一种快速筛选转抗霜霉病基因甜瓜种子的方法研究

The Screening of Transgenic Plants Resistant to Downy Mildew Disease in Melon

以用花粉管通道技术转入抗霜霉病基因At2的甜瓜T1代转基因种子为试材,采用组培技术得到膨大的外植体,利用PCR技术对组培得到的外植体进行检测,以期尽快筛选出阳性植株。结果表明:在甜瓜外植体培养过程中,筛选到的种子最佳灭菌条件为72℃、干灭4h,抑菌效果最好的培养基组分是MS无机盐+B5维生素+2mg/L 6-BA+7g/L琼脂+500mg/L头孢噻肟钠+1.1g/L甲基托布津;对1 100个转抗霜霉病基因At2的T1代种子外植体进行PCR检测,阳性率为4.5‰。

Taking T1 melon of transgenic plant seed that transgened into At2 downy mildew resistant gene though pollen- tube pathway technology as material,swollen explants was gotten through tissue culture technology. Then PCR analysis was performed on the explant,which were obtained from transgenic seed to screen out positive transgenic plant rapidly. The result indicated that the optimal sterilization condition was 72℃ for 4 h and the optimal medium components with antifungal effect was MS salts＋B5 vitamins＋2 mg/L 6-BA＋7 g/L agar ＋500 mg/L cefotaxime＋1. 1 g/L thiophanate- methyl,during the culture processes of melon explants. 1 100 T1 generation seed explants transformed with downy mildew resistance genes At2 were detected using PCR technique,and the positive rate was 4. 5 ‰.