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İletiTarih: 26 May Per, 2016 3:07 pm    İleti konusu: Alıntıyla Cevap Gönder

http://www.cost873.ch/_uploads/_files/m_Bourrain_InVitroPropagationWalnut.pdf
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İletiTarih: 26 May Per, 2016 3:08 pm    İleti konusu: Alıntıyla Cevap Gönder

http://www.euroavia-ge.com/index_htm_files/aclimatization.pdf
http://journals.usamvcluj.ro/index.php/horticulture/article/viewFile/8456/7181
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İletiTarih: 26 May Per, 2016 3:09 pm    İleti konusu: Alıntıyla Cevap Gönder

https://hal.archives-ouvertes.fr/hal-00882513/document
Micropropagation of hybrid walnut trees (Juglans nigra
x Juglans regia) through culture and multiplication of
embryos
D. Cornu, C. Jay-Allemand
To cite this version:
D. Cornu, C. Jay-Allemand. Micropropagation of hybrid walnut trees (Juglans nigra x
Juglans regia) through culture and multiplication of embryos. Annales des sciences foresti`eres,
INRA/EDP Sciences, 1989, 46 (Suppl), pp.113s-116s. <hal>
HAL Id: hal-00882513
https://hal.archives-ouvertes.fr/hal-00882513
Submitted on 1 Jan 1989
HAL is a multi-disciplinary open access
archive for the deposit and dissemination of scientific
research documents, whether they are published
or not. The documents may come from
teaching and research institutions in France or
abroad, or from public or private research centers.
L’archive ouverte pluridisciplinaire HAL, est
destin´ee au d´epˆot et `a la diffusion de documents
scientifiques de niveau recherche, publi´es ou non,
´emanant des ´etablissements d’enseignement et de
recherche fran¸cais ou ´etrangers, des laboratoires
publics ou priv´es.
Micropropagation of hybrid walnut trees
(Juglans nigra x Juglans regia) through culture
and multiplication of embryos
D. Cornu C. Jay-Allemand
Am6lioration des Arbres bresForestiers, Forestiers, INRA, Ardor Ardon, 45160 Olivet, France
Introduction
Walnuts are very valuable trees for nuts
and wood production, but we see a reduction
in the number of logs available for
veneer. Although Persian walnut (Juglans
regia) and black walnut ((J. nigra) can be
used, hybrids between these two species
have better growth and wider adaptability.
Breeding programs are in progress at
INRA (Institut National de la Recherche
Agronomique, Bordeaux and Orl6ans) and
they need efficient methods of vegetative
propagation. Micropropagation was established
in 1984 for ’Paradox’ (Driver and
Kuniyuki, 1984) and recently for Persian
walnut (McGranahan et al., 1988). Previous
works have shown strong effects of
ageing and rejuvenation (Jay-Allemand et
al., 1988), latent contamination, low reactivity
of buds or meristems on the establishment
of mature selected clones. On
the contrary, with very juvenile material,
such as embryos, it is possible to avoid
these problems (Jay-Allemand and Cornu,
1986; Heile-Sudholt et al., 1986).
The purpose of this study was: 1) to
estimate the ability of hybrid progeny to be
propagated by micropropagation, and 2)
to improve culture factors acting on shoot
development.
Materials and Methods
This study used half-sib hybrid nuts (J. nigra
(no. 23) x J. regia) supplied by E. Germain
(INRA, Bordeaux), collected in September
1987. Embryonic axes (48) were isolated under
sterile conditions and then introduced in vitro
into the medium defined by McGranahan et al.
(1987). Three main steps have been determined:
1) elongation of epicotyls and buds during 3
wk of darkness followed by 2 wk of 16 h of light at 28°C; 2) multiplication by transferring nodes
from elongated shoots or clusters of buds every 2 wk (16 h light, 28°C); 3) rooting (not presented
in this paper).
Two kinds of solidifying compounds (DifcoBacto
agar, 6 g/I, and Gelrite, 2.3 g/I) in 750 ml
jars were compared. Then, instead of one
transfer onto a fresh medium after 2 wk of culture,
the addition of about 2 cm of a new liquid
medium without transfer was studied (double
phase system, Viseur, 1987).
The number of shoots (>5 mm) was determined
for each clone at the 3rd and 8th transfers,
