Regulation Of Capsaicin By Pepper MYB Transcription Factor Based On Transcriptome Sequencing

Capsaicin is a hot pepper in the placenta,synthesis of important secondary metabolites is an important index,evaluation of pepper fruit quality traits in the field of medical,food,agriculture,military,etc all play the important role,but in the market,chili used to appear the situation of demand is less than the supply,the main reason lies in the yield of Capsaicin is too low,is limited by environmental conditions,the production cost,traditional breeding ways hardly developed high hot high yield of new varieties,so at the molecular level,through the research of Capsaicin synthesis mechanism,which affect the key gene of the content of Capsaicin It is of great significance to find a way to improve the accumulation of capsaicin in this field.Of this study was based on the devil pepper transcriptome sequencing analysis,four control gene screening differences,including three from MYB family,1 from WRKY family,based on the genetic variations in different varieties of pepper and fruit varieties of the same pepper quantity analysis,expressed different organs in the regulation of gene expression of regularity and reveal the differences,the correlation between by cloning and bioinformatics analysis,using VIGs and Bifc method to study the effect of difference control gene for capsaicin content,the results are as follows.1.Three differentially up-regulated genes(CcMYB76,CcMYB171,and Cc WRKY107)were screened by transcriptome sequencing,and one differentially down-regulated gene(CcMYB143)was detected.Based on keyword screening on the cloud platform,CcMYB76 and Cc WRKY107 were speculated to be related to capsicin,while CCWRKY107 was related to anthocyanin.2.The fluorescence quantitative expression analysis in fruits,leaves and stems of six varieties of pepper showed that the expression levels of CcMYB76,CcMYB171,CcMYB143 and Cc WRKY107 in fruits were significantly higher than those in leaves and stems,and all four transcription factors played a role in fruits of pepper.The expression of CcMYB171 in mature fruits of three types of pepper showed little correlation with capsaicin regulation.3.By using the method of virus induced gene silencing silence silent CcMYB76 in pepper fruit,CcMYB143,Cc WRKY107 gene after five structure on the determination of capsaicin synthesis pathway gene expression,and the content of capsaicin,when difference regulation genes have been silent,compared with the control group,the content of capsaicin are decreased,which when silence Cc WRKY107 capsaicin content decrease up to about 1.5 times,structural gene will also change the corresponding raised or lowered,These results indicate that CcMYB76,CcMYB143 and Cc WRKY107 all regulate the expression of structural genes during capsaicin synthesis and are transcription factors that affect capsaicin content.4.BIFC method transfer CcMYB76,CcMYB143 and Cc WRKY107 tobacco,when CcMYB76 and Cc WRKY107 turn,observed in the cell membrane and nucleus yellow fluorescent signal,two transcription factors in the nucleus and happen interactions on the cell membrane and CcMYB76 Cc WRKY107 similar expression in the fruit,and silent Cc WRKY107,the expression of CcMYB76 exponentially decline,speculation Cc WRKY107 transcription factor itself directly involved in the regulation of capsaicin It also indirectly regulates capsaicin synthesis by acting on CcMYB76.