西瓜NCED基因的鉴定及其对枯萎病和脱落酸的表达模式分析  

Identification of Watermelon NCED Genes and Analysis of Its Expression Pattern Against Fusarium Wilt and Abscisic Acid

作　　者：姚依秀 李玉林 何艳军 范敏[2] 高杰[1] Yao Yixiu;Li Yulin;He Yanjun;Fan Min;Gao Jie(College of Forestry and Horticulture,Xinjiang Agricultural University,Urumqi,830052;Institute of Vegetables,Zhejiang Academy of Agricultural Sciences,Hangzhou,310021)

机构地区：[1]新疆农业大学林学与园艺学院,乌鲁木齐830052 [2]浙江省农业科学院蔬菜研究所,杭州310021

出　　处：《分子植物育种》2022年第5期1393-1405,共13页Molecular Plant Breeding

基　　金：国家重点研发计划项目(2018YFD0100703);国家自然科学基金面上项目(31572145,31772332);新疆维吾尔自治区园艺学重点学科建设项目(2016-10758-37);浙江省自然科学基金青年科学基金项目(LQ18C150003);浙江省农业新品种选育重大科技专项子课题(2016C02051-4-2);浙江省农业科学院青年人才培养项目(2018R23R08E03)共同资助。

摘　　要：脱落酸(abscisic acid,ABA)是植物生长发育和逆境胁迫所必需的一种重要的植物激素,而9-顺式-环氧类胡萝卜素双加氧酶(NCED)是催化植物ABA合成的关键限速酶,在植物对干旱、高盐及病原菌等逆境的响应过程中发挥十分重要的作用。然而,目前还没有报道对西瓜中的ClNCED基因进行分离和鉴定。本研究在西瓜基因组中利用生物信息学技术共鉴定出10条ClNCED基因,预测并分析了这10条ClNCED基因的基因结构、染色体位置、亚细胞定位等基本信息及在西瓜不同组织器官、ABA和枯萎病处理下的表达模式。亚细胞定位预测发现大部分西瓜ClNCED蛋白主要被定位在叶绿体和细胞质中。利用不同物种的NCED蛋白进行进化分析发现西瓜ClNCED蛋白聚类在5个亚族中。亚族Ⅳ是西瓜特异亚族,只包含一对串联重复基因ClNCED09和ClNCED10编码的蛋白。在所有西瓜ClNCED基因中,只有一半有内含子,内含子数由5~12不等。另外,我们还利用荧光定量PCR(RT-qPCR)对ClNCED基因的时空表达模式进行分析,发现一半的西瓜ClNCED基因为特异表达基因,并且大部分在雄花中优势表达。此外,我们从ClNCED基因的启动子中发现了许多重要的顺式作用元件,这些元件与激素和逆境相关,对西瓜ClNCED基因在喷施ABA及接种尖孢镰刀杆菌后的表达水平进行分析,发现绝大部分ClNCED基因能响应ABA处理,同时还鉴定到多个响应枯萎病的ClNCED基因,这也为今后西瓜ClNCED基因的功能鉴定及相关的抗病分子育种提供重要的理论依据。Abscisic acid(ABA)was a hormone signal in the plant response to various stresses.9-cis-epoxycarotenoid dioxygenase(NCEDs)control a key regulatory step in the process of ABA biosynthesis in higher plants.NCEDs in plants played important roles in response to biological and abiotic stresses such as drought,high salinity and pathogens.However,it has not been reported to identify and study NCED genes in watermelon.In this study,a total of 10 ClNCED genes were identified in watermelon genome by bioinformatic methods,and their gene structure,chromosome location,subcellular location,protein structure,evolutionary relationship,promoter cis-acting elements,and expression patterns were predicted and analyzed.Most of the watermelon ClNCED proteins were predicted to located in chloroplast and cytoplasm.Phylogenetic tree constructed by NCED proteins from different species showed that watermelon NCED proteins in plants were clustered in five subfamilies.SubfamilyⅣwas a watermelon-specific subfamily that only contained a pair of proteins encoded by a pair of tandem duplicated genes ClNCED09 and ClNCED10.Only half of the ClNCEDs have introns and the number of introns ranges from 5 to 12.In addition,quantitative PCR(RT-qPCR)was used to analyze the spatio-temporal expression patterns of ClNCEDs.The results found that most of the ClNCEDs in watermelon were tissue-specific genes and most of them were predominantly expressed in male flowers.Meanwhile,numerous hormone-and stress-related elements were identified from ClNCED promoters.And RT-qPCR results found that the vast majority of ClNCEDs could respond to ABA treatment,as well as identified some ClNCED genes responsive to watermelon fusarium wilt.This provided an important theoretical basis to explore the function of NCEDs and watermelon disease resistance by molecular breeding.

关 键 词：西瓜 9-顺式-环氧类胡萝卜素双加氧酶(NCED) 脱落酸(ABA) 基因表达 进化分析 

分 类 号：S436.5[农业科学—农业昆虫与害虫防治]