柑橘AOS1-2的克隆及其响应溃疡病菌侵染的表达分析  

Cloning and Expression Analysis of CsAOS1-2 in Responding to Citrus Canker Disease

作　　者：张乐欢 邹昌玉 王兆昊 杨雯 邹修平[1] 何永睿[1] 陈善春[1] 龙琴 ZHANG Lehuan;ZOU Changyu;WANG Zhaohao;YANG Wen;ZOU Xiuping;HE Yongrui;CHEN Shanchun;LONG Qin(National Citrus Engineering Research Center,Citrus Research Institute,Southwest University/Chinese Academy of Agricultural Sciences,Chongqing 400712,China)

机构地区：[1]西南大学/中国农业科学院柑桔研究所,国家柑桔工程技术研究中心,重庆400712

出　　处：《园艺学报》2023年第6期1355-1367,共13页Acta Horticulturae Sinica

基　　金：国家自然科学基金青年科学基金项目(32202425);中央高校基本科研业务费专项资金资助项目(SWU120059);国家现代农业产业技术体系建设专项资金项目(CARS-26)。

摘　　要：对柑橘中茉莉酸(Jasmonic acid,JA)生物合成途径中的关键限速酶丙二烯氧化物合酶AOS家族基因进行注释,确定其受溃疡病菌Xanthomonas citri subsp. citri(Xcc)诱导的表达模式。从甜橙数据库中共注释出3个AOS家族成员,其中CsAOS1-2对溃疡病菌响应最强烈;CsAOS1-2在‘晚锦橙’(甜橙,易感溃疡病)和‘金弹’(金柑,抗溃疡病)中均受病原菌诱导,但在‘晚锦橙’中表达量上调幅度远高于金弹;CsAOS1-2全长2 656 bp,开放阅读框1 599 bp,编码532个氨基酸,分子量为59 499.81,为非分泌性蛋白,其349~510 aa为典型的P450功能域;Cs AOS1-2的表达无组织特异性;‘晚锦橙’和‘金弹’的AOS1-2启动子均含有参与植物激素和逆境应答的顺式作用元件,但在数量和类型上存在差异;烟草瞬时转化亚细胞定位分析显示CsAOS1-2定位在叶绿体中。CsAOS1-2强烈响应柑橘溃疡病菌的侵染,推测其与柑橘溃疡病敏感性具有密切的关系。The AOS genes,encoding the key rate limiting enzyme in the jasmonic acid(JA)biosynthetic pathway in citrus,are annotated,and their expression patterns in response to Xanthomonas citri subsp.citri(Xcc)were determined by qRT-PCR.Based on the reference genome databases of sweet orange(Citrus sinensis),three AOS genes(CsAOS1-1,CsAOS1-2 and CsAOS1-3)were detected in citrus.Gene expression analysis showed that CsAOS1-2 is the most responsive to Xcc,compared with the other two AOS genes.The expression of CsAOS1-2 was increased by Xcc in both‘Wanjincheng’orange(Citrus sinensis Osbeck,canker-susceptible cultivar)and‘Jindan’(Fortunella crassifolia Swingle,canker-resistant cultivar).But its expression level induced by Xcc in‘Wanjincheng’orange was significantly higher than that in‘Jindan’.A 2656 bp long sequence of CsAOS1-2 was amplified from Wanjincheng orange.Bioinformatics analysis showed that CsAOS1-2 had a 1599 bp open reading frame which encodes a 59499.81 protein with 532 amino acids.CsAOS1-2 is a non-secretory protein and the 349-510 aa is a typical P450 domain.The similar expression levels of CsAOS1-2 were found in leaf,stem,rind and seed tissues.The AOS1-2 promoters of‘Wanjincheng’orange and‘Jindan’contained multiple cis-acting elements involved in plant adversity and hormone responses,but there were differences in their number and type.Subcellular localization analysis showed CsAOS1-2 located in tobacco chloroplasts.The study suggested that CsAOS1-2 strongly respond to Xcc infection and is closely related to susceptibility to citrus canker in citrus.

关 键 词：柑橘 溃疡病 茉莉酸 限速酶 AOS 克隆 表达 

分 类 号：S666[农业科学—果树学]