花生胚小叶的离体再生及筛选压力选择

In vitro plant regeneration from leaflet of peanut and its sensitivity to selection agent

以花生胚小叶为外植体优化离体再生条件,通过对再生苗进行筛选,为外源基因的遗传转化提供实验依据。研究表明,不同基因型胚小叶的再生能力差异显著,在供试基因型中以中花8号不定芽诱导率和成苗率最高,且外植体状态最好。中花8号胚小叶再生的最佳诱导培养基为MSB(MS无机盐+B5有机成分)+6-BA(4.5mg/L)+NAA(1mg/L)+AgNO3(2mg/L),最佳诱导培养时间为21d。对中花8号再生苗进行筛选浓度实验,草丁膦(PPT)为20mg/L时,可筛选出抗性苗;300mg/L的卡那霉素(Km)可用于较小幼苗筛选,而对较大幼苗则需将Km的浓度提高至400mg/L以上。

In order to develop a tissue culture system for peanut leaflet, the regeneration condition was optimized and plantlets sensitivity to selective agent was studied. Results showed significant differences on plant regeneration between different peanut genotypes, and Zhonghua 8 was more suitable for plant regeneration. MSB （MS mineral salt and B5 organic component） ＋ 6 - BA 4.5mg/L ＋ NAA lmg/L ＋ AgNO3 2mg/L was the best induction medium for Zhonghua 8 leaflet regeneration, and the induction time was 21d. Glufosinate ammonium （PVT,20mg/L） could be used for selecting Zhonghua 8 regenerated plantlets, 300mg/L kanamyein （Kin） could be used for selection of small seedlings and the concentration above 400mg/L would be more suitable for selection of larger seedling.