst, petioles of leaves visible, twig has started
lengthening, leaves are fully expanded
(numbers in brackets correspond to the 5-step scale for smaller plants)
17
1
2
3
Photos: INRA
Pros and Cons
a) Pros
The scoring system covers the total range of developmental stages described, including
shoot elongation.
It is quick and simple.
b) Cons
Timing of observation is important to record maximum variation in the trial.
References
CUNDALL EP, CAHALAN CM, CONNOLLY T (2003) Early results of ash (Fraxinus excelsior L.) provenance
trials at sites in England and Wales. Forestry 76: 385–400.
DOUGLAS GC, PLIURA A, DUFOUR J, MERTENS P, JACQUES D, FERNANDEZ-MANJARES J, BUITEVELD J, PARNUTA G,
TUDOROIU M, CURNEL Y, THOMASSET M, JENSEN V, KNUDSEN M, FOFFOVÁ E, CHANDELIER A, STEENACKERS M,
2013: Common Ash (FRAXINUS EXCELSIOR L.) In: L.E. Pâques (ed.), Forest Tree Breeding in Europe:
Current State-of- the-Art and Perspectives, Managing Forest Ecosystems 25, DOI 10.1007/978-94007-6146-9_9.
VITASSE Y, DELZON S, DUFRÊNE E, PONTAILLER J-Y, LOUVET J-M, KREMER A, MICHALET R, 2009: Leaf phenology
sensitivity to temperature in European trees: Do within-species populations exhibit similar
responses? Agricultural and forest meteorology 149: 735 – 744.
BAKYS R, VASAITIS R, SKOVSGAARD JP, 2013: Patterns and Severity of Crown Dieback in Young Even-Aged
Stands of European Ash (FRAXINUS EXCELSIOR L.) in Relation to Stand Density, Bud Flushing
Phenotype and Season. Plant Protection Science Vol. 49 (3): 120-126.
JOUVE L, JACQUES D, DOUGLAS GC, HOFFMANN L, HAUSMAN J-F, 2007: Biochemical characterization of early
and late bud flushing in common ash (Fraxinus excelsior L.) Plant Science 172: 962–969.
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3.14 Poplars specific protocols
Core group: Marijke Steenackers (VLAGEW),
Sven MG de Vries (ALTERRA),
Catherine Bastien (INRA),
Alain Berthelot (FCBA)
Lorenzo Vietto, Fulvio Ducci (CRA),
Volker Schneck, Mirko Liesebach (vTI now THÜNEN)
3.141 Phenology
3.141.1 Bud burst
In springtime, when air temperatures rise to optimal levels, the buds swell and new leaf
tissue pushes out of the protective scales. This process is called bud burst or bud break. The
objective of the assessment of the earliness of vegetative budbreak is an indirect selection
for resistance to spring late frosts.
Description of the assessment
Bud break is evaluated on terminal shoot at two or three years in nursery test with a 6
classes score. The most important stage associated to frost damages is stage 3 (see scoring
scheme). Good correlation between nursery stage and adult stage in plantation have been
observed.
Bud break earliness is assessed at two year old trees in nursery test on terminal buds of main
stem or codominant shoots (coppice). When done at only one date, the assessment of bud
break with this 6 classes score is optimum when about 50 % of trees could be scored 3 or
more. Optimal evaluation of bud break earliness is obtained with an average score collected
on three successive dates (date 1: 25 % of trees have reached score 3 or more, date 2 : 50 %
of trees have reached score 3 or more, date 3 : 75 % of trees have reached score 3 or more.
Scoring scheme
Scale values
1
2
3
4
5
6
Text
Dormant bud completely enveloped by the scales (= perulae)
Bud swelling with scales slightly diverging showing a narrow yellow margin;
presence of one or more droplets of balsam
Bud sprouting, with tips of the small leaves emerging out of the scales
Buds completely opened with leaves still clustered together; scales still present
Leaves diverging with their blades sill rolled up; scales may be present or absent
Leaves completely unfolded (but smaller in size than mature ones); lengthening
of the axis of the shoot evident; scales absent
19
1
2
3
4
5
6
Pro and con
This protocol is suitable to measure quickly a large number of trees.
Reference clones
Budflush phenology
Standard clones
Early budflush
Flevo, I-214, Soligo
Medium budflush
Alcinde,Catherine aanvullen
Late budflush
Koster, Dorskamp, Blanc du Poitou, Triplo
References
CASTELLANI E, FRECCERO V, LAPIETRA G, 1967: Proposta di una scala di differenziazione delle gemme
fogliari del pioppo utile per gli interventi antiparassitari. Giorn Bot Ital 1967, 101: 355-360.
TUROK J, LEFÈVRE F, CAGELLI L, DE VRIES SMG, (compilers), 1996: Populus nigra Network. Report of the
second meeting, 10 – 12 September 1995, Casale Monferrato, Italy. Rome, Italy: International
PlantGenetic Resources Institute, 26 pp.
3.141.2 Bud set
Definition
The objective of the assessment bud set lateness is an indirect selection for resistance to
early frosts in autumn. Date of bud set is evaluated on terminal shoot at two years in nursery
test with the following 7-classes-score (protocols B or C). The most important stage is stage
1,5; stage 2,5 correspond to cessation of elongation. Length of photoperiod and average
temperature are known to influence jointly date of growth cessation and date of bud set.
20
Date of bud set is evaluated on terminal shoot at two years in nursery test with the 7classes-score (protocols B or C) given below. The most important stage is stage 1,5; stage 2,5
correspond to cessation of elongation. Length of photoperiod and average temperature are
known to influence jointly date of growth cessation and date of bud set.
When done at only one date, the assessment of bud set with this 7-classes-score is optimum
when about 50 % of trees could be scored 1.5 or less. Optimal evaluation of date of bud set
is obtained with an average score collected on three to five successive dates.
Scoring scheme
Scale values Text
1 (0 in figure) Apical bud red brown
2 (0,5)
F1 fully stretched comparable to older leaves apical but of older leaves;
apical bud of fully closed between green and red
3 (1)
F1 fully stretched with ‘juvenile aspect’;
F2 comparable to older leaves apical bud not fully closed
4 (1,5)
F2 fully stretched transition to bud structure
5 (2)
F2 fully stretched no bud structure
6 (2,5)
2 rolled-up young leaves at equal height
7 (3)
>2 rolled-up young leaves
No more
rolled
leaves
Presence
of rolled
leaves
21
22
Pro and con
These protocols are suitable to measure quickly a large number of trees.
References
ROHDE A, STORME V, JORGE V, GAUDET M, VITACOLONNA N, FABBRINI F, RUTTINK T,ZAINA G, MARRON N, DILLEN
S, 2011: Bud set in poplar - genetic dissection ofa complex trait in natural and hybrid populations.
New Phytol 189: 106-121.
RHODE A, BASTIEN C, BOERJAN W, SEAN T, 2011: Temperature signals contribute to the timing of
photoperiodic growth cessation and bud set in poplar.Tree Physiol 31 (5): 472-482.
3.142 Melampsora laric-populina
Melampsora larici-populina is the most common and dangerous Eurasian rust, and requires
and alternate host „Larix“ to complete its life cycle. Uredinia, telia and basidia are formed on
poplar, spermogonia and aecia are formed on larch. Uredinia commonly develop on the
lower surface of leaves, giving rise to yellow or necrotic spots and flecks on the upper
surface. Severely affected trees are defoliated prematurely, reducing growth potential and
predisposing them to environmental stresses and invasion by secondary damaging agents.
Early leaf fall also increases the risk for winter injury, dieback and infection by secondary
pathogens. The disease is generally controlled by planting tolerant cultivars or clones to the
local populations of rust. The potential evolution of new rust pathotypes underscores the
critical need of monitoring poplar rust populations.
Different evaluation tests are used to evaluate susceptibility to Melampsora larici-populina:
(1) qualitative resistance tests under artificial inoculation with known reference strains;
(2) quantitative resistance tests on leaf disks inoculated with known reference strains and
(3) susceptibility tests in field experiments under natural inoculation associated to
characterization of the pathogen population.
(1) Protocol for qualitative resistance tests under artificial inoculation
The objective of qualitative resistance tests is to identify qualitative resistances inherited
from P. deltoides and overcome by at least one of the eight known M. larici-populina
virulences. Leaf disks collected on disease-free plants are inoculated artificially at high
density (40000 spores/ml) with a solution of mono-uredinial strains of known combination
of virulences (or pathotype). A minimum of 6 leaf disks sampled from the 5 th to the 8th
unrolled leaf below the apex are inoculated per tested genotype. After inoculation, leaf disks
are maintained in climatic chamber at a temperature between 17°C and 20°C with a 16hours photoperiod. The response observed 14 days after inoculation is presence/absence of
23
uredinia or any leaf decoloration or any leaf necrosis. A genotype is considered as
susceptible to the strain studied when a minimum of one leaf disk presents sporulated
uredinia. A genotype is considered as resistant to the strain studied when all the inoculated
leaf disks are free of any symptoms.
The M. larici-populina strains are chosen in order to represent the eight known virulences.
For each inoculation test and each strain, the following differential genotypes are used to
confirm the pathotype of the strain used:
Type botanique
P. × canadensis
P. × canadensis
P. candicans
P. × canadensis
P. x generosa
P. × generosa
P. deltoides, P. × generosa
P. × generosa
P. ×x generosa
Differential Clones
Robusta
Ogy
Aurora, Grimminge
Brabantica
Unal
Rap
87B12, 84B09
Beaupré
Hoogvorst,hazendans
List of Mlp virulences for which a
compatible response oft he
differential clone is observed
1,2,3,4,5,6,7,8
1
2
3
4
5
6
7
8
(2) Protocol for qualitative resistance tests under artificial inoculation
The objective of quantitative resistance tests is to rank poplar genotypes for their ability to
limit infection by M. larici-populina in absence of qualitative resistance. The three
components of quantitative response associated to pathogen fitness are measured on 3 cm
diameter excised leaf disks inoculated with a reference mono-uredinial strain. The leaf disks
are floated upside down on distilled water in polycarbonate 6-well cell culture plates. The
inoculum suspension concentration was 30 mg/liter : 3 mg urediniospores in 100 ml distilled
water added with 0.5 mg agar. Sprayed suspension volume per leaf surface ratio was
20 ml/m2. The disks were kept for 14 days under growth chamber controlled conditions
(17 °C, 16 h photoperiod, 30 E.m-2.s-1).
The following three components of quantitative resistance are observed on each leaf disk :

the latent period, measured on a half a day basis, which corresponds to the date when
the first sporulating uredinia can be observed.

the number of sporulating uredinia 13 days after inoculation

The average uredinia size score (USscale) 14 days after inoculation. This score uses a 1 to
5 scale defined according to the size variability observed in the experiment. To obtain
an objective assessment of uredinia size, 25 leaf disks per score (75 for the whole
tests) are sampled for image analysis. Average uredinia surface in mm2 is measured
and calculated on all sporulating uredinia of the inoculated leaf disk.
24
(3) Protocol to evaluate susceptibility to Melampsora larici-populina leaf rust under
natural infection in field tests
Susceptibility to Melampsora larici-populina leaf rust under natural infection can be
evaluated by two protocols, dependent on the time of the year: protocol A) the maximum
infected leaf score and protocol B) the global sanitary tree score.
Protocol A:
The maximum infected leaf score quantifies the infection level of the most infected leaf
on the terminal shoot. This observation is done quite early, between end of June and midJuly when the infection could be considered as homogeneous in the field experiment. At
this time of infection, the most infected leaves are still on the trees.
Protocol B:
The global sanitary tree score quantifies the damages (infection, necrosis, leaf fall and
even plant death) at the plant level. This observation is done late in season, between midAugust and mid-September just before growth cessation.
Pros and cons
The protocols are easy to carry out on a large number of trees per time unit (several
hundreds per day).
The global sanitary tree score could take into account other damages than those directly
linked to Melampsora larici-populina infection.
Reference genotypes
The following standard clones with known levels of susceptibility to Melampsora laricipopulina to populations including the eight known virulences need to be included in field
experiments:
Susceptibility level to Mlp
in field experiements in
presence of all virulences
Very low susceptible
Low susceptible
Susceptible
Very susceptible
Standard clones
Alcinde, Lena, Bakan, Skado, Soligo, Triplo, Villafranca
87B12, Dvina, I-214, Flevo, Dorskamp, Koster, Fritzi Pauley,
Trichobel, Grimminge, Vesten
Robusta, Ogy, Brabantica, Unal, 84B09, Blanc du Poitou, I-45/51,
Raspalje, Vereecken
Aurora, Rap, Beaupré, Hoogvorst
It is important to characterize the pathogen population responsible of the natural infection
observed in field tests. This characterization is obtained by molecular analysis to confirm the
pathogen species and by isolation and pathotype determination of about 100 distinct
uredinia sampled on different leaves of the ‘Robusta’ standard clone.
