棉花GaLMI1-like基因功能及其启动子表达分析  

Function and Promoter Expression Analysis of GaLMI1-like Gene in Cotton

作　　者：杨小贝 陈二永 李成伟 YANG Xiaobei;CHEN Eryong;LI Chengwei(College of Life Science and Technology,Henan Institute of Science and Technology,Henan Engineering Research Center of Crop Genome Editing,Xinxiang 453003,China;College of Biological Engineering,Henan University of Technology,Zhengzhou 450001,China)

机构地区：[1]河南科技学院生命科技学院,河南省粮食作物基因组编辑工程技术研究中心,河南新乡453003 [2]河南工业大学生物工程学院,河南郑州450001

出　　处：《华北农学报》2022年第3期27-36,共10页Acta Agriculturae Boreali-Sinica

基　　金：河南省自然科学基金青年基金项目(105020220029);河南科技学院2017年博士科研启动费项目(201010617014)。

摘　　要：LMI1基因是叶片锯齿状结构发育调控的关键基因。为了研究棉花鸡脚叶发育的机理,通过PCR扩增技术从A基因组棉花亚洲棉石溪亚1号中克隆出GaLMI1-like基因及其启动子序列,大小分别为681,1439 bp。结构域分析发现,GaLMI1-like蛋白含有与陆地棉中同源基因一样的homeobox结构域,进一步构建了GaLMI1-like基因过表达载体p6MYC-GaLMI1-like,转化拟南芥后验证了GaLMI1-like基因具有调控叶片缺刻表型发育的功能。对启动子序列进行顺式作用元件分析,发现其除了具有CACA-box和TATA-box等基本作用元件外,还具有光响应及根、茎和叶肉特异性表达相关元件。构建了GaLMI1-like启动子的GUS融合表达载体并转化拟南芥,GUS染色结果显示,该启动子能够驱动GUS基因在根中柱、茎和叶片中表达,其中在叶片中染色较深。上述结果表明,GaLMI1-like基因具有调控缺刻叶形成的功能,且此调控棉花叶形发育的功能是通过GaLMI1-like启动子调控其在叶片中强表达实现的。LMI1 is a key gene in the regulation of serrated leaf development.In order to study the mechanism of okra leaf development in cotton,we cloned GaLMI1-like gene and GaLMI1-like promoter by PCR amplification technology from Shixiya 1,which is an Asia cotton species of a genome.The sequence lengths were 681,1439 bp,respectively.Domain analysis found that GaLMI1-like contained a homeobox domain.Then,we constructed a reorganization overexpression vector p6MYC-GaLMI1-like.The transgenic Arabidopsis showed incised leaf shape when GaLMI1-like was overexpressed in Arabidopsis.Moreover,we analyzed the cis-acting elements of the promoter sequence of GaLMI1-like.It was found that it had basic acting elements,such as CACA-box and TATA-box,and also had photoresponsive element,root,stem and mesophyll-specific expression elements.GUS fusion expression vector of GaLMI1-like promoter was constructed and transformed into Arabidopsis thaliana.GUS staining results showed that the promoter could drive GUS gene expression in root column,stem and leaf,and the expression was especially high in leaf.These results indicated that GaLMI1-like had the function of regulating the formation of incised leaves,and its function was realized by the strong expression of GaLMI1-like regulated by GaLMI1-like promoter in cotton leaves.

关 键 词：棉花 GaLMI1-like基因 叶形 启动子 顺式作用元件 转基因拟南芥 GUS染色分析 

分 类 号：S562[农业科学—作物学]