复色菊花青素生物合成关键基因探究OA

[Objective]This study aims to identify key genes involved in the anthocyanin biosynthesis path-way of multicolored Chrysanthemum morifolium,thereby providing a molecular basis for elucidating the mecha-nisms underlying floral pigmentation patterning.[Method]Takes the multicolored Chrysanthemum morifolium'Huasayaogun'as research object,floral coloration was quantified by using the Royal Horticultural Society Co-lour Chart(RHSCC)in combination with a colorimeter,and the total anthocyanin and flavonoid contents were determined.Comparative transcrip tome sequencing and analysis were performed on distinct color sectors of the same ray florets across different developmental stages.Genes with P＜0.05 and log2|fold change|≥1.5 were identified as differentially expressed genes(DEGs)and subsequently subjected to GO(Gene Ontology)and KEGG(Kyoto Encyclopedia of Genes and Genomes)functional enrichment analyses.Structural genes and MYB transcription factors associated with anthocyanin biosynthesis were screened and identified based on func-tional annotation results,and quantitative real-time PCR(RT-qPCR)was used for validation.[Result]The multicolored Chrysanthemum morifolium'Huasayaogun'was characterized by a combination of purple-red and white color sectors.Stage S2 is the critical phase for pigmentation in the purple variegated regions,where antho-cyanin accumulation and spatial distribution are the key determinants of purple spot formation.Transcriptomic analysis revealed a total of 4 779 differentially expressed genes(DEGs)between the purple(PS2)and white(WS2)sectors during Stage S2,with 2 506 DEGs upregulated and 2 273 DEGs downregulated in the purple sectors in comparison with the white sectors.The GO enrichment analysis revealed that during the S2 stage,3 533 differentially expressed genes(DEGs)were annotated to three functional categories:biological process,molecular function,and cellular component.Within the biological process category,the subcategory cellular pro-cess was annotated with 1 629 genes.KEGG pathway enrichment analysis revealed that DEGs were signifi-cantly enriched in major functional categories,including metabolism,genetic information processing,and organis-mal systems,based on the KEGG database classification hierarchy.Based on integrated analysis of KEGG anno-tations,flavonoid biosynthesis and anthocyanin biosynthesis pathways were identified as key pathways for floral pigmentation.By analyzing the flavonoid and anthocyanin biosynthesis pathways during the critical stage of floral pigmentation,six key genes:F3'H,FLS,DFR,ANS,MYB1,and MYB3,were identified as being closely asso-ciated with pigment synthesis.RT-qPCR validation results were consistent with the transcriptomic analysis.[Conclusion]F3'H,DFR,ANS,MYB1,and MYB3 were identified as key positive regulators driving anthocy-anin accumulation and subsequent purple spot formation.