大豆(Glycine max)SUT4基因的克隆、表达及抗逆功能的鉴定  

Cloning and Expression of Soybean(Glycine max)SUT4 and Identification of Stress Resistance Functions

作　　者：张引鹤 周永刚 高红桃 刘伟灿 李海燕 Zhang Yinhe;Zhou Yonggang;Gao Hongtao;Liu Weican;Li Haiyan(Engineering Research Center for Bioreactor and Drug Development(Ministry of Education),College of Life Sciences,Jilin Agricultural University,Changchun,130118;College of Tropical Crops,Hainan University,Haikou,570228)

机构地区：[1]吉林农业大学生命科学学院,生物反应器与药物开发教育部工程研究中心,长春130118 [2]海南大学热带作物学院,海口570228

出　　处：《分子植物育种》2022年第17期5578-5587,共10页Molecular Plant Breeding

基　　金：国家自然科学基金项目(32001464,321QN182)资助。

摘　　要：以大豆‘吉育72’为材料,利用PCR克隆技术,克隆大豆蔗糖转运蛋白GmSUT4基因,对其进行生物信息学分析,并利用实时荧光定量PCR方法检测GmSUT4在大豆不同组织及不同胁迫处理下的相对表达量。同时利用酵母系统对GmSUT4基因功能进行初步鉴定。GmSUT4基因全长5672 bp,CDS区1518 bp,编码505个氨基酸,具有6个外显子,位于2号染色体Chr.2:40653016~40658687位置。预测该基因编码的蛋白质分子量大小为54.17kD,等电点为9.04,具有12个跨膜区域。通过序列比对以及进化分析发现,GmSUT4属于SUT4分支,且与其他物种SUT4家族成员高度保守,属于低亲和力蔗糖转运蛋白。另外,组织特异性结果显示,Gm SUT4在大豆的各组织中均有表达,在根中的表达量最高,在叶中表达量最低。通过对水培幼苗进行胁迫处理发现,该基因在干旱、盐和盐碱胁迫下表达量均明显上调。进一步利用酵母突变株SUSY7/ura3对其功能进行验证,同时发现该基因对干旱、盐和盐碱逆境均有响应,且在盐和盐碱胁迫下表型更为明显。GmSUT4基因作为胁迫相关基因可能在大豆逆境中发挥着重要作用。Using soybean’Jiyu72’as material,the soybean sucrose transporter GmSUT4 was cloned by PCR cloning technology,and the bioinformatics analysis was performed on it.The real-time fluorescent quantitative PCR method was used to detect GmSUT4 in different soybean tissues and under different stress treatments.At the same time,the yeast system was used to analyze the function of GmSUT4 gene.The GmSUT4 gene has a total length of 5672 bp and a CDS region of 1518 bp,which encodes 505 amino acids,has 6 exons,and is located at the Chr.2:40653016~40658687 position of chromosome 2.It is predicted that the molecular weight of the protein encoded by this gene is 54.17 kD,the isoelectric point is 9.04,and it has 12 transmembrane regions.Through sequence alignment and evolutionary analysis,it is found that GmSUT4 belongs to the SUT4 branch,and is highly conserved with other species of SUT4 family members,and belongs to a low-affinity sucrose transporter.In addition,the results of tissue-specific real-time fluorescence quantitative PCR showed that GmSUT4 was expressed in all tissues of soybean,with the highest expression in roots and the lowest expression in leaves.Through stress treatment of hydroponic seedlings,it was found that the expression of this gene was significantly up-regulated under drought,salt and saline-alkali stress.Furthermore,GmSUT4 was transformed into yeast mutant(SUSY7/ura3)for functional verification.The results showed that GmSUT4 participated in drought,salt and saline-alkali stress.Besides,the phenotype of overexpression line is more obvious in response to salt and saline-alkali stress.As a stress-related gene,GmSUT4 may play an important role in soybean stress.

关 键 词：大豆 蔗糖转运蛋白基因GmSUT4 功能 逆境 

分 类 号：S565.1[农业科学—作物学]