Expression And Regulation Analysis Of Key Genes In Resveratrol Pathway In Grape(Vitis Vinifera L.)Under Different Stresses

Grape(Vitis vinifera L.),a vine of the genus Vitis in the family Vitaceae,is widely grown in the world.It is also one of the important cash crops due to highly nutritious fruits and wine.However,the products of grapevines are affected by various diseases during its growing and developmental period,which could result in reduced grape yield and quality.Previous studies had found the close relationship between the content of resveratrol and its derivatives(resveratrol glycosides,grapein,rosewood stilbene)in grapes and the disease resistance of grape tree.Therefore,studying the expression regulation mechanism of key genes in the resveratrol metabolism pathway is of great significance for the increasing content of resveratrol and its derivatives in grape and for the new variety development to resist disease.The main results in this study were listed at the following.1.Expressing patterns of the resveratrol pathway genes in grape.The transcriptome data of grape variety Pinot Noir under 6 stress conditions were downloaded from NCBI SRA Database to compare the expressing patterns of the differentially expressed genes(DEGs)in grape under different stresses.The results showed that the DEGs under different stresses were significantly enriched in 5 profiles,among which STS genes were only present in 2 profiles.The up-regulated fold of DEGs under different stresses was in the turns of ABA<DM <Wound <UV?C in the Profile 34,while it was ABA<Wound<UV?C<DM in Profile 16.Two PAL genes were up-regulated by ABA treatment,while 31 STS and 12 PAL genes were induced by UV?C.This result indicated that the higher abundance of key resveratrol pathway genes was induced by UV?C than ABA treatment.Fifteen resveratrol pathway genes were up-regulated respectively by DM and T39+DM stresses,but they were not induced to differentially express under T39 treatment.Nine common resveratrol pathway genes were up-regulated respectively by DM and Wound stresses.However,7 up-regulation and 5 down-regulation resveratrol pathway genes were induced by DM but not Wound stresses,while 3 up-regulation and 7 down-regulation resveratrol pathway genes were induced by Wound but not by DM stresses.These results indicated that different resveratrol pathway genes were involved in different response to different stress conditions.2.Development of a tool for the prediction of transcription factors.In order to screen the transcription factors(TFs)for the target genes,the information of the conserved sites in the up-stream 2K sequence,the PCC values of the candidate transcription factor and its target gene,the TATA-box near the binding site and the results from Plant TFDB were used to develop a new predictors for plant TFs,Plant TF-predictor,which could be accessed at http://bioinformatics.fafu.edu.cn/Plant TF-predictor.In this tool,the users only need to input the gene ID and screening threshold on the web interface to get the screening TFs for plant genes.The predicting accuracy of this tool was evaluated by the experimentally identified TF data.The result showed that the accuracy of Plant TFpredictor was 83%,indicating that Plant TF-predictor was efficient to predict TFs for plant genes.3.Screening the transcription factors for key genes in the resveratrol pathway in grape.Plant TF-predictor was applied to predict TF for the key genes in the resveratrol pathway in grape.The results showed that Vv MYB14?Vv MYB15 ? Vv MYB80 might be the TFs for Vv STS and Vv PAL genes,Vv MYB14 and Vv MYB15 might regulate the transcription of Vv C4H2 genes,etc.Moreover,16 STS genes in Pattern 34 had the conservation sequence,GTTGGTGAGA,which was the DNA-binding domain for MYB TFs.On the contrary,2 STS genes in Pattern 16 owned this conservation sequence.This was consistent with the above results of different up-regulated fold of resveratrol pathway in these 2 patterns.4.Revealing the upstream signal transducting genes for the resveratrol pathway in grape.The PCC value was used to screen the signaling related genes for resveratrol synthesis in grape.The genes involved in ABA,JA,SA,IAA and STKs trans-membrane signal transduction showed highly correlated with STS genes.PCC between 3 STKs genes,including VIT?219s0027g00390,VIT?217s0000g04380 and VIT?204s0023g01080,and 23 STS genes in Profile 34 and 16 was over 0.9,indicating that these 3 STKs genes transduct the stress signal to STS genes and induced them up-regulation to resist stresses.However,the STS genes in Profile 34 were regulated by the interaction of b HLH with Vv MYB14 or Vv MYB15,while STS genes in Profile 16 were regulated by WRKY,interpretating that STS genes in these 2 pattern might response to different stress condition.In this paper,bioinformatic methods,such as DEGs analysis,transcription factor prediction and genes correlation analysis were applied to explore the expression regulated mechanism of key genes in the resveratrol pathway in grape.The results in this paper would provide new insight in the functional mechanism of resveratrol pathway genes in grape.