大豆Hsp20基因家族生物信息学分析及高温高湿胁迫响应的分析

Bioinformatics Analysis of Soybean Hsp20 Gene Family and Its Response to High Temperature and Humidity Stress

热激蛋白(Hsps)家族中的小热激蛋白(Hsp20)在保护植物免受非生物胁迫方面发挥至关重要的作用,为探究大豆Hsp20家族基因的特性及其对高温高湿胁迫的响应,对大豆进行了全基因组分析,本研究采用生物信息学方法对进化关系、保守结构域、染色体位置、启动子顺式调控元件和组织特异性表达情况进行预测分析,并采用同位素标记相对和绝对定量(iTRAQ)技术对湘豆3号和宁镇1号两个大豆品种在高温高湿胁迫下不同时间的蛋白表达量进行了分析。结果表明:在大豆基因组中共查找出33个GmHsp20基因,不均匀分布在16条染色体上并根据系统发育分析和亚细胞定位结果分为11个亚族。序列分析表明GmHsp20蛋白表现出较高的结构保守性,含有典型的ACD结构域,32个GmHsp20基因(96.96%)没有内含子,或只有1个内含子。启动子分析表明,GmHsp20基因含有大量与胁迫相关的顺式作用元件,其中MYB数目最多(110)个。基因组织特异性表达预测分析结果显示,除GmHSP22.6外,其余基因在各种组织和器官中均有表达。其中GmHSP18.5、GmHSP23.7、GmHSP15.2基因在所有组织中均高度表达。蛋白定量分析结果表明,与对照相比,宁镇1号和湘豆3号两个品种中,经高温高湿胁迫处理后,GmHSP18.5、GmHSP23.7、GmHSP26.0B、GmHSP17.8、GmHSP25.95个蛋白均上调表达,并且在两个品种间表达趋势基本一致,表明这些小热激蛋白家族成员可能参与了大豆对高温高湿胁迫的应答反应过程。本研究结果为进一步研究大豆小热激蛋白GmHsp20家族成员的功能提供有价值的信息。

Soybean is an important economic crop in the world.However,Soybean is often subjected to various stresses during its growth and development,which seriously affects its yield and quality.As a member of the heat shock protein family,small heat shock protein(Hsp20)plays an important role in protecting plants from abiotic stress.In order to explore the characteristics of soybean Hsp20 family genes and their responses to high temperature and humidity stress,the whole genome analysis was carried out,and the evolutionary relationship,conservative domain,chromosome position,promoter cis regulatory elements and tissue specific expression were predicted and analyzed using bioinformatics methods.The protein expression of Xiangdou 3 and Ningzhen 1 soybean varieties under high temperature and high humidity stress at different times was analyzed by using isotope labeled relative and absolute quantification(iTRAQ)techniques.The results showed that 33 GmHsp20 genes were identified in soybean genome,distributed unevenly on 16 chromosomes and divided into 11 subgroups according to phylogenetic analysis and subcellular localization results.Sequence analysis showed that the GmHsp20 protein was highly conserved in structure and contained a typical ACD domain.Most of the GmHsp20 genes(96.96%)had no introns or only one intron.Promoter analysis showed that the GmHsp20 gene contained a large number of cis acting elements related to stress,with the largest number of MYBs(110).The prediction of tissue specific expression of genes showed that all genes except GmHSP22.6 were expressed in various tissues and organs.Among them,GmHSP18.5,GmHSP23.7 and GmHSP15.2 were highly expressed in all tissues.Quantitative proteomics results showed that GmHSP18.5,GmHSP23.7,GmHSP26.0B,GmHSP17.8 and GmHSP25.9 were up-regulated and expressed in both Ningzhen 1 and Xiangdou 3 varieties after high temperature and high humidity stress treatment compared to the control,and the expression trends were essentially the same between the two varieties.This suggests that these members of the small heat shock protein family might participate in the response process of soybean to heat and humidity stress.The results of this study provide valuable information for further investigation of GmHsp20 function.