甘薯蔓割病(Fusarium oxysporum f.sp.batatas)相关基因IbMAPKK9的克隆与特性分析  

Cloning and characterization of IbMAPKK9 gene associated with Fusarium oxysporum f.sp.batatas in sweet potato

作　　者：靖小菁 杨新笋[2] 靳晓杰 刘意 雷剑[2] 王连军[2] 柴沙沙[2] 张文英[1] 焦春海[2] JING Xiao-Jing;YANG Xin-Sun;JIN Xiao-Jie;LIU Yi;LEI Jian;WANG Lian-Jun;CHAI Sha-Sha;ZHANG Wen-Ying;JIAO Chun-Hai(College of Agriculture,Yangtze University,Jingzhou 434025,Hubei,China;Food Crops institute,Hubei Academy of Agricultural Sciences,Wuhan 430064,Hubei,China;Horticulture College,Hainan University/Key Laboratory for Quality Regulation of Tropical Horticultural Plants of Hainan Province,Haikou 570228,Hainan,China)

机构地区：[1]长江大学农学院,湖北荆州434025 [2]湖北省农业科学院粮食作物研究所,湖北武汉430064 [3]海南大学园艺学院/海南省热带园艺作物品质调控重点实验室,海南海口570228

出　　处：《作物学报》2023年第12期3289-3301,共13页Acta Agronomica Sinica

基　　金：国家重点研发计划项目(2019YFD1001300,2019YFD1001305);财政部和农业农村部国家现代农业产业技术体系建设专项(甘薯,CARS-10)资助。

摘　　要：促丝裂原活化蛋白激酶(MAPK)级联是生物体内一种重要的信号转导途径,广泛存在于植物中。MAPKKs位于该级联反应通路中间,对信号传递起到关键作用,目前在甘薯中少有报道。基于前期的转录组学分析,本研究克隆出1个与甘薯抗病相关的基因IbMAPKK9。生物信息学分析表明,IbMAPKK9包含一个987 bp开放阅读框(open reading frame,ORF),编码328个氨基酸,具有1个蛋白激酶家族保守结构域(PF00069),IbMAPKK9蛋白以α螺旋和无规则卷曲为主,启动子区包含多种激素(茉莉酸甲酯、乙烯、脱落酸、赤霉素、水杨酸)相关及胁迫响应元件。进化分析表明,IbMAPKK9蛋白与三叶裂薯、日本牵牛花、番茄和马铃薯亲缘关系较近。亚细胞定位显示IbMAPKK9蛋白定位于细胞核。实时荧光定量PCR发现IbMAPKK9在甘薯根、茎、叶和叶柄中均表达,并响应甘薯蔓割病侵染。瞬时表达分析结果表明,IbMAPKK9引起5个与水杨酸合成途径及信号转导途径相关的基因在48 h内表达上调,推测IbMAPKK9通过介导水杨酸信号途径影响植物的抗性。本研究可为进一步解析甘薯IbMAPKK9的生物学功能提供理论依据。Mitogen-activated protein kinase(MAPK)cascades are important signaling modules in all organisms,which widely exist in plants.Mitogen-activated protein kinase kinase(MAPKKs)are located in the middle of the cascade reaction pathway and play the key roles in signal divergence.The disease resistance gene IbMAPKK9 was screened and cloned based on transcriptome data in sweetpotato,which contained a 987 bp open reading frame,encoding 328 amino acids and 1 domain(PF00069).Its protein mainly consisted ofα-helixes and random coils.Multiple hormones(methyl jasmonate,ethylene,abscisic acid,gibberellin,and salicylic acid)and stress-related response elements were detected in the promoter region.Homologous protein comparison showed that IbMAPKK9 was closely related to ItMAPKK9-like(XP_031110493.1),InMAPKK9-like(XP_019189439.1),SlMAPKK9(NP_001234595.1),and StMAPKK9(XP_006363984.1).Besides,IbMAPKK9 gene encoded nuclear localization protein.The relative expression levels revealed that IbMAPKK9 was expressed in roots,stems,leaves,and petioles in response to Fusarium oxysporum f.sp batatas(Fob)infection.Transient expression analysis showed that IbMAPKK9 induced up-regulation of 5 genes related to salicylic acid synthesis pathway and signal transduction pathway within 48 hours,suggesting that IbMAPKK9 affected plant resistance by mediating salicylic acid signal pathway.This study can provide a theoretical basis for further analysis of the biological function of IbMAPKK9.

关 键 词：甘薯 IbMAPKK9 蔓割病 基因克隆 亚细胞定位 瞬时表达 

分 类 号：S435.31[农业科学—农业昆虫与害虫防治]