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杜鹃红山茶CaCPI基因的克隆及过量表达提高烟草植株的抗虫性

DOI: 10.7668/hbnxb.2015.05.010, PP. 57-64

Keywords: 杜鹃红山茶,基因克隆,虫害,表达分析,转基因烟草

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Abstract:

绝大多数观赏山茶花属于设施栽培植物,因环境湿热易造成病虫害出现,导致观赏价值降低,影响种植效益。为了提高抗虫性,根据山茶同源序列设计特异性引物,利用同源克隆和3',5'-RACE技术,从杜鹃红山茶嫩叶组织中克隆出半胱氨酸蛋白酶抑制剂(Cysteineproteinaseinhibitor,CPI)基因,命名为CaCPI,基因全长579bp,开放阅读框306bp,编码101个氨基酸,分子量为11.078kDa,等电点pI=6.72。利用实时荧光定量PCR方法对杜鹃红山茶根、茎、叶中该基因的表达特性进行分析,结果表明,CaCPI基因在叶中的表达量较高,其次为茎,在根中表达量较低,比较显示叶的表达量分别约为茎的1.53倍,根的1.61倍。转基因烟草分析表明,过量表达CaCPI基因后,转基因烟草中目的基因表达量提高了55.84~174.83倍,并且转基因植株抗蚜虫能力获得提高,接虫5d后植株上的蚜虫绝大多数死亡,累积死亡率约为90.75%,高达野生型的5.14倍。

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