Multiple Shoot Regeneration Study on Three Varieties of Grape vine (Vitis vinifera L.) from shoot tip and nodal culture

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Addis Ababa University


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



Vitis vinifera L, shoot initiation, root initiation, Cytokinin, Auxin