| Ginger(Zingiber officinale)is a perennial herb of the Zingiberaceae family that grows abundantly in tropical and subtropical regions.This plant is renowned for its medicinal properties and as a source of food,making it a valuable commodity with high economic and culinary worth.Gingerol is the main flavor substance of ginger,and also the main functional factor of various functional activities in ginger.It has broad application prospects in fields such as medicine,food,seasoning,cosmetics,and hygiene.,However,the differences in gingerol content among different varieties are still unclear.However,some key enzymes in curcumin and gingerol biosynthesis pathway in ginger have not been identified.In this study,gingerol was extracted from ginger and its content was determined by high performance liquid chromatography,the difference of gingerol content among ginger varieties was statistically analyzed,and the key enzyme PKS and transcription factor GRAS in gingerol synthesis pathway were analyzed by bioinformatics and Functional verification.The research results of this study are summarized as follows:1.Study on Gingerol Content in Ginger Germplasm ResourcesA total of 24 gingerol components were extracted from ginger and their content was determined.Different species contain different components,and the content of different components is also different.By combining multiple methods and perspectives to analyze the detection results,differential metabolites are accurately excavated,and a ginger flavor component evaluation model is established based on the detection analysis results,providing guidance for the planting and production of ginger.2.Functional verification of Key Genes for Gingerol SynthesisIn the early stage,the research team analyzed the ginger genome,obtained the curcumin synthesis pathway,and inferred that the key genes PKS(PKS1,PKS7)and AOR were synthesized at the end of the curcumin synthesis pathway.PKS1 and PKS7 had high homology with CURS and DCS sequences respectively.Synthesize target gene fragments of PKS1,PKS7,and AOR,construct them onto PET-28 prokaryotic expression vectors,and successfully purify PKS1,PKS7,and AOR proteins through nickel column affinity chromatography.The substrates feruloyl coenzyme A and malonyl coenzyme were reacted with genes in sequence,and the reaction results were measured by high performance liquid chromatography for the peak value of curcumin and gingerol.The presence of curcumin was detected by chromatography in the reaction products of feruloyl coenzyme A and malonyl coenzyme A with PKS1 and PKS7.It provides a basis for the application of curcumin,and also provides a reference for the analysis of key genes for gingerol synthesis in ginger.3.Analysis of GRAS gene family in GingerThe differential expression genes(DEGs)of ginger rhizome,root and stem at five development stages were identified by transcriptome analysis.According to the co expression network of structural and transcriptional genes involved in gingerol biosynthesis,multiple transcription factors are associated with gingerol biosynthesis.We selected GRAS gene family for in-depth analysis.This study identified 66 GRAS genes by searching the entire genome sequence of ginger.In summary,the composition and content of gingerol in ginger were determined,and the results were observed and analyzed.It was found that 6-gingerol is the main component of gingerol,with the highest content compared to other components of gingerol;Based on the results of PCA and HCA,all varieties can be clearly separated,indicating significant differences in gingerol content among the 10 ginger varieties.This provides a reference for future research on the synthesis mechanism of gingerol and the selection of materials.In the Functional verification of the key gene for gingerol synthesis,it was found that the selected enzyme had catalytic activity and successfully catalyzed the substrate reaction to produce curcumin;At the same time,the catalytic step of curcumin in ginger is different from the catalytic step of curcumin in ginger.It is generated by two genes assisted by step reaction with substrate;To verify the role of PKS enzyme in the synthesis of curcumin,and provide reference for further research on the synthesis of gingerol;However,there has not been significant progress in the catalytic reaction of gingerol,and further research and exploration are needed in the future.A detailed analysis of the transcription factor GRAS gene in ginger was conducted,and it was found that gingerol expressed differently in different tissues and also played a role in stress resistance,providing gene reserves for further research on the role of transcription factors in gingerol synthesis. |
