紫肉甘薯及其突变体花青素积累差异的比较转录组分析  

Comparative transcriptome analysis on anthocyanin accumulation differences in purple-fleshed sweetpotatoes and their mutants

作　　者：后猛[1] 李臣 宋炜涵 张允刚[1] 李强[1] KOU Meng;LI Chen;SONG Wei-han;ZHANG Yun-gang;LI Qiang(Xuzhou Institute of Agricultural Sciences in Jiangsu Xuhuai District/Sweetpotato Research Institute,Chinese Academy of Agricultural Sciences/Key Laboratory of Biology and Genetic Breeding of Sweetpotato,Ministry of Agriculture and Rural Affairs,Xuzhou 221131,China;School of Life Science,Jiangsu Normal University,Xuzhou 221116,China)

机构地区：[1]江苏徐淮地区徐州农业科学研究所/中国农业科学院甘薯研究所/农业农村部甘薯生物学与遗传育种重点实验室,江苏徐州221131 [2]江苏师范大学生命科学学院,江苏徐州221116

出　　处：《江苏农业学报》2022年第2期313-325,共13页Jiangsu Journal of Agricultural Sciences

基　　金：国家重点研发计划项目(2019YFD1001300、2019YFD1001304);国家现代农业产业技术体系项目(CARS-10)。

摘　　要：紫肉甘薯因其块根中富含紫色花青素而得名。本研究以2组栽培种紫肉甘薯及其淡黄肉突变体为材料,基于甘薯转录组测序(RNA-seq)数据进行生物信息学分析。以|log_(2) FC|≥1且错误发现率(FDR)<0.01作为筛选标准,鉴定差异表达基因(DEGs),对DEGs进行GO功能分类和KEGG代谢通路分析,并采用实时荧光定量PCR(qRT-PCR)方法对测序结果进行验证。样品经转录组测序后,均获得超过8 Gb的高质量数据,拼接组装共得到164427条Unigenes。在2组甘薯材料及其突变体中分别检测到2262个和1546个DEGs,其中有244个DEGs是2组甘薯材料共有的。GO功能分析发现,DEGs主要富集于苯丙氨酸解氨酶活性、着色、类黄酮生物合成过程、含花青素化合物生物合成过程等。KEGG通路分析发现,DEGs主要分布在苯丙素类生物合成、苯丙氨酸代谢、类黄酮生物合成和花青素生物合成等代谢通路。随机选择27个与花青素生物代谢相关的候选基因进行实时荧光定量PCR验证,表达模式与测序结果一致。将2组材料中候选基因的表达水平与花青素含量进行相关性分析,结果表明,花青素生物合成相关的候选基因PAL-1、CHS、ANS、CCoAOMT、GST-2、ABC-1、bHLH和WRKY-2的相对表达量与花青素积累量的相关性达到显著或极显著水平。表明,2个甘薯突变体材料块根薯肉紫色的缺失是由花青素的合成、转运和相关转录因子表达水平的降低共同引起的。通过转录组测序可以从中挖掘一些与花青素代谢相关的候选基因,为研究其在生物合成途径中的调控机制奠定基础。Purple-fleshed sweetpotato(PESP)is famous for the rich storage of purple anthocyanins in its storage root.In this study,two groups of PFSP cultivated species and their mutants with light yellow flesh were used as materials,and bioinformatics analysis was conducted based on transcriptome sequencing(RNA-seq)data of sweetpotatoes.|log_(2)FC|≥1 and FDR(False discovery rate)<0.01 were used as screening standards to identify differentially expressed genes(DEGs).Gene ontology(GO)functional classification and KEGG metabolic pathway analysis of DEGs were performed,and real-time quantitative PCR(qRT-PCR)method was used to verify the sequencing results.After transcriptome sequencing of the samples,more than eight Gb data with high quality were obtained for each sample,and a total of 164427 Unigenes were obtained by splicing and assembling.2262 and 1546 DEGs were detected respectively in two groups of sweetpotato materials and their mutants,among which 244 DEGs were shared between two groups of sweetpotato materials.Results of GO functional analysis showed that,DEGs mainly enriched in phenylalanine ammonia-lyase activity,pigmentation,flavonoid biosynthesis process and anthocyanin-containing compound biosynthesis process.Results of KEGG pathway analysis showed that,DEGs mainly distributed in metabolic pathways such as phenylpropanoid biosynthesis,phenylalanine metabolism,flavonoid biosynthesis and anthocyanin biosynthesis.In addition,27 candidate genes related to anthocyanin biological metabolism were selected randomly for real-time quantitative PCR verification,and their expression patterns were found to be consistent with the sequencing results.Results of correlation analysis on gene expression level and anthocyanin content revealed that,relative expression levels of anthocyanin biosynthesis related candidate genes such as PAL-1,CHS,ANS,CCoAOMT,GST-2,ABC-1,bHLH and WRKY-2 were in significant or extremely significant correlation with anthocyanin accumulation.The results indicated that,the deficiency of purple color in the

关 键 词：紫肉甘薯 转录组测序 花青素代谢 差异表达基因 

分 类 号：S531[农业科学—作物学]