苹果乙烯信号转导相关基因MdEIL1的克隆及功能鉴定  

Molecular Cloning and Functional Characterization of the Ethylene Signaling Related Gene MdEIL1 in Apple

作　　者：苏玲[1] 王淑惠[1] 郝玉金[1] 王小非[1] 由春香[1] 

机构地区：[1]山东农业大学园艺科学与工程学院,作物生物学国家重点实验室,农业部黄淮地区园艺作物生物学与种质创制重点实验室,山东泰安271018

出　　处：《园艺学报》2016年第3期409-418,共10页Acta Horticulturae Sinica

基　　金：国家重点基础研究发展计划(‘973’)项目(2013CB945103);国家自然科学基金项目(31272142,31471854)

摘　　要：以苹果‘嘎拉’(Malus×domestic‘Royal Gala’)为试材,分离了乙烯信号转导相关的MdEIL1基因(基因序列号:MDP0000423881)。序列分析表明,该基因包含长为1 980 bp完整的开放阅读框,编码658个氨基酸的蛋白。进化树分析表明MdEIL1属于EIN3/EIL转录因子家族蛋白。定量分析显示,MdEIL1基因在苹果的根、茎、叶、花和果实中有不同程度的表达,而且它的表达也受到乙烯、生长素和细胞分裂素的诱导。通过原核表达试验体外诱导MdEIL1蛋白成功,为后续蛋白功能鉴定奠定了基础。MdEIL1基因在拟南芥中异位表达,增强其乙烯响应,在暗处表现为下胚轴变短,并且AtERF1的表达量上升,显示转基因植株对乙烯的敏感性提高。An ethylene signal relative transcription factor named MdEIL1 was cloned from Malus × domestic‘Royal Gala＇. Sequence analysis showed that MdEIL1 contained 1 980 nucleotides,which encoded a protein containing 658 amino acids. Phylogenetic tree analysis showed that MdEIL1 belonged to EIN3/EIL transcription factor family proteins. q RT-PCRs analysis indicated that MdEIL1 has different levels of expression in apple＇s roots,stems,leaves,flowers and fruits,and was induced by ethylene,auxin and cytokinin hormones hormones in plants. We obtained the recombinant MdEIL1 protein by Prokaryotic expression,which was essential for protein functional identification in vitro. Ectopic expression of MdEIL1 in Arabidopsis can partially enhance ethylene response,which exhibited the short hypocotyl in darkness and expression of AtERF1 was improved in the MdEIL1 overexpressing transgenic lines.

关 键 词：苹果 MdEIL1 表达分析 原核表达 功能鉴定 

分 类 号：S661.1[农业科学—果树学]