and the number of bud-clusters and
elongated shoots (>15 mm) at the 10th, 11th,
l2th and 13th transfers.
Results
The 48 clones which were cultivated
under the same conditions show great
variability in their bud-cluster development
and shoot elongation. After 3 and 8 transfers,
we obtained a normal distribution of
clones (Fig. 1A and iB). Ranking of some
clones changed during this time but stabilized
after the 8th transfer. Eight of the
best clones were selected for bulk propagation.
They were characterized by good
development of leaves and elongation of
shoots. The production of buds and
shoots during 3 transfers is summarized in
Table I. In 6 wk, the number of bud-clusters
multiplied by 1.5. An average of 60
shoots, usable for rooting, were produced
every 2 wk to 100 bud-clusters.
After 2 transfers, the development of
clones and particularly callus formation
increased significantly (1% level) in the
Gelrite (Table 11). If liquid medium did not
increase the mean number of elongated
shoots, the E’longation of those shoots
was significantly higher (Table III).
Discussion and Conclusion
The studies have shown that the micropropagation
of juvenile walnut depends
upon many factors. At a general level,
characteristics of the medium are im-
portant. Results obtained with Gelrite
confirm our previous observations when
we lost all material growing on agar (unpublished
data). Many different hypotheses
have been proposed to explain the
influence of agar. These include the presence
of inhibitors, rate of diffusion of molecules
and variability in the availability of
water. This last effect could be associated
with the positive action observed with
liquid medium. Chun et al. (1986) obtained
better results with poplar in liquid medium
than with a gelified one. Nevertheless,
some cases of vitrification appear after
long-term culture in liquid medium. On the
contrary, Viseur (1987), avoided vitrification
in pear and increased bud production
by adding liquid medium. All these
phenomena should be connected with the
metabolism of phenolic compounds, lignification
or ethylene. With our system on
walnut, studies are and will be conducted
in these fields to determine the more important
medium factor.
McGranahan et aG (1988) recommended
for Persian walnut micropropagation a
1 wk transfer interval for gelified medium.
According to our results, and from a practical
point of view, results presented here
clearly illustrate that some of the very
expensive transfer work can be avoided
by adding liquid medium to cultures. Finally,
the great variability between clones,
also observed by Heile-Sudholt et aL
(1986), could limit the interest of bulk
micropropagation. Complementary research
is needed to determine if the best
clones for micropropagation are also the
best for field plantations.
References
Chun Y.W., Hall R.B. & Stephens L.C. (1986)
Influence of medium consistency and shoot
density on in vitro proliferation of Populus alba
x P. grandidentata. Plant Cell Tissue Cult. 5,
179-185
Driver J.A. & Kuniyuki A.H. (1984) In vitro propagation
of paradox walnut rootstock. HortScience
19, 507-509
Heile-Sudholt C., Huetteman C.A., Preece J.E.,
Van Sambeek J.W. & Gaffney G.R. (1986) In
vitro embryonic axis and seedling shoot tip culture
of Juglans nigra L. Plant Cell Tissue Cult.
6, 189-197
Jay-Allemand C. & Cornu D. (1986) Culture
in vitro d’embryons isol6s de noyer commun
(Juglans regia L.). Ann. Sci. For. 43, 189-
198
Jay-Allemand C., Cornu D. & Macheix J.J.
(1988) Biochemical attributes associated with
rejuvenation of walnut tree. Plant Physiol. Biochem.
26, 139-144
McGranahan G., Driver J.A. & Tulecke W.
(1987) Tissue culture of Juglans. In: Cell and
Tissue Culture in Forestry, Vol. 3, (Bonga J.M.
& Durzan D.J., eds.), Martinus Nijhoff, Dordrecht,
pp. 261-271
McGranahan G., Leslie C.A. & Driver J.A.
(1988) In vitro propagation of mature Persian
walnut cultivars. HortScience 23, 220
Viseur J. (1987) Micropropagation of pear,
Pyrus communis L., in a double phase culture
medium. Acta Hartic. 212, 117-124
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