灯盏花中灯盏乙素生物合成相关基因转录组分析  

Transcriptome analysis of genes involved in scutellarin biosynthesis in Erigeron breviscapus

作　　者：闫亚哲 闻豪 杜江 杨勇 刘正杰[1,3] 林春[1,3] 文国松[1,3] 毛自朝[1,3] YAN Ya-zhe;WEN Hao;DU Jiang;YANG Yong;LIU Zheng-jie;LIN Chun;WEN Guo-song;MAO Zi-chao(College of Agronomy and Biotechnology,Yunnan Agricultural University,Kunming 650201,China;Yunnan Bio-Valley Pharmaceutical Co.,Ltd.,Kunming 650503,China;Center of Improvement and Utilization of Characteristic Resource Plants,Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南农业大学农学与生物技术学院,昆明650201 [2]云南生物谷药业股份有限公司,昆明650503 [3]云南农业大学小宗作物研究中心,昆明650201

出　　处：《西南农业学报》2022年第6期1318-1324,共7页Southwest China Journal of Agricultural Sciences

基　　金：云南生物谷药业股份有限公司项目(KX141611000)。

摘　　要：【目的】通过转录组测序(RNAseq)筛选灯盏乙素生物合成相关基因,为筛选灯盏花优质种质资源提供理论基础。【方法】通过HPLC测定灯盏花叶片中灯盏乙素含量,筛选其中有显著差异的4份样品,以1份低灯盏乙素含量叶片样品作为对照(CK)与3份高灯盏乙素含量样品同时进行RNAseq。测序完成后分析筛选差异表达基因(Differential expressed genes,DEGs),并对DEGs进行Gene Ontology(GO)和Kyoto Encyclopedia of Genes and Genomes(KEGG)功能富集分析。再选择表达量较高的参与类黄酮生物合成途径的DEGs进行qRT-PCR验证其表达模式。【结果】样品S3和S5灯盏乙素含量极显著高于样品S4(CK),样品S6灯盏乙素含量显著高于样品S4(CK)。高灯盏乙素样品S3、S5、S6与低灯盏乙素样品S4(CK),两两比较分别鉴定出2143个、1968个和1801个DEGs。3组差异比较组合的交集有490个共同基因,其中277个在高灯盏乙素含量样品中上调表达,另外213个在低灯盏乙素含量样品中上调表达,差异表达基因KEGG富集分析分别有13、14、14条基因在类黄酮生物合成途径。其中类黄酮代谢下游的基因查尔酮异构酶(CHI)、黄烷酮3-羟化酶(F3H)和类黄酮3’-羟化酶(F3’H)在高灯盏乙素样品中上调表达,参与咖啡酸酯途径的莽草酸/奎宁酸羟基肉桂酰转移酶(HCT)和咖啡酰辅酶A-O-甲基转移酶(CCoAMT)基因在低灯盏乙素样品S4(CK)中上调表达。qRT-PCR对5个DEGs表达模式的检测结果与RNAseq结果一致,CHI(Eb_V2_03181)、F3H(Eb_V2_40771)、F3’H(Eb_V2_04241)在高灯盏乙素样品中上调表达,HCT(Eb_V2_4655)、CCoAMT(Eb_V2_20841)在低灯盏乙素样品中上调表达。【结论】灯盏乙素生物合成涉及多个基因,与花青素途径及咖啡酸酯途径密切相关。【Objective】In this study,RNAseq was used to screen genes related to scutellarin content,which would provide a theoretical basis for screening high-quality germplasm of Erigeron breviscapus.【Method】The content of scutellarin in E.breviscapus leaves was determined by HPLC;And 4 samples with significant differences in scutellarin were screened.One low scutellarin control sample(CK)and 3 high scutellarin samples were subjected to RNAseq simultaneously.After sequencing,differentially expressed genes(DEGs)were screened;and GO and KEGG functional enrichment analyses were performed on DEGs.Then,DEGs with higher expression levels involved in the flavonoid biosynthesis pathway were selected for qRT-PCR to verify the expression.【Result】The content of scutellarin in samples S3 and S5 was extremely significantly higher than that in sample S4(CK),and the content of scutellarin in sample S6 was significantly higher than that in sample S4(CK).There were 2143,1968 and 1801 DEGs in samples S3,S5,S6 with high scutellarin and the sample S4(CK)with low scutellarin,respectively.Among them,277 genes were up-regulated in high-scutellarin-content samples,and 213 genes were up-regulated in low-scutellarin-content samples.Among them,the genes chalcone isomerase(CHI),flavanone 3-hydroxylase(F3H)and flavonoid 3’-hydroxylase(F3’H)downstream of flavonoid metabolism were up-regulated in the high-scutellarin samples.The shikimate/quinate hydroxycinnamoyl transferase(HCT)and caffeoyl-CoA O-methyltransferase(CCoAMT)involved in the caffeinate pathway were up-regulated in the low-scutellarin samples.The results of qRT-PCR for 5 DEGs expression patterns were consistent with the results of RNAseq.CHI(Eb_V2_03181),F3H(Eb_V2_40771),F3’H(Eb_V2_04241)were up-regulated in high-scutellarin samples,and HCT(Eb_V2_4655)and CCoAMT(Eb_V2_20841)were up-regulated in low-scutellarin samples.【Conclusion】Scutellarin biosynthesis involves multiple genes,which are closely related to the anthocyanin pathway and the caffeate pathway.

关 键 词：灯盏花 灯盏乙素 转录组 表达分析 QRT-PCR 

分 类 号：R979[医药卫生—药品]