以黄瓜(Cucumis sativus)子叶和子叶节为外植体, 利用农杆菌(Agrobacterium tumefaciens)介导法建立黄瓜遗传转化体系并进行优化, 将南方根结线虫(Meloidogyme incognita)寄生相关基因的RNA干扰载体导入黄瓜, 通过潮霉素(Hyg)浓度梯度筛选得到99株潮霉素抗性植株。经PCR和Southern-blot检测, 获得10株目的基因以单拷贝形式整合的转基因黄瓜, 子叶和子叶节的Southern-blot阳性率分别为13.9%和6.9%。该研究建立并优化了RNA干扰载体导入黄瓜的高效遗传转化体系, 成功获得了转基因黄瓜植株。
The RNAi vector of the target gene was transfected into cucumber (Cucumis sativus) cotyledons and cotyledon nodes by Agrobacterium tumefaciens-mediated transfection. A total of 99 regeneration plants were hygromycin-resistant. PCR and Southern-blot analysis revealed that target gene was integrated into the genome of 10 regenerated plants. The transformation efficiency in the cucumber cotyledon and cotyledon node was significantly increased from 4.1% to 13.9% and 3.1% to 6.9%, respectively, when using different hygromycin concentrations in the selection cultivation. We have optimized the genetic transformation system with an RNA interference vector in cucumber and successfully obtained transgenic cucumber plants.
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