水稻EIN3/EIL转录因子家族克隆与表达分析  

Cloning and expression analysis of EIN3/EIL transcription factor family in rice

作　　者：高熊枫 李杰 朱佳美 寿惠霞[1] GAO Xiongfeng;LI Jie;ZHU Jiamei;SHOU Huixia(College of Life Sciences,Zhejiang University,Hangzhou 310058,China)

机构地区：[1]浙江大学生命科学学院,杭州310058

出　　处：《植物生理学报》2022年第8期1475-1484,共10页Plant Physiology Journal

基　　金：浙江省重点研发计划(2021C02057)。

摘　　要：乙烯是植物体内的重要激素,除了参与调控植物生长发育与各种胁迫响应以外,还参与调控植物的缺铁响应。EIN3/EIL转录因子家族被认为参与植物乙烯信号转导。前期本课题组报道了水稻(Oryza sativa) bHLH型转录因子OsbHLH156是一个响应缺铁的关键调控因子,酵母单杂交结果显示EIN3/EIL转录因子家族能与OsbHLH156启动子结合。为了进一步研究EIN3/EIL转录因子家族在水稻铁稳态中的作用,本研究对水稻EIN3/EIL家族6个转录因子进行生物信息学分析;通过农杆菌(Agrobacterium tumefaciens)介导的烟草(Nicotiana tabacum)瞬时表达,观察水稻EIN3/EIL转录因子家族的亚细胞定位,并利用实时荧光定量PCR技术分析不同组织中水稻EIN3/EIL转录因子家族编码基因的表达水平以及缺铁条件下的表达水平。结果表明,与拟南芥(Arabidopsis thaliana)乙烯调控核心转录因子AtEIN3相比,水稻EIN3/EIL转录因子家族在基因结构上具有保守性,OsEIL1、OsEIL2与AtEIN3在进化树上位于同一分支,蛋白相似性最高。亚细胞定位结果显示除OsEIL6以外的其他5个成员均定位于细胞核。组织表达模式分析显示6个水稻EIN3/EIL基因主要在根茎叶中表达。缺铁条件表达模式分析显示OsEIL2、OsEIL3受缺铁诱导。本研究为水稻EIN3/EIL转录因子家族的进一步功能研究奠定基础。Ethylene is an important hormone in plants,which not only participates in regulating plant growth and development and various stress responses,but also regulates iron deficiency responses in plants.The EIN3/EIL transcription factor family is thought to be involved in plant ethylene signaling.Previously,our group found that the rice(Oryza sativa) bHLH-type transcription factor OsbHLH156 is an essential regulator for iron deficiency response.A yeast-one hybridization showed that the EIN3/EIL transcription factor family can bind to the OsbHLH156 promoter.To understand the role of EIN3/EIL transcription factor family in regulating iron homeostasis in rice,we conducted bioinformatics analysis of the six rice EIN3/EIL transcription factors.Through Agrobacterium tumefaciens–mediated transient expression in tobacco(Nicotiana tabacum),the subcellular localization of the rice EIN3/EIL transcription factors was investigated.The expression levels of rice EIN3/EIL transcription factor family genes in different tissues and the expression levels under iron deficiency were analyzed by real-time quantitative PCR.Results showed that compared with the Arabidopsis thaliana ethylene-regulated core transcription factor AtEIN3,the rice EIN3/EIL transcription factor family was conserved in gene structure.OsEIL1 and OsEIL2 are located in the same branch with AtEIN3 on the phylogenetic tree,with the highest protein similarity.Subcellular localization results showed that all members except OsEIL6 were localized in the nucleus.Tissue expression analysis showed that the Os EIN3/EIL transcription factors were mainly expressed in rhizomes and leaves.The expression of OsEIL2 and OsEIL3 were induced by iron deficiency.This study provides a basis for further functional studies of the rice EIN3/EIL transcription factor family.

关 键 词：水稻 乙烯 EIN3/EIL 转录因子 表达模式 

分 类 号：S511[农业科学—作物学]