| Xerocomus spadiceus(Fr.)Quel.belongs to Basidiomycotina,Hymenomycetes,Agaricales,Boletacea,Xerocomus.Mainly distributed in Yunnan,Sichuan,Guangxi,Guizhou and other places in China,growing solely or socially in pine and oak forest during summer and autumn and belong to ectomycorrihizal fungi.The fruit bodies are media to large,it become blue when wounded,so called popularly "red while see hand"or "red onion" in local areas,and it is extremely common large variety of wild mushroom in trading market.X.spadiceus tastes delicious,has unique flavor,is rich in mineral elements,possess the feature of high protein and low fat,and is a kind of rare mountain delicacy.Modern medical research shows that polysaccharides,alkaloids in wild boletus have various pharmacological activity on anti-inflammatory,anti-oxidation,anti-virus and anti-tumor.Therefore,boletus has high medicinal value.Most species of the boletus are symbiotic with pine and oak,thus in addition to individual rotten boletus species,they still can't carry out artifical domestication and cultivaton.At present X.spadiceus for market sales are collected from the wild,but with the destruction of the ecological environment and man-made excessive picking in recent years,the price of boletus rose year after year,which has reached 600-1000 yuan/kg,with the products in short supply,it is sustainable use.Thus it is imminent to strengthen the basic research of artifical domestication of X.spadiceus.Now the main cruxes about mycorrhizal edible fungi such as boletus being difficult to artificial culture are:basic research of them is weak,for example,the nutrition,metabolism and physiological mechanisms of natural conditions are still blank;although could be induced mycelia successfully under pure culture conditions,but they grow slowly,differentiated primordia difficultly,and couldn't carry out the whole life history.Therefore,most of the current research focused on the classification,optimization of mycelia media and extration of the active ingredients of mycelia fermentation and so on.Through the work accumulation of our laboratory in recent years,the mycelia and primordia of X.spadiceus has been induced successfully,and primordia are the most important period in the process of growth and development of edible fungi.Therefore,based on the preliminary discussion on media of mycelia and primordia,differential metabolites among three growth and development period of X.Spadiceus were studied emphasically in this paper.The studies above provide important basis for enriching further the artificial domestication theories of boletus,having a good understanding of growth,nutrition and metabolic mechanisms of mycelia and primordia,revealing the mystery of the growth and development of fruit bodies etc.The main contents and conclusions of this study are as follows:The influence on the growth of mycelia and primordia of X.spadiceus with different carbon