25
Scoring schemes
Protocol A: Maximum infected leaf score (1-8); end of July (mid-infection)
Source: INRA-FCBA
Scale values
8
7
6
5
4
3
2
1
6
Text
No uredinia
Few difficult to detect
Uredinia easy to detect but not joined
Joined uredinia covering less than 10 % of the leaf area
Joined uredinia covering between 10 % and 25 % of the leaf area
Joined uredinia covering more than 75 % of the leaf area
5
4
3
2
1
Protocol B: global sanitary tree score (1-7); mid-August till mid-September
Source: INRA-FCBA, INBO
Scale value
7
6
5
4
3
2
1
Description
0 uredinia on the plant (qualitative resistance)
Few uredinia hard to detect
Many uredinia per leaf but no decoloration neither necrotic zones
Many leaves infected but still green. Limited decolorated or necrotic zones,
no defoliation
Numerous leaves highly infected with decoloration and necrotic areas, significant
defoliation but between 1/2 and 1/3 of green leaves on the top of the plant.
Most of leaves have decolorations and necrotic areas, important defoliation (30%50%), less than ¼ of remaining leaves are still green.
Important defoliation (>50 %), no more significant elongation, only few remaining
leaves on the top
26
References
DOWKIW A, HUSSON C, FREY P, PINON J, BASTIEN C, 2003: Partial resistance to Melampsora larici-populina
leaf rust in hybrid poplars: genetic variability in inoculated excised leaf-disk bioassay and
relationship with complete resistance. Phytopathology 93, 421-427.
PINON J, BERTHELOT A, FABRE B, 2011. Comportement des cultivars de peuplier envers la rouille. Revue
Forestière française, LXIII, 333-346.
OSTRY M, RAMSTEDT M, NEWCOMBE G, STEENACKERS M, 2014: Diseases of poplars and willows. In:
ISEBRANDS JG, RICHARDSON J, (eds.). FAO-book Poplars and Willows: Trees for Society and the
Environment, Chapter 8 (pp.443-458)
3.143 Marsonia brunea
Marssonina brunnea is a damaging foliar pathogen of poplar. Severe attacks on susceptible
clones can cause premature defoliation, resulting in significant growth reduction, dieback,
and predisposition to secondary organisms and abiotic stress. Marssonina brunnea causes
dark brown, circular to angular leaf spots that usually measure about 1 mm in diameter. On
highly susceptible clones, lens-shaped lesions develop on petioles and current-year shoots.
Damage can be especially severe where poplar species and clones are selected without
regard to their susceptibility to the disease.
This protocol has been adapted from the international protocol defined in 1975 by the
International Poplar Commission and optimized by INRA-FCBA. A special field experiment is
established in nursery for a minimum of three years of observation at very high plantation
density (0.4 to 0.6 m in the line, 1.5 to 2m between lines) to favor contamination. It is also
recommanded to plant at regular spacing a very susceptible reference clone such as
Magister Géant or I-214 or Tardif de Champagne. To favor contamination, infected leaves
can be attached on the very susceptible reference clone. Between three and five reference
clones from the following list are included in the test:
Botanic type
P. deltoides
P. × canadensis
Very susceptible
reference clones
Lena
Magister géant,
I-45/51, Tardif de
Champagne
P. trichocarpa
P. × generosa
Other interspecific hybrids
Susceptible reference
clones
Lux
I-214, Robusta, Brenta,
Mella, Triplo, Lambro,
Soligo, Blanc du Poitou
Fritzi Pauley
Resistant/Tolerant
reference clones
Dvina
Dorskamp, Polargo,
Lux
Hazendans,
Grimminge, Raspalje
Taro
Source: INRA-FCBA
27
Response to contamination is scored twice during the growing season: once early in June
and once mid August just before significant leaf fall.
Scoring scheme
On the main stem of each plant, all leaves are scored with the following scoring system:
Score
Description
4
Leaf free of disease
3
Between 1 and 11 acervulae on the leaf
2
Between 11 and 100 acervulae on the leaf
1
More than 100 acervulae on the leaf
Relative frequencies of the 4 leaf scores at an individual tree level are then calculated to
estimate genotype susceptibility to Marssonina brunnea.
Pros and cons of the described protocol
This protocol is very detailed but not suitable to measure quickly a large number of trees.
3.144 Xanthomonas populi
The bacterium Xanthomonas populi is one of the main poplar pathogens throughout much
of Europe.
Susceptibility of clones is tested by artificial inoculation on two years old plants in the field.
Xanthomonas populi isolates used for inoculation can be obtained from the collection CFBP
at INRA-Angers (FR) or from INBO collection, Geraardsbergen (BE). Isolates are reactivated
and multiplicated at 20°C in Petri dishes filled with a LPGA medium. The inoculation
suspension is adjusted to 108-109 cells per ml just before inoculation.
Two or three transversal incisions are made on tree main stem between 0,80 and 1,20m
(about 10mm large and 2mm long). 25 µl of the inoculation suspension is deposited on the
fresh wound. The inoculation is done in early September, in the period just before closing of
the buds.
Response to inoculation is measured one and two years after inoculation at each inoculation
point thanks to two different traits:
 Longitudinal canker extension (mm)
 Girdling Index score (0-5):
28
Score
6
5
4
3
2
1
Description
No symptom
canker is developed along less than 20 % of stem circumference
canker is developed along between 20 % and 40 % of stem circumference
canker is developed along between 40 % and 60 % of stem circumference
canker is developed along more than 60 % of stem circumference
total girdling of tree stem with associated mortality of the apex
The following reference clones are used for comparison:
Botanic type
Very susceptible
to race1
P. x generosa
S.6-2
Susceptible to race1
Boelare, Raspalje, Beaupré
P. trichocarpa
P. x canadensis
Donk, I-45/51
Resistant to race1
S.3-31
Fritzi Pauley
Blanc du Poitou,
Dorskamp
I-214, Koster
Pros and cons of the described protocol
Selection for susceptibility to Xanthomonas populi is a long-term test, as it is done at 1 and 2
years after artificial inoculation on 2-year-old trees.The protocol itself is easy to perform.
References
NÈSME X, STEENACKERS M, STEENACKERS V, PICARD CH, MÉNARD M, RIDÉ S, RIDÉ M, 1993: Differential hostpathogen interactions among clones of poplar and strains of Xanthomonas populi pv.populi. The
American Phytopathological Society 84 (1), 101-107.
29
3.15 Wild Cherry specific protocols
Core group: Fulvio Ducci, Roberta Proietti (CRA),
Fréderique Santi (INRA),
Joukje Buiteveld (ALTERRA),
Bart de Cuyper, Marijke Steenackers (VLAGEW)
3.151 Phenology
Definition
Phenology is an important aspect of adaption. It is the study of the timing of periodic
phenomena such as bud break, flowering, growth initiation, growth cessation, water flows,
cambial and physiological activity, especially as related to seasonal changes in temperature,
photoperiod, etc. (Nanson 2004).
Phenology traits are conditioned by biological and environmental factors (chilling
requirements, forcing temperatures, seasonal changes, etc.) necessary for launching the
processes, but they are also under strong genetic control. They are important selection
criteria as they might condition stem form in the case of stress, frost damage (mostly late
frost), but they also affect growth potential in relation to the growth season length. Then
they are directly related to the growth and to the tree architecture.
Experimental data from already performed surveys show the adaptive meaning of this group
of specific traits. Indeed, clones and provenances are strongly influenced in phenology by
their origin. Phenology traits are therefore practically relevant indicators of the adaptability
and adaptedness of forest trees and are useful to study the effect to global change. A series
of assessments on the same individuals over more years allows conclusions about the
climate development. It might be useful to identify extreme standard materials which could
be used as references in trials to alert for phenology observations. Preferably it should be
material shared by as many test sites as possible.
3.151.1 Flushing (bud break)
Definition
Bud break is the period spanning from dormant bud up to shoot elongation. It is conditioned
by internal (genetic) and environmental factors (chilling requirements and forcing
temperatures).
A subjective scoring system is commonly used. But in any case, due to the size of trees,
different methods need to be adopted depending on whether young or adult trees have to
be monitored.
In young trees (up to 2.5 m), observation usually concerns the terminal buds, while in adult
and taller trees (more than 2.5 m) the terminal bud of lateral branches or the same portion
of the crown should be monitored. Attention should be also paid to the acropetal or
basipetal sense of phenology in the crown. I.e., in wild cherry, phenology proceeds in an
30
acropetal way. That means that the crown should be ideally divided in 3 sectors to keep the
same method in all the monitored trees (Figure 3.15.1).
Figure 3.15.1: For phenological surveys in adult trees, the crown should be ideally divided into 3
sectors.
The monitoring has to be performed in each sector and located in the same aspect. A
phenological stage of a sector is reached when 50 % of buds show the characteristics
describing a specific score.
A sufficient number of trees to perform statistical analysis must be surveyed. When clones
are considered, the number of plants per clone can be sensibly reduced to 3 – 5, while when
progenies or provenances are monitored, the number can vary between 20 and 30 or more
trees.
Based on scientific questions and on funding resources, monitoring can be either conducted
following a regular timing or just occasionally. The most common criteria are:
- twice a week during all the flushing period. Each tree is monitored at each assessment;
- at a given periodicity (days/week) during the flushing period by assessing only trees having
reached a given score or all trees at different scores;
- only once or twice during the flushing season when distances are an important limiting
factor for surveys.
The monitoring season should start soon enough to capture early stage and end late enough
to catch the last stage. Each flushing assessment is to be carried out on the same day, or
with a difference of one day maximum.
31
To reduce errors due to a subjective attribution of scores, the same staff should be involved
in the survey.
Monitoring can be carried out either in controlled or semi controlled environments (growth
chamber, greenhouse) or in field (nursery, plantation). Anyway in field trials and plantations
in general it is suggested that the monitoring start at least two years later in order to reduce
the acclimation effect. The observation will stop when all trees have reached the last stage.
Scoring scheme
The scoring system is the same for either in young and adult trees, but in the first case the
assessment is carried out on the apical bud, while in the second one it is on the branch apical
buds within a sector. A 5-step-scale is considered (Table 3.15.1; Figures 3.15.2 and 3.15.3).
Table 3.15.1. 5-step-scale used for wild cherry flushing monitoring
Scale values
1
2
3
4
5
Text
Buds not active, scales brown and closed (dormant winter bud)
Buds expanding, scales start to separate
Bud-burst (first green is visible)
Leaves are flushing
Leaves are fully expanded and new sprouts are in elongation
Figure 3.15.2: 5-step scale for young trees
32
Figure 3.15.3: 5-step scale for adult trees
For P. avium an integrated approach for phenology assessment can be also considered that
combines bud break, flowering and ripening of fruits (DUCCI et al. 2012). Scores used can be
compared with phenological growth stages defined by BBCH scale (MEIER 1997, MEIER et al.
2009) for stone fruit (Table 3.15.2 and Figure 3.15.4).
33
Table 3.15.2: BBCH Principal growth Stage 0 (sprouting/Bud development) and Stage 1 (leaf
development) and BBCH-identification keys of stone fruit.
BBCH growth stage
00
03
10
11
19
Text
Dormancy: leaf buds and the thicker inflorescence buds closed and covered
by dark brown scales
End of leaf bud swelling: scales separated, light green bud sections visible
First leaves separating: green scales slightly open, leaves emerging
First leaves unfolded, axis of developing shoot visible
First leaves fully expanded
Figure 3.15.4: BBCH score scale for stone fruit. Scores from 00 to 31will be considered in this
framework.
This scoring system can be expensive in terms of time/personnel (a person is on average
able to sample about 200 trees/day according to the size of trees) and it is suggested to be
applied within collections, with special attention to reference clones.
34
References
DUCCI F, DE CUYPER B, PÂQUES LE, PROIETTI R, WOLF H, (Compilers), 2012: Reference protocols for
assessment of trait and reference genotypes to be used as standards in international research
projects. Adaptive traits: Protocol nr. 8 – Definitions/recommendations; Protocol nr. 8.7 and
Protocol nr. 8.12. Ed. CRA SEL - Arezzo, Italy: p. 82.
DUCCI F, (ed), 2005: Monografia sul Ciliegio Selvatico (Prunus avium L.) [Monograph on wild cherry
(Prunus avium L.)]. CRA – Istituto Sperimentale per la Selvicoltura, Arezzo, Italy, 126 p. ISBN 88901923-0-5
DUCCI F, SANTI F, 1998: The distribution of clones in managed and unmanaged populations of wild
cherry (Prunus avium). Can J For Res 27: 1998–2004.
