玉米冷响应基因ZmRanBP1遗传转化分析  

Genetic Transformation Analysis of Maize(Zea mays)Cold Response GeneZmRanBP1

作　　者：马宁 苏胜忠[1] 单晓辉[1] 刘宏魁[1] 李世鹏[1] 李贺 韩俊友[1] 原亚萍[1] 吴颖 Ma Ning;Su Shengzhong;Shan Xiaohui;Liu Hongkui;Li Shipeng;Li He;Han Junyou;Yuan Yaping;Wu Ying(College of Plant Science,Jilin University,Changchun,130062;College of Horticulture Gardening,Tianjin Agricultural University,Tianjin,300380)

机构地区：[1]吉林大学植物科学学院,长春130062 [2]天津农学院园艺园林学院,天津300380

出　　处：《分子植物育种》2020年第17期5724-5729,共6页Molecular Plant Breeding

基　　金：国家转基因生物新品种培育重大专项(2014ZX080030-5B);吉林省重大科技招标专项(20160203005NY)共同资助。

摘　　要：中国东北地区春季温度较低,玉米容易受冷害影响,导致萎蔫,甚至死亡,最终减产,所以提高玉米适应冷胁迫的能力十分重要。本研究利用RT-PCR的方法从抗冷玉米自交系‘W9816’中成功克隆冷响应基因ZmRanBP1(RanBP1 domain containing protein)并构建过表达载体pCAMBIA-3301-35S-Zm RanBP1。通过农杆菌介导法获得了3个转基因拟南芥株系,逐代自交均获得T3代纯合拟南芥株系。对转基因拟南芥株系、突变体、野生型拟南芥进行耐冷及耐盐表型鉴定及比价分析,结果发现突变体对逆境胁迫的耐受力更强,具有较高的发芽率。利用农杆菌介导法分别侵染玉米‘Y423’的幼胚和Ⅱ型愈伤,阳性率分别为3.09%和4.15%。在拟南芥中进行逆境胁迫表型鉴定,并转入玉米自交系,可用于抗冷转基因玉米新种质的创制与遗传改良研究。Since the low temperature in spring in the northeast of China,maize is easily affected by the cold injury which causes wilting,plant death and yield reduction.Therefore,it is particularly important to improve low temperature resistance of maize.The cold response gene ZmRanBP1(RanBP1 domain containing protein)was successfully cloned by PT-PCR from the cold-resistant inbred line'W9816'.The expression vector pCAMBIA-3301-35S-ZmRanBP1 was constructed by double enzyme digestion method.Transgenic Arabidopsis thaliana strains were obtained by Agrobacterium-mediated method,and homozygous Arabidopsis thaliopsis strains of T3 generation were obtained by self-crossing.The cold-resistant and salt-resistant phenotype analysis of A.thaliana were tested.The results showed that the mutants had stronger tolerance to cold-and salt-stress and possessed higher germination rate.After Agrobacterium-mediated infection of immature embryo and callus of maize,the obtained positive rate was 3.09%and 4.15%,respectively.In this case,the adversity stress phenotype identification is carry out in the A.thaliana,and the corn inbred line is transferred into the corn inbred line and can be conducive to generating new germplasm creations and improving the cold-resistant transgenic corn.

关 键 词：玉米(Zea mays) 冷响应 ZmRanBP1 农杆菌介导法 

分 类 号：S513[农业科学—作物学]