玉米光敏色素A1基因(ZmPHYA1)在棉花中的转化及分子鉴定  

Transformation and molecular identification of maize phytochrome A1 gene(ZmPHYA1)in cotton

作　　者：马燕斌 王霞[2] 李换丽 王平[3] 张建诚 文晋 王新胜 宋梅芳[3] 吴霞 杨建平[4] MA Yan-Bin;WANG Xia;LI Huan-Li;WANG Pin;ZHANG Jian-Cheng;WEN Jin;WANG Xin-Sheng;SONG Mei-Fang;WU Xia;YANG Jian-Ping(Institute of Cotton Research,Shanxi Agricultural University(Shanxi Academy of Agricultural Sciences),Yuncheng 044000,Shanxi,China;Department of Life Science,Yuncheng University,Yuncheng 044000,Shanxi,China;Beijing Radiation Center,Beijing 100875,China;College of Agronomy,Henan Agriculture University,Zhengzhou 450002,Henan,China)

机构地区：[1]山西农业大学(山西省农业科学院)棉花研究所,山西运城044000 [2]运城学院生命科学系,山西运城044000 [3]北京市辐射中心,北京100875 [4]河南农业大学农学院,河南郑州450002

出　　处：《作物学报》2021年第6期1197-1202,共6页Acta Agronomica Sinica

基　　金：山西省重点研发计划子课题(201703D211007-3);国家自然科学基金项目(31201253,31871709);国家转基因生物新品种培育重大专项(2016ZX08010-003);山西省农业科学院应用基础研究计划(YGJPY2007);运城学院博士科研启动项目(YQ-2017008)资助。

摘　　要：为评价玉米ZmPHYA1基因在棉花种质资源改良中的价值,本研究利用农杆菌介导法在陆地棉(Gossypium hirsutum)R15材料中进行了玉米ZmPHYA1基因的遗传转化。经过愈伤组织诱导、抗性愈伤筛选、体细胞分化诱导后获得棉花转基因再生植株。通过田间草铵膦除草剂筛选鉴定抗性植株,并利用PCR扩增其草铵膦抗性基因和目的基因ZmPHYA1进行分子鉴定,发现阳性植株对草铵膦除草剂具较好抗性,并可扩增到256 bp的草铵膦抗性基因和217 bp ZmPHYA1基因的特异条带。进一步通过免疫印迹检测表明,3个不同转基因株系中外源ZmPHYA1基因可正常表达约170 kD大小的蛋白,且在不同组织中该外源蛋白均可正常表达。此外,对转基因植株的不同农艺性状分析表明,转基因株系株高明显低于受体对照,而铃重和纤维长度等性状无明显差异。本研究成功获得具有草铵膦抗性和外源ZmPHYA1基因的棉花新种质材料,为进一步利用光敏色素基因创新种质资源提供了材料来源。In order to evaluate the potential value of maize phytochrome A1 gene(ZmPHYA1)in the improvement of cotton germplasm resources,we transferred it into upland cotton(Gossypium hirsutum L.)R15 via Agrobacterium tumefaciens-mediated transformation with glufosinate-resistance gene as selection marker.The regenerated cotton plants were obtained through callus induction,antibiotic resistance screening and differentiation induction.After screening the regenerated plants by the herbicide glufosinate ammonium in the field,PCR detection confirmed that both the target bands,including 256 bp of the glufosinate gene and 217 bp band of ZmPHYA1 gene,were detected in the homozygous transgenic plants.In addition,the exogenous ZmPHYA1 protein of about 170 kD was also checked by immuno-blot in three transgenic cotton lines.The results showed that the specific proteins could be detected in different tissues,including leaves,flowers and stems in the transgenic Line 9.The plant height of transgenic Line 9,Line 14,Line 41 were significantly shorter than that of the wild type,while the differences of other yield-related agronomic traits were not observed between the transgenic lines and the wild type.In this study,new cotton germplasms with glufosinate resistance and ZmPHYA1 gene were successfully obtained,which provided a material source for further utilization of phytochrome gene to innovate germplasm resources.

关 键 词：陆地棉 玉米光敏色素A1 蛋白表达 草铵膦抗性 

分 类 号：S562[农业科学—作物学]