紫花苜蓿转录因子基因MsAP_(2)的克隆及转化  

Cloning and Transformation of MsAP_(2) Gene in Medicago sativa

作　　者：石欣玥 尚骁尧 周玲芳 张铁军 晁跃辉[1] SHI Xin-yue;SHANG Xiao-yao;ZHOU Ling-fang;ZHANG Tie-jun;CHAO Yue-hui(College of Grassland Science of Beijing Forestry University,Beijing 100083)

机构地区：[1]北京林业大学草业与草原学院,北京100083

出　　处：《生物技术通报》2021年第12期13-21,共9页Biotechnology Bulletin

基　　金：国家自然科学基金面上项目(31772656)。

摘　　要：紫花苜蓿(Medicago sativa)为重要的豆科牧草植物,研究紫花苜蓿转录因子基因MsAP_(2)的功能,为探究MsAP_(2)的信号转导网络提供理论指导和材料基础,也为紫花苜蓿生物技术育种提供一定的参考和借鉴。运用RT-PCR技术克隆MsAP_(2),利用DNA重组技术构建3302-3flag-AP_(2)植物表达载体,并利用农杆菌介导法对紫花苜蓿进行遗传转化,对再生植株进行转基因鉴定、基因表达量分析、内源激素的测定(脱落酸、细胞分裂素、生长素和赤霉素)和表型鉴定。结果表明,成功获得转MsAP_(2)紫花苜蓿植株,与野生型相比,转基因植株的MsAP_(2)表达量和激素水平均发生明显改变,转基因植株呈现早衰性状,叶片形态和大小发生改变,根系生长受到抑制,分枝数发生改变。Alfalfa(Medicago sativa)is an important leguminous forage plant.Studying the function of MsAP_(2) provides theoretical guidance and material basis for exploring the signal transduction network of MsAP_(2),and also provides some reference for alfalfa biotechnology breeding technology.The MsAP_(2) was cloned by RT-PCR,and the 3302-3flag-AP_(2) plant expression vector was constructed by DNA recombination technology.The M.sativa was transformed by Agrobacterium-mediated method,and the regenerated plants were identified by transgene detection,gene expression analysis,determination of endogenous hormones(abscisic acid,cytokinin,auxin and gibberellin)and phenotype identification.The results showed that the MsAP_(2) transgenic M.sativa plants were successfully obtained.Compared with the wild type,the MsAP_(2) gene expression and hormone levels of transgenic plants were significantly changed.The transgenic plants demonstrated premature senescence,the leaf shape and size changed,the root growth was inhibited and the number of branches changed.

关 键 词：紫花苜蓿 AP_(2)/ERF转录因子 MsAP_(2) 转基因 

分 类 号：S541.9[农业科学—作物学]