Standardization of Rapid In Vitro Regeneration Technique in Vitis vinifera cv. Manjari Naveen us ing Lateral Bud

Abstract

In vitro propagation is a commercial technology of tissue culture used in this era for plant regeneration around world. In the present investigation efficacious, rapid in vitro protocol for plant regeneration was developed using lateral buds in Vitis viniferacv. Manjari Naveen on Murashige and Skoog (MS) medium amended by cytokinins and auxins. 0.1 % HgCl2 was used to sterilize explants for 10 min and ascorbic acid (0.1 g/L) plus citric acid (0.15 mg/l) were used for minimization of browning in medium. Sterilized explants were cultured on MS media containing different concentration of BAP (0.1, 0.3, 0.5, 1.0 mg/ml and control) for shoot induction. The 0.5 mg/l BAP results in maximum shoot length (3.40 cm), high number of shoots (1.96) and low percentage of contamination for initiation. Afterwards, multiplication was obtained with the use of different concentration of cytokinins and auxins i.e., 0.1 mg/l BAP + 0.1 mg/l IBA, 0.5 mg/l BAP + 0.1 mg/l IBA, 1.0 mg/l BAP + 0.1 mg/l IBA, 1.5 mg/l BAP + 0.1 mg/L IBA. Control treatment was maintained without growth hormones. Among these, the maximum shoots multiplication was obtained on MS medium containing 0.5 mg/l BAP + 0.1 mg/l IBA. So, the protocol using MS medium containing combination of 0.5 mg/l BAP+0.1 mg/l IBA was standardized and used successfully for grapevine propagation. Therefore, use of auxins and cytokinins @ 0.5 mg/l BAP for shoot initiation, 0.5 mg/l BAP + 0.1 mg/l IBA for multiplication and 2.5 mg/L IAA for rooting can be exploited for in vitro propagation at commercial level in grapes.