基于干旱胁迫转录组信息的蚕豆ASPAT基因家族分析  

Analysis of ASPAT gene family based on drought-stressed transcriptome sequencing in Vicia faba L.

作　　者：魏正欣 刘昌燕[2] 陈宏伟[2] 李莉[2] 孙龙清 韩雪松 焦春海[2] 沙爱华[1] WEI Zheng-Xin;LIU Chang-Yan;CHEN Hong-Wei;LI Li;SUN Long-Qing;HAN Xue-Song;JIAO Chun-Hai;SHA Ai-Hua(College of Agriculture,Yangtze University/Engineering Research center of Ecology and Agricultural Use of Wetland,Ministry of Education,Jingzhou 434025,Hubei,China;Hubei Academy of Agricultural Sciences Institute of Food Crops/Hubei Provincial Key Laboratory of Germplasm Innovation and Genetic Improvement of Food Crops,Wuhan 430064,Hubei,China)

机构地区：[1]长江大学农学院/湿地生态与农业利用教育部工程研究中心,湖北荆州434025 [2]湖北省农业科学院粮食作物研究所/粮食作物种质创新与遗传改良湖北省重点实验室,湖北武汉430064

出　　处：《作物学报》2023年第7期1871-1881,共11页Acta Agronomica Sinica

基　　金：国家重点研发计划项目(2019YFD1001303/2019YFD1001300);财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-08);湖北省农业科技创新中心项目(2021-620-000-001-01)资助。

摘　　要：蚕豆(Vicia faba L.)是一种重要的食用豆作物,干旱会导致蚕豆产量降低。鉴定蚕豆耐旱基因,有利于通过分子标记辅助育种和基因工程手段培育抗旱品种。在前期研究中,我们通过转录组测序鉴定出了一类天冬氨酸转氨酶基因(ASPAT)响应蚕豆干旱胁迫。ASPAT催化天冬氨酸(ASP)和α-酮戊二酸的可逆转氨反应,生成草酰乙酸和谷氨酸,是参与植物体代谢的关键酶。本文基于转录组测序数据,通过生物信息学方法全基因组鉴定了蚕豆ASPAT基因家族成员,并分析了各基因家族成员的理化性质、亚细胞定位、基因结构、蛋白结构域、保守基序、系统进化、蛋白质互作和基因表达模式。结果显示,蚕豆转录参考基因组中共有8个ASPAT基因家族成员,包括3个真核型ASPAT(AAT)基因和5个原核型ASPAT(PAT)基因,分别定位于线粒体和叶绿体。系统发育分析表明,蚕豆ASPAT蛋白主要可分为2个亚族Iα和Iβ。Iα为AAT蛋白,包括VfASPAT1~VfASPAT3。Iβ为PAT蛋白,包括VfASPAT4~VfASPAT8。蚕豆ASPAT家族成员均含有motif1、motif3、motif9,且具有相似的基因结构和共同的蛋白结构域Aminotran-1-2。VfASPAT蛋白互作网络分析表明该家族成员可能参与逆境胁迫代谢过程。转录组和荧光定量PCR分析表明该家族8个成员在干旱处理16h和64h表达模式不同,除VfASPAT4和VfASPAT6外其他基因表达量都呈上调趋势,表明VfASPAT基因可能对干旱胁迫有正向调控作用。本文研究结果为进一步分析蚕豆ASPAT的抗旱功能,应用ASPAT提高蚕豆抗旱性奠定了基础。Fava beans(Vicia faba L.)is an important food bean crop,and drought can lead to lower fava bean yields.The identification of drought tolerance genes of fava beans is beneficial to breed the drought-resistant varieties through molecular marker-assisted breeding and genetic engineering.In a previous study,we identified a class of aspartate aminotransferase genes(ASPAT)in response to faba bean drought stress by transcriptome sequencing.ASPAT catalyzes the reversible reaction of transamination between aspartate andα-oxoglutarate to generate oxaloacetate and glutamate,and plays a key role in plant metabolism.Based on transcriptomic sequencing data,ASPAT members were identified in genome-wide level.The physicochemical properties,subcellular localizations,gene structures,protein domains,conserved motifs,phylogeny,protein interaction,and gene expression patterns of each member were analyzed.The results indicate that eight ASPAT genes were identified from the transcriptomes,which included three eukaryotype ASPAT(AAT)genes and five prokarytoype ASPAT(PAT)genes with mitochondria and chloroplast location,respectively.Phylogenetic analysis showed that ASPAT could be divided into two subfamilies Iαand Iβ.Iαis AAT protein including VfASPAT1–VfASPAT3,and Iβis PAT protein consisting of VfASPAT4–VfASPAT8.ASPAT family members of Vicia faba contain motif 1,motif 3,and motif 9,and had similar gene structures and common protein domain aminotran-1-2.VfASPATs protein interaction network suggested that the members might be involved in stress metabolism.Transcriptome and qRT-PCR indicated that he expression patterns of the eight members were different at 16 hours and 64 hours under drought stress.All the members were up-regulated with the exception of VfASPAT4 and VfASPAT6,implying that VfASPATs may have a positive regulation effect on drought stress.In this study,eight ASPAT genes of Vicia faba were identified,and the structures,distribution,evolutionary relationship,and the relative expression patterns of ASPAT gene family memb

关 键 词：蚕豆 天冬氨酸转氨酶 基因家族 干旱胁迫 基因表达 

分 类 号：S643.6[农业科学—蔬菜学]