Abstract: To investigate the expression of key enzymes affecting the composition of anthocyanins in the transformation stage of grape peel of Vitis amurensis. With the young leaves of Vitis amurensis as the test material, the genomic DNA of Vitis vinifera was extracted and the full length sequence of PAL (GenBank accession number:MH045991) was obtained by homologous cloning, and conducted a biological information analysis. The Q-PCR method was used to analyze the expression levels of 8 grapevine skins at different pigmentation stages. The results showed that the DNA sequence of the PAL gene was 1 763 bp in length and had a 1 671 bp open reading frame (ORF) encoding 556 amino acids. The molecular weight was 61.07 ku and the isoelectric point was 5.76, which was a stable protein. The whole polypeptide chain had a transmembrane helix structure with no signal peptide. Molecular phylogenetic tree analysis showed that the homology was higher with European and Asian grape species; The expression pattern of PAL gene in the 8 stages of the grape rind of P. villosa. The PAL gene was analyzed in the last 4 periods. With irregular expression levels, the PAL gene was hardly expressed during both the 50% color shift and 100% color shift. The regulation of PAL gene expression was regulated by many factors. It changed regularly in the early stage of growth and development of Vitis vinifera. In the last 4 periods, the expression level changes irregularly. PAL gene was in 50% color rotation and 100% color rotation. Almost no expression, which may be affected by some kind of inhibition of PAL enzyme activity factors.

Key words: Vitis amurensis, Phenylalaninammo-nialyase, Gene cloning, Sequence analysis

CLC Number: 

• Q78

• S663.03