杜仲种子无菌苗的获得及染色体加倍技术研究

Achievement of Tube-cultured Plantlets from Seeds and Technique of Chromosome Doubling of Eucommia ulmoides

研究了杜仲种子试管苗萌发和无菌苗培育技术,以及种子染色体加倍技术,提出了以种子为材料通过试管萌发进行染色体加倍的技术方法：杜仲成熟种子剥去种皮,用清水反复冲洗,经500 mg.L-1的赤霉素浸泡24 h后,置于浓度为0.3%的秋水仙素中浸泡48 h,再于70%酒精消毒30 s,1 g.L-1HgCl2表面消毒12 min,无菌水冲洗3~4次,切去少许胚芽和胚根端胚乳后胚根向下接入无激素的MS培养基中培养。在组织培养条件下,种子的萌发率最高可达100%,且无菌苗生长良好;多倍体诱变率最高达到76.09%。本试验最终获得多倍体变异植株424株。

Based on the germination and breeding of tube-cultured plantlets of Eucommia ulmoides,the technology of chromosome doubling of seeds was conducted.The methodology of in vitro chromosome doubling using seeds was as follows：stripping the mature husk of E.ulmoides,rinsing repeatedly with tap water,immersing in 500 mg·L-1 gibberellin for 24 h,then soaking seeds for 48 h in 0.3% colchicine solution.The disinfection time was 30 s with 70% alcohol and 12 mins with 1 g·L-1HgCl2.After rinsed three to four times with sterile water,truncated endosperms were placed into hormone-free Ms medium.The highest germination rate from seeds was up to 100% in tissue culture,with the superior quality as well;the highest rate of the polypoid induction was 76.09%.Introduced polyploid plants with a number of 424 were obtained finally.