柑橘CitCEP基因家族的鉴定及对逆境和激素的响应  

Identification of Citrus CitCEP Genes and Their Transcriptional Response to Stress and Hormone Treatments

作　　者：潘小婷 张静 葛廷 马岩岩 邓烈[1] 何绍兰[1] 易时来[1] 郑永强[1] 吕强[1] 谢让金[1] PAN XiaoTing;ZHANG Jing;GE Ting;MA YanYan;DENG Lie;HE ShaoLan;YI ShiLai;ZHENG YongQiang;LüQiang;XIE RangJin(Citrus Research Institute,Southwest University-Chinese Academy of Agricultural Sciences/National Engineering Research Center for Citrus Technology,Chongqing 400712)

机构地区：[1]西南大学柑桔研究所/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：《中国农业科学》2018年第16期3147-3158,共12页Scientia Agricultura Sinica

基　　金："十二五"国家科技计划(2014BAD16B02-1);中央高校基本科研业务费专项资金(XDJK2016B022)

摘　　要：【目的】基于全基因组鉴定分析柑橘CEP基因(CitCEPs)家族成员,了解各成员的分类进化关系,研究各基因成员在不同组织中的表达特异性以及对激素和非生物胁迫的响应,为进一步研究CEP基因家族的生物学功能打下基础。【方法】利用BLASTp基于Phytozome数据库鉴定柑橘CEP家族成员,采用GSDS、Prot Scale Tool、EXPASY、CLUSTALX、MEGA6.0、plant CARE、Cello等软件绘制家族成员基因结构图;分析预测蛋白的相对分子质量与等电点等理化性质;构建系统进化树及亚细胞定位预测等,利用实时荧光定量PCR技术(q RT-PCR)检测柑橘CEP基因家族各成员在‘资阳香橙’(‘Citrus junos Ziyang’)不同处理下的瞬时表达情况。【结果】柑橘CEP基因家族由11个成员组成,其中CitCEP10和CitCEP11含1个内含子,其余成员均无内含子;该基因家族成员均含有SPGV/IGH保守结构域序列,其预测蛋白均为亲水性蛋白,亲水性最强的氨基酸其值为3.711,亲水性最弱的氨基酸其值为-2.778;亚细胞定位预测结果显示该基因家族成员位于细胞不同位置,其中CitCEP1、CitCEP2、CitCEP4、CitCEP5、CitCEP6、CitCEP8定位于细胞外,CitCEP3和CitCEP7在细胞外和线粒体中均存在,CitCEP9位于细胞核中,CitCEP10在细胞质膜上,而CitCEP11位于线粒体和细胞核中;聚类分析发现,该家族成员分布于不同的分支中,分别与拟南芥CEP不同成员聚在一起,表明柑橘CEP成员间具有不同生物功能。表达分析显示,CitCEP2主要在茎、叶和子叶中表达,CitCEP3主要在根和子叶中表达,而CitCEP10和CitCEP11主要在果皮中表达,其余成员在上述组织中表达极低或不表达,体现了不同成员间组织特异性的差异。在干旱条件下,CitCEP2表达下调,CitCEP3、CitCEP10和CitCEP11的相对表达量均逐渐上调;而在盐胁迫下,CitCEP2、CitCEP10的响应模式与其所对应的干旱胁迫类似,CitCEP3和CitCEP11则呈现先升后降趋势。在乙烯(ETH)、�【Objective】The objectives of this study are to identify citrus CEP genes(CitCEPs)based on citrus whole genome sequences,understand the phylogenetic relationship among CitCEPs as well as their expression specificity in different tissues and response to hormone and abiotic stress,and to lay a foundation for further study on the biological function of CitCEPs.【Method】The citrus CEP genes were identified by BLASTp based on Phytozome database.Gene structure,phylogenetic tree,relative molecular mass,isoelectric point and other physical and chemical properties of CitCEP members were analyzed using GSDS,ProtScale Tool,EXPASY,CLUSTALX,MEGA6.0,plant CARE and Cello.Real-time quantitative PCR(qRT-PCR)was used to detect the expression levels of CitCEPs under different treatments of‘Citrus junos Ziyang’.【Result】The CitCEPs consists of 11 members,of which CitCEP10 and CitCEP11 contain 1 intron,and the other members have no intron.All CitCEPs contain conserved domain sequences(SPGV/IGH),and belong to hydrophilic protein.The magnitude of the most hydrophilic amino acid is 3.711 and that of the weakest hydrophilic amino acid is-2.778.Subcellular localization prediction showed that 11 CitCEPs were located in different positions of the cell.Among them,CitCEP1,CitCEP2,CitCEP4,CitCEP5 CitCEP6 and CitCEP8 were all located in extracellular.CitCEP3 and CitCEP7 existed in both extracellular and mitochondria.CitCEP9 and CitCEP10 were located in nucleus and plasma membrane,respectively.CitCEP11 was located in both mitochondria and nucleus.The phylogenetic analysis showed that the CitCEP members together with Arabidopsis thaliana CEPs were clustered into 3 groups,which indicated that the CitCEP members had different biological functions.Expression analysis showed that CitCEP2 was mainly expressed in stem,leaf and cotyledon,CitCEP3 was mainly expressed in root and cotyledon,while CitCEP10 and CitCEP11 were mainly expressed in pericarp,and the other members expressed very low or no expression in the above tissues,reflecting the

关 键 词：柑橘 CitCEP基因家族 基因表达 激素 胁迫 

分 类 号：S666[农业科学—果树学]