DUCCI F, DE ROGATIS A, PROIETTI R, 2009: ‘Clonal replicated provenances’ performance and clone
selection techniques. (Leopold Poljakovic-Pjnik and Albina Tarjan Tobolka Eds.) Proceedings of the
international Conference “Forestry in achieving Millennium Goals”, Inst. of Lowland Forestry and
Environment, Novi Sad: 93–103.
MEIER U: BBCH (Stone fruits), http://www.bba.de/veroeff/bbch/bbcheng.pdf
MEIER U, 1997: Growth stages of Mono- and Dicotyledonus Plants. BBCH Monograph, Blackwell
Wissenschafts-Verlag Berlin Wien.
MEIER U, BLEIHOLDER H, BUHR L, FELLER C, HACK H, HESS M, LANCASHIRE PD, SCHNOCK U, STAUSS R, VAN DEN
BOOM T, WEBER E, ZWERGER P, 2009: The BBCH system to coding the phenological growth stages of
plants – history and publications. J. Kulturpflanz 61: 41–52.
NANSON A, 2004: Génétique et amelioration des arbres forestiers. Le Presses Agronomiques de
Gembloux, A.S.B.L., Belgique: 712 p. (ISBN 2-87016-070-4).
1.151.2 Leaf senescence
Definition
Leaf senescence corresponds to the period when progressive changing of leaf color is
observed. Leaf color can be characterized by high heritability, with special regard to clonal
materials. Under local conditions relatively homogeneous color can be an important source
of information for recording adaptive variation.
Spatial micro-environmental variations, extreme environmental events (e.g., drought), the
plant health conditions (bacterial/fungal spots), the sampled leaf amount and
acropetal/basipetal senescence (thus the crown has to be divided into 3 sectors) represent
sources of measurement error.
For leaf senescence (yellowing) a subjective scoring system allows trees to be classified
according to a grid representing different stages of tree yellowing. Observation is best done
on small trees like in a nursery (but transplantation shock could be a source of error) or in
field experiments (with trees of heights up to 3-4 m). Generally the whole crown of trees is
observed.
35
Scoring scheme
Assessment of senescence via a scoring system is generally used. It is a very subjective
method. The organization of the survey is the same as for flushing. The senescence progress
within the crown is acropetal, starting the first color variation from the bottom towards the
tip. So, even in this case observations have to be divided among the three crown sectors or
focused on only one of them.
The BBCH system allows a subjective evaluation of color phases to be avoided and this
system is again suggested. The BBCH growth Stage 9 (Senescence, beginning of dormancy)
for stone fruit is reported in Table 3.15.3.
Table 3.15.3: Principal growth BBCH Stage 9: Senescence, beginning of dormancy.
BBCH growth stage
91
92
93
95
97
99
Text
Shoot growth completed; foliage still fully green
Leaves begin to discolor
Beginning of leaf fall
50% of leaves discolored or fallen
All leaves fallen
Harvested product (not to be considered in the framework of T4F)
A further possibility to evaluate senescence is offered by the use of colorimeters. These
allow yellowing to be assessed via quantitative methods. At the clone level these traits can
be highly heritable (0.67 on average for sensu lato heritability). Data can be managed as
quantitative traits using a Portable Colorimeter Microflash v4.0. (methodology is described
in wood color sheet).
3.152 Wood color
Definition
Wood color is an aesthetic trait, for which special attention is placed in wild cherry and in
broadleaves because wood is used to make valuable furniture and cabinetry. Then the
evaluation of wood color is useful for both the characterization of reproductive materials
and for the wood industry.
Assessment
Quantitative parameters of the physical components of the color are evaluated using
colorimeters, portable tools currently adopted by the paint or tissue industries for
standardizing their production. Colorimeters using standard illuminants (mainly white light
illuminants), and recording the reflexed light components, are able to measure basic
parameters with very good approximation. Data can be managed as quantitative traits
according to usual statistic methods.
36
Wood color, both of hardwood (duramen) and soapwood can be measured using:
1) destructive method, on circular sections of the trunk at different heights (at the base, at
breast height and at 3-m);
2) non-destructive method , taking cores at the same heights from trees about 15-20 years
of age (or > 20cm DBH).
Figure 3.15.5: Colorimeter Portable Microflash v4.0 and colorimetric coordinates
The Colorimeter Portable Microflash v4.0 (Figure 3.15.5) provides data relating to the
colorimetric coordinates of the 3 system CIEL * a * b *:
• L* - Luminance: is the variation of the color component along the black to white axis (z,);
• a*: is the variation of the color component along the green to red axis (x);
• b*: is the variation of the color component along the blue or yellow axis (y).
Other parameters can be derived from the above mentioned ones, such as: white index
(expressing the darkness/whiteness of the sample), C* (chroma/saturation; expressing the
intensity of the color measured) and h (taint angle; expressing the height of the sample in
the color space).
Methods
The color measurements (BOURGOIS et al. 1991) are carried out in the CIE- L*a*b* colour
space using spectrophotometers with a standard illuminant D65 and 10° circular
illumination.
The color coordinates are calculated from the reflection data on the sample surface. In this
color range, a colour is defined by its Cartesian chromatic coordinates: lightness L* (varies
from 0 - black to 100 - white) and a* and b* (these coordinates define the chromatic plane;
negative values of a* indicate green, while positive values indicate red; negative values of b*
indicate blue, while positive values indicate yellow). The difference in chromaticity is defined
as: ΔC = [(Δa*) 2+ (Δb*) 2]1/2.
The CIE L*a*b* system produces a fair correlation with the visual perceptions (CHRISMENT
2000).
Concerning the color development during irradiation, it is useful to present the data in the
CIE L*C*h* system which is more uniform (closer to the psycho-sensory assessment). The
37
color development can be derived simply from the CIE L*a*b* system by changing the
Cartesian coordinates to cylindrical ones in the chromatic plane [a*, b*]. The Chroma
(concentration or degree of color saturation) is defined then by C* = (a*2 +b*2)½ and the
hue (saturation angle) h = arctan (b*/a*).
The mean values of chromatic coordinates are calculated from measurements at different
points on the radial surface of the sample (30x30 x10 mm, LxTxR).
Tools/Equipment: quantitative or relative data can be recorded with spectrophotometers by
different brands, i.e.,:
- Spectrocolorimeter Datacolor ®Portable Microflash v4.0, with a measurement opening of 3
mm,
- Konica Minolta spectrophotometer CM-2600 ®with D65 illuminant, a standard angle of 10°
and a measurement opening of 8 mm.
References
BOUGOIS J, JANIN G, GUYONNET R, 1991: La mesure de couleur: une méthode d'étude et d'optimisation
des transformations chimiques du bois thermolysé. Holzforschung 45 (5): 377-382.
CHRISMENT A, 2000: Color by the numbers. CD ROM Datacolor International, 3C-Conseil.
DUCCI F, GERMANI A, JANIN G, PROIETTI R, SIGNORINI G, 2006: Clone selection for wild cherry (Prunus
avium L.) with special reference to some traits used. In: BOZZANO M, RUSANEN M, ROTACH P, KOSKELA
J, (compilers). EUFORGEN Noble Hardwoods Network, Report of the sixth (9-11 June 2002, Alter
do Chão, Portugal) and seventh meetings (22-24 April 2004, Arezzo, Italy). International Plant
Genetic Resources Institute, Rome, Italy: 53-60.
FIORAVANTI M, SIGNORINI G, DUCCI F, 2009: Prunus avium L.: uno studio sull'ereditabilità del colore del
legno. [Prunus avium L.: a study on the heritability of the wood color]. Act of the III Selvicolture
National Congress, Taormina (ME), 2008 October, Vol. 3: 1498-1503.
EL BAKALI I, YAGI S, MERLIN A, DEGLISE X, 2011: A Screening Study of Natural Colour of Wood from
Different Geographical Regions. Research Journal of Forestry 5: 162-168.
10.3923/rjf.2011.162.168
JANIN G., 1987: Mesure de la couleur: Interet forestier et industriel. Ann. Sci. For., 44: 455-472.
JANIN G, GONCALEZ J, ANANIAS R, CHARRIER B, DILEM A, DA SILVA F, DILEM A, 2001: Aesthetics appreciation
of wood colour and patterns by colorimetry: Part 1. Colorimetry theory for the cielab system.
Madears Ceincia Technol. 1: 3-11.
38
3.16 Broadleaves non-species specific protocols
Core group (broadleaves species leader):
Mirko Liesebach (vT now THÜNENI; beech),
Joukje Buiteveld (ALTERRA; common ash),
Fulvio Ducci, Roberta Proietti (CRA; wild cherry),
Marek Rzonca (IBL; oak),
Marijke Steenackers (VLAGEW; poplar),
Sven M.G. de Vries (ALTERRA; poplar)
The non-species specific traits for broadleaves describe the quality. These traits have an
effect on the economic value of the final product. The final product is the trunk, particularly
the lower part of the trunk.
3.161 Straightness
Definition
A non-species specific trait for broadleaves is straightness. Stem straightness is under
genetic control and can be affected by environmental conditions.
Results from progeny trials show large differences in the frequency of straightness classes.
Scoring straightness should be started when the trial has a mean height of 5 m. From then
on straightness can be assessed without an upper height limit.
The assessment has to be concentrated on about ¾ of the tree height beginning from the
ground. That means the straightness in the crown is not taken into the assessment. The
significance of the assessment increases with the age which is correlated with the height of
the tress.
In case of forked stems, only the trunk below the deepest forking point is evaluated.
The assessment can be carried out during the whole year, particularly in older trials.
39
Scoring schema
A 5-step-scale is recommended:
Scale values
Text
5
absolutely straight stem
4
fairly straight (in one direction slightly crooked)
3
slight to moderate bends in different directions
2
moderate to strong bends
1
no straight stem
5
4
3
2
1
a) Pros
There is good experience in using the protocols. In several investigations a simplified version
(only 3 steps were assessed: 1, 3, 5).
b) Cons
There is a slight risk that the straightness of a thinner tree will be classified better than those
of a thicker tree.
References
KLEINSCHMIT J, SVOLBA J 1995: Intraspecific variation of growth and stem form in Quercus robur and
Quercus petraea. In: Proceedings of the workshop, Brussels (BE), 15-16 June 1994. European
Commision DG XII-E.2: 217-238
HOFMANN M, 2014: Hornbeam-accompanying tree species with perspective? Results of a provenance
trial in Lower Saxony. Landbau forsch. Appl Agric Forestry Res 64: 99-106.
40
3.162 Forking
Definition
A non-species specific trait for broadleaves is forking. Forking is can be caused both under
genetic control and as damage to the terminal bud affected by environmental conditions.
Forking can result by biotic and abiotic effects, for example frost events and deer browsing,
respectively. Results from progeny trials show differences in the frequency of forking classes.
Compared to a ramicorn, a fork has two leaders of equal importance in thickness and length,
while a ramicorn is a branch thicker than mean branches, but thinner and usually shorter
than the main stem.
Presence versus absence of a fork is the most straightforward method. According to the
height at which a fork appears, the effect can be judged detrimental or not.
Scoring forking should be started when the trial has a mean height of 5 m. From then on
forking can be assessed without any height limit.
Scoring schema
For trees up to a mean height of 2 m of a trial series a 5-steps-scale is recommended:
Scale values
Text
5
No fork
4
Branch (ramicorn; thinner than a fork) with a similar angle
3
Fork(s) only in upper half of the tree height
2
Fork(s) only in the lower half of the tree height
1
Forks developed both in upper and in lower half of the tree height
½h
5
4
3
2
1
41
When felling a tree with a fork, there is a risk that the trunk below the fork will tear. The
parts above the forking point are thinner than the trunk below. This means the economic
value of the trunks is smaller than a continuous thick trunk from the ground to the crown.
An additional counting of the forks is therefore unnecessary.
a) Pro
There is experience in using the protocol to assess forking.
b) Con
It is sometimes difficult to distinguish between the reasons of forking, in particular when a
tree has one fork. In the case of many forks in a tree, a genetic effect is presumable.
A change from a fork to a ramicorn is fluent.
References
NW-FVA (not published) cited in PÂQUES L 2009: European survey on assessment methods of
morphological traits. Treebreedex Doc. 916
3.163 Branch habit
3.163.1 Branch angle
Definition
Branch angle is important for the wood quality and has an effect on economic value. A steep
branch is longer in wood then a plain growing branch. Between a steep growing branch and
the stem, water can accumulate and can cause rotting.
Branch angle is defined as the angle between a given branch and the main stem axis. The
angle is measured at the insertion point.
Practical assessment: dividing the angle between a rectangular growing branch and the stem
into 3 angle classes with the same size.
The scoring schema can be used at any age but preferable beyond the juvenile stage when
the tree architecture stabilizes. For the assessment the mean branch angle class of all
branches should be regarded.
