柑橘4个WRKY转录因子基因的克隆及其响应柑橘溃疡病菌侵染的表达分析  

Cloning and Expression Analysis of Four Citrus WRKY Genes Responding to Xanthomon asaxonopodis pv.citri

作　　者：周鹏飞[1] 贾瑞瑞[1] 陈善春[1] 许兰珍[1] 彭爱红[1] 雷天刚[1] 李强[1] 陈敏[1] 白晓晶[1] 邹修平[1] 何永睿[1] 

机构地区：[1]西南大学/中国农业科学院柑桔研究所,重庆400712

出　　处：《园艺学报》2017年第3期452-462,共11页Acta Horticulturae Sinica

基　　金：中央高校基本科研业务费重大项目(XDJK2014A018);国家科技支撑计划项目(2012BAD19B06);国家现代农业产业技术体系建设专项资金项目(CARS-27)

摘　　要：以纽荷尔脐橙和四季橘为试验材料,基于溃疡病菌诱导的柑橘转录组数据库,利用PCR方法克隆获得4个WRKY家族基因Cs WRKY22、Cs WRKY50、CsWRKY72-1和CsWRKY72-2的cDNA全长序列。序列分析结果表明,这4个基因的cDNA全长分别为1 123、1 312、1 809和2 208 bp,开放阅读框长度分别为921、480、1 809和1 767 bp,各编码306、159、602和588个氨基酸。氨基酸序列和结构分析显示,这4个基因属于第Ⅱ类WRKY蛋白。进化树分析显示,所克隆的4个柑橘WRKY蛋白与可可、葡萄等WRKY蛋白亲缘关系较近。亚细胞定位显示,所克隆的4个WRKY基因定位于细胞核,与亚细胞定位预测相符。对纽荷尔脐橙和四季橘中4个基因受溃疡病菌诱导的表达分析表明,CsWRKY22参与寄主的感病反应,而CsWRKY50参与抗溃疡病的免疫应答反应。柑橘溃疡病菌能抑制CsWRKY72-1和CsWRKY72-2介导的寄主基础免疫。水杨酸(SA)和茉莉酸甲酯(MeJA)处理表明,4个Cs WRKY基因均不参与SA和MeJA介导的寄主抗病和抗逆反应。Based on the information of transcriptome database derived from Xanthomon asaxonopodis pv. citri （Xac） infected citrus, the cDNA sequences of WRKY22, WRKY50, WRKY72-1 and WRKY72-2 were cloned from Newhall （Citrus sinensis Osbeck） and Calamondin. Sequence analysis showed that the full length ofcDNAs were 1 123, 1312, 1809 and 2208 bp, with 921, 480, 1809 and 1767 bp open reading frames （ORF）, respectively. Those cDNAs encode 306, 159, 602 and 588 amino acids, respectively. Analysis of amino acid sequences and protein structures indicated that the 4 CsWRKYs belonged to the group II WRKY proteins. The results of phylogenetic analysis showed that the 4 Cs WRKY genes had a close relationship with TcWRKY and VvWRKY family from Theobroma cacao and I,~tis vinifera, respectively. Subcellular localization analysis indicated that the 4 CsWRKY proteins were located in the nucleus. The expression of these genes in Newhall navel orange and Calamondin after Xac inoculation revealedthat WRKY22 was involved in the susceptible response of the host, while WRKY50 was involved in the immune response of Xac resistance. Xac inhibited the WRKY72-1- and WRKY72-2- mediated host basal immunity. Results of SA and MeJA treatments indicated that none of the 4 WRKY genes was involved in the disease and stress resistance of host mediated by SA and MeJA.

关 键 词：柑橘 CsWRKY 基因克隆 表达分析 

分 类 号：S666[农业科学—果树学]