海岛棉H268 PPR蛋白家族基因鉴定及SNP/InDel分析  

Identification and SNP/InDel analysis of H268 PPR protein family genes in Gossypium barbadense

作　　者：李枝玲 孔祥军[1] 郑杰[1] 韦美玲 李斌[1] 周瑞阳[1] LI Zhi-ling;KONG Xiang-jun;ZHENG Jie;WEI Mei-ling;LI Bin;ZHOU Rui-yang(College of Agriculture,Guangxi University,Nanning 530004,China)

机构地区：[1]广西大学农学院,南宁530004

出　　处：《南方农业学报》2020年第11期2614-2625,共12页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(31571719)。

摘　　要：【目的】通过对海岛棉不育系H276A及其育性恢复材料H268进行三角状五肽重复蛋白(PPR)家族基因鉴定及比对分析,获得序列存在差异的PPR基因,为后续育性恢复基因(Rf)的发掘提供理论基础。【方法】通过生物信息学分析海岛棉PPR基因,并对其进行亚细胞定位预测与功能注释分析;采用Illumina HiSeq 4000高通量测序技术对海岛棉H276A及H268对PPR蛋白家族基因进行转录组测序,将筛选获得的PPR蛋白与其他植物具有育性恢复的PPR蛋白进行氨基酸序列同源比对并构建系统发育进化树。【结果】鉴定获得912个PPR基因,其中有846个在海岛棉H276A和H268间存在序列差异,且分别存在7226个SNP差异位点和301个InDel差异位点,其中作用于线粒体的PPR基因有2143个SNP差异位点和134个InDel差异位点。转录组测序结果显示,共检测到表达基因数量为68197个,其中已知基因56311个、新基因11886个,可满足后续数据分析。GO功能注释结果显示,PPR基因主要富集于生物学过程,富集于分子功能的PPR基因最少。50.0%PPR蛋白定位于线粒体,29.0%PPR蛋白定位于叶绿体,0.9%PPR蛋白定位于细胞核,20.0%PPR蛋白定位于胞外,0.1%PPR蛋白定位于细胞质膜。xp_016708712(LOC107923023)和xp_016710013(LOC107924193)与多个具有育性恢复功能的PPR蛋白氨基酸序列相似性达33%,说明二者亲缘关系较近,且亚细胞定位于线粒体。【结论】xp_016708712和xp_016710013 2个PPR蛋白可能与海岛棉不育系H276A育性恢复相关,可用于发掘海岛棉CMS的Rf基因。【Objective】Through the identification of pentatricopeptide repeat protein PPR protein family genes and comparative analysis of gene sequence between male sterile line H276 A and fertility restoration material H268 of Gossypium barbadense,the PPR protein coding genes with sequence difference was obtained.It provided a theoretical reference for the further mining of fertility restoration(Rf)genes.【Method】The PPR genes of G.barbadense were identified by bioinformatics,and subcellular location prediction and gene function annotationanalysis were performed,then further highthroughput sequencing technology was used to identify the PPR protein family genes in G.barbadense H276 A and H268.Homologous alignment in amino acid sequencewas performed between the identified PPR protein and PPR protein with fertility restore genes,then the phylogenetic tree was constructed.【Result】912 PPR protein coding genes were identified,846 among them were different in sequence between H276 A and H268,7226 SNP differential loci and 301 InDel differential loci were obtained in total.Among them,PPR genes acting on mitochondria contained 2143 SNP differential loci and134 InDels differential loci.Transcriptome sequencing results showed that a total of 68197 genes were detected,including56311 known genes and 11886 new genes,which could meet the needs of subsequent data analysis.The results of GO functional classification showed that PPR genes were mainly enriched in biological processes and the least in molecular functions.50.0%of PPR protein was located in mitochondria,29.0%in chloroplast,0.9%in nucleus,and the remaining20.1%in plasma membrane and extracellular structure,0.1 in cellular branch membrane.The results of homology alignment showed thatboth of the homology between xp016708712(LOC107923023),xp016710013(LOC107924193)and several identified PPR protein amino acids with fertility restoration function were more than 33%,which indicated that they were closely related,and both of them were located in mitochondria.【Conclusion】

关 键 词：海岛棉 PPR蛋白家族基因 转录组测序 生物信息学分析 SNP INDEL 

分 类 号：S562[农业科学—作物学]