Cloning And Interaction Analysis Of CaAFYB And CaAJMYB Genes With Pun1 In Pepper

Pepper(Capsicum annuum L.)is an annual or perennial herb of Capsicum in Solanaceae.It is rich in vitamin C and capsaicin and has rich nutritional value.Capsaicin is an important character to determine the quality of capsaicin,but current studies have shown that capsaicin biosynthesis is closely related to environment,genotype and cultivation conditions,which leads to the fact that natural capsaicin is far from meeting the market demand of capsaicin.So capsaicin,which is pure natural,pollution-free and has many biological functions,has broad prospects for development.Qin and Kim completed genome sequencing of pepper in 2014.In capsaicin biosynthesis pathway,the expression of acyltransferase gene Pun1 plays a decisive role in pepper fruit spicy.The study on the regulation of Pun1 gene plays an important role in comprehensively analyzing the regulation mechanism of capsaicin biosynthesis pathway,improving the quality of pepper fruit and increasing its commercial value.In this paper,two MYB transcription factor genes of Capsicum MYB family were cloned and sequenced,bioinformatics analysis,subcellular localization and interaction with Pun1 gene.The results are followed:1.In this study,two MYB family transcription factors gene Ca AJMYB and Ca AFYB were cloned from pepper by PCR.The full length of Ca AJMYB is 1096 bp,encoding 262 amino acids.There is one MYB domain located in the nucleus of the transcription factor amino acid sequence from 14 to 61 and from 67 to 112,respectively.The full length of Ca AFYB is 1128 bp,encoding 340 amino acids.There is one MYB domain located in the cell wall from 13 to 60 and 66 to 109 of the amino acid sequence,respectively.R2R3-MYB transcription factors refer to MYB transcription factors that contain two different MYB domains.It is inferred that Ca AFYB and Ca AJMYB transcription factors belong to the R2R3-MYB subfamily.2.The promoter of acyltransferase gene Pun1 was amplified by PCR.Its full length is 982 bp.AD target vectors p Lac Zi-Pun1 and BD report vectors p GAD424-AJMYB and p GAD424-AFMYB are constructed.The vector is transferred into yeast YM4271 by yeast single hybridization technology.The interaction between the two transcription factors and Pun1 gene is verified by color reaction.Ca AJMYB has stronger interaction ability.