桑树MnGATA7基因克隆及干旱胁迫下的表达分析  

Cloning and expression analysis of MnGATA7 gene in mulberry under drought stress

作　　者：刘江 刘刚[1] 代洁 曾益春[1] 危玲[1] 姚永权[1] 刘三梅 吴一磊 郑继川[1] 李永远[1] 黄盖群[1] LIU Jiang;LIU Gang;DAI Jie;ZENG Yichun;WEI Ling;YAO Yongquan;LIU Sanmei;WU Yilei;ZHENG Jichuan;LI Yongyuan;HUANG Gaiqun(Sericultural Research Institute,Sichuan Academy of Agricultural Sciences,Nanchong,Sichuan 637000,China)

机构地区：[1]四川省农业科学院蚕业研究所,四川南充637000

出　　处：《植物生理学报》2024年第12期1737-1746,共10页Plant Physiology Journal

基　　金：国家重点研发计划专项(2023YFD1600900);四川省财政自主创新专项(2022ZZCX085);四川省科技计划项目(2024YFCY009);四川省农业科学院“1+9”揭榜挂帅科技攻关项目(1+9KJGG004);国家现代农业产业技术体系四川蚕桑创新团队项目(SCCXTD-2024-17);南充市研发资金项目(23YYJCYJ0041)。

摘　　要：GATA转录因子对植物的生长发育过程、生物胁迫及非生物胁迫、叶绿素合成以及碳、氮代谢等生理活动发挥着重要作用。本研究以桑树幼苗为材料,成功克隆了桑树MnGATA7基因序列,并对其进行生物信息学分析、原核表达、Western blot及干旱胁迫下在各组织中的表达等探究。结果显示,MnGATA7序列开放阅读框长933 bp,编码310个氨基酸,蛋白分子量为34.25 kDa,理论等电子点为4.76,且定位于细胞核中。原核表达及Western blot分析显示,成功构建了MnGATA7基因的原核表达载体,并在体外实现了重组蛋白体的表达与纯化。组织表达分析表明,MnGATA7基因在桑树各组织中的表达模式存在显著差异,且随着干旱胁迫时间的延长,在根和表皮中的相对表达量呈现先增后降的趋势,在叶片中是呈现逐渐下调的趋势。综合表明,桑树MnGATA7基因可能参与了桑树干旱胁迫的生理响应。GATA transcription factors are crucial for various physiological activities such as plant growth and development,chlorophyll synthesis,carbon and nitrogen metabolism,as well as biological and abiotic stress responses.In this study,MnGATA7 gene sequence was successfully cloned from mulberry seedlings,and its bioinformatics analysis,prokaryotic expression,Western blot analysis and expression in various tissues under drought stress were performed.The results showed that the open reading frame length of MnGATA7 sequence was 933 bp,encoding 310 amino acids,the molecular weight of the protein was 34.25 kDa,the theoretical isoelectronic point was 4.76,and it was located in the nucleus.Prokaryotic expression and Western blot analysis showed that the prokaryotic expression vector of MnGATA7 gene was successfully constructed in this study,and the recombinant proteome was expressed and purified in vitro.Tissue expression analysis showed that there were significant differences in the expression patterns of MnGATA7 gene in various tissues of mulberry,and with the prolongation of drought stress,the relative expression in roots and epidermis showed a trend of first increasing and then decreasing,and in leaves was a trend of gradual down-regulation.These results indicate that MnGATA7 gene may be involved in the physiological response of mulberry to drought stress.

关 键 词：桑树 MnGATA7克隆 原核表达 非生物胁迫 

分 类 号：S888.2[农业科学—特种经济动物饲养] Q943.2[农业科学—畜牧兽医]