Cloning of MYB Family Gene MawuCPC of Magnolia wufengensis and Analysis of Its Role in Flower Color Regulation
Magnolia wufengensis is a rare ornamental tree species endemic to China.Its flower color variation types are extremely rich.There are a series of flower color variation varieties such as deep red,pink and white.How-ever,the molecular mechanism of flower color regulation is still unclear.In order to explore the molecular mecha-nism of flower color regulation of Magnolia wufengensis,MawuCPC,a key candidate gene for flower color regulation of Magnolia wufengensis,was cloned from Magnolia wufengensis.Phylogenetic analysis,amino acid sequence struc-ture analysis,subcellular localization experiment,yeast two-hybrid experiment and luciferase experiment were car-ried out.The expression level of MawuCPC in different flower color varieties was compared and analyzed by Real-time Quantitative PCR.The results showed that the coding region of MawuCPC gene was 258 bp,encoding 85 amino acids.Phylogenetic analysis showed that MawuCPC was clustered with CPC members of Arabidopsis thaliana and other species in the same evolutionary branch,which further supported that MawuCPC belonged to the CPC family of Magnolia wufengensis.Sequence structure analysis further showed that MawuCPC protein had only a conserved R3 domain and a bHLH binding motif.The result of subcellular localization experiment showed that MawuCPC pro-tein was specifically localized in the nucleus.Yeast two-hybrid and luciferase assays showed that MawuCPC and MawuPAP1 could interact with the bHLH family protein MawubHLH1 of Magnolia wufengensis,and the binding of MawuCPC to MawubHLH1 could seriously inhibit the enhancement expression of MawuPAP1 and MawubHLH1 on the promoter of the structural gene MawuDFR gene.Real-time Quantitative PCR results showed that the expression level of MawuCPC was the highest in the white flower variety JB,followed by the pink flower variety Jiaoyan,and the lowest in the red flower variety Jiaohong 1.The above experimental results indicate that in Magnolia wufengen-sis,MawuCPC may also play its role in flower color regulation by competing with PAP1/2 protein to bind bHLH pro-tein,and maybe the difference in the expression level of this gene promotes Magnolia wufengensis to produce ex-tremely rich flower color variation types.
Magnolia wufengensisMawuCPCPCRYeast two-hybridExpression regulationFlower color
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