陆地棉细胞色素P450基因GhP450-94C1黄萎病抗性功能验证  

Functional Verification of GhP450-94C1 that a Verticillium wilt Resistant Gene in Gossypium hirsutum L.

作　　者：胡子曜 李秀青 代培红[1] 雷建峰[2] 柳建飞 赵燚 邓嘉辉 刘超[1] 刘晓东[1] 李月[1] HU Ziyao;LI Xiuqing;DAI Peihong;LEI Jianfeng;LIU Jianfei;ZHAO Yi;DENG Jiahui;LIU Chao;LIU Xiaodong;LI Yue(College of Life Sciences,Xinjiang Agricultural University,Urumqi 830052,China;College of Agronomy,Xinjiang Agricultural University,Urumqi 830052,China)

机构地区：[1]新疆农业大学生命科学学院,新疆乌鲁木齐830052 [2]新疆农业大学农学院,新疆乌鲁木齐830052

出　　处：《华北农学报》2022年第6期72-81,共10页Acta Agriculturae Boreali-Sinica

基　　金：国家自然科学基金项目(32160494)。

摘　　要：为了探究陆地棉细胞色素P450基因GhP450-94C1在棉花黄萎病反应中的生物学功能,为棉花黄萎病抗性基因挖掘及抗病育种奠定基础,通过转录组筛选,克隆获得一个细胞色素P450基因GhP450-94C1,利用生物信息学的方法分析该基因的理化性质,通过荧光定量PCR技术分析GhP450-94C1在黄萎病侵染下的表达模式,利用病毒诱导的基因沉默(VIGS)技术初步探究其在棉花抗黄萎病反应中的生物学功能。结果表明,克隆获得一个陆地棉细胞色素P450基因GhP450-94C1,其开放阅读框(ORF)为1503 bp,编码一个含500个氨基酸的酸性、亲水、不稳定的跨膜蛋白,分子式为C_(2597)H_(4025)N_(691)O_(725)S_(22),分子质量为57.23 ku,定位于内质网膜,含有一个P450结构域;有86.45%的概率存在信号肽;二级结构预测显示,该蛋白含有24个α-螺旋和8个β-折叠;该基因响应黄萎病菌侵染,抑制其表达后植株对黄萎病菌的敏感性增强。初步证明GhP450-94C1是棉花黄萎病抗性反应的一个正向调控因子。To explore the biological function of cotton cytochrome P450 gene GhP450-94C1 in cotton Verticillium wilt response,and lay a foundation for cotton Verticillium wilt resistance gene mining and disease resistance breeding.A cytochrome P450 gene,GhP450-94C1,was cloned through transcriptome screening.The physicochemical properties of the gene were analyzed by bioinformatics methods.The expression pattern of GhP450-94C1 under Verticillium wilt induction was analyzed by Real-time quantitative polymerase chain reaction(qRT-PCR).Virus-induced gene silencing(VIGS)technology was used to preliminarily explore its biological function in cotton resistance to Verticillium wilt.The main results were as follows:upland cotton cytochrome P450 gene,GhP450-94C1,was obtained by cloning.The open reading frame(ORF)was 1503 bp,encoding an acidic,hydrophilic and unstable transmembrane protein with 500 amino acids.The molecular formula was C_(2597)H_(4025)N_(691)O_(725)S_(22),with a molecular weight of 57.23 ku,which was located in the endoplasmic reticulum membrane and contained a P450 domain.There was 86.45%probability of signal peptide;the secondary structure prediction showed that the protein contained 24α-helixes and 8β-sheets.This gene responds to Verticillium wilt infection,and after inhibiting its expression,the sensitivity of plants to Verticillium wilt is enhanced.GhP450-94C1 is a positive regulator of cotton resistance to Verticillium wilt.

关 键 词：棉花黄萎病 GhP450-94C1 基因克隆 病毒诱导的基因沉默 

分 类 号：Q78[生物学—分子生物学] S562.03[农业科学—作物学]