紫花苜蓿atpA基因的克隆及表达模式分析  

Cloning and Expression Analysis of atpA Gene in Medicago sativa

作　　者：李莉萍 李旭漉 张晓捷 谷丽丽 张博[1] LI Li-ping;LI Xu-lu;ZHANG Xiao-jie;GU Li-li;ZHANG Bo(Western Arid Region Grassland Resources and Ecology Key Laboratory,Xinjiang Key Laboratory of Grassland Resources and Ecology,College of Grassland Science,Xinjiang Agricultural University,Urumqi,Xinjiang 830052,China)

机构地区：[1]西部干旱荒漠区草地资源与生态教育部重点实验室,新疆草地资源与生态重点实验室,新疆农业大学草业学院,新疆乌鲁木齐830052

出　　处：《草地学报》2022年第6期1413-1421,共9页Acta Agrestia Sinica

基　　金：国家自然科学基金(31660687);新疆维吾尔自治区社厅留学人员科技活动择优资助项目(2016);新疆农业大学博士后流动站;新疆农业大学研究生创新项目(XJAUGIR2021023)资助。

摘　　要：叶绿体atpA基因位于ATP合酶的CF_(1)上,编码α亚基,是光合作用不可缺少的关键基因。本研究利用RT-PCR技术,从‘新疆大叶’苜蓿(Medicago sativa L.‘Xinjiang Daye’)中克隆出atpA基因,并进行生物信息学分析,采用实时荧光定量PCR技术进行基因表达模式检测。结果显示,MsatpA基因编码510个氨基酸,相对分子质量为55.71 KD,等电点为5.22。MsAtpA蛋白含有多个磷酸化位点且无跨膜结构和信号肽,二级结构中α-螺旋占比最高;亚细胞定位预测该基因编码的蛋白位于叶绿体中,与同为豆科苜蓿属的黄花苜蓿(Medicago falcata)、蒺藜苜蓿(Medicago truncatula)的AtpA氨基酸序列同源性较高。表达模式检测结果显示,MsatpA基因在叶中的表达量最高,根中最少;结荚期的基因表达量显著高于其他发育时期;MsatpA基因响应低温、高温、盐和渗透胁迫应答,且呈现出不同的表达特点。研究结果为紫花苜蓿atpA基因功能研究奠定基础。The chloroplast atpA gene is a key indispensable gene for photosynthesis,which encodes ATP synthaseαsubunit and is located on chloroplast ATP synthase CF_(1).atpA gene was cloned from Medicago sativa L.‘Xinjiang Daye’by RT-PCR,and bioinformatics analysis was carried out in our research.Then,expression pattern of this gene was detected by quantitative Real-time PCR.The results showed that MsatpA gene encoded 510 amino acids with the relative molecular weight of 55.71 KD and the isoelectric point of 5.22.MsatpA protein contained multiple phosphorylation without transmembrane structure and signal peptide.In the secondary structure of the protein,theαhelix accounted for the highest proportion.The subcellular localization prediction showed that the protein encoded by the gene was located in chloroplast,and its amino acid sequence had high homology with Medicago falcata and Medicago truncatula which all belonged to Leguminosae Medicago genus.The results of gene expression pattern showed that the relative expression level of MsatpA gene was the highest in leaves and the lowest in roots,and the gene expression level in pod setting stage was significantly higher than that in other development stages.MsatpA gene responded to low temperature,high temperature,salt and osmotic stress,and showed different expression characteristics.The results laid the foundation for the functional study of ATP synthaseαsubunits in Medicago sativa.

关 键 词：ATPA 紫花苜蓿 基因克隆 生物信息学分析 基因表达 

分 类 号：S812.6[农业科学—草业科学]