Acta Agrestia Sinica ›› 2019, Vol. 27 ›› Issue (4): 789-796.DOI: 10.11733/j.issn.1007-0435.2019.04.001

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Cloning and Expression Analysis of MsLEA4 Promoter from Medicago Sativa

JIA Hui-li1,2, SHI Yong-hong2, WANG Xue-min3, WANG Yun-qi2, WU Xin-ming2, LIU Jian-ning2, FANG Zhi-hong2, ZHANG Yan2, DONG Kuan-hu1   

  1. 1. College of Animal Science and technology, Shanxi Agricultural University, Taigu, Shanxi Province 030801, China;
    2. Animal Husbandry and Veterinary Institute, Shanxi Academy of Agricultural Sciences, Taiyuan, Shanxi Province 030032, China;
    3. Institute of Animal Science, Chinese Academy of Agricultures Sciences, Beijing, 100193, China
  • Received:2019-01-07 Revised:2019-06-11 Online:2019-08-15 Published:2019-09-26

紫花苜蓿MsLEA4启动子的克隆及表达分析

贾会丽1,2, 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   

  1. 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
  • 通讯作者: 董宽虎
  • 作者简介:贾会丽(1980-),女,山西临猗人,副研究员,博士研究生,主要从事牧草种质资源与育种研究,E-mail:jiahuili333@126.com
  • 基金资助:
    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

Abstract: 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.

Key words: Medicago sativa, MsLEA4 promoter, Adversary stress, Cis-element

摘要: 为了深入研究紫花苜蓿(Medicago sativaMsLEA4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。

关键词: 紫花苜蓿, MsLEA4启动子, 逆境胁迫, 顺式作用元件

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