谷子萌发吸水期关键代谢途径的筛选与分析  

Screening and Analysis of Key Metabolic Pathways in Foxtail Millet During Different Water Uptake Phases of Germination

作　　者：余爱丽[1] 赵晋锋[1] 成锴[1] 王振华[1] 张鹏 刘鑫[1] 田岗[1] 赵太存[1] 王玉文[1] YU AiLi;ZHAO JinFeng;CHENG Kai;WANG ZhenHua;ZHANG Peng;LIU Xin;TIAN Gang;ZHAO TaiCun;WANG YuWen(Millet Research Institute,Shanxi Academy of Agricultural Sciences,Changzhi 046011,Shanxi)

机构地区：[1]山西省农业科学院谷子研究所,山西长治046011

出　　处：《中国农业科学》2020年第15期3005-3019,共15页Scientia Agricultura Sinica

基　　金：国家现代农业产业技术体系(CARS-06-13.5-A23);山西省农业科学院科研项目(YGG17021和YCX2018206)。

摘　　要：【目的】谷子适应性强,抗旱耐瘠,是起源于中国的重要作物。通过转录组测序技术分析谷子萌发不同吸水期的转录组差异,以期获得谷子萌发过程中的差异表达基因,寻找调控谷子萌发的重要代谢途径和代谢物。【方法】以晋谷20为材料,构建谷子萌发过程中开始快速吸水期、滞缓吸水期和重新大量吸水期的cDNA文库,进行转录组分析;采用K-Means开展基因表达聚类分析;利用DESeq筛选差异表达基因;通过COG、GO、KEGG等对差异表达基因进行功能注释;利用KEGG富集挖掘不同吸水期调控种子萌发的关键代谢途径和关键基因;并采用qRT-PCR验证其可靠性;用HPLC分析关键代谢物含量。【结果】转录物测序分析获得谷子萌发开始快速吸水期、滞缓吸水期和重新大量吸水期覆盖整个基因组的基因表达谱,共获得33643个基因,识别9个具有不同表达模式的共表达基因簇。比较种子萌发的开始快速吸水期与滞缓吸水期、滞缓吸水期与重新大量吸水期、开始快速吸水期与重新大量吸水期,分别筛选出3893、4612和8472个差异表达基因。KEGG富集分析表明,3个比较的差异表达基因都显著富集到phenylpropanoid biosynthesis、phenylalanine metabolism、starch and sucrose metabolism代谢途径;开始快速吸水期与滞缓吸水期、开始快速吸水期与重新大量吸水期的差异表达基因还显著富集到plant hormone signal transduction途径。并且3个比较中富集到phenylpropanoid biosynthesis和phenylalanine metabolism代谢途径的差异表达基因数都最多,其中过氧化物酶基因(peroxidase)比例最高。通过qRT-PCR对4个苯丙烷生物合成途径相关基因的分析表明,其表达趋势与转录组分析结果基本一致,其中,4-香豆酸-CoA连接酶3(4-coumarate-CoA ligase 3)在谷子种子中存在已形成mRNA,萌发吸水过程中呈先下调后上调再下调的表达趋势。苯丙烷类相关代谢物含量分析显示,�【Objective】Foxtail millet(Setaria italica L.)is a hardy cereal and known for its tolerance against drought and barrenness,which originated in China.In this study,the transcriptome of foxtail millet was analyzed in germinating seeds during different uptake of water phases,which is to get a lot of differently expressed genes,and find the important metabolic pathways,key genes and metabolites of regulating germination in foxtail millet.【Method】The cDNA libraries of Jingu 20 were constructed in germinating seeds during rapid initial uptake of water,plateau phase,further increase in water uptake for the transcriptome analysis.The cluster analysis of gene expression was performed by K-Means.Differential expression analysis used DESeq.The functional annotations of differentially expressed genes were obtained by using COG,GO,KEGG,and so on.The key metabolism pathways and genes that regulated germination in foxtail millet were found during different water uptake phase through KEGG enrichment of DEGs.The reliability of sequencing results was confirmed by qRT-PCR.The contents of metabolites were assayed by HPLC.【Result】The genome-wide gene expression profile was obtained by RNA-sequencing during rapid initial uptake of water,plateau phase and further increase in water uptake.A total of 33643 genes were expressed.Nine co-expression clusters with distinctive patterns were identified.There were 3893,4612 and 8472 DEGs in rapid initial uptake of water and plateau phase,plateau phase and further increase in water uptake,rapid initial uptake of water and further increase in water uptake,respectively.The KEGG pathway enrichment analysis showed that these DEGs of the three comparisons were significantly associated with phenylpropanoid biosynthesis,phenylalanine metabolism,starch and sucrose metabolism.The DEGs between rapid initial uptake of water and plateau phase,between rapid initial uptake of water and further increase in water uptake were also enriched in plant hormone signal transduction.And the number of genes en

关 键 词：谷子 种子萌发吸水期 转录物组分析 苯丙烷生物合成途径 苯丙氨酸代谢途径 

分 类 号：S515[农业科学—作物学]