抗菌肽基因转化厚皮甜瓜的研究

以厚皮甜瓜FY1的子叶为试材,研究不同激素组合和配比对甜瓜植株再生的影响,并用农杆菌介导法将银杏抗菌肽基因Gnk2-1转化甜瓜。结果表明:(1)甜瓜子叶的最佳愈伤组织及不定芽诱导培养基为MS+2.0mg.L-1 6-BA+0.1mg.L-1 IAA,出愈率和出芽率均达95%以上;最佳芽伸长培养基为MS+0.2mg.L-1 KT;最佳生根培养基为1/2MS+0.1mg.L-1 IAA。(2)卡那霉素对甜瓜外植体的生长和分化有明显的抑制作用,适宜的筛选压力为100mg.L-1。(3)将子叶预培养2d,在农杆菌菌液浓度OD600值为0.5左右时,侵染外植体10min左右,共培养3d,转化效率最高。(4)经PCR鉴定获得了抗性植株,抗病性鉴定显示,转基因植株对枯萎病的抗性有所增强,发病迟缓。

Preliminary Study of Muskmelon(Cucumis melon L.) by Transferring Antibacterial Peptide Gene

In this research,the cotyledon explants of muskmelon FY1 were used as the experimental material to study the influence of different hormone combination on the regeneration system of the melon.Then we investigated the agrobacterium-mediated transformation method to transfer antibacterial peptide gene Gnk2-1 into melon.The results of the study showed:(1)that the optimal medium for the melon cotyledon callus and adventitious bud induction was MS+2.0 mg·L-1 6-BA+0.1 mg·L-1 IAA,and their inductivity were both above 95%;the most suitable medium for the buds to lengthen was MS+0.2 mg·L-1 KT;the best medium for root induction was 1/2MS+0.1 mg·L-1 IAA.(2)Kanamycin had an apparent inhibition to the growth and the differentiation of melon explants,the appropriate selection pressure was 100 mg·L-1.(3)The cotyledons were precultured for 2 d,when the bacterial concentration is about 0.5 OD600,the explants were infected for around 10 min,then the explants were cultured with the bacterial for 3 d,and the conversion efficiency was the highest.(4)The resistant plant was obtained by PCR identification and the enhanced resistance to Fusarium wilt was observed in transgenic plants.