紫花苜蓿MsLEA4启动子的克隆及表达分析  

Cloning and Expression Analysis of MsLEA4 Promoter from Medicago Sativa

作　　者：贾会丽 石永红[2] 王学敏[3] 王运琦[2] 吴欣明[2] 刘建宁[2] 方志红[2] 张燕[2] 董宽虎[1] JIA Hui-li;SHI Yong-hong;WANG Xue-min;WANG Yun-qi;WU Xin-ming;LIU Jian-ning;FANG Zhi-hong;ZHANG Yan;DONG Kuan-hu(College of Animal Science and technology,Shanxi Agricultural University,Taigu,Shanxi Province 030801,China;Animal Husbandry and Veterinary Institute,Shanxi Academy of Agricultural Sciences,Taiyuan,Shanxi Province 030032,China;Institute of Animal Science,Chinese Academy of Agricultures Sciences,Beijing,100193,China)

机构地区：[1]山西农业大学动物科技学院,山西太谷030801 [2]山西省农业科学院畜牧兽医研究所,山西太原030032 [3]中国农业科学院北京畜牧兽医研究所,北京100193

出　　处：《草地学报》2019年第4期789-796,共8页Acta Agrestia Sinica

基　　金：现代农业产业技术体系建设专项(CARS-34);山西省重点研发计划重点项目(201703D211002-9-2);山西省农业科学院雁门关农牧交错带专项(YCX2017D2107,YCX2018102)资助

摘　　要：为了深入研究紫花苜蓿(Medicago sativa)MsLEA4基因的功能,本试验采用染色体步移技术从紫花苜蓿基因组中扩增出MsLEA4启动子序列,构建GUS植物超表达载体,并根据序列分析结果对紫花苜蓿进行相应逆境胁迫。结果表明:紫花苜蓿MsLEA4启动子含有响应脱落酸(abscisic acid,ABA)调控的顺式作用元件ABRE,响应赤霉素(gibberellin,GA)调控的顺式作用元件P-box,参与胚乳合成的顺式作用元件GCN4和Skn-1,以及涉及光调控和光周期的顺式作用元件;外源ABA、GA、持续光照以及黑暗均能诱导MsLEA4基因的表达;GUS基因只在拟南芥花和荚果中表达。因此,MsLEA4基因能参与紫花苜蓿的逆境调控,与该基因启动子序列中所含的一系列顺式作用元件相关。本研究为进一步探索MsLEA4基因在紫花苜蓿逆境胁迫、光调控以及组织特异性表达中的作用奠定了基础。In order to further analyze the function of MsLEA4,we cloned MsLEA4 promoter sequence from alfalfa(Medicago Sativa)genome by genomic walking technique,constructed MsLEA4 promoter::GUS transient expression vector,and also conducted corresponding stresses on alfalfa according to the promoter sequence analysis.The results showed that:MsLEA4 promoter contained the basic elements TATA-box and CAAT-box,included abscisic acid(ABA)responsive elements(ARBE),gibberellin(GA)responsive elements(P-box),and cis-elements(GCN4,Skn-1)involved in endosperm synthesis.In addition to this,the promoter sequence also contained 6 cis-elements related to light regulation and a cis-element related to circadian rhythm.ABA,GA,continuous light and darkness all can induce MsLEA4 expression;the GUS gene was expressed only in flowers and siliques of Arabidopsis thaliana.Therefore,MsLEA4 can participate in the regulation of alfalfa stress responses,which is related to a series of cis-elements contained in the promoter.This study laid a foundation for further exploring the role of MsLEA4 in stresses,light regulation and tissue specific expression of alfalfa.

关 键 词：紫花苜蓿 MsLEA4启动子 逆境胁迫 顺式作用元件 

分 类 号：S5517[农业科学—作物学] Q78[生物学—分子生物学]