Mining Of Volatile Oil Metabolites And Related Genes From Wild Alpinia Oxyphylla In Haina

Alpinia oxyphylla Miq.is also called yizhiren,yizhizi and zaiyuzi.It is a genus of ginger in the ginger family,and is one of the four famous southern medicines in China.As early as in the Classic of the Materia Medica,it is written that Alpinia oxyphylla Miq.seeds are used to treat spermatorrhea,deficiency and leakage,and urinary dripping,which are all signs of kidney qi not being solid.It is said that Alpinia oxyphylla Miq.have the effect of benefiting the spleen and stomach,regulating vital energy,and replenishing kidney deficiency and slippage.The main active ingredient,volatile oil,has various medicinal effects such as antibacterial,cardiac,analgesic and anticancer.It is also an important raw material for many health products,traditional Chinese medicine and food,and has a wide range of economic and medicinal values and a long history of application.The content of various compounds in the volatile oil of Alpinia oxyphylla Miq.as an active medicinal component of Alpinia oxyphylla Miq.is related to factors such as ecological factors of the origin of Alpinia oxyphylla Miq.,collection sites,processing and extraction methods.So far,the main compounds found in Alpinia oxyphylla Miq.are terpenoids,phenols and sterols,among which the characteristic components are terpenoids,which have been clinically proven to have pharmacological effects such as neuroprotection,memory enhancement,antibacterial and anti-inflammatory,improvement of respiratory function,cardiac stimulation and sedation and hypnosis.In this study,the metabolites of Baoting County,Hainan Province,were isolated and identified using UPLC-MS/MS metabolomics,and a BSA-seq extreme hybrid pool was constructed based on the results of volatile oil analysis,combined with second-generation high-throughput sequencing technology to survey the Alpinia oxyphylla Miq.genome and to mine the target trait genes related to volatile oil metabolites.The contents and the research results obtained in this thesis are as follows.Through comparative metabolome analysis of 20 selected wild Alpinia oxyphylla Miq.fruit samples,a total of 239 metabolites were identified,including 67 terpenoids,46 esters,22 aromatic hydrocarbons,20 ketones,15 alcohols,8 aldehydes,8 hydrocarbons,6 amines,3 halogenated hydrocarbons,1 phenol,1 acid,1 nitrogenous compound,1 sulfurous compound,36 heterocyclic compounds and 3 other compounds.The experimental results showed that the chemical composition also differed significantly between the 20 groups,with a total of 195 compounds within the 20 groups.The four groups of 01-F,26-F,31-F and 32-F contained the most non-shared differential metabolites,mainly terpenoids and oxygenated derivatives.One unique differential terpene metabolite,(+)-camphor,was also found in 20 groups of samples.In this study,the main components of the Alpinia oxyphylla Miq.volatile oil were mostly terpenoids and oxygenated derivatives such as esters,and KEGG enrichment analysis showed that there were 17 metabolites enriched in different metabolic pathways,and these metabolites were enriched in 8 different KEGG metabolic pathways.At the same time,according to the compound content,6,6-dimethyl-bicyclo[3.1.1]heptane-2-ene-2-formaldehyde,citral B,(-)-β-Diterpenes and laurenes may play a more important role in the concentration of metabolic pathways.Three BSA-seq extreme hybrid pools were constructed based on the content of volatile oil metabolites in Alpinia oxyphylla Miq.fruits,and the Alpinia oxyphylla Miq.reference genome was surveyed in combination with second-generation high-throughput sequencing technology to evaluate the double-end sequencing data and finally assembled to obtain a Alpinia oxyphylla Miq.reference genome of about 1.3G in size and 1.05%heterozygosity maximum.Based on this,the BSA data were further re-matched to the obtained Alpinia oxyphylla Miq.reference genome survey,and finally 1,336,372 SNP loci and 188,502 InDel loci were obtained,and finally,three groups of candidate genes that may be involved in the regulation of volatile oil metabolites in the DNA extreme mixing pool were analyzed,of which the first group obtained four genes;The second group obtained 16 genes;The third group obtained 8 genes,and 28 related genes were selected from the three groups..In this study,we screened and analyzed the metabolites and major metabolic pathways of Alpinia oxyphylla Miq.fruits from different origins to reveal the metabolic differences among different populations of wild Alpinia oxyphylla Miq.fruits.The low coverage sequence of the whole Alpinia oxyphylla Miq.genome was obtained by BSA analysis combined with second generation sequencing,which can lay the foundation for the subsequent deep sequencing of the whole Alpinia oxyphylla Miq.genome,and at the same time,the target trait genes related to volatile oil metabolites were mined,which can be used for It also provides a theoretical basis for the subsequent germplasm analysis,medicinal function study and application of the volatile oil of Alpinia oxyphylla Miq.