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棉花学报  2015 

棉花GhSKIP35基因的克隆与表达分析

DOI: 10.11963/issn.1002-7807.201502003, PP. 111-117

Keywords: 棉花,大丽轮枝菌,SKIP35,qRT-PCR,VIGS

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Abstract:

以中植棉KV1SSH文库为基础,从陆地棉中植棉KV1中克隆出来泛素途径SKIP35基因,命名为GhSKIP35。该基因的开放阅读框全长为1863个核苷酸,编码620个氨基酸,含有1个ANK保守结构域。进化分析表明,GhSKIP35蛋白与甜橙(Citrussinensis)的SKIP35蛋白相似性最高。qRT-PCR分析显示在接种大丽轮枝菌V991菌系后初期GhSKIP35基因表达量激增,12h便可达到顶峰,24~48h逐渐下降,推测它在陆地棉抗黄萎病早期防卫反应中起重要作用。利用VIGS(Virusinducedgenesilencing)技术,在中植棉KV1中成功地沉默GhSKIP35基因;对沉默棉株接种大丽轮枝菌V991鉴定显示,其病情指数为50.75,而野生型的中植棉KV1和转化空载体棉株的病情指数分别为9.38和11.54。GhSKIP35基因沉默后,中植棉KV1对黄萎病的抗性丧失,表明GhSKIP35基因在陆地棉抗黄萎病的过程中起重要作用。

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