摘要
高产是玉米育种的重要目标,籽粒大小是决定籽粒产量的重要因子。基于水稻中控制籽粒大小的OsGS5基因编码序列,利用同源克隆方法,获得了玉米ZmGS5基因,其cDNA全长为1695bp,开放阅读框1491bp,编码496个氨基酸,含有Peptidase-S10结构域、1个信号肽和1个丝氨酸羧肽酶(serine carboxypeptidases,SCP)类蛋白所特有的催化活性中心,这些都与SCP家族结构特点相符,也与OsGS5相关研究结果相同;磷酸化位点分析结果表明,ZmGS5含有丝氨酸、苏氨酸和酪氨酸等蛋白激酶识别位点。qRTPCR分析显示ZmGS5在雄穗和叶片中的表达量较高,而在胚及胚乳中的表达量相对较低。除此之外,采用农杆菌介导法,建立了拟南芥的遗传转化体系,得到纯合转基因株系,T3代转基因拟南芥种子千粒重为0.0169g,较野生型千粒重(0.0139g)高。
High yield is an important goal of the maize breeding,grain size is a major determinative factor of high yield.In this study,based on the method of homology cloning,the OsGS5 sequence was used as a template to obtain the ZmGS5 gene.The sequencing result revealed that the full length cDNA of ZmGS5 was 1695bp.The Open Reading Frame(ORF)encoding 496 amino acid was 1491bp.The ZmGS5 protein contained conservative structure domain Peptidase-S10,a signal peptide,an active site of the serine carboxypeptidases,which were all conformed to the structure of serine carboxypeptidase family and also conformed to the OsGS5 related research results.Phosphorylation locus analysis indicated that the protein contained recognition loci of Ser,Thr,Tyr kinases.The result of real-time quantitative PCR showed that ZmGS5 had a much higher expression level in tassel and leaves,and the lower expression level in embryo and endosperm.According to agrobacterium-mediated method,we established genetic transformation system in Arabidopsis and obtained homozygous transgenic lines.The 1000-seed weight of T3 Arabidopsis(0.0169g)was higher than that of wild type(0.0139g).
作者
刘晓丽
韩利涛
魏楠
申飞
蔡一林
Liu Xiaoli;Han Litao;Wei Nan;Shen Fei;Cai Yilin(Department of Food Engineering,Henan Vocational College of Agriculture,Zhengzhou 451450,Henan,China;Maize Research Institute,Southwest University,Chongqing 400715,China)
出处
《作物杂志》
北大核心
2021年第1期16-25,共10页
Crops
基金
河南农业职业学院科研创新团队项目(HNACKT-2020-05)
河南农业职业学院科研创新团队项目(HNACKT-2019-02)。
关键词
ZmGS5
基因克隆
丝氨酸羧肽酶
分子特性分析
遗传转化
ZmGS5
Gene cloning
Serine carboxypeptidases
Molecular characteristics analysis
Genetic transformation
