玉米冷响应基因ZmCyp40克隆及其遗传转化  

Cloning and Genetic Transformation of Cold Response Gene ZmCyp40 in Maize

作　　者：郭春艳 刘宏魁[1] 吴颖[1] 单晓辉[1] 苏胜忠[1] 李世鹏[1] 李贺 韩俊友[1] 原亚萍[1] 

机构地区：[1]吉林大学植物科学学院,长春130062

出　　处：《分子植物育种》2018年第2期434-439,共6页Molecular Plant Breeding

基　　金：国家转基因生物新品种培育重大专项(2014ZX0800305B);吉林省科技发展计划项目(20160203005NY)共同资助

摘　　要：从抗冷玉米自交系W9816中克隆冷响应基因ZmCyp40,对该基因进行序列分析,发现其编码389个氨基酸,与高粱中的同源基因亲缘关系较近。qRT-PCR分析玉米自交系W9816低温(4℃)处理0 h、3 h、6 h、12 h、24 h和48 h后其根、茎、叶中基因ZmCyp40表达情况,发现ZmCyp40在玉米不同部位冷响应表达趋势不同,但均受冷诱导表达。构建ZmCyp40过表达载体,采用农杆菌介导法转化玉米优良自交系Y423的幼胚和愈伤。PCR检测转化植株,ZmCyp40阳性植株率为5.9%。qRT-PCR分析发现部分阳性植株中ZmCyp40基因的表达量有极显著的升高,可用于进一步的功能分析及玉米抗冷性状的遗传改良。ZmCyp40,the cold response gene was cloned from the cold resistance maize inbred line W9816 and the gene sequence of ZmCyp40 was analyzed,the result of which showed that it encoded 389 amino acids and was closely related to homologous genes in sorghum.The expressions of ZmCyp40 in roots,stems and leaves of maize inbred lines W9816 treated at low temperature（4℃） for 0 h,3 h,6 h,12 h,24 h and 48 h were analyzed by qRTPCR,and the results found that the expression trend of cold response of ZmCyp40 in different parts of maize were different,but all of those were cold induced expression.ZmCyp40 overexpression vector was constructed and the immature embryo and callus of maize inbred line Y423 were transformed by Agrobacterium tumefaciensmediated method.Conversion plants were detected by PCR and the positive rate of ZmCyp40 was 5.9%.qRT-PCR analysis indicated that the expression of ZmCyp40 gene significantly increased in some positive plants,which could be used for further functional analysis and genetic improvement of cold resistant characters of maize.

关 键 词：玉米 冷胁迫 ZmCyp40基因 农杆菌介导法 遗传转化 

分 类 号：Q943.2[生物学—植物学] S513[农业科学—作物学]