سال انتشار: ۱۳۸۵
محل انتشار: دومین سمپوزیوم بین المللی تکنولوژی و بیولوژی زعفران
تعداد صفحات: ۸
E Darvishi – Agricultural Biotechnology research Institute Karaj Iran
R Zarghami – Department of Plant Breeding College of Agriculture and Natural Resources University of Tehran Karaj 31587 Iran
C.A Mishani, – Department of Plant Breeding College of Agriculture and Natural Resources University of Tehran Karaj 31587 Iran
M Omidi – Department of Plant Breeding College of Agriculture and Natural Resources University of Tehran Karaj 31587 Iran
In order to determine the best hormone treatment for nonembryogenic and embryogenic calli induction, apical meristem of young corms derived from saffron (Crocus sativus L.) were cultured in solid LS medium supplemented with different concentration of two auxin (NAA and 2,4-D) and three cytokinin (BAP, Kin and 2ip) at 22°C in darkness. Statistical analysis using Kruskal-Wallis Test showed that treatment containing 2 mg/L NAA and BAP with highest Mean Rank had the beat effect on induction of nonembryogenic callus and treatment containing 1 mg/L 2, 4-D and BAP had the best effect on induction of embryogenic callus. The results obtained from counting number and viability of protoplasts isolated from embryogenic calli with homeocytometer and Trypan Blue under the enzyme solution consisting of MS medium with 0.1 % (w/v) Pectolyase Y-23, 1 % Cellulose R-10, 1% Deriselase, 0.1 % MES (2-N-morpholino ethane sulfonic acid), and 0.3 M mannitol at pH 5.7 on a rotary shaker (200 rpm) at 25 °C in darkness for 4 time-term treatment (1.5, 3, 4 & 5 hour) indicated that after 3 h enzyme treatment, 40×105protoplasts per 1 mL of suspension with 98 % viability obtained that were the best time- term treatment.Finally, in order to protect and maintain the viability of fragile protoplasts, Calsium-alginats beads were used for their immobilization.