Cloning and Expression Analysis of MsLEA4 Promoter from Medicago Sativa

doi:10.11733/j.issn.1007-0435.2019.04.001

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

Cloning and Expression Analysis of MsLEA4 Promoter from Medicago Sativa

JIA Hui-li^1,2, SHI Yong-hong², WANG Xue-min³, WANG Yun-qi², WU Xin-ming², LIU Jian-ning², FANG Zhi-hong², ZHANG Yan², DONG Kuan-hu¹

1. 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. 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

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 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。

关键词: 紫花苜蓿, MsLEA4启动子, 逆境胁迫, 顺式作用元件

CLC Number:

S551+7
Q78

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. Cloning and Expression Analysis of MsLEA4 Promoter from Medicago Sativa[J]. Acta Agrestia Sinica, 2019, 27(4): 789-796.

贾会丽, 石永红, 王学敏, 王运琦, 吴欣明, 刘建宁, 方志红, 张燕, 董宽虎. 紫花苜蓿MsLEA4启动子的克隆及表达分析[J]. 草地学报, 2019, 27(4): 789-796.

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Cloning and Expression Analysis of MsLEA4 Promoter from Medicago Sativa

紫花苜蓿MsLEA4启动子的克隆及表达分析

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