棉花茉莉酸氧化酶基因GhJOX2参与抗黄萎病的功能分析  

Functional analysis of a jasmonate-induced oxygenase gene GhJOX2 involved in Verticillium wilt resistance in cotton

作　　者：刘盟盟 赵佩 陈伟[2] 桑晓慧 赵云雷 王红梅[2] 吴立强[1] LIU MengMeng;ZHAO Pei;CHEN Wei;SANG XiaoHui;ZHAO YunLei;WANG HongMei;WU LiQiang(Hebei Base of State Key Laboratory of Cotton Biology,Hebei Agricultural University,Baoding 071001,China;State Key Laboratory of Cotton Biology,Institute of Cotton Research of the Chinese Academy of Agricultural Sciences,Anyang 455000,China)

机构地区：[1]河北农业大学,棉花生物学国家重点实验室河北基地,保定071001 [2]中国农业科学院棉花研究所,棉花生物学国家重点实验室,安阳455000

出　　处：《中国科学：生命科学》2022年第4期523-533,共11页Scientia Sinica(Vitae)

基　　金：河南省自然科学基金(批准号:202300410548);中央级公益性科研院所基本科研业务费专项(批准号:1610162021005)资助。

摘　　要：茉莉酸信号通路在植物发育和抵御生物和非生物胁迫中发挥着重要作用.茉莉酸的持续积累抑制了植物生长,因此茉莉酸诱导的氧化酶(JASMONATE-INDUCED OXYGENASEs,JOXs,又称茉莉酸氧化酶)将JA羟基化为12-羟基JA,对植物生长至关重要.本研究鉴定到一个棉花抗黄萎病相关的茉莉酸氧化酶(JOX2)基因,GhJOX2.基因克隆分析表明,该基因在抗病品种和感病品种中的CDS序列全长均为1074 bp,编码357个氨基酸,但两个品种在第226位氨基酸(位于2OG-Fe(Ⅱ)结构域内部)上存在差异,分别为亮氨酸和苯丙氨酸.基因表达分析表明,GhJOX2在叶中高量表达,并且在抗病品种中的表达量显著低于感病品种;黄萎病菌和外源激素(茉莉酸和乙烯)处理均能诱导该基因表达.病毒诱导的基因沉默(virus-induced gene silencing,VIGS)实验结果表明,沉默植株对大丽轮枝菌感染表现出更高的耐受性,同时沉默植株体内茉莉酸信号通路和乙烯信号途径相关基因的表达量在接菌后呈现不同程度的上调表达趋势.总之,GhJOX2沉默提高棉花对黄萎病的抗性,推测该基因可能是通过调节茉莉酸信号通路相关基因的表达来调控棉花对黄萎病的抗性.Jasmonic acid signaling pathway plays an important role in plant development and resistance to biological and abiotic stresses.The persistent accumulation of JA inhibits plant growth,and the hydroxylation of JA to 12-hydroxy-JA by JASMONATE-INDUCED OXYGENASEs(JOXs,also named jasmonic acid oxidases)is therefore vital for plant growth.In this study,the function of a cotton jasmonate-induced oxygenase(JOX2)gene,GhJOX2,which was detected to be associated with Verticillium wilt(VW)in our previous study,was analyzed.The results showed that the CDS sequence of GhJOX2 was 1074 bp and it encoded a polypeptide of 357 amino acids.There was one amino acid difference in the GhJOX2 protein sequence between the resistant cultivar ZZM2 and susceptible cultivar J11,i.e.,the 226th amino acid located in the 2OG-Fe(Ⅱ)domain was leucine(L)in the resistant cultivar ZZM2but phenylalanine(F)in the susceptible cultivar J11.Expression analysis showed that GhJOX2 was highly expressed in leaves,and the expression was significantly induced by V.dahliae and exogenous hormones.Virus-induced gene silencing(VIGS)experiments demonstrated that silencing of GhJOX2 significantly enhanced the resistance to V.dahliae in cotton,and the expression levels of related genes in jasmonic acid pathway were up-regulated in the VIGS plant compared to the control after inoculation.We concluded that Gh JOX2 silencing improved cotton resistance to Verticillium wilt by regulating the expression levels of related genes in jasmonic acid pathway in cotton.

关 键 词：棉花黄萎病 茉莉酸氧化酶(JOX) VIGS QRT-PCR 

分 类 号：S435.621.2[农业科学—农业昆虫与害虫防治]