The Regulation Of PbUFGT1 Gene In ‘redzaosu’ Pear

In this study, ‘Zaosu’ pear and its red mutant ‘Red zaosu’ were taken as materials. The promoter sequence of PbUFGT1 gene which participated in the biosynthesis of anthocyanin was cloned. The transcription factors which have R2 and R3 domain or bHLH domains were searched in the scope of the pear genome. Then, these transcription factors were compared with the amino acid sequences of MYBs and bHLHs, which involved in the biosynthesis of anthocyanin in other species. We used the software online to analysis the cis-acting elements in the promoter sequence. In order to establish the regulation model of PbUFGT1, Y1 H assay and Y2 H assay were conducted. The main results are as follows:1. The cds sequence of PbUFGT1 was taken as probe and its flanking sequence was found. Then, a 2197 bp of upstream regulation sequence was obtained and there were no difference between ‘Zaosu’pear and its red mutant. The cis-acting elements in the promoter sequence of PbUFGT1 were predicted though the bioinformatic analysis using online software. Then some elements, that involved in light responsiveness, temperature responsiveness, plant growth hormone responsiveness, MYB and bHLH recognition sites were identified.2. According to the database of pear genomic, the genes, which have R2-R3 domain or bHLH domain, were found out and aligned to MYBs and bHLHs, which involved in the biosynthesis of anthocyanin in other species. Then the Real-time quantitative PCR were conducted and five MYB transcription factors: 1-2s1, 1-2s5, 2-1, 4-3, 7-3 and one bHLH transcription factor: b1, which expressed higher in ‘Red Zaosu’ were isolated.3. There are some MYB and bHLH recognition sites in the promoter sequence of PbUFGT1. In order to verify whether or not these transcription factors could regulate the expression of PbUFGT1, we established the regulation model between the transcription factors and the promoter using the Y1 H assay. The results show that 1-2s1、2-1、4-3 and b1 could bind to the promoter of PbUFGT1 and it indicated that these transcription factors could regulate the expression of PbUFGT1.4. To further expound the regulation model of PbUFGT1, we conducted the Y2 H assay and the result showed that 2-1 and 4-3 could interact with b1. All the results indicated that PbUFGT1 was regulated by multiple transcription factors and the high expression of these transcription factors stimulated the expression of PbUFGT1 and resulted in a large amount accumulation of anathocyanin.