Cloning of PvWRI1 transcription factor and analysis of its function in oil synthesis of Plukenetia volubilis
WRI1 is a kind of transcription factors with APETALA2 domain,which is essential for the oil bio-synthesis.In this study,the genomic sequence of PvWRI1 was cloned from fresh leaves of Plukenetia volubilis and analyzed by bioinformatics methods.Through the construction of overexpression vector,PvWRI1 gene was trans-formed into tobacco to obtain transgenic positive tobacco plants.Wild type and transgenic mature seeds were har-vested and the fatty acid components were determined by gas chromatography.The expression level of downstream fatty acid pathway genes of PvWRI1 was analyzed by real-time PCR.The DNA sequence of PvWRI1 was 5072 bp,the coding region is 1312 bp,encoding 436 amino acids,and including 7 exons and 6 introns.Hydrophobicity analy-sis showed that the protein was hydrophilic.Domain analysis showed that the protein contained two AP2/EREBP structural regions and four inherently disordered protein regions,and the secondary structure analysis showed that the protein was relatively conserved.Phosphorylation predicts the characteristics of N-terminal chloroplast trans-port peptides.Amino acid homology analysis showed that the AP2 domain had high homology in several species,and the phylogenetic tree showed that PvWRI1 had the highest homology with WRI1 of castor.According to the analysis of fatty acid components,the contents of stearic acid and linolenic acid increased significantly,by 16.8%and 7.5%,respectively.The total oil content of transgenic seeds increased from 12.84%to 24.06%,and it was up to 1.87 times.At the same time,the linoleic acid/linolenic acid balance ratio decreased by 7.8%.RT-qPCR results showed that overexpression of PvWRI1 gene led to the up-regulation of fatty acid pathway genes,in which WRI1 gene was most significant,and FATA,KASI and FAD3 genes were significantly up-regulated.Therefore,as a crucial regulator in fatty acid biosynthesis,PvWRI1 plays a guiding effect on improving oil content and fatty acid balance in oil plants.
Plukenetia volubilisPvWRI1gene cloningoverexpressiondetermination of fatty acidsex-pression quantity
万方数据
维普
