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Abstract

Abstract The aim of this research was to optimize protocol for micropropagation and synthetic seeds production of C. sativus. From 15-day-old in vitro plantlets, bulblets and microshoots were removed as explants in order to encapsulate them. Best culture media for producing multiple shoots was MS medium fortified with BAP + Kinetin (3+0.4) and for rooting BAP+NAA (3+3 mg/L) among all the media tested. Optimal concentration for encapsulation was found to be 3% sodium alginate with 100 mM calcium chloride. Encapsulated beads were stored at different temperatures i.e. 4, 20 and 25ºC for different days i.e. 0, 7, 15 and 30 days. It was observed that 4°C was optimum temperature for storage. To evaluate the regeneration potential of encapsulated beads, same media of micropropagation i.e. BAP+ NAA + Kinetin (3+3+0.4 mg/L) was used. The beads inoculated right after their formation was designated as control exhibited the highest germination rate (100%) and the synthetic seeds kept at 4ºC for 7 days showed 94% potential. On the other hand, seeds retained for 15 days showed 82% potential and seeds that were kept for 30 days had the lowest germination rate (70%). In control, maximum root length was 20.3a±0.58 cm while shoot number and shoot length was 4.66a±0.61 and 7.34a±0.56 cm. During 7 days storage, maximum shoot length was noted as 6.55ab±0.61 cm and shoot number was 3.33a±0.57 while root length and number was found to be 18.3a±0.54 cm and 2.63a±0.57, respectively. As storage times increased the frequency of synthetic seed conversion to plantlets gradually reduced. In vitro grown plantlets were acclimatized in green house. The results indicates novel encapsulation technique can potentially act as a valuable substitute for the preservation and rapid propagation of saffron. Key words: Synthetic seeds, micropropagation, encapsulated beads, bulblets, microshoots, Plant growth regulators

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