大豆幼胚子叶胚性悬浮细胞系的建立与次生胚诱导

ESTABLISHMENT OF EMBRYOGENIC SUSPENSION WITH IMMATURE COTYLEDONS OF SOYBEAN (Glycine max(L.)Merr.) AND INDUCTION OF SECONDARY EMBRYOS

本研究选用黑龙江省 5个大豆品种的幼胚子叶 ,建立了胚性悬浮培养体系 ,并由悬浮体系增殖产生次生胚。系统探讨了培养基组成和浓度对体细胞胚诱导和悬浮培养物生长的影响 ,以及悬浮体系继代时间与胚成熟率和再生率的关系。结果表明 ,高效体细胞胚诱导培养基为 :MSB +4 0mg/L 2 ,4 -D + 6 %蔗糖。高效球型胚增殖培养基为 10mg/L2 ,4 -D + 1/2MS氮源 + 5mM谷氨酰胺 + 5mM天门冬酰胺。培养 8周后的次生胚成熟率和再生率显著提高 ,到第 12周分别达6 3%和 35 % 。

With 5 soybean cultivars in Heilongjiang province, somatic embryos were induced with immature cotyledons and secondary embryogenesis systems were established through suspension proliferation of embryogenic cultures. We studied systematically the influence of component and concentration of medium on somatic embryo induction and suspension culture proliferation. The results show,high efficient medium of inducing somatic embryos is MSO+40mg/L 2,4-D +6%sucrose. High efficient medium of global embryos proliferation is 10mg/L 2,4-D+1/2 MS nitrogen+5mM glutamine+5mM asparagines. The percentage of embryos mature and regeneration of secondary embryos after cultured for 8 weeks were significantly improved,and reached 63% and 35% respectively at the 12 week,after then,the increased rate declined.