玉米ZmDHN15基因在烟草中的遗传转化及抗冷性分析  

Genetic Transformation and Chilling Resistance Analysis of Maize ZmDHN15 Gene in Tobacco

作　　者：陈楠楠 王春来 蒋振忠 焦鹏 关淑艳[2,3] 马义勇[2,3] CHEN Nan-nan;WANG Chun-lai;JIANG Zhen-zhong;JIAO Peng;GUAN Shu-yan;MA Yi-yong(College of Life Sciences,Jilin Agricultural University,Changchun 130118;College of Agronomy,Jilin Agricultural University,Changchun 130118;Joint International Research Laboratory of Modern Agricultural Technology,Ministry of Education,Jilin Agricultural University,Changchun 130118)

机构地区：[1]吉林农业大学生命科学学院,长春130118 [2]吉林农业大学农学院,长春130118 [3]吉林农业大学现代农业技术教育部国际合作联合实验室,长春130118

出　　处：《生物技术通报》2023年第4期259-267,共9页Biotechnology Bulletin

基　　金：吉林省科技发展计划项目(20220202008NC)。

摘　　要：玉米起源于亚热带,是喜温作物,易受低温胁迫的影响,脱水素(dehydrin,DHN)作为胚胎发育晚期丰富蛋白(late embryogenesis abundant protein,LEA)Ⅱ家族成员,是一类在植物非生物胁迫中发挥重要功能的蛋白。本研究克隆获得ZmDHN15基因,使用生物信息学手段,实时荧光定量PCR等技术对该基因的基本特性、组织表达特性进行分析,并进行植物过表达载体的构建及烟草的遗传转化,对T2代阳性植株进行抗冷性功能验证。结果表明,ZmDHN15基因全长1442 bp,共编码290个氨基酸,分子量为31.44 kD,理论等电点为6.05,是亲水性非跨膜蛋白,具有脱水素家族特有保守结构域。RT-qPCR分析表明ZmDHN15基因的在玉米叶片中表达量较高且在冷胁迫条件下表达量增加;获得T2代转基因烟草植株9株;在冷胁迫下转基因烟草与野生型相比,萌发率提高1.40倍,根长提高1.58倍,其叶片萎蔫程度更低,脯氨酸含量、丙二醛含量和过氧化物酶活性分别降低41.17%、28.47%和23.33%,可溶性糖含量、氧化物酶活性和超氧化物歧化酶活性分别升高58.97%、47.85%和47.53%,H_(2)O_(2)和O_(2)^(-)积累量分别减少34.78%和47.00%。综上所述,过表达ZmDHN15基因可以有效提高烟草植株对冷胁迫的耐受性,为进一步研究ZmDHN15基因在玉米中的功能奠定基础。Maize(Zea mays L.)originated in the subtropical zone,is a thermophilic crop,and is susceptible to low temperature stress.Dehydrin,group 2 members of the late embryogenesis abundant protein(LEA),are a class of proteins that play an important role in plant abiotic stress.In this study,the ZmDHN15 gene was cloned and obtained,and the basic characteristics and tissue expression characteristics of the gene were analyzed using bioinformatics methods,real-time fluorescent quantitative PCR and other techniques,and the construction of plant overexpression vector and the genetic transformation of tobacco were carried out.T2 generation plants were used to verify their cold resistance.The results showed that the ZmDHN15 gene was 1442 bp in length,encoded a total of 290 amino acids,with a molecular weight of 31.44 kD and a theoretical isoelectric point of 6.05.It was a hydrophilic non-transmembrane protein with a conserved domain unique to the dehydratin family.RT-qPCR analysis showed that the ZmDHN15 gene was highly expressed in maize leaves and increased under cold stress conditions.Nine T2 generation transgenic tobacco plants were obtained.Compared with the wild type,the germination rate of transgenic tobacco increased by 1.40 times,the root length increased by 1.58 times,the wilting degree of its leaves was lower,the content of proline,malondialdehyde and peroxidase reduced by 41.17%,28.47%and 23.33%,respectively.Soluble sugar content,oxide activity and superoxide dismutase activity increased by 58.97%,47.85%and 47.53%,respectively,and the accumulation of H_(2)O_(2)and O_(2)^(-)decreased by 34.78%and 47.00%,respectively.In conclusion,overexpression of ZmDHN15 gene can effectively improve the tolerance of tobacco plants to cold stress,which lays a foundation for further research on the function of ZmDHN15 gene in maize.

关 键 词：玉米 ZmDHN15基因 脱水素 抗冷 异源表达 

分 类 号：S513[农业科学—作物学]