大豆GmAMS基因及其启动子的克隆和表达分析  

Cloning and Expression Analysis of GmAMS Gene and Its Promoter in Soybean

作　　者：金玲[1] 张浩[1] 王松明 李强[1] 丁先龙 杨守萍[1] JIN Ling;ZHANG Hao;WANG Song-ming;LI Qiang;DING Xian-long;YANG Shou-ping(Soybean Research Institute,Nanjing Agricultural University/National Center for Soybean Improvement/Key Laboratory for Biology and Genetic Improvement of Soybean(General),Ministry of Agriculture/State Key Laboratory of Crop Genetics and Germplasm Enhancement/Jiangsu Modem Crop Production Collaborative Innovation Center,Nanjing 210095,China)

机构地区：[1]南京农业大学大豆研究所/国家大豆改良中心/农业部大豆生物学与遗传育种重点实验室(综合)/作物遗传与种质创新国家重点实验室/江苏省现代作物生产协同创新中心

出　　处：《大豆科学》2019年第6期889-897,共9页Soybean Science

基　　金：国家重点研发计划(2016YFD0101500,2016YFD0101504);中央高校基本科研业务费专项资金(KYT201801);长江学者和创新团队发展计划(PCSIRT_17R55)

摘　　要：bHLH(basic Helix-Loop-Helix)转录因子在植物雄配子发育中具有重要的调控作用。为探究bHLH转录因子在大豆花发育中的作用,以大豆细胞质雄性不育系NJCMS5A的花芽cDNA和叶片DNA为模板,通过RT-PCR克隆得到具有典型bHLH结构域的GmAMS基因及其启动子区域,并进行生物信息学分析和时空表达分析。生物信息学分析结果显示GmAMS基因的编码区序列全长为1 716 bp,编码571个氨基酸。亚细胞定位预测GmAMS位于细胞核中。荧光定量分析结果显示,相对于保持系NJCMS5B,在不育系NJCMS5A的花芽中GmAMS基因的表达水平显著下调。组织化学染色结果表明GmAMS启动子驱动的GUS蛋白主要集中在转基因拟南芥的幼小花药中表达。研究结果为进一步探究GmAMS在大豆花发育中的生物学功能和调控机制奠定了基础。The basic Helix-Loop-Helix(bHLH) transcription factor plays an important role in the regulation of male gamete development in plants. In order to explore the role of bHLH transcription factor in the flower development of soybean, the transcription factor GmAMS gene with typical bHLH domain and its promoter were cloned by RT-PCR using cDNA from the flower bud and DNA from the leaf of soybean cytoplasmic male sterile line NJCMS5 A as the template. Bioinformatics analysis and spatiotemporal expression analysis of GmAMS gene were conducted. The results of bioinformatics analysis showed that the coding region sequence(CDS) of GmAMS gene was 1 716 bp in length, encoding 571 amino acids. Subcellular localization prediction results showed that GmAMS was localized in the nucleus. Real-time fluorescence quantification analysis showed that the expression level of GmAMS gene in the flower bud of NJCMS5 A was significantly downregulated compared with the maintainer line NJCMS5 B. Histochemical staining results showed that the GUS protein driven by GmAMS promoterwas mainly expressed at the early anther development stage.The above results provided the foundation for further study on the biological function and regulation mechanism of GmAMS gene in the flower development of soybean.

关 键 词：大豆 GmAMS基因 启动子 基因克隆 表达分析 

分 类 号：R73[医药卫生—肿瘤]