苹果CIA2基因克隆及其在烟草中的初步功能验证  

Cloning of Malus domestica CIA2 Gene and Its Preliminary Functional Verification in Tobacco

作　　者：马志[1] 王海波[2] 阴启忠[2] 李林光[2] 李金英[1] 吴林[1] 王颖[1] Ma Zhi;Wang Haibo;Yin Qizhong;Li Linguang;Li Jinying;Wu Lin;Wang Ying(College of Horticulture,Jilin Agricultural University,Changchun,130118;Shandong Institute of Pomology,Tai'an,271000)

机构地区：[1]吉林农业大学园艺学院,长春130118 [2]山东省果树研究所,泰安271000

出　　处：《分子植物育种》2022年第21期6994-7002,共9页Molecular Plant Breeding

基　　金：山东省自然科学基金项目(ZR2017YL025);山东省重点研发计划项目(2018GNC113018);泰安市科技发展计划项目(2017NS0095)共同资助。

摘　　要：为研究CIA2基因在低温胁迫下的功能及表达模式,本研究利用RT-PCR技术,从苹果‘71-3-150’组培苗中克隆得到CIA2基因,该基因编码234个氨基酸,属于CCT家族。对CIA2编码氨基酸的同源性与系统进化分析显示,CIA2所编码蛋白与白梨、桃、日本樱花中的同源蛋白的同源性都达到了86%以上,与白梨的CIA2亲缘关系较近。在4℃低温诱导处理下,苹果组培苗叶片中CIA2基因的表达量均高于常温对照组,且在24 h处理下表达量最高。将CIA2基因通过农杆菌介导法转入烟草中,对转基因植株进行低温诱导处理。在4℃低温条件下,转基因烟草的丙二醛含量、电导率低于野生型植株,可溶性糖含量高于野生型植株。研究结果表明苹果CIA2基因与植株的抗寒性相关,CIA2基因在提高植物抗寒性方面具有重要作用。In order to study the function and expression pattern of CIA2 gene under low temperature stress,we cloned CIA2 gene from apple’71-3-150’tissue culture seedlings by RT-PCR technology.The CIA2 gene encoded234 amino acids and belonged to CCT family.The homology and phylogenetic analysis of the amino acids encoded by CIA2 showed that the homology of the protein encoded by CIA2 was more than 86%with the homologous proteins of Pyrus bretschneideri,Amygdalus persica,Cerasus yedoensis,indicating that CIA2 was closely related to the CIA2 of Pyrus bretschneideri.Under low temperature induction at 4℃,the expression level of CIA2 gene in apple tissue culture leaves was higher than that in normal temperature control group,and the expression level was the highest at 24 h treatment.The CIA2 gene was transferred into tobacco by Agrobacterium-mediated method,and the transgenic plants were subjected to low temperature induction treatment.At 4℃,the soluble sugar content and conductivity of transgenic tobacco were lower than that of wild-type plants,and the soluble sugar content of transgenic tobacco was higher than that of wild-type plants.The results showed that CIA2 gene of apple was related to the cold resistance of plant,and CIA2 gene played an important role in improving the cold resistance of plant.

关 键 词：苹果 CIA2基因克隆 生物信息学分析 表达模式 转基因 

分 类 号：S572[农业科学—烟草工业] S661.1[农业科学—作物学] Q943.2[生物学—植物学]