42
Scoring schema
Scale values
Text
1
<30° (steep)
2
30°-60°
3
60°->90° (plain)
1 (<30°)
2 (30°-60°)
3 (>60°)
Pro and con of the described protocols
a) Pros
The assessment can be carried out easily.
b) Cons
There is the danger of overestimation in the case of concentrating on a single branch, which
is wrongly in focus. The reason for such a branch might be environmental.
3.163.2 Branch thickness
Definition
Branch thickness is important for the wood quality and has a direct effect on the economic
value. A thin branch is better for the wood quality. The cutting point is restored soon after
natural and artificial pruning.
43
Direct measurement is very time consuming. Therefore a 3-step scoring is recommended.
The trait branch thickness is provided as a relation of branch thickness to stem diameter.
To get a feeling for branch thickness it could be helpful to measure a number of branches.
Scoring schema
3-step scale
Scale values
Text
1
Øbranch / Østem <= 0.3 (fine branches)
2
0.3 < Øbranch / Østem < 0.6 (moderate branches)
3
Øbranch / Østem >= 0.6 (thick branches)
a) Pros
Having the feeling it is easy to assess.
b) Cons
Time is needed to get the feeling for evaluation. An underestimation of thicker trees can
happen because thinner trees give the impression of having thinner branches.
44
3.2 Conifers
3.21 Norway spruce specific protocols
Core group: Luc Harvengt (FCBA),
Jan Kowalczyk, Marek Rzońca (IBL),
Egbert Beuker (METLA now LUKE),
3.211 Flushing
Definition
Bud break and bud set are among the most heritable traits of Norway spruce (Picea abies [L.]
Karst.). Provenance tests show a great variability between populations in flushing.
Provenances originating from higher elevations flush earlier than provenances from lower
elevations. Also provenances originated in the northern part of the distribution area need a
lower amount of temperature sum to flush and therefore, flush earlier than those originated
in the south. Genotype's ranking for bud break is also very stable across sites and years.
Bud break should be assessed separately for each tree in the field.
Part of the crown to be assessed: Preferably the top of the crown (light crown) should be
visible from one observation point. If this is not possible, then the middle part of the crown
is also acceptable. The same part of the crown should be considered for subsequent
phenological observations throughout the whole flushing period, as well as for subsequent
years.
Scoring schema
5-step scale
Scale values
1
2
3
4
5
Text
Dormant winter bud
Starts flushing (first green is visible at the tip)
First needles visible (“brush stage”)
New shoot app. 3 cm
New shoot > 4 cm
45
1
2
3
4
(Photos: M. Liesebach, vTi)
5
a) Pros:
The described protocol is very easy to carry out on a large number of trees per time unit.
b) Cons:
When observed once in a test, it is not always easy to get at the right time to have a good
flushing differentiation between the genetic units.
References
SCHMIDT-VOGT H, 1987: Die Fichte. Band I [Norway spruce. Vol. 1]. P. Parey, Hamburg, Berlin: 647 pp.
46
3.22 Douglas-fir specific protocols
Core group:
Jean-Charles Bastien (INRA),
Daniel Michaud (FCBA),
Jan Kowalczyk (IBL),
3.221 Phenology
Definition
Bud break and bud set are among the most heritable traits of Douglas-fir (Pseudotsuga
menziesii [Mirb.] Franco). Provenance tests show a great variability between natural
populations for bud flushing date. Interior provenances, which generally originate from high
elevation, flush earlier than coastal provenances, whereas in the coastal part of the range,
flushing date is mainly driven by the latitude of the geographic origin. Bud break and bud set
are very important adaptive traits, related to the tolerance to abnormal climatic events that
could occur in the growing environment. Very easy to score on a large number of (young)
trees per time unit, these phenology traits are very useful to indirectly assess frost hardiness
and, in some extent the stem form (e.g., forking). Genotype's ranking for bud break and bud
set are also very stable across sites and years.
3.221.1 Flushing
Description
Flushing is recorded on each tree separately. The assessment method depends on whether
the terminal bud is visible or not.
Scoring scheme
When terminal bud is visible, a score is given to each tree according to the development
stage of the terminal bud.
Bud break on terminal bud (5-step score)
Scale values
Text
1
Dormant bud
2
Bud enlarges, slightly greenish appearance
3
Bud scales start to burst; needle points are visible
4
The needle bundle stretches up to twice the size of the bud size; in general the
needles are still tight to the shoot; only rarely are needles around the bud scales
splayed out
5
Needle bundle starts to loosen, shoot still enlarges; the young shoot looks like a
paint brush
47
Stage 1
Stage 2
Stage 4
Stage 3
Stage 5
When the observation is made only once in the growing season, ideal time of recording is
when around half of the trees have reached Stage 3. Flushing is better assessed when it is
possible to record it repeatedly (e.g., beginning and end of flushing season during a given
year or over two different years).
When possible, an alternative (and more accurate) method consists to score 1 or 0 for each
tree on a regular time basis (2 to 3 times a week), when the terminal bud reaches Stage 3.
This method enables bud break to be expressed in days or degree-days (from a given origin).
When terminal bud is not visible (total height above 3m), a score is given to each tree
according to the estimated percentage of crown buds having reached the development
Stage 3 described above.
Bud break on the whole tree (5-step score)
Scale values
Text
1
All buds dormant or just enlarged
2
1-25 % buds reach Stage 3 or more
3
4
5
76–100 % buds reach Stage 3 or more
48
a) Pros:
The protocols described above are very easy to carry out on a large number of trees per time
unit (several thousand per day).
They enable a very efficient screening of genotypes for late frost resistance and, indirectly
for lowering forking defects.
b) Cons:
For a given trial, bud flushing assessment has to be done in the shortest time possible
(ideally within a day).
When observed once in a test, it is not always easy to get the right time to have a good
flushing differentiation between the genetic units.
References
ACEVEDO‐RODRIGUEZ R, VARGAS‐HERNANDEZ JJ, LOPEZ‐UPTON J, et al., 2006: Effect of geographic origin and
nutrition on shoot phenology of Mexican Douglas‐Fir (Pseudotsuga sp.) seedlings. Agrociencia
(Montecillo), 40 (1): 125‐137.
ADAMS WT, BASTIEN JC, 1994: Genetics of second flushing in a French plantation of coastal Douglas‐fir.
Silvae Genetica, 43 (5/6): 345‐352.
BAILEY JD, HARRINGTON CA, 2006. Temperature regulation of bud‐burst phenology within and among
years in a young Douglas fir (Pseudotsuga menziesii) plantation in western Washington, USA. Tree
Physiology, 26 (4): 421‐430.
CHEN Z, CLANCY KM, KOLB TE, 2003: Variation in budburst phenology of Douglas‐fir related to western
spruce budworm (Lepidoptera: Tortricidae) fitness. Journal of Economic Entomology, 96 (2): 377‐
387.
LAVADINOVIC V, KOPRIVICA M, ISAJEV V, 2004: Phenological characters of Douglas‐fir provenances in
Serbia. Silva Balcanica, 4 (1): 89‐94.
OLSZYK D, WISE C, VAN ESS E, et al., 1998: Phenology and growth of shoots, needles, and buds of
Douglas‐fir seedlings with elevated CO2 and (or) temperature. Canadian Journal of Botany, 76
(12): 1991‐2001.
3.221.2 Bud set
Description
Bud set assessment enables to indirectly evaluate hardiness to early fall frosts. This trait
should be assessed on terminal bud (young trees) only.
49
Scoring schema
Bud set on terminal bud (5-step score)
Scale values
Text
1
Terminal shoot is completely green, terminal bud is only allusively differentiated
2
Terminal bud is very small and visible between the terminal needles; terminal
shoot is mainly green, sometimes slightly red
3
Terminal bud is conical and brown coloration starts; terminal shoot starts to
lignify, cork cells are produced
4
Terminal bud is about 2‐3 mm, the brown color is more intense; bark appears
mainly grey/greenish
5
Terminal bud is well developed and dark green; terminal shoot is lignified, that
means the bark is greenish‐grey, and appear slightly teared
a) Pros:
The above described protocol is easy to carry out on a large number of trees per time unit
(several thousand per day).
When possible, it is suggested to assess bud set 2 or 3 times during the growth cessation
period in order to better discriminate genotypes.
50
b) Cons:
Bud set assessment requires a minimum training before starting evaluation on a large
number of trees.
This observation is generally more time consuming than flushing; therefore it is highly
recommended to assess this trait at nursery stage rather than in the field.
References
PETKOVA K, RUETZ W, POPOV EMIL, et al., 2008: Testing of American, German and Bulgarian Douglas-fir
progenies in experimental plantations in Bulgaria Austrian Journal of Forest Science 125 (2): 135156.
ST CLAIR JB, HOWE GT, 2007: Genetic maladaptation of coastal Douglas-fir seedlings to future climates.
Global Change Biology 13 (7): 1441-1454.
3.222 Frost Hardiness
Definition
Frost damage is frequently observed in young plantations. In "frost-prone" sites, damage
occurs more frequently than on milder sites and can be severe, leading to mortality, reduced
growth and poor stem form. The greatest risk of cold injury occurs in the fall, before shoots
are fully hardened for the winter or in the spring when shoots begin, or are about to begin,
active growth.
In the field, cold hardiness can be assessed after damaging frost events. However, in order to
get a good statistical precision, it is important to ensure that incidence of natural frost injury
is not sporadic across field test site and that frost damage can be separated from other
causes of injury (e.g., drought, disease, insect). If not, artificial freezing tests on detached
shoots should be preferred.
Description
The following protocol is to be applied on a single tree basis.
Subjective 5-step score:
Scale values
Text
1
Whole plant frozen to death
2
Terminal shoot frozen and dead, and lateral shoot moderately damaged
3
Severe damage on lateral shoots only
4
Little damage on lateral shoots only
5
No damage visible
51
a) Pros:
The above protocol is very fast to implement on a large number of trees in nursery or in field
tests.
This evaluation is complementary to bud flushing or bud set assessment to screen
genotype's cold tolerance.
b) Cons:
Frost hardiness assessment requires occurrence of a frost event (generally unpredictable)
Frost Damage can sometimes be compounded with disease or insect damage.
References
BRAUN,H, 1998: Results of hybridization in Douglas fir (Pseudotsuga menziesii). Beiträge für die
Forstwirtschaft, 22 (1): 1-7.
HAWKINS BJ, STOEHR M, 2009: Growth, phenology, and cold hardiness of 32 Douglas-fir full-sib families.
Canadian Journal of Forest Research 39 (10): 1821-1834.
O'NEILL GA, ADAMS WT, AITKEN SN, 2001: Quantitative genetics of spring and fall cold hardiness in
seedlings from two Oregon populations of coastal Douglas-fir. Forest Ecology and Management
149 (1/3): 305-318.
3.223 Needle cast
Definition
Douglas-fir Swiss needle cast (Phaeocryptopus gaeumannii), is a foliar disease which
originates from North America. It induces heavy needle losses, which generally start with the
oldest ones. In case of severe attack, only last year's needles persist. This disease is favoured
by a wet climate during the summer season or in wind-protected environments. Damages
are observed mainly in young plantations (up to 15 years) and reveal most often mineral
alimentation problems (weak growth, shallow soils, etc.). Although less heritable than
growth traits, foliage traits appear to be reasonable indicator of Douglas-fir disease
tolerance and can help screen for families that show tolerance to Swiss needle cast.
52
Description
The following protocol is to be applied on a single tree basis.
Subjective 5-step score:
Scale values
Text
1
Very severe infestation (76 % - 100 % needle loss)
4
Severe infestation (51 % - 75 % needle loss
3
Medium infestation (26 % - 50 % needle loss)
2
Very limited infestation (1 % - 25 % needle loss)
1
No infestation (0 % needle loss)
a) Pro:
Quick and low cost evaluation method, which can be completed with an evaluation of foliage color
(from 1= yellow o 3 = dark green)
b) Con:
This needle retention evaluation method does not enable to separate tolerance and resistance to
Swiss needle cast.
References
HOOD IA, KIMBERLEY MO, 2005: Douglas fir provenance susceptibility to Swiss needle cast in New
Zealand. Australasian Plant Pathology 34 (1): 57-62.
JOHNSON GR, 2002: Genetic variation in tolerance of Douglas fir to Swiss needle cast as assessed by
symptom expression. Silvae Genetica 51 (2/3): 80-86.
TEMEL F, JOHNSON GR, ADAMS WT, 2005: Early genetic testing of coastal Douglas-fir for Swiss needle
cast tolerance. Canadian Journal of Forest Research 35 (3): 521-529.
53
3.23 Scots pine specific protocol
Core group: Egbert Beuker (METLA now LUKE),
Volker Schneck, Mirko Liesebach (vTI now THÜNEN)
3.231 Lophodermium needle cast
Definition
The needle cast disease caused by the fungus Lophodermium seditiosum is the most serious
foliage disease of Scots pine (Pinus sylvestris) in the temperate zone. In some years heavy
infections can cause severe defoliation to young pine stands over large areas and a reduction
of tree growth. Therefore, the search for resistant or less susceptible populations or
individuals is of great significance.
