芒遗传转化过程中选择压力的确定

以芒(Miscanthus sinensis)胚性愈伤组织为材料,研究了植物遗传转化过程中常用的潮霉素(Hyg)、遗传霉素重硫酸盐(G418)和除草剂(Basta)等3种筛选剂,不同浓度下的不同培养时间对愈伤组织生长、不定芽分化以及再生植株生根的抑制作用。实验结果表明,3种筛选剂均表现出很强的抑制效果。使用75mg/L G418筛选培养20d,50mg/L Hyg筛选20d或20mg/L Basta筛选40d可完全抑制愈伤组织的生长;使用60mg/L G418,40mg/L Hyg或10mg/L Basta筛选培养约20d可完全抑制愈伤组织分化出不定芽;而使用60mg/L G418,50mg/L Hyg或10mg/L Basta筛选培养20d可完全抑制再生植株的生根。3种筛选剂中以Basta对芒植株再生各个阶段的抑制效果最为明显,Hyg的抑制效果次之,G418抑制效果相对不明显。

Determination of the selecting pressure for genetic transformation of Miscanthus sinensis

The effects of Hygromycin,Geneticin and Basta under different concentrations on callus growth,bud differentiation and seedling rooting of a bioenergy crop,Miscanthus sinensis,were studied.The results indi-cated that at the callus subculture stage,the concentration of 65~75 mg/L G418 culturing for 20 days,40~50 mg/L Hyg for 20 days or 10~20 mg/L Basta for 40 days could inhibit callus growth completely;at bud differentiation stage,the concentration of 60 mg/L G418,40 mg/L Hyg or 10 mg/L Basta culturing for 20 days could completely restraint bud differentiation from calli;and at rooting stage,the concentration of 60 mg/L G418,50 mg/L Hyg or 10 mg/L Basta culturing for 20 days can repress root regeneration from seedlings absolutely.Basta was the most stringent selecting agent for M.sinensis,Hyg the next,and followed by G418.