Multiple Shoot Regeneration Study on Three Varieties of Grape vine (Vitis vinifera L.) from shoot tip and nodal culture
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Date
2010-11
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Addis Ababa University
Abstract
Grape-vine (Vitis vinifera L.) is a perennial deciduous woody vine which is
cultivated all over the world. Conventional breeding of grape vine is impeded by
long juvenile periods and inbreeding depression. The objectives of this study
were to establish a protocol for in vitro micropropagation of selected grape vine
varieties. Preliminary studies were undertaken in order to optimize the duration
of sterilization of the explants and the culture used to initiate shoots. Shoot tips
of variety Chenin blanc was cultured in MS medium supplemented with 0.5
mg/l after sterilizing in 1% NaOCl to determine the optimum duration of
sterilization. For shoot initiation experiment, shoot-tip and nodes of the three
varieties, namely, Ugni blanc, Chenin blanc and Canonannon were cultured in
Murashige-Skoog medium supplemented with five different concentrations of
BAP including the BAP free. Various experiments were carried out to optimize
shoot multiplication using MS medium supplemented with different
combination and concentrations of BAP with or without IBA. To optimize root
induction different concentrations of IAA were used. Data such as number of
shoots and root, shoot height, root length and node number were recorded and
analyzed. Sterilization of explants using 1% of NaOCl was found to be optimum
for 7 minutes duration. Cheninblanc showed high percentage of survival rate
(96%) followed by Ugniblanc and Canonannon (88%) at 0.5 mg/l BAP. Among
the different concentrations and combination of PGRs, a maximum mean
number of shoots 7.2, 6.7, 6.1 was achieved at 1mg/l BAP combined with 0.1
mg/l IBA for Chenin blanc, Canonnanon and Ugni blanc respectively. The
shoot length and node number of all varieties showed a decreasing trend when
the concentration of BAP was increased, except for Canonannon, in terms of
node number. Number of shoots, node and shoot height showed significant
variability (p=0.05) at different level of BAP or BAP combined with IBA. All
varieties induced root on MS medium without IAA or supplemented with
different concentration of IAA. Among these rooting culture, however, MS
medium with higher concentration (2 and 4 mg/l) of IAA were found to be the
best for root induction in all varieties. The plantlets were acclimatized in the
glasshouse and survival percentage was 92% for Chenin blanc followed by
78.6% and 73.9% for Ugni blanc and Canonannon, respectively. Thus, the
achievements of this study will play a big role in the grape vine culture program.
Key words: Vitis vinifera L., shoot initiation, root initiation, Cytokinin, Auxin
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Keywords
Vitis vinifera L, shoot initiation, root initiation, Cytokinin, Auxin