The assessment of needle cast can easily be recorded at younger plants up to a height of 1 m
(average height of a trial series). The whole plant has to be observed. The assessment has to
be carried out in spring.
Scoring schema
Scale values
1
2
3
4
5
Text
Very severe infection, all needles brown or dropped off
Medium to severe infection, 50-100 % brown needles
Medium infection, 10-50 % brown needles
Weak infection, few brown needles
No infection
54
1. Very severe infestation, all 2. Medium to severe
needles brown or dropped
infestation, 50-100 % brown
off
needles
3. Medium infestation, 10-50
% brown needles
Photos: S. ŚLUSARSKI - IBL
4. Weak infestation, few
brown needles
5. No infection
The scheme is to easy-to-use. Also the small size of the trees is an advantage for the
assessment.
Sometimes it can be difficult to distinguish the damage by Lophodermium seditiosum from
other sources of browning of needles. For the statistical analysis it also has to be taken into
account that the distances between the single values of the scale are not equal. So analysis
of variance and estimation of variance components are not possible.
55
References
STEPHAN BR, KRUSCHE D, 1986: Genetic variation of resistance to Lophodermium needle cast in Scots
pine progenies of intraprovenance and interprovenance crossings. In Peterson GW (ed.): Recent
research on conifer needle diseases. USDA, Forest Service, GTR WO-50: 28-34.
3.232 Pine twisting rust
Definition
The pine twisting rust is caused by the fungus Melampsora pinitorqua. It causes severe
damage to the shoots of Scots pine seedlings and young trees, resulting in growth reduction,
malformation or even death, especially of seedlings. The pine twisting rust is very common
in the northern temperate and boreal areas, where also European aspen (Populus tremula
L.), which is the alternate host of the pathogen, is common. The pathogen enters the pine
seedlings in spring and early summer during shoot elongation. The occurrence of pine
twisting rust varies largely between years, due to its strong dependence upon various
weather factors. There is large genetic variation in the resistance of Scots pine against the
pine twisting rust, both within and between populations.
Damage by the pine twisting rust can be distinguished most clearly at the end of the
summer, when the effected parts of the shoots show resin secretion and become black. The
assessments should be made at a young stage, up to a height of about 1 meter, assessing the
whole tree. Due to large annual variation in the appearance of the disease, assessments
should be made during years when the disease is common or, alternatively, repeated
assessments should be made over several years.
Scoring schema
Stem deformation by Melampsora pinitorqua:
Scale values
0
1
Text
present
lack
56
(Photos: ANTTI POUTTU, Luke)
The scheme is easy to use. Also the small size of the trees is an advantage for the
assessment. Especially when not assessed at the right time, the damage may be confused
with other shoot deformations, such as those caused by insects.
References
ANDERSSON B, DANELL, O, 1997: In Pinus sylvestris resistance to pine twist rust associated with fitness
costs or benefits? Evolution 51: 1808–1814.
MATTILA U, 2005. Probability models for pine twisting rust (Melampsora pinitorqua) damage in Scots
pine (Pinus sylvestris) stands in Finland. Forest Pathology 35: 9–21.
QUENCEZ C, BASTIEN C, 2001: Genetic variation within and between populations of Pinus sylvestris l.
(Scots pine) for susceptibility to Melampsora pinitorqua Rostr. (pine twist rust). Heredity 86: 36–
44.
57
3.24 Mediterranean pines specific protocol
Core group: Eduardo Notivol (INIA),
Allain Bailly (FCBA),
Fulvio Ducci (CRA)
Mediterranean pines are not special from the point of view (need) of protocols
standardization. For all generic traits (according to a simple ontology mainly grouped in
growth and biomass, phenology, pests and diseases resistance and reproduction), the
assessing protocols should be the same that used in others pines or conifers.
The differential focus for this group of species is not “how” but “for what”, and this is
because the main purpose of these assessments is related to specific adaptation to the
particular Mediterranean environment. When conservation and landscape protection is
considered more interesting than wood production the interpretation of the results from
measurements are different even when the protocols are the same.
For forest trees, adaptation is important for any kind of environment but when species have
to face up to global change, some environments are more susceptible than others to the
consequences of that change. In this context there are two groups of traits related with
resilience facing drought stress and perturbances, mostly fire.
3.241 Drought
For drought tolerance a good compilation of proxy assessment is included at the end of the
document. Drought tolerance is not a trait and it is dependent not only on the plant but on
the environment. In addition measuring drought tolerance is not straightforward.
Drought tolerance for the same genotype varies with the soil characteristics, air
temperature, water availability, light intensity and their regimes. The interpretation of
results from measurements of mortality, soil water content, photosynthetic parameters,
transpiration, CO2 concentration, water potential, water use efficiency assessed by means of
isotopic discrimination, phytoregulators concentrations, terpene acids, or other by-products
from metabolism, etc. can be used for this purpose but without a standardized protocol due
to the different possibilities and framework of testing.
3.242 Flowering
Regarding reproduction, flowering and fructification assessments are not different for the
species group. Protocols for cones and flower counting are not different for the
Mediterranean pines compared with other species while other traits related to reproduction
58
like cone serotiny or masting can be highly relevant for Mediterranean pines. Unfortunately
there are not agreed protocols for these traits (but see HERNANDEZ-SERRANO, 2014).
Apart from multinomial calibrated scales (0 to 3; 0 to 5 similar to Scots pine or Norway
spruce), a continuous variable can be obtained by visually counting strobili (male or female)
in a fixed time (for example, 15 seconds), following protocols used in Californian oaks (KOENIG
et al. 2009). It has been used with good results in several ongoing papers (HERNANDEZ et al.,
SANTOS DEL BLANCO et al., CLIMENT et al.). A manual click counter helps homogeneity among
observers and observations.
3.243 Polycyclism
Polycyclism (the ability for a plant to produce several flushes in the same growing season)
can be contemplated as vegetative or reproductive. In Aleppo pine (Pinus halepensis), but
also occasionally in maritime pine (Pinus pinaster), multiple female cone cycles can be
observed. Reproductive polycyclism can be recorded as presence / absence in each tree or
rate of reproductive polycyclisms compared to a fixed number of reproductive shoots.
59
Quantitative way of assessment is made by means of counting the number of twigs and
number of cycles for rates calculation (PARDOS 2003, GIRARD et al. 2011).
3.244 Bark thickness
Bark can be a paramount trait in protective strategies against forest fires. This trait (bark
thickness) is recorded by means of bark calipers with no special requirements. Assessment
of this trait is usually made by cross measurements in two directions (N-S and E-W).
References
CLIMENT JM, CHAMBEL MR, LÓPEZ R, MUTKE S, ALÍA R, GIL L, 2006: Population divergence for heteroblasty
in the Canary Island pine (Pinus canariensis, Pinaceae). American Journal of Botany 93(6): 840–8.
CLIMENT JM, SAN-MARTÍN R, CHAMBEL MR, MUTKE S, 2011: Ontogenetic differentiation between
Mediterranean and Eurasian pines (sect. Pinus) at the seedling stage. Trees - Structure and
Function (25): 175–186.
CLIMENT J, MARTÍN-SANZ R, SANTOS DEL BLANCO L , CHAMBEL MR, NOTIVOL E, 2014: Conference Paper:
Genetic, environmental and ontogenetic effects on cone serotiny in Aleppo pine (Pinus halepensis
Mill.) MEDPINE5. 5th International Conference on Mediterranean Pines; 09/2014
CLIMENT JM, et al., (in prep): Population differentiatidon for life history traits in Aleppo pine (Pinus
halepensis Mill.)
CLIMENT JM, et al. (in prep): Divergence between subspecies of European black pine (Pinus nigra
Arnold) on life history traits.
ESPELTA JM, ARNAN, X, RODRIGO A, 2011: Non-fire induced seed release in a weakly serotinous pine:
climatic factors, maintenance costs or both? Oikos 120 (11): 1752–1760.
60
GIL L, LÓPEZ R, GARCÍA-MATEOS Á, GONZÁLEZ-DONCEL I, 2009: Seed provenance and fire-related
reproductive traits of Pinus pinaster in central Spain. International Journal of Wildland Fire 18 (8):
1003.
GIRARD F, VENNETIER M, OUARMIM S, CARAGLIO Y, MISSON L, 2011: Polycyclism, a fundamental tree growth
process, decline with recent climate change: the example of Pinus halepensis Mill. in
Mediterranean France. Trees-Structure and Function, DOI: 10.10: 1–12.
HERNÁNDEZ, et al. (in prep.): Genetic structure of cone serotiny in Aleppo pine (Pinus halepensis Mill.)
KOENIG WD, KNOPS JMH, CARMEN WJ, SAGE RD, 2009: No trade-off between seed size and number in the
valley oak Quercus lobata. The American Naturalist 173 (5): 682–8.
MAGINI E, 1969: The heritability of the stem form in Pinus pinaster Ait. 2ème Consultation Mondiale
sur l’Amélioration des arbres forestiers. Washington FO-FTB 3/8: 351-360.
MARTÍN-SANZ R, NOTIVOL E, SANTOS-DEL-BLANCO L, SAN MARTÍN R, CHAMBEL MR, CLIMENT J . Phenotypic
plasticity and allometric effects of cone serotiny in a Mediterranean conifer (In prep.)
PARDOS M, CLIMENT J, GIL L, PARDOS JA, 2003: Shoot growth components and flowering phenology in
grafted Pinus halepensis Mill. Trees Struct Funct 17: 442-450
SANTOS DEL BLANCO et al., (in prep.): Quantitative genetics of male and female reproduction in pines.
61
3.25 Larch specific protocol
Core group: Luc Pâques (INRA),
Volker Schneck (vTI now THÜNEN)
3.251 Phenology
Definition
Bud burst and bud flushing are the most critical parts of phenology in larch as they are
directly related to the risk of late frost damage. They are also a clear signal of re-start of
growth of primary meristems. A large variability at population and individual levels is
observed for that trait.
Way of assessment recommended
For young trees, that is up to a height the terminal bud is easily observable (2-3 m),
observation of bud flushing can be limited to the terminal bud. In this case, at each date of
observation, a score is given following the scoring-scale.
Observation can be done weekly if the kinetics of flushing is of interest; if not, it can be done
at only one date when 50 % of the trees are at Stage 3.
62
Score
For older and taller trees, that is when total height prevents an easy observation of the
terminal bud (above 3 m), the international BBCH scoring system will be favoured and has
been adapted to larch. The observation considers all vegetative buds of the whole crown.
Scores range from 0 up to 31. Scores 0 to 10 consider the phase up to the first buds reached
score 5; scores 11 to 19 evaluate the proportion (from 10 % to 90 %) of buds having reached
stage 5; score 31 corresponds to the stage when bud flushed is achieved and first twig
elongation is observed. As for young trees, observation can be done weekly or at only one
date. Trees are considered flushed at the date when they reached BBCH-score 15.
63
Code
BBCH
Stage
0
1
3
7
9
leafing
leafing
leafing
leafing
leafing
All vegetative buds at stage 0 (dormant)
Most of buds have reached stage 1
10
leafing
First buds have reached stage 5
11
leafing
10% of buds with score 5
12
leafing
13
leafing
14
leafing
15
leafing
16
leafing
17
leafing
18
leafing
19
leafing
31
growth
Start of twigs elongation
Description
Most of buds have reached stage 2
Most of buds have reached stage 3-4
3.252 Canker
Definition
Larch canker due to Lachnellulla willkommii is the most common disease observed in
European larch, especially in alpine populations, but Sudetan and Central Poland populations
are not exempt.
Canker produces deformation observable on branches and on stem, including at some
stages, resin flow and presence of fructification. In the worse cases, girdling of the stem
might be complete.
64
Method of assessment
Observations can usually start when trees are around 10 years old, rarely before except in
case of artificial inoculation.
- Artificial inoculation :
Inoculation is done through insertion of a calibrated sorgho straw infected by Lachnellulla
willkommii, in a hole drilled by a machine. A nearby hole is performed for control. Young
trees (2 years old) and young organs (1-2 years-old branches) can be used (SylvestreGuinot and Delatour, 1983).
Assessment is usually done 6 months after inoculation by measurement of the length
(mm) of the canker, the appreciation of the proportion of the twig girdled (ex. 1/3) and by
the notation of resin flow and of fructification (presence/absence).
- Observation in nature
The severity of the disease is appreciated through the presence (1) vs absence (0) of
symptoms (0) on stem or branches. The frequency of trees of a given genotype presenting
canker is then calculated. (SCHOBER 1985).
