摘要
目的探究蒙古黄芪Astragalus membranaceus三萜皂苷的合成基因及参与后加工修饰的CYP、UGT基因。方法对两、三年生蒙古黄芪进行转录组测序和生物信息学分析。结果Illumina HiSeq平台测序共得到42.22 Gb数据,Trinity软件拼接得到369790条Transcripts和336068条Unigene,与非冗余蛋白质序列(non-redundant protein sequence database,Nr)、基因本体论(gene ontology consortium,GO)、同源蛋白质簇(cluster of orthologous groups of proteins,egg NOG/COG)、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)、Swiss-Prot、Pfam等数据库比对注释,结果显示蒙古黄芪与植物鹰嘴豆Cicer arietinum同源序列最多。KEGG分析表明蒙古黄芪差异基因主要富集在萜类骨架生物合成、二萜类生物合成、氨基酰基-tRNA生物合通路中,进一步从萜类骨架生物合成通路中鉴别出114条差异表达Unigene,包含15种关键酶基因,其中三年生蒙古黄芪hexPS表达均显著上调,ACAT、HMGCS、HMGCR、MVK、MVAK2、MVD、IDI、FDPS、GGPS1、FNTB、STE24、STE14、FCLY、DHDDS有上调也有下调表达的Unigene。进一步挖掘蒙古黄芪三萜皂苷生物合成后修饰基因,发现8条CYP差异表达关键酶基因,分别为acyP、PPID/CYPD、PPIH/CYPH、CYP26A、CYP51、CYP84A、CYP61A、cypD_E/CYP102A/CYP505;2条UGT差异表达关键酶基因HUGT、SUGT1。结论两、三年生蒙古黄芪的皂苷合成基因和CYP、UGT基因表达存在显著差异,这些基因共同参与黄芪皂苷的合成、加工与修饰。
Objective To investigate the synthesis genes of triterpenoid saponins from Astragalus membranaceus var.mongholicus and the CYP and UGT genes involved in post-processing modification.Methods Transcriptome sequencing and bioinformatics analysis were performed on 2 and 3 years old Mongolian Astragalus.Results Illumina HiSeq platform was used for sequencing,and 42.22 Gb of data were obtained.After Trinity software stitching,369790 Transcripts and 336068 Unigene Transcripts were obtained.Compared with non-redundant protein sequence database(NR),Gene Ontology Consortium(GO),Cluster of Orthologous Groups of proteins(eggNOG/COG),Kyoto Encyclopedia of Genes and Genomes(KEGG),Swiss-Prot and PFAM databases,the results showed that Astragalus mongholicus and chickpea(Cicer arietinum Linn.)had the most homologous sequences.KEGG analysis showed that astragalus differential genes were mainly enriched in the terpene skeleton biosynthesis pathway,diterpene biosynthesis pathway and aminoacyl-trNA biosynthesis pathway.Further,114 differentially expressed Unigene including 15 key enzyme genes were identified from the terpene skeleton biosynthesis pathway.The expression of hexPS was significantly up-regulated in three-year-old Mongolian astragalus.ACAT,HMGCS,HMGCR,MVK,MVAK2,MVD,IDI,FDPS,GGPS1,FNTB,STE24,STE14,FCLY and DHDDS were up-regulated and down-regulated in Unigene.After further mining of astragalus triterpenoid saponin biosynthetic modifier genes,eight key CYP genes were found to be differentially expressed,which were acyP,PPID/CYPD,PPIH/CYPH,CYP26A,CYP51,CYP84A,CYP61A,cypD_E/CYP102A/CYP505.Two UGT differentially expressed key enzyme genes HUGT and SUGT1.Conclusion There are significant differences in the expressions of saponin synthesis genes,CYP and UGT genes in 2-and 3-year-old astragalus.These genes are involved in the synthesis,processing and modification of astragalus.
作者
王祥
王育朋
安佳
王七龙
井亚江
黄建萍
张岗
彭亮
颜永刚
WANG Xiang;WANG Yu-peng;AN Jia;WANG Qi-long;JIN Ya-jiang;HUANG Jian-ping;ZHANG Gang;PENG Liang;YAN Yong-gang(Shaanxi Engineering Research Center for Application and Development of Chinese Medicine in Qinling Mountains/Key Laboratory of“Qin Medicine”Research and Development,Shaanxi University of Chinese Medicine,Xianyang 712046,China;Shaanxi Haitian Pharmaceutical Co.,Ltd.,Xianyang 712000,China)
出处
《中草药》
CAS
CSCD
北大核心
2023年第3期915-925,共11页
Chinese Traditional and Herbal Drugs
基金
内蒙古自治区科技重大专项(2019ZD005)
国家中医药管理局全国名老中医药专家传承工作室建设项目(国中医药人函[2019]41号)
陕西中医药大学“秦药”品质评价与资源开发学科创新团队项目(2019-QN01)。
关键词
蒙古黄芪
转录组
差异表达基因
黄芪皂苷
生物合成途径
Astragalus membranaceus(Fisch.)Bge.var.mongholicus(Bge.)Hsiao
transcriptome
differentially expressed genes
astragalus saponin
biosynthetic pathway
