PJB-2019-945
EFFICIENT STERILIZATION PROTOCOL AND SUCCESSFUL CLONAL PROLIFERATION OF SPANISH SAFFRON (CROCUS SATIVUS L.) IN VITRO
Asif Khan
Abstract
Saffron is derived from the elongated dried stigmas of saffron (Crocus sativus L.) flowers, an autumn-flowering herbaceous plant that is propagated solely through clonal corm multiplication. The rate of natural propagation of corms is very low; hence an alternative in vitro culture method like micropropagation is urgently needed. In the present study, corms were surface sterilized with various sterilization protocols and cultured on medium fortified with several concentrations of different phytohormonal combinations for ensuring bud sprouting, multiple shoots proliferation and cormlet production. It was observed that 2-3 min treatment of corms with 70% ethyl alcohol and washing in 40% clorox and 0.2% HgCl2 for 5 min each ensured clean aseptic viable cultures (93.33%). MS enriched with BAP (4 mg/l) in combination with IAA (1 mg/l) resulted in maximum buds sprouting response (100%) followed by highest number of multiple shoot primordia (5.60) from the base of sprouted buds. Further sub culturing of shoot primordia in BAP (6 mg/l) and IAA (0.5 mg/l) was superior for achieving multiple shoots proliferation. Significantly, maximal cormlet number (4.33) and cormlet weight (0.58 g) was attained in MS medium supplemented with GA3 (4 mg/l) and 4% sucrose. However, media containing 2% sucrose and GA3 (2 mg/l) gave highest root number (5.2) and root length (13.10 cm) per root formed explants. These observations could be used to produce pathogen-free corms of saffron and other related species.
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