Wild cherry - Treebreedex

Wild cherry (Prunus avium L.)
micropropagation for
plantation establishment
Szczygieł K. and Wojda T.
Forest Research Institute
Dept. of Genetics and Forest Tree
Physiology
Prunus avium L. is one of the most important
hardwood trees,with valuable wood, suitable for
producing timber for cabinet making, furniture,
parquet floors, musical instruments.
It has a wide natural distribution in Poland.
The map of natural range distribution of wild
cherry in Poland (Borowicz i Gostyńska-Jakuszewska 1974)
MEAN DIAMETER AND HEGIHT OF WILD
CHERRY AT THE AGE OVER 50 (DATA
30.06.2003)
Age (years)
51-60
Nr of stand
units
297
Mean DBH
(cm)
30,5
Mean height
(m)
18,5
61-70
133
34,4
19,6
71-80
50
36,4
20,1
over 80
31
39,5
21,4
The genetic improvement programmes for this
species have been started in 1991. Up to now there
are selected 244 of plus trees located in all regions
of Poland, and 2 seed orchards in Forest District
Świdnik and Dynów have been established
(plus tree in Forest
Dist. Lwówek Śląski)
(plus tree
in Forest
Dist.
Gdańsk –
age-80,
diameter
46cm,
heigh-28m)
For breeding and improvement purposes the
method of wild cherry micropropagation by
organogenesis of selected clones was
applied, as well as for obtaining planting
stock through the nursery system and into
the plantation (production plantation
forest).
The present studies were started in 2007,
and they have been aimed at applying tissue
culture method to obtain the planting stock
of selected genotypes for plantation
establishment, and to conserve the best
clones of P. avium in clone archive.
• Explants: The current year 2-5 cm long shoots or
shoots with developing buds from early spring 2007
and 2008, were collected as primary explants for
clones regeneration. Also, the new cultures were
obtained from leaves (secondary explants) excised
from shoots proliferating in vitro. The explants
were taken from 20 clones of 2 years old seed
orchards established in Forest District Świdnik
and Dynów
•
Sterilization: First, the shoots were washed in
water with a drop of 20% Tween, then sterilized
in solutions containing 0.1% of mercuric chloride or
in 13% of sodium hypochlorite for 10 minutes
each, and followed by three time washing in sterile
distilled water
• Initiation of organogenesis: The sterilized
plant material was transferred on the MS
(Murashige and Skoog 1962) basal medium,
supplemented with: 200 mg·l-1 glutamine,
200 mg·l-1 casein hydrolysate (Mala
personal com.), 2 mg·l-1 BAP (Tang et al.
2002) and 0.1 mg·l-1 IBA, and 30 g·l-1 of
sucrose.
• The pH of all media were adjusted to 5.6.
• Multiplication of adventitious buds and
shoots: Performed in the same medium as
above mentioned but supplied with lower
BAP concentration (1 mg·l-1) or with the
same (2 mg·l-1).
• Shoot elongation medium: With the use of
basal medium, without growth hormones or
supplied with 1g·l-1 of activated charcoal
(for 3-4 weeks).
•Rooting of micropropagated shoots:
After removal of the lower leaves and small
shoots, the shoots of 1-2 cm long were
rooted on MS medium with 100 mg·l-1 of
glutamine, 100 mg·l-1 of casein
hydrolysate, 2-3 mg·l-1 of IBA, 20 g·l-1 of
sucrose (for 3-4 weeks)
The cultures were maintained in the growth
chambers at the temp. 23°C during day and
18°C at night, 16 h day (light intensity 30
µmol m-2sec-1).
Adaptation of plants to growth and development
under natural conditions:
• transferring the jars with rooted shoots to
greenhouse for 10-14 days,
• transplantation of rooted shoots to natural
substrate (mixture of peat, vermiculite and perlit
50:25:25%) and cultivation for 2 weeks under high
(80-90%), then after 2 weeks gradually lower (6070%) air humidity,
• early spring adaptation outside the greenhouse for
2-3 months,
• transplantation to the nursery for 2 years and
measurements of height growth increment and
phenology of buds in spring and in autumn,
• establishing from obtained plant material the
research area in Forest District Świdnik and clone
archive in the Arboretum in Syców
Primary results and conclusions
• the spring current year 1-2 cm long shoots were
more suitable as a source of explants then flushing
buds
• the young leaves obtained by explant multiplication
were very useful as a secondary explants for
future proliferation
• the sterilization of explants in 13% sodium
hypochlorite for 10 minutes was not successful,
and the material from the forest needed stronger
disinfection in 0.1% HgCl2
• most explants from the seed orchard in Forest
District Dynów showed infection by fungi and
bacteria, the only 4 from 10 clones were possible
to disinfect
• from all explants (10 clones from seed orchard in
Forest District Świdnik) the regeneration have
been obtained
•depending on explant genotype clones revealed
different ability of regeneration – between and inside
provenance (Świdnik and Dynów), and also they
needed individual concentration of BAP in the basal
medium for multiplication. Addition 2 mg·l-1 of BAP
to the MS medium was optimal for most of clones
•the media MS0 (i.e. without growth hormones) or
supplied with activated charcoal were efficient for
shoot elongation before rooting
Influence of BAP concentration in MS medium on
fresh weight increment of wild cherry shoots
•root induction and growth appeared after 3-4 weeks
of shoot cultivation on medium with IBA, however the
spontaneous growth of roots on MS0 medium have
been observed. For most of clones the optimal
concentration of IBA was 3 mg·l-1. The percentage
of rooting ability was in the range of 40-90%
Influence of IBA concentration on rooting percentage
of 3 clones of wild cherry
• after 2 months of acclimatization on May 2008, 712
seedlings were planted into nursery. After the first
vegetation season (May-October), in autumn 2008, the
measurements of growth increment (20-42cm) and
observations of seedlings with high survival of 87.1% were
made. In this autumn, we also planted a new part of
seedlings (300) and we still produce the new ones
• in autumn of 2009, the research area with the
micropropagated clones which are growing in the nursery will
be established
Thank you
for your
attention !