Transcriptome analysis of Aucklandiae lappa from different growth years and mining genes related to sesquiterpenes synthesis
[Objective]Sesquiterpene is the main active component of Aucklandiae lappa.By analyzing the sesquiterpene content and tran-scriptome differences in the roots of A.lappa at different growth years,the study aimed to explore the related genes for sesquiterpene synthe-sis,providing theoretical references for further research on functional genes and active ingredient synthesis in A.lappa.[Method]In October,the roots of 2-year-old and 3-year-old healthy A.lappa plants from the same plot were collected as materials,and the sesquiterpene content was determined by high performance liquid chromatography(HPLC),and a cDNA library was constructed.Transcriptome sequencing was per-formed using the IIllumina NovaSeq 6000 high-throughput sequencing platform,and bioinformatics analysis was used to analyze the original sequencing data.The gene expression results were verified by qRT-PCR technology.[Result](ⅰ)The results of HPLC showed that the ses-quiterpene content was higher in 3-year-old A.lappa and increased with the extension of growth years.(ⅱ)After quality control screening,a total of 40 Gb Clean base was obtained by transcriptome sequencing,and 90 912 Unigenes were obtained after splicing,with an average length of 1081 bp and N50 of 1637 bp.After comparing with 7 databases,the results showed that a total of 61 268 Unigenes obtained annotation in-formation,with the highest number of homologous sequences between A.lappa and Cynara cardunculus L.(ⅲ)827 DEGs were screened based on the standard that the absolute value of log2 multiple of FPKM value was greater than 1 and the error detection rate was less than 0.05.Among them,61 DEGs had upregulated expression levels and 766 DEGs had downregulated expression levels in 3-year-old A.lappa.509 DEGs were annotated in 109 GO items,and 12 of them were significantly enriched.Among 12 significantly enriched GO items,3 groups were in molecular function,4 groups were in biological process,and 5 groups were in cell components.509 DEGs were enriched into 109 GO items,with 12 significantly enriched,including 3 groups in molecular function,4 groups in biological process,and 5 groups in cell compo-nents.KEGG enrichment analysis showed that 287 DEGs were annotated into 178 metabolic pathways,mainly enriched in ribosome,Alzheimer's disease,cGMP-PKG signaling pathway and PPAR signaling pathway.According to the annotation information of DEGs,7 key en-zyme genes involved in sesquiterpene biosynthesis pathway were excavated,including 3 DXS,1 MECPS and 3 GERD,which were downregu-lated in the expression level of 3-year-old A.lappa.(ⅳ)Four candidate genes were verified by qRT-PCR technology,and the validation results were consistent with transcriptome data.[Conclusion]There are significant differences in the content of sesquiterpene substances and the ex-pression of sesquiterpene biosynthesis genes between 2-year-old and 3-year-old A.lappa.The excavated genes related with sesquiterpene bio-synthesis provide a basis for elucidating molecular mechanism of sesquiterpene biosynthesis and molecular breeding of excellent varieties in A.lappa.
万方数据
维普