Scoring schema
Scale values
1
0
Text
Disease present on stem or branches
Disease absence
Method recommended:
Artificial inoculation has the main advantage to allow a much earlier screening of trees and
in a relatively fast way (6 months). Compared to observation in natural conditions, the
inoculum source is controlled but limited, applied at a high concentration and through manmade damage (hole), which might amplify the response.
65
Reference
SYLVESTRE-GUINOT G, DELATOUR C, 1983: Possibilités d’appréciation de la sensibilité du genre
Larix au Lachnellulla willkommii (Hartig) Dennis par inoculation artificielle. Ann.Sci.For. 40:
337-354.
SCHOBER R, 1985: Neue Ergebnisse des II.Internationalen Lärchenprovenienzversuches von
1958/59 nach Aufnahmen von Teilversuchen in 11 europäischen Ländern und den U.S.A.
Schriften aus der Forstlichen Fakultät der Univ. Göttingen, Band 83.
3.253 “Drought” cracks
Definition
Cracks are sometimes visible along larch stem. Their origin is not well known (growth speed,
drought?) but is more frequent in drier sites (shallow soil, southern aspect). The defect may
be considerable as the cracks usually turn around the stem over one to several meters and
can be as deep as the pith. After a few years, the crack might be filled up through healing,
which makes the defect less easily observable. But an attentive observation allows it to be
detecte through the resulting cicatrix.
All species of larch are susceptible, but more frequently this defect is observed on European
larch than on hybrid and finally on Japanese larch.
Way of assessment recommended
The observation requires a close look at stem and to turn around it to have a full vision as
the cracks are not necessarily oriented and are usually hidden by branches.
The best season is during winter time when needles have fully fallen off.
The defect seems to appear at a juvenile stage when trees are 10-15 cm diameter thick at
BH, which seems a critical stage.
66
Score
A simple scoring system, namely presence/absence is used.
References
CAZAUX JP, CHEVALIER R, GILBERT JM, GINISTY C, 1994: Le mélèze hybride en plantation - résultats
provisoires sur 17 sites en France. (Hybrid larch plantations - provisional results from 17 sites in
France). Etudes du CEMAGREF: Série Gestion des Territoires 12: 149-162.
67
3.26 Conifer non-species specific protocol
Core group (conifer core group leaders):
Luc Pâques (INRA; Larch),
Luc Harvengt (FCBA; Norway spruce),
Jean-Charles Bastien (INRA; Douglas-fir),
Egbert Beuker (METLA now LUKE; Scots pine),
Eduardo Notivol (INIA; Mediterranean pines)
3.261 Stem straightness
Definition
Stem deformation includes several defects: basal sweep, lining, bending, crookedness,
twisting. Their origin can be genetic (due to delayed lignification, production of compression
wood?) or accidental. The trunk aspect may change over time with a more or less rapid
apparent correction due to the accumulation of growth layers.
It is recommended to separately evaluate these different defects for a better description of
stem form and a separation of accidental from genetic effects.
Stem sinuosity or stem crookedness (vs Stem straightness) aim to assess the external quality
of the stem in connection to local deviations from the main tree axis, present along the
stem, in any direction. It also includes twisting defects common in some species (larch).
Severity of deviations is more or less important: in some cases, the defects disappear
externally after a few years, but in the worse cases, they remain over years and are highly
detrimental to wood quality. These should be distinguished stem deformation due to
intrinsic causes (like due to lignification problems) and stem defects clearly resulting from an
68
identified biotic (e.g., Rhyacionia buoliana in pines) or abiotic (such as frost/drought damage
to terminal buds) factors. In the latter case, a rupture of direction at a given annual
increment is observed.
In larch species, stem crookedness is clearly a major defect which might be accidental but
which has overall a strong genetic control.
Age: For most studies, evaluation should be done when trees are enough developed and
better express defects, that is when trees are between (5)-10-15-(20) years old. Afterwards,
stem correction through diameter enlargement hides defects which nevertheless remain
within the log.
Season: Observation should be done during the winter time, when the stem is fully visible,
that is in absence of needles.
Assessment: it is recommended:
1) to note separately the different defects such as basal sweep, lining, bending and
stem sinuosity,
2) to use the scoring scale in an absolute manner (agreed by participants in Leuven)
which means that a given score must have the same meaning across different sites;
3) to limit thus observation of crookedness to the part of the stem above 1.30 m (to
avoid basal sweep) and avoid the last 2 annual increments: in some species, this part
of the stem is heavily influenced by local weather conditions (wind) and corrections
of the stem appear in next season(s).
Before starting observation:
4) to walk through the plantation and find reference trees for each score, these trees
will be marked and referred to several times if needed during the assessment. Picture
of these trees should be taken and archived for further documentation.
During the observation:
5) to observe trees in two perpendicular planes, so to have a better vision of defects,
6) a given block will be assessed by only one observer, so that observer effects will be
included in block effects,
7) in case of doubt between two scores, use the smaller one.
According to available resources, it is recommended to have notation by two independent
observers.
69
Scoring scheme
The following scoring system is proposed: it combines a count of crooks along the stem
together with an appreciation of the severity of the crooks.
Scores 5 and 4 include what can be seen as ‘straight’ trees: defects in Score 4 are weak
enough to suppose that they will disappear with time and have no or little effect on wood
quality;
Scores 1 and 2 include ‘crooked’ trees: the defect is so severe that it is supposed to remain
over time and strongly affect wood quality (purge needed).
This scoring system can be used in two different ways:
1) in an “absolute” manner, which means that a given score will have the same
meaning across different sites;
2) in a “relative” manner: which means to adapt the scoring scale to each site.
Pros and cons
The advantage of the “absolute” manner is that it allows a real comparison of results over
sites (and ages); the inconvenience being that in a given site, the distribution of scores will
be asymmetric.
The advantage of the “relative” manner is that it allows normalising the distribution of
scores; but it prevents an objective comparison among sites. Comparison could only be done
through genotype ranking with a loss of information.
At the Leuven workshop (September 2009), participants agreed that the most important
factor in international trial evaluation was the possibility to compare genotypes’ behaviour
across sites and the absolute way was retained.
70
References
KEIDING H., 1966: Aim and prospects of taek breeding in Thailand. Nat. His. Bull. Siam. Soc. 21 (1 & 2)
3.262 Basal sweep
Definition
Basal sweep is a trunk deformation observed at the base of the tree. It can be of genetic
origin or accidental (in connection with a poor plantation, poor weed control or wind
damage, snow creepage on steep slope). Some species, like maritime pine, larch, etc., are
highly susceptible to this defect
Assessment
A subjective scoring (1/0: presence /absence of the defect) might be enough in most cases.
When a more objective evaluation is needed, we recommend the following procedure:
1.30 m
The longer horizontal distance (d) between the deformed tree and a vertical line up to 1.3 m
will be measured to evaluate the basal sweep. The vertical line is materialised by a pole
standing at the base of the stump and maintained vertical (plumb line, or ). The measure (d)
is expressed in centimeters.
Recommendation
The deformation is observed in the plan where the defect is the most severe and up to 1.3
m.
Pros and cons
Several other methods exist as shown in next figure.
71
Advantage
Inconvenient
Method 1
Most rapid & simple
But not always 2 points
of contact with trunk
Method 2
Adapted to all forms of
basal sweep and more
precise
Need a pole with a
spirit level and a
mobile gauge to
measure deviation
Method 3
Apparently convenient
method
- needs 2 measures (150 cm
and HT),
- difficulties if weed at the
base of the tree - difficulties
to measure HT with enough
precision
Reference
MAGINI E; 1969: The heritability of the stem form in Pinus pinaster Ait. 2ème consultation mondiale sur
l’amélioration des arbres forestiers. Washington FO-FTB 3/8. 351-360.
3.263 Forking
Definition
Forking is considered when the main stem axis splits (usually in conifers) into 2 stems of
equal importance (similar diameter). Other types of split of the main axis can occur when,
for example, a vigorous branch takes an up-right position: in this case, the second axis is
usually of a smaller diameter: it is then called ‘ramicorn’.
Forking can be genetically inherited and/or be accidental due to wind, snow, frost, birds,
etc., damages. Re-iteration of the defect is possible.
Assessment
It is recommended to adopt different protocols according to the age/size of trees:
For young trees (< 15 m), scoring simply identifies trees with or without a fork (1/0).
For older trees (> 15 m), the assessment combines the position of forking occurrence and
the number of forks:
72
No fork
Position: relative to tree
height
Upper third
Fork
Middle third
Lower third
Score relative to position
4
3
2
1
Number of axis (forks)
1
n1
n2
n3
Forking index
Example: number of axes
Forking index
= (score x 10)/nber of axis
1
40
2
15
2
10
2
5
Recommendation
For species with usually no forking re-iteration, the score relative to forking position may be
suitable.
For species with usual forking re-iteration, it is recommended to combine both records
(position + number) into a forking index: the highest index being for non-forking trees and
for trees with a lower number of forks at a higher position.
3.264 Branching habit
Definition
Branching is commonly evaluated through 3 main parameters including branch thickness,
branch angle and branch number of primary branches along the trunk. A high density of
branches, coarse branches, and branches with an acute angle towards the trunk are indeed
considered to downgrade overall stem quality and affect internal wood properties (due to
73
knots size, angle, density). Thin, flat angle branches at a low density are commonly searched
for by breeders.
3.264.1 Branch angle
A 4-scale subjective scoring system is used judging the angle made by branches to the main
stem: the flatter the angle (close to 90° to the trunk), the better the branching.
Scale values Text
4
branch in the 1st quarter (angle between 67° and 90°angle close to 90°)
3
branch angle in the 2nd quarter (between 45° and 67°)
2
branch angle in the 3rd quarter (between 23° and 45°)
1
branch angle in the 4th quarter (below 23°)
The rationale of this scoring system is that an observer can quite easily figure out the 4
quarters of the angle between the trunk and a perpendicular horizontal line; firstly by
dividing the 90° into two parts and then again into two parts. So the observer starts to
position the branch in the first (45-90°) or in the second half (0-45°), and then respectively
by dividing it in two, in the 1st or 2nd quarter or in the 3rd or 4th quarter.
Recommendation:
Assessment is done at the age of 10 years after plantation, provided total tree height
exceeds 6 m. The height (or whorl) at which branching angle is to be assessed will be
decided within each trial network.
74
For some species, at certain ages and sites, branches can hang down at more than 90°. An
additional score (5) could be added to take this pattern into consideration.
For some species, branches can start from the trunk at a certain angle and then flatten
onwards. Angle from the closest part of the branch to the trunk should be then considered.
3.264.2 Branch thickness
A 4-scale subjective scoring system is recommended: it is made more objective by relating
primary branch sizes (diameter) to stem diameter.
Step
Description
4
branch diameter less than ¼ stem diameter
3
branch diameter between 1/3 and ¼ stem diameter
2
branch diameter between 1/2 and 1/3 stem diameter
1
branch diameter above ½ stem diameter
Recommendation
Assessment is done at the age of 10 years after planting, provided total tree height exceeds
8 m. In any case, the branches should be easily visible from beneath the trees.
The position of the whorl where branch thickness will be assessed (or the height of
branches) will be decided within each trial network.
3.264.3 Branch number
The total number of primary branches (above a certain diameter) is an important feature of
branching but its consequences on stem and wood quality are also influenced by the way
branches are distributed along the trunk.
A two-fold scoring system is suggested combining number and distribution of branches:
75
Score 1: distribution of branches along the stem
1 = not in whorls
2 = in whorls
Score 2: number of branches along the stem or in whorls
1 = many branches
2 = moderate number of branches
3 = few branches
Position
Number of branches
Not in whorls
1
Many
1
Moderate
2
Few
3
In whorls
2
Comments
- For some species like larch, branches are not organised in neat whorls so that the
distribution score may be useless. In such cases, only the score for branch number will
be used.
- If operators are able to identify which branching distribution is more favourable (in
relation to wood quality, …), one could consider combining scores so that the ‘best’
position and low branches numbers give the highest score (i.e., Low number in whorls =
3 x 2 = 6).
Recommendation
- The height (or whorl) at which branching number is to be assessed will be decided
within each trial network.
- It might be recommended to fix the minimum diameter above which branches are
counted.
76
4. Trait-oriented common protocols
4.1 Wood quality
Manuel Touza Vazquez (CIDTG) (comp.)
There is an increasing interest regarding how to determine wood properties in standing
trees with non-destructive techniques (NDTs). This is of particular interest to breeders as
these new possibilities may be introduced into breeding programs, allowing, at the same
time, the preservation of the original trees.
One of the best known examples is the recent application of acoustic methods for estimating
the mechanical properties (modulus of elasticity) in standing trees.
Other available technologies include penetrometers (pylodin), drilling resistance methods
(resistograph), measurement of longitudinal surface growth strains with strain gauge devices
(as the one developed by CIRAD-Forêt), etc.
