绿豆乙醇脱氢酶基因生物信息学分析及其在镉胁迫下的表达特性变化  

Bioinformatics analysis on alcohol dehydrogenase genes in Vigna radiata and changes of their expression characteristics under cadmium stress

作　　者：程斌 张创娟 杨乐 冷艳[1] 李师翁[1] CHENG Bin;ZHANG Chuangjuan;YANG Le;LENG Yan;LI Shiweng(School of Biological and Pharmaceutical Engineering,Lanzhou Jiaotong University,Lanzhou 730070,China)

机构地区：[1]兰州交通大学生物与制药工程学院,甘肃兰州730070

出　　处：《植物资源与环境学报》2022年第2期10-21,共12页Journal of Plant Resources and Environment

基　　金：国家自然科学基金资助项目(31760110)。

摘　　要：基于绿豆〔Vigna radiata(Linn.)R.Wilczek〕基因组注释文件鉴定乙醇脱氢酶基因(ADHs),并采用生物信息学方法分析了绿豆ADHs基因结构及其编码的氨基酸序列的理化特性和系统关系;运用转录组和酶学方法分析了Cd胁迫条件下绿豆幼苗根、茎和叶片中ADHs基因的表达量和ADH酶活性的变化。结果表明:绿豆基因组含有15个ADHs基因,依次命名为VrADH 1至VrADH 15;15个VrADHs基因的序列长度为237~1603 bp,其中,仅VrADH 10基因包含2个内含子和2个外显子,其他VrADHs基因分别包含7~9个内含子和7~10个外显子。15个VrADHs的氨基酸序列中,除VrADH1和VrADH10外,其他VrADHs均含有10个motif。15个VrADHs的氨基酸残基数为78~444,相对分子质量8840~48390,等电点pI 4.83~pI 8.22,亲水性系数-0.259~0.090;其中,仅VrADH10属于短链脱氢酶,其他VrADHs均属于中、长链脱氢酶;仅VrADH10含有ADH_N结构域,其他VrADHs均含有ADH_N和ADH_zinc_N结构域,且VrADH1还含有ADH_zinc_N_2结构域。NJ系统发育树将15个VrADHs及大豆〔Glycine max(Linn.)Merr.〕的31个GmADHs和拟南芥〔Arabidopsis thaliana(Linn.)Heynh.〕的8个AtADHs分为3个分支,其中绿豆与大豆的ADHs亲缘关系更近。转录组和酶活性分析结果显示:VrADH 7、VrADH 8和VrADH 10基因在根、茎和叶片中均未表达,而其他VrADHs基因均不同程度表达,且表达水平随生长时间而异;在根、茎和叶片中相对表达量最高的基因分别为VrADH 1、VrADH 3和VrADH 14。经100μmol·L^(-1) Cd胁迫处理后,多数VrADHs基因的相对表达量较对照上调,且部分基因的相对表达量与对照差异显著(P<0.05)。经Cd胁迫处理后绿豆幼苗根、茎和叶片中ADH酶活性总体上均高于对照。综合分析结果表明:绿豆幼苗根、茎和叶片中VrADHs基因的表达特性存在明显差异,部分VrADHs基因的表达呈现组织特异性;Cd胁迫处理后绿豆幼苗ADH酶活性的升高与某些VrADHs基因的表达水平上调有关,表明这些VAlcohol dehydrogenase genes(ADHs)were identified based on the genome annotation file of Vigna radiata(Linn.)R.Wilczek,and the genetic structures of ADHs of V.radiata and the physicochemical properties and systematic relationships of their encoded amino acid sequences were analyzed by using bioinformatics method;the changes of expression levels of ADHs and activities of ADH enzyme in roots,stems,and leaves of V.radiata seedlings under Cd stress condition were analyzed by using transcriptome and enzymology methods.The results show that there are 15 ADHs in V.radiata genome,which are named as VrADH 1 to VrADH 15 in sequence;the sequence length of 15 VrADHs are 237-1603 bp,in which,only VrADH 10 contains 2 introns and 2 exons,and the other VrADHs contain 7-9 introns and 7-10 exons.Among the amino acid sequences of 15 VrADHs,all VrADHs except VrADH1 and VrADH10 contain 10 motifs.The number of amino acid residues of 15 VrADHs are 78-444,the relative molecular masses are 8840-48390,the isoelectric points are pI 4.83-pI 8.22,and the hydrophilic coefficients are-0.259-0.090;in which,only VrADH10 belongs to short-chain dehydrogenase,and the other VrADHs belong to medium or long-chain dehydrogenases;only VrADH10 contains ADH_N domain,the other VrADHs all contain ADH_N and ADH_zinc_N domains,and VrADH1 also contains ADH_zinc_N_2 domain.The 15 VrADHs,31 GmADHs of Glycine max(Linn.)Merr.,and 8 AtADHs of Arabidopsis thaliana(Linn.)Heynh.are grouped into three branches by NJ phylogenetic tree,in which,the relationships of ADHs between V.radiata and G.max are closer.The analysis results of transcriptome and enzyme activities show that VrADH 7,VrADH 8,and VrADH 10 are not expressed in roots,stems,and leaves,while the other VrADHs are all expressed at different degrees,and the expression levels vary with the growth time;genes with the highest relative expression levels in roots,stems,and leaves are VrADH 1,VrADH 3,and VrADH 14,respectively.The relative expression levels of most VrADHs are up-regulated compared with the control afte

关 键 词：绿豆 乙醇脱氢酶基因 转录组 镉胁迫 表达特性 

分 类 号：Q75[生物学—分子生物学] Q948.113