柑橘Rboh家族鉴定及其对激素和柑橘溃疡病的响应  

Identification of Rboh Family and the Response to Hormone and Citrus Bacterial Canker in Citrus

作　　者：秦秀娟 祁静静 窦万福 陈善春[1] 何永睿[1] 李强[1] QIN XiuJuan;QI JingJing;DOU WanFu;CHEN ShanChun;HE YongRui;LI Qiang(Citrus Research Institute,Southwest University/Chinese Academy of Agricultural Sciences,Chongqing 400712)

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：《中国农业科学》2020年第20期4189-4203,共15页Scientia Agricultura Sinica

基　　金：国家重点研发计划(2018YFD1000306);中央高校基本科研业务费(SWU115025);广西科技重大专项(桂科AA18118046-6);国家现代农业产业技术体系建设资金(CARS-26)。

摘　　要：【目的】活性氧(reactive oxygen species,ROS)是植物抗病反应中重要的信号分子,而Rboh是参与活性氧产生的关键酶之一。本研究通过鉴定和分析柑橘全基因组中Rboh基因家族生物信息学特性、生物胁迫相关植物激素和溃疡病菌(Xanthomonas citri subsp. citri,Xcc)侵染诱导下表达模式,为研究Rboh基因家族与柑橘抗溃疡病过程的相关性,进一步研究和利用柑橘Rboh基因打下基础。【方法】利用过氧化物酶数据库RedOxiBase和柑橘(甜橙)CAP数据库获得柑橘Rboh家族序列信息;利用生物信息学软件对CsRboh进行理化性质、系统进化关系、染色体定位、基因结构、蛋白功能结构域、保守基序和启动子元件系统分析。通过实时荧光定量PCR(qRT-PCR)分析植物激素水杨酸(salicylic acid,SA)、茉莉酸(jasmonic acid,JA)、脱落酸(abscisic acid,ABA)和溃疡病菌处理下CsRboh表达模式。【结果】鉴定得到7个柑橘Rboh家族成员(CsRboh01—CsRboh07),其编码蛋白包含784—946个不等的氨基酸残基,等电点(PI)分布在8.67—9.40,主要定位于细胞膜结构和细胞质。根据系统进化树可将柑橘Rboh家族划分为I—IV 4个亚家族。CsRboh家族基因不均匀分布在甜橙的5条染色体,均含有典型的呼吸爆发NADPH氧化酶结构域(NADPHOx)、铁还原酶跨膜组件(Ferricreduct)、FAD结合结构域(FADbinding)和铁还原酶NAD结合结构域(NADbinding)4个功能结构域。CsRboh家族各基因的启动子区域含不同激素响应元件,数目和类型各有差异。qRT-PCR结果显示CsRboh基因家族各成员可响应激素和溃疡病菌诱导,在不同激素和溃疡病菌处理下感病品种晚锦橙和抗病品种四季橘中表达模式具有差异。结合系统进化树聚类分析、同源基因功能研究及其启动子区域顺式作用元件分析发现,CsRboh02、CsRboh04和CsRboh06在抗、感两个品种中受柑橘溃疡病菌诱导表现出明显不同的表达趋势和表达量。【结论【Objective】Rboh is the main producer of reactive oxygen species(ROS), which is the key molecule involved in plant disease resistance signaling. To clarify the relationships between CsRboh family and citrus bacterial canker(CBC), the Rboh family of Citrus sinensis was identified and the bioinformatics characteristics as well as the expression patterns induced by biotic stress-related plant hormones and Xanthomonas citri subsp. citri(Xcc) were analyzed. It is also aimed to lay the foundation for further research of citrus Rboh genes in molecular breeding.【Method】Firstly, the RedOxiBase database and the citrus(sweet orange) CAP database were used to identify the citrus Rboh family sequences from the whole genome of C. sinensis. Then, series of bioinformatics tools were used to perform analysis of physical and chemical properties, phylogenetic relationships, chromosomal localizations, gene structures, protein functional domains, the conserved motifs and promoter elements systematically. Finally, q RT-PCR was used to analyze the expression patterns of CsRbohs induced by biotic stress-related plant hormones(salicylic acid, SA;jasmonic acid, JA;abscisic acid, ABA) and Xcc.【Result】A total of 7 members of the Rboh family in C. sinensis genome were identified(CsRboh01-CsRboh07). These genes encoded proteins contain 784-946 amino acid(AA) residues with isoelectric points(PI) ranging from 8.67 to 9.40. CsRbohs mainly distribute on the cell membrane system and cytoplasmic. The CsRbohs family can be divided into I to IV sub-groups based on the phylogenetic tree. These 7 genes are unevenly distributed on 5 sweet orange chromosomes. Each of the CsRbohs contains 4 typical functional domains, namely the respiratory burst NADPH oxidase domain(NADPH_Ox), the ferric reductase like transmembrane component(Ferric_reduct), the FAD binding domain(FAD_binding) and the ferric reductase NAD binding domain(NAD_binding). The promoter regions of Cs Rboh family genes contain different hormone response elements with different numbers

关 键 词：柑橘溃疡病 Rboh家族 柑橘溃疡病菌 水杨酸 茉莉酸 

分 类 号：S436.66[农业科学—农业昆虫与害虫防治]