In other cases, traditional lab techniques together with X-Ray densitometry, fractometers,
etc., allow the determination of an increasing number of wood properties (density,
sapwood-heartwood ratio, early-wood-late-wood ratio, bending strength, compression
strength…) in increment cores.
NIR techniques also allow the determination of an increasing number of wood properties
from wood cores and/or wood power.
Due to the previous reasons, a questionnaire was drawn up on non-destructive methods for
assessing wood properties in standing trees and distributed by CIDTG in September 2014 to
WP2 and WP11 partners.
The questionnaire was based on the outcome of a previous survey carried out in the frame
of WP11 and was structured into four parts (General overview of available techniques and
equipment; Acoustic tools; Near infrared Spectroscopy and Innovation, and future trends).
The objective of the survey was to gain information on existing equipment and protocols for
determining wood properties in standing trees as well as to identify gaps and future trends.
For the purpose of the questionnaire, only wood properties determined on standing trees or
small wood samples (as a wood core or wood power) may be considered as non-destructive
as they will allow the preservation of the individual tree.
Twelve partners from ten countries responded to the WP2 survey (Partners 1-INRA [FR], 3ALTERRA [NL], 6-BOKU [AT], 8-CIDGT [PO], 9-CNR [], 10-CRA [], 12-FCBA [FR], 18-IICT [], 20INNVENTIA [], 24-METLA (now LUKE) [FI], 26-UGent [BE] and 28-vTI (now THÜNEN) [DE]).
The survey was structured into four parts:
Part I. General overview of available techniques and equipment
Part II. Acoustic tools
Part III. NIR
Part IV. Innovation and future trends
77
The responses were analysed in October 2014 and presented at the 3rd Annual meeting in
Tulln (AT). The main results are discussed below.
4.11 General overview of available techniques and equipment
Amap was created of the available techniques that are employed and/or being evaluated for
determining wood properties in standing trees. At the same time, several institutions are
evaluating the future use of the same techniques.
Project partners from ten countries responded to the WP2 survey
BOKU
CIDGT
CNR
IVALSA
CRA
FCBA
IICT
INNVENTia
INRA
METLA
(LUKE)
WUR
vTI*
U Gent
Pylodin
Resistograph
Acoustic
methods
Wood cores
Trad. Lab.
Wood cores
X-Ray densit.
Wood cores
NIR
Wood cores
Colorimeter
Rigidimeter
for MoE
*now THÜNEN
Codes employed:
Employing the technique
Evaluating the future use
Most experience in other fields (timber construction)
Only four technique, comprise the 80 % of the total that are being employed for determining
wood properties in standing trees.
78
Wood cores / Traditional lab. Procedures
Wood cores / X-Ray densitometry
Pylodin
Wood cores and NIR
Resistograph
Acoustic methods
Wood cores / Colorimeter
Rigidimeter for MoE
Wood density in standing trees is mainly determined employing a wood core. Traditional
volumetric methods and X-Ray densitometry are being employed in similar percentages.
Wood core and volumetric method
Wood core and X-Ray densitometry
Penetrometer (pylodin)
Wood core and NIR
The next graph points out how the different techniques agree with the expectations of the
users (1 Absolutely agree, 0,6 Rather agree, 0,3 Rather disagree, 0 Absolutely disagree).
Wood cores / Traditional laboratory
procedures
Pylodin
Resistograph
Wood cores / X-Ray densitometry
0
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
79
Half of the institutions that answered the survey are evaluating new techniques for
determining wood properties in standing trees. NIR, resistograph and acoustic methods
comprise 80% of the techniques being evaluated
3.5
3
Number of institutions
2.5
2
1.5
1
0.5
0
Resistograph
Wood cores and Acoustic methods Wood cores / XNIR
Ray densitometry
Pylodin
4.12 Acoustic tools
A map of the equipment being used, as well as the species tested.
Equipment
Director ST 300
Microsecond Timer
IML Hammer
Hitman
Ultrasonic Timer
CIDGT
x
x
x
Species tested
Pinus pinaster
Picea abies
Pinus taeda
Pseudotsuga menziesii
Eucalyptus spp
Populus spp
CIDGT
x
FCBA
x
INNVENTIA
(x)
x
FCBA
x
INNVENTIA
x
x
x
(x)
(x)
80
4.13 Near Infrared Spectroscopy (NIR)
Six of the twelve partners that answered the questionnaire are using NIR and two others are
planning to do in the future.
More than ten years
BOKU
FCBA
IICT
INNVENTIA
x
x
x
x
Between 5-10 years
CNR
IVALSA
INRA
x
x
Planning to do so in the
future
METLA
(LUKE)
U Gent
x
x
Together with traditional properties such as chemical composition or density, NIR is being
used increasingly in new applications such as wood identification, durability, surface
weathering, etc.
6
Number of institutions
5
4
3
2
1
0
Chemical
composition
Mechanical
properties
Physical
properties
Wood
identification
Durability
Surface
weathering
The partners were asked to select whether or not the following statements hold true for
them (1 Absolutely agree, 0.66 Rather agree, 0.33 Rather disagree, 0 Absolutely disagree).
NIR may save money and time in the determination of wood properties
It is better to work with wood cores for having an idea of the radial variation of the property
NIR will have an increasing use in the forestry sector
It is better to work in the laboratory with a properly seasoned sample than try to determine a
quick reference value in the field
NIR seem the most promising technique for determining new wood properties in standing trees
NIR equipment is expensive and require a well-trained, staff so its use will be restricted to a
reduced number of research institutions
0.94
0.94
0.83
0.78
0.66
0.61
81
The answers pointed out a clear agreement that NIR may save money and time in the
determination of wood properties, and as well that it is better to work with wood cores to
obtain an idea of the radial variation of the property.
4.14 Innovation and future trends
The next graph points out the main interests in employing NDTs in standing trees (1
Absolutely agree, 0.6 Rather agree, 0.3 Rather disagree, 0 Absolutely disagree).
All the partners absolutely agree that their main interest in using NTDs it a better
understanding of the effect of sylviculture on wood properties as well as to improving the
traditional breeding programs introducing new criteria.
Look for the genetic gain associated with
individual trees
Look for applications of the timber
Look for an early selection of the best
provenances/clones
A better understanding of the effect of
silviculture on wood properties
Improve the traditional breeding programs
introducing new criteria
0.6 0.65 0.7 0.75 0.8 0.85 0.9 0.95
1
1.05
The next graph shows the future trends regarding the use of NDTs in standing trees (1
Absolutely agree, 0.6 Rather agree, 0.3 Rather disagree, 0 Absolutely disagree).
There is a clear agreement that cooperation and the definition of common protocols are
essential for the development of NTD´s.
82
In 10 years my organization will determine additional
properties in standing trees
The information obtained from a wood core may give
an overall idea of the whole tree
The number of wood properties that may be
determined with NDTs in standing trees it is still very
reduced
In 10 years, new properties will be determined with
NDTs in standing trees
Cooperation and definition of common protocols, it
would be essential for the development of NDTs
0
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
1
The following is a list of innovative methods that have been recently developed by the
partners for the assessment of wood properties in standing trees:
• CIDGT: Tension wood in standing Eucalyptus globulus
• CIDGT: Acoustic methods for determining MoE in young trees (work in progress)
• CNR-IVALSA: X-ray tomography (density distribution and 3D maps)
• CNR IVALSA: Hyperspectral imaging (variation of wood properties in space, spectral
maps)
• IICT: Physical and chemical characterization using core sampling
• INNVENTIA: Radial variations in spiral grain for some hardwood species (work in
progress with larch)
• INRA: Rigidimeter on larch and Douglas fir
• U GENT: Microdensitometrical analysis of long wood cores with micro-CT
The partners were asked if they were interested in testing any particular wood property that
cannot be predicted nowadays. The following table shows the answers.
BOKU
Vulnerability to drought stress
Complete characterization of material –
multisensory approach
Retrospective assessment of annual
growth
Hydraulic conductivity using small
branches in Pinus pinaster
Whole-timber wood homogeneity and
performance during wood drying
Chemical composition and heat value
Spiral grain, Heartwood/sapwood limits
NIR models for the constitutive stilbene
content in Scots pine heartwood or
induced content in other tissues
CNR
IVALSA
FCBA
IICT
INNVENTIA
INRA
METLA
(LUKE)
vTI
(THÜNEN)
x
x
x
x
x
x
x
x
83
References
ALVES A, SANTOS A, ROZENBERG P, PÂQUES L, CHARPENTIER JP, SCHWANNINGER M, RODRIGUES J, 2012: A
common NIR-based PLS-R model for the prediction of basic wood density of Pinus pinaster and
Larix × eurolepis. Wood Science and Technology 46: 157–175 (DOI: 10.1007/s00226-010-0383-x).
ALVES A, SANTOS A, SILVA-PEREZ D, RODRIGUES J, PEREIRA H, SIMÕES R, SCHWANNINGER M, 2007: NIR PLSR
model selection for Kappa number prediction of Maritime pine Kraft pulps. Wood Science and
Technology 41: 491–499.
ALVES A, SCHWANNINGER M, PEREIRA H, RODRIGUES J, 2006: Calibration of NIR to assess lignin composition
(H/G ratio) in Maritime pine wood using analytical pyrolysis as the reference method.
Holzforschung 60: 29-31.
ALVES A, SCHWANNINGER M, PEREIRA H, RODRIGUES J, 2006: Analytical pyrolysis as a direct method to
determine the lignin content in wood. Part 1: Comparison of Py-lignin with the Klason lignin
method. Journal of Analytical and Applied Pyrolysis 76: 209-213.
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globulus wood extractives content by NIR-based PLS-R models: Comparison between extraction
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86
4.2 Drought resistance
Maria Cristina Monteverdi (CRA) (comp.)
Introduction
The report of the IPCC in 2007 confirms that in the last decades the climate is changing,
significantly affecting the natural resources on Earth. Several studies have shown how
climate change may affect the thermal regimes and precipitation, they are changing very
important physiological processes of ecosystems on our planet. According to the IPCC report
(2007), although the effect space-time climate change is still partially uncertain, it seems
certain that the adaptability / resilience of ecosystems, including forests, is at serious risk.
Hence the importance of studying the adaptive biodiversity in relation to climate change at
the inter - intraspecific of forest species, in order to identify adaptive traits useful for the
sustainable management of genetic resources. Most climate predictive models show
increased temperature trends of, and decreased precipitation trends with, a consequent
increase of evaporation rate. So the drought stress is one of important abiotic factor that
affect natural ecosystems, also forest ecosystems. Therefore, to assess the adaptive traits at
inter – intraspecific levels, they play an important role to identify early selection parameters,
suitable for new breeding programs of forest reproductive material, able to tolerate the
drought condition maintaining a good productivity. All of these tools help to produce forest
reproductive material suitable for a sustainable forest ecosystem management.
The purpose of this work is to find out a reference method for a joint evaluation that could
be identified and shared by scientific community. In particular, the purpose is to obtain
information on existing methods to assess drought resistance on their quantitative,
qualitative, and operational significance and to identify eventual gaps. A “Survey on different
methods to assess drought resistance and tolerance and related adaptive traits” was created
and sent to participants:
Some authors defined (LARCHER 1980, TESCHE, 1992) the drought resistance, such as the ability
of a plant to ensure its life processes, as keeping its water balance positive during a period of
drought as long as possible. In the following, the term of drought resistance will be used as a
generic term to describe the ability of tree plants to tolerate, resist, and avoid different
levels of water stress.
87
STRESS RESISTANCE (LEVITT 1972)
Stress avoidance
Stress avoidance/escape
(ability to avoid stress)
Tey are characters
incorporated into the
genotype, but they are not
really resistant plants.
Examples:
- Ability of some plants to lose
their leaves in the case od
water stresss
- effimer plants of the dessert
who make the life cycle only
when it rains.
Tolerance (real resistance to stress)
Avoidance of strain to stress
For example Drought stress
-Es: maintaining cell turgor
through osmotic
adjustments or cell wall
elasticity
Tolerance of strain caused by
stress
Sample: desert plants that
reach extremely low values of
water potential and continue
the work (the presence of a
little water in the tissues, but
the ability to maintain active
metabolism with internal
intrinsic mechanisms. For
example greater quantity of
water bound to the plasma
membrane.)
4.21 Methodology and Evaluation System
The survey is structured into three main parts:
A. Overview of parameters, methods, and species investigated.
B. Past experiences in drought resistance and tolerance studies (participation to
projects, and scientific papers).
C. Evaluation of methods according to the following criteria:
 objectiveness;
 capacity building;
 simplicity;
 destructiveness;
 cost.
4.22 Preliminary results
Participation of involved Institutions
Until now, ten Institutions from nine countries answered the questionnaire on a total of 20
participant Institutions on WP2+WP3+Wp11 (Table 4.2.1). Only 10 of 20 participant
institutions answered the questionnaire, of these only 4 of 10 are studying or studied
drought resistance.
