The Transcriptome Sequencing Of Vitis Amurensis Peel And The Clone And Analysis Of PAL Gene

Vitis amurensis peel is rich in anthocyanins,and its color changes mainly depend on the composition and content of anthocyanins.This test took Vitis amurensis peel in different maturation periods as test matreial and adopt high-throughput sequencing technology to do transcriptome analysis;this test cloned the full length genome of the key gene VamPAL of anthocyanin synthesis with normal PCR method and do bioinformatic analysis;this test analyzed the gene expression of 8 maturation periods of Vitis amurensis peel with Real-time Quantitative PCR technology.The main research result are as fellow:The RNA-Seq analysis was carried out on the Vitis amurensis peel that before color transformed,half color transformed,and after color transformed and the results were:the total amount of differentially expressed genes of before color transformed and half color transformed was 2986,the amount of differentially expressed genes with GO annotation was 2632,the amount of COG funcutional annotation was 1982,the amount of DEGs that involved in phenylpropane biosynthesis pathway was 14,the amount of DEGs that involved in flavonoid biosynthesis pathway was 14;the total amount of differentially expressed genes of before color transformed and after color transformed was 3841,the amount of differentially expressed genes with GO annotation was 3390,the amount of COG funcutional annotation was 2532,the amount of DEGs that involved in phenylpropane biosynthesis pathway was 27,the amount of DEGs that involved in flavonoid biosynthesis pathway was 14.Between these two different comprision periods,5 PAL genes were annotated in common.The test results clarified the changes of related genes in Vitis amurensis peel color transform process and made the foundation for futher exploration.This test cloned the full length genome of the VamPAL gene with PCR technology.The full length genome showed that with complete ORF,VamPAL gene full length 1764 bp,encoding 557 amino acids,belongs to phenylalanine ammonia-lyase supergene family,the protein encoded by the gene neither have a signal peptide nor contain a transmembrane domain.The overall performance is hydrophilic,and it is a stable hydrophilic protein.The phylogenetic tree analysis showed the highest homology with the Eurasian grape.This test analyzed the gene expression of 8 maturation periods of Vitis amurensis peel with Real-time Quantitative PCR technology.The results showed that the expression amount of VamPAL gene in 8 maturation periods has a certain regularity,and the expression amount from the four weeks after flowering to the mature stage showed an overall increasing trend.The expression amount of VamPAL gene was extremely low in half color transformed and after color transformed period.The conclusion was consistent with the transcriptome sequencing results.