Coloning Of Pingguoli Chi And F3H Genes And Their Expression In Fruit Peel Before And After Bag Removal

This experiment adopts the RT-PCR technique successfully cloned two key enzymes (CHI、F3H) in Pingguoli and analysis related bioinformations. Then monitoring their expressing characteristics by real-time fluorescent quantitative PCR. The results are as follows:1. Designed a pair of primers of each gene by the conservative district and got the CDS of CHI and F3H by amplification. The results show that the length of CHI is702bp while F3H is1095bp, they code234and364amino acids respectively.2. The results of blast show that:comparing other representative plants on Genbank with Pingguoli the bases similarities of Pingguoli CHI and F3H are around90%, some get98%above. For example, the CHI gene in Pingguoli, it got a98%similarity with Pyrus communis and96%with Pyrus serotina; the F3H gene in Pingguoli got a99%similarity with Pyrus communis and98%with Malus pumila. The phylogenetic analysis manifest that the CHI gene in Pingguoli is closest to Pyrus communis in evolution and Pyrus serotina take the second place; the F3H in Pingguoli is closest to Pyrus communis, strawberry take the second place and it also has close relationship with Pyrus serotina.3. The real-time fluorescent quantitative PCR result indicate that bagging could suppress CHI gene’s expression, after solution bag CHI gene expression will reach a higher level in a short time and then drop down, after this stage, the expression of the gene reach up again. It indicate the CHI gene is closely related to the cyanine nucleoside synthesis in Pingguoli; Bagging could also suppress F3H gene expression, after solution bag F3H gene’s expression will reach a peak level in a short time and keep stable, during the cyanine nucleoside synthesis stage, the expression of the gene don’t shows a dramatic change, this suggest that the F3H gene don’t have a direct relationship with cyanine nucleoside synthesis. It maybe play a coordinating role with other genes in cyanine nucleoside synthesis.