88
Table 4.2.1: Participating institutions involved with the questionnaire, the number of those
interested and those that have not answered.
Figure 4.2.2: Results of participation
The geographic distribution of the institutions that answered the questionnaire shows two
distinct groups: those involved in the assessment of drought resistance (Mediterranean
countries), and those not involved in the study of drought resistance (northern Europe
countries).
89
Figure 4.2.3: Geographical distribution of participant institutions that answered to drought resistance
questionnaire. The red color indicates the countries involved on drought resistance studies, the blue ones the
countries currently not involved on drought resistance studies.
Table 4.2.2: Countries, institutions, and drought resistance interest
Countries
Austria
Netherlands
Spain
Spain
Italy
UK
Germany
France
Finland
Belgium
Respondent institutions
BOKU - Vienna
ALTERRA - Wageningen
INIA ACoruna
CITA - Aragon
CRA SEL
vTI (now THÜNEN)
FCBA
METLA (now LUKE)
VLAGEW
Drought resistance interest
Yes
Yes
Yes
Yes
Yes
No
No
No
No
No
Overview of parameters, methods, and species investigated
From the questionnaire, it appears that the forest species of interest about the drought
resistance are the following: Scots pines, black pines, Mediterranean pines, Norway spruce,
aspen, beech, ash, Pubescent oak, rowan, maple, oaks, Douglas fir, wild cherry, and silver fir.
Table 4.2.3 shows the main objectives of the study declared by involved institutions, with
the relative parameters investigated, forest species, and methods used.
90
Objectives
Parameters studied
Species
investigated
Method
Evaluation of drought
effects on biomass
production
Air moisture, soil moisture,
physiological drought
Norway spruce
Seedlings cultivation in growth chambers
under different air drought conditions
Individual survival
Scots pine, back pine,
Mediterranean pines
Norway spruce,
aspen,
beech,
ash,
pubescent oak,
rowan,
maple
Watering restriction in nursery /greenhouse
/phytotron, weighing method, TDR probes
Bench top dehydration,
air injection,
acoustic emission testing
P50 (the applied air pressure causing
50 % loss of conductivity),
80 % loss of conductivity),
RWL50 (the applied air pressure
necessary to cause 50 % loss of
relative water content)
Dry weight ration (leaf, stem, root)
Water Use Efficiency (WUE)
Oven and balance (dry at 70 °C for 72 h)
Mass spectrometer to measure carbon
isotopic composition (δ13C)
Gas exchange analysis, portable gas
exchange analyser (CIRAS-2, PPSystem)
Portable fluorescence monitoring system
(Hansateck Instruments, FMS 2)
Photosyntetic parameters (A; gs; T)
To study the
adaptation to climate
change
Chlorophyll fluorescence of
photosystem II (PSII)
Fv/Fm ratio,
φPSII (quantum yield of PSII
photochemeistry),
qP (photochemical quenching),
NPQ (non photochemical quenching)
Photosynthetic pigments
concentration (Chla, Chlb,
xanthophylls cycle)
Antioxidant enzyme activities
Proline production
Phytohormone production
Xylem anatomy
Root morphology
Hydraulic architecture
50 % loss of conductivity)
Water relations at cell level
Phenological plasticity
Cambial phenology
Carbon isotope discrimination ∆13C
(structural carbon and soluble
sugars), WUE
HPLC (High Pressure Liquid Chromatograph)
Oaks,
Mediterranean pines,
Douglas fir,
wild cherry,
silver fir
Enzyme extraction and spectrophotometric
assay
Proline extraction and determination (BATES
et al. 1973 ) spectrophotometer
Liquid chromatography coupled to tandem
mass spectrometry (DURGBANSHI et al. 2005)
Microscopic method
Digital camera, root system analysis
software WinRHIZO 4,0
Evaporative flux method (COCHARD et al.
1996) balance, Scholander pressure
chamber, sliding microtome
Cavitron technique (for cavitation
vulnerability curves (COCHARD et al. 2005))
Free transpiration method and sap
expression methods, Scholander pressure
chamber
Scoring system, phenologycal camera
(Twingscapes TimelapseCam)
Microscopic method
Extraction methods (BRUGNOLI et al. 1988)
measures carbon isotopic composition
(δ13C), dendro-isotopic analysis
Table 4.2.3: Main objectives of the study declared by involved institutions, with the relative
parameters investigated, forest species, and methods used
It can be noted overall, that most of the parameters used for the assessment of drought
resistance are physiological parameters (64 %), while the quantitative, morphological and
phenological parameters are used less, 14 % to 14 % - 9 % respectively.
91
Past experiences in drought resistance and tolerance studies (participation to projects, and
scientific papers)
The analysis of the results obtained from the questionnaire, indicates that of the institutions
that declared they work or have worked on drought resistance, 3 out of 5 have experience in
drought resistance and tolerance measurements prior to the year 2000, while 3 out of 5
started studies on drought resistance between 2000 and 2005.
Four institutions out of 20 are still studying drought resistance aspects, 1 out of 20 stopped
the scientific activities on drought resistance, while 5 out of 20 not actually involved in
drought resistance assessments, and more than 11 out of 20 did not answer the
questionnaire (Figure 4.2.4).
Most of the institutions are involved in international projects and networks, while only a
small part is only involved in national programs and projects on drought resistance
assessment.
Figure 4.2.4: Distribution of institutions (or groups) in those that are still studying drought resistance
aspects, those that have stopped working, those that declared they do not work on drought
resistance, and those that have not replied.
Most of the institutions said they don't have specialized staff. About half of institutions have
acquired the technical/scientific knowledge on drought resistance by learning by
themselves, while a smaller part conducted postgraduate and undergraduate studies,
specialized commercial courses in other countries in related subjects.
Methods evaluation
Objectiveness of methods
The partners involved in the questionnaire indicated 21 different methods for assessing
drought resistance. All methods are described in the literature, and almost all are
standardized methods according to existing protocols.
92
Capacity building
Only 3 out of 21 methods below are easily applicable in the field. Approximately half of
below methods require voluminous equipment, and only a few (2 out of 21) need for special
infrastructure. Almost all of the below methods (86%) allow statistically valid information to
be acquired in a short time. On 21 methods about half allow measurements of multiple
parameters to be collected at the same time.
Destructiveness
Most of the methods identified are not destructive.
Table 4.2.4: Estimation of destructiveness and/or invasiveness of methods
Method
Bench top dehydration
Incremental reaction to reduce air moisture
Watering restriction in nursery/greenhouse/phytotron
Dry weight ratio
Stable isotope analysis
Photosynthetic pigments analysis (Chla; Chlb; xantophylis cycle)
Antioxidant enzymes activities
Proline and phytohormone production
Xylem anatomy
Root morphology
Hydraulic architecture
Pressure-volume curves
Cavitation curves
Gas exchange method
Scoring system, phonological camera
Cambial activity / microscopic method
Extraction of soluble sugars methods (BRUGNOLI et al. 1988), measures
carbon isotopic composition (δ13C), dendro-isotopic anylysis
Invasive
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
Yes
No
No
Yes
No
Destructive
Yes
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
No
Costs
The cost analysis is currently in progress due to the absence of direct information.
4.23 Conclusions
Currently we can only deduce that the physiological traits are the most investigated to
assess the drought resistance. This is probably because of the complexity of information that
can be provided, but also for the capacity building of the methods used for their assessment.
Normal are methods that allow statistically valid information to be acquired in a short time
93
and are often able to provide simultaneous measurement of multiple parameters. Moreover
they are generally not destructive methods.
In any case, due to the low number of questionnaires filled out, the results obtained until
now do not allow a reference method for a joint evaluation to be identified and shared by
scientific community. So these results cannot be considered definitive. In order to get a valid
result, the questionnaire should also be sent to the remainder of the project participants and
integrated all with the results obtained in TREEBREEDEX project. Otherwise, it could be a
good solution to take as reference the results obtained in the TREEBREEDEX project about
the reference method for drought resistance assessments.
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99
5. Reference genotypes
Luc Pâques (INRA) (comp.)
Definition
In the course of the TREEBREEDEX project, the need to share common reference genotypes
for main forest tree species was noted several times. This might be useful for several
research purposes.
The concept “genotypes” encompasses different types of genetic units: seed stands, seed
orchards, full-sib progenies, half-sib progenies and clones. According to research needs, at
least three different types of reference genotypes have been identified, directly linked to
genetics & breeding activities or more broadly to ecological studies.
Type 1 ‘control genotypes’: in the course of their breeding programme, breeders evaluate
their own genotypes and candidate new varieties (for example for certification) relative to
some controls towards which they rank their genotypes and estimate genetic gains.
Comparison of gains is rarely possible at international level because of this lack of common
references.
Type 2 ‘signal genotypes’: evaluation of some traits is complex because either it relies on
subjective scoring (e.g., stem straightness, branching) or it is done relative to known
genotypes within a restricted collection of genotypes available in a country (e.g. phenology,
disease resistance). ‘Signal’ genotypes would be useful as references along scoring scales
(straight vs crooked/ early flushing vs late flushing/ resistant vs sensitive, etc). They would be
unavoidable for international evaluation and later on for comparison of results.
Type 3 ‘widespread common genotypes’: for some ecological studies, in particular to
monitor climate changes impact (phenotypic plasticity study; phenology modelling; pest &
disease epidemics monitoring; etc), the creation of a dense grid of sites with the same
genotypes, across contrasted ecological conditions through Europe, would be highly
beneficial. To be feasible, the creation of this network should be done in a soft, low-cost
way: for example in any new field trial, some few trees (how many?) from various genotypes
of some species could be systematically planted, additionally to the main experiment.
Further description will be given below. To be successful, conditions are that:
a) we are able to identify such genotypes for a given set of species of commercial interest,
b) we are able to make them available to the scientific community,
c) we agree to use them as much systematically as possible.
Objective
The objective of this task is to come up with a roadmap to build up this “reference
genotype” provider service and hopefully to implement it as far as we can for a couple of
species.
Steps include:
100
- identification at national levels of existing genotypes with adequate properties and/or of
possibilities to identify such genotypes: collection of this information, agreement on a list
of suitable ‘reference genotypes’,
- identification of national (infra-)structures which could potentially offer this ‘sharing’
service (seed storage facilities, veg. prop. facilities (nurseries, tissue culture/ somatic
embryogenesis lab, etc.)): these structures would have the responsibility to
store/preserve the material (seed storage, cryoconservation, rootstock hedges…), to
mass-produce it (collect seed lot or buy it, veg. prop. clones) and to disseminate it
according to needs. Most probably more than one structure/species is wished.
- identification of technical, legal and financial conditions to reliably establish such service.
Description of reference genotypes
The nature, the characteristics and the use of these reference genotypes will vary according
to their objective.
Nature of genotypes
Characteristics
Use
Conditions
Type1: ‘Control’
Variable according to
the species breeding
strategy and its level of
advancement:
- synthetic varieties
from seed orchard (ex.
for conifers)
- clones (ex. for vegprop. based strategy:
ex. poplar, wild cherry)
- selected seed stands
(i.e., starting
programme)
Genotypes of a given
species, the best
known material
‘Regionalised’
(according to biogeographic zones)
material could be
needed according to
species (ex. Norway
spruce)
In international and
national genetic trials
(e.g. evaluation,
certification trials)
- mandatory in
international trials
- free but
recommended in
Type2: ‘Signal’
Preferably clones
But some other
material (FS…) might
be necessary when
veg. prop. is not/hardly
possible
Type3: ‘Widespread
Preferably clones
But some other
material might be
necessary when veg.
prop. is not/hardly
possible (FS
progenies….?)
Genotypes of a given
species; contrasting
pairs of genotypes for
given set of traits
representing extreme
scores on evaluation
scale
‘Regionalised’ material
could be needed
according to species
Genotypes from
several species; from
different bioclimates
across Europe
In international and
national genetic trials
of all types
Potentially in all new
(genetic) trials to be
established, whatever
the species tested
Recommended
- mandatory in
international trials
- free but
recommended in
101
Volume of plants
national trials
Like any other genetic
units tested in the trial
for a given species
national trials
A few (2-5?) ramets /
clone or sibs / family
(<10?) for a given
species
A few (5-10?) trees
from each species,
whatever the main
species tested
Remarks:
Whatever the types of reference genotypes,
1) genotypes should be (as) stable (as possible) in their genetic composition (collection only
in good flowering years, mixture of crops from several yrs, …) and their genetic conformity
guaranteed. Reference DNA samples could be stored at the Repository Centre (AIT) for
control.
2) the most straightforward references to implement this service will be genotypes already
mass-produced (from seed orchards, commercially propagated clones), certified and
commercially available: these should be probably favoured whenever they fit the above
requirements (like for type 1 and perhaps type 3 genotypes above).
102

D2.1 Common protocols and reference standards for

Transcription

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