芝麻子叶基因转化受体系统的建立

Establishment of an Acceptor System for Gene Transformation in Sesame (Sesamum indicum L.) Cotyledon

试验以芝麻栽培种豫芝11号为材料,对子叶丛生芽诱导及植株再生体系以及子叶受体材料对卡那霉素、潮霉素和头孢霉素的天然耐性进行了研究,建立了芝麻子叶基因高频转化受体系统。结果表明,适宜的诱芽培养基为MS+5.0mg/L6-BA+1.0mg/LIAA+5.0mg/LAgNO3+1.0mg/LABA+3%蔗糖+0.8%琼脂;6-BA对子叶膨大起决定作用,ABA、IAA则是诱导丛生芽高频发生的关键因子,AgNO3对丛生芽发生有显著的促进作用。在芝麻子叶膨大出芽期、芽增殖期及试管苗生根期等遗传转化的三个重要时期,适宜的卡那霉素筛选浓度分别为70mg/L、70mg/L、20mg/L,潮霉素筛选浓度分别为40mg/L、60mg/L、10mg/L。在农杆菌介导的芝麻遗传转化中,头孢霉素使用浓度以500mg/L为宜,抑菌效果好且不影响转化受体生长。

To establish the acceptor system for gene transformation in sesame cotyledon, perfect culture medium for bud inducement from sesame cotyledon and plantlet regeneration has been investigated, as well as the endurance points of explants and plantlets on selective agents, i. e., Kanamycin, Hygromycin and Cefotaxime, have been studied deeply. Results showed that suitable differentiation medium was MS containing 5.0 mg/L 6-BA, 1.0 mg/L IAA, 5.0 mg/L AgNO3, 1.0 mg/L ABA, 3% sucrose and 0.8% agar. 6-BA would determine the cotyledon expansion. Moreover, being the key factors for shoot regeneration, ABA and IAA could induce high frequency of adventitious bud formation, and AgNO3 could accelerate remarkable multiple shoots regeneration. During three important stages of cotyledon expansion and bud formation, adventitious bud proliferation, and plantlet root formation in the genetic modification system for Sesamum indicum L., appropriate concentration of Kanamycin are 70 mg/L, 70 mg/L and 20 mg/L, Hygromycin are 40 mg/L, 60 mg/L, 10 mg/L, respectively. In the genetic modification system by A. tumefaciens infection, 500 mg/L Cefotaxime could be chosen, by which A. tumefaciens strain growth could be inhibited when needed, yet sesame explants and shoot regeneration would not impaired.