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通过人参Defensin基因在拟南芥中的过表达,研究其在人参锈病菌胁迫过程中的抗性作用。采用RT-PCR技术克隆人参Defensin基因,其全长为240 bp,编码80个氨基酸,预测其蛋白分子量为12.091 1 ku。通过农杆菌介导浸花法将人参Defensin基因转到拟南芥中,经过抗性筛选及分子鉴定获得转基因拟南芥。以野生型拟南芥作对照,分析植株对锈病菌的抗性作用。结果表明:经锈病菌胁迫5 d,转基因拟南芥的生长状态明显优于野生型拟南芥,Defensin基因在转基因拟南芥中的表达量极显著高于野生型对照。人参Defensin基因对人参锈病菌具有一定的抗性作用。
Through the overexpression of ginseng Defensin gene in Arabidopsis thaliana, the resistance of the defensin gene to stress during the rust of the ginseng rust was studied. The full-length cDNA of Defensin was cloned by reverse transcription-polymerase chain reaction (RT-PCR). The total length of the cDNA was 240 bp and encoded a protein of 80 amino acids. The predicted protein molecular weight was 12.091 1 ku. Transgenic Arabidopsis thaliana was obtained by Agrobacterium tumefaciens-mediated infusion of Defensin gene into Arabidopsis. After resistance screening and molecular identification, Arabidopsis thaliana was obtained. The wild-type Arabidopsis thaliana was used as a control to analyze the plant’s resistance to rust. The results showed that the growth of transgenic Arabidopsis thaliana was significantly better than that of wild-type Arabidopsis thaliana under 5 d stress, and the expression of Defensin gene in transgenic Arabidopsis was significantly higher than that of wild-type control. Ginseng Defensin gene has some resistance to the rust of ginseng.