Cloning and expression analysis of PyCAD genes in Pinus yunnanensis
[Objective]The paper aimed to comprehend the expression characteristics of cinnamyl alcohol dehydrogenase from Pinus yun-nanensis and its involvement in lignin biosynthesis.[Method]The relative concentrations of 18 specific small molecules within the lignin pathway were assessed in the stems,buds,and needles of one-year-old P.yunnanensis.Additionally,the CAD genes of P.yunnanensis were identified,cloned and bioinformatics analyzed by comprehensive analysis of transcriptome data.RT-qPCR was employed to examine their expression patterns across various tissues.[Result]The findings revealed that among the 18 lignin pathway molecules,the relative con-tent of 5-O-caffeoylshikimic acid,5-O-p-coumaroyl shikimic acid,and caffeic acid were higher in the buds,while the relative content of 3-O-p-coumaroyl quinic acid,p-coumaric acid,ferulic acid,coniferylaldehyde,L-phenylalanine,coniferyl alcohol,p-coumaryl alcohol,L-(-)-tyrosine,sinapinaldehyde were higher in the stems.In addition,the relative content of coniferin,p-coumaraldehyde,caffeic aldehyde,and syringin were higher in the needles.Four members of the CAD family were screened from the transcriptome of P.yunnanensis,and phy-logenetic analysis showed that PyCAD-3 of P.yunnanensis was homologous to AtCAD1,and PyCAD-1,PyCAD-2 and PyCAD-4 were closely related to CAD of Pinus,AtCAD4 and AtCAD5.The results of correlation analysis showed that the PyCAD-1 and PyCAD-2 were closely relat-ed to small molecules in the lignin pathway upstream and downstream.The open reading frame of PyCAD-1 and PyCAD-2 was 570 and 1083 bp,respectively,and the molecular weights of PyCAD-1 and PyCAD-2 were 20.38 and 39.30 kDa,respectively.The results of RT-qPCR showed that the relative expression of PyCAD-1 was the highest in bud and tender stem,and PyCAD-2 was the highest in tender stem.[Con-elusion]It is speculated that PyCAD-2 may be directly involved in lignin biosynthesis,while PyCAD-1 may have some functional redundancy.The study provides a theoretical basis for studying CAD genes family and genes related to the lignin metabolism pathway of P.yunnanensis.
万方数据
维普
