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半夏凝集素基因克隆及其对桃蚜的抗性研究

DOI: 10.7685/j.issn.1000-2030.2010.02.009, PP. 45-50

Keywords: 半夏凝集素,基因克隆,转基因烟草,抗性,桃蚜

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Abstract:

研究了半夏凝集素(Pinelliaternataagglutinin,PTA)基因对桃蚜(Myzuspersicae)的抗性。从半夏幼叶中提取总RNA,采用RT-PCR法分离克隆出PTA1基因cDNA片段,该基因在GenBank中登录号为DQ092435。序列分析表明该cDNA片段全长为810bp,编码269个氨基酸组成的蛋白,具有单子叶植物甘露糖凝集素基因家族的特征,含有3个由4个氨基酸(QDNY)组成的甘露糖专一结合位点,与报道的天南星科凝集素基因氨基酸序列的同源性在72.7%~92.9%。构建了35S启动子控制下的PTA1基因的植物表达载体pBI121PTA1,该载体含有抗卡那霉素的选择标记基因(nptⅡ),利用根癌农杆菌介导法转化烟草获得抗卡那霉素的植株。PCR和半定量RT-PCR分析表明PTA1基因已经整合到植物基因组中并在转录水平上得到了有效表达。抗蚜虫鉴定表明转基因植株有效地抑制了蚜虫的群体增长率,平均抑制率达77.02%,表明该基因对同翅目害虫的抗虫基因工程有重要的应用价值。

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