Identification of Key Genes in the Flavonoid Synthesis Pathway of Meconopsis integrifolia Based on WGCNA
[Objective]Flavonoids are known for their anti-inflammatory,anti-cancer,and antibacterial properties,and are one of the main medicinal components of Meconopsis integrifolia.By analyzing the spatial metabolome information and transcriptome data from various parts(roots,stems,leaves and petals)of M.integrifolia,the key genes regulating the flavonoid synthesis could be excavated.This research could provide valuable insights into the mechanism underlying flavonoids synthesis in M.integrifolia,paving the way for genetic breeding aimed at enhancing flavonoid content.[Method]Transcriptome sequencing was conducted on the root,stem,leave and petal of M.integrifolia to analyze the distribution of flavonoids across different parts using spatial metabolomic data.The weighted gene co-expression network analysis(WGCNA)was employed to identify key modules and key genes closely related to flavonoid synthesis.To validate the reliability of the transcriptome data,12 genes were selected for qRT-PCR analysis.[Result]Flavonoids accumulation in M.integrifolia varied across different parts,with petals being the primary site of accumulation,where 8 main flavonoids were identified.Transcriptome sequencing revealed a total of 20 085 expressed genes,among which 286 genes expressed were exclusively expressed in flowers,showing 3.6-4.2 times more expression than that in other plant parts.Using WGCNA to categorize highly expressed differential genes,a total of 14 co-expression modules were identified,and the key modules,including MEturquoise and MEgreen,were significantly associated with 8 main flavonoids(P＜0.05).KEGG analysis demonstrated that the genes within these two modules were primarily enriched in metabolism-related pathways,with some genes enriched in pathways related to flavonoid synthesis.MEturquoise and MEgreen comprised 18 and 6 genes related to flavonoid synthesis,respectively,and screened 14 core structural genes(5 CHSs,2 HIDHs,2 CCoAOMTs and FLS,CYP75B1,CHI,HCT,and CYP73A)and one transcription factor HB2,which were predominantly highly expressed in petals or stems.The consistent gene expression trends between qRT-PCR and transcriptome data were observed,which showed that the analysis results derived from the transcriptome data were reliable.[Conclusion]The accumulation of flavonoids and gene expression patterns in different organs of M.integrifolia varied significantly,and 14 core structural genes and one transcription factor were screened to be closely related to the accumulation of flavonoids across different organs.These genes might play a key role in regulating the synthesis and differential accumulation of flavonoids in different organs of M.integrifolia.
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