Development of Cryoconservation Protocol for In-vitro Shoot Bases of Allium ampeloprasum L. for Long-term Conservation

Abstract

Critical factors for the development of cryo conservation protocol for Allium ampeloprasum L. conserved in the in-vitro Genebank (IVGB) at ICAR-NBPGR of India were investigated. Shoot bases (1.0 x 1.5 mm) excised from 8-wk-old shoot cultures grown on growth media (MS + 0.1 mg/l NAA + 0.02 mg/l 2iP + 0.3 M sucrose) and pregrown at 22/ 5ºC for 16/8 hours alternate temperature regime, precultured on MS medium supplemented with 0.3 M sucrose at standard culture conditions (SCC) for 16 hours followed by loading solution treatment for 60 minutes, PVS2 dehydration for 40 minutes were successfully cryoconserved using vitrification and droplet-vitrification technique. Droplet-vitrification technique improved post-thaw regrowth by ~58.33% as compared to vitrification. Thus, a standardized protocol was used for cryobanking of shoot bases of A. ampeloprasum