Technique for rapid in vitro multiplication of Vitis vinifera L. cultivars

Mass multiplication through in vitro means is desired in newly released grape varieties and a protocol developed through repetitive two-node micro-cutting technique is proposed. Culture could be initiated from nodal segments collected from field-grown vines of grape cultivars Pusa Urvashi and Pusa Navrang in Murashige and Skoog medium supplemented with 2.0 or 4.0 mg l(-1) BAP + 0.2 mg l(-1) NAA. The sprouted shoots were successfully proliferated and rooted on half-strength MS medium supplemented with 2.0 mg l(-1) IBA + 200 mg l(-1) activated charcoal. The elongated shoots in each subculture could be then used for multiplication on the same medium after 6-7 weeks, by excising them into two-node micro-cuttings, which ranged in number from seven to 12 for the two cultivars. The rooted plantlets (after three weeks of inoculation) obtained on the above medium were successfully hardened in glass jars filled with peat: soilrite (1:1) or delayed in the same medium up to 6-7 weeks for further multiplication in a repetitive manner. A complete protocol has been developed for multiplication of 85,000 to 119,000 plantlets per annum, for a small-scale tissue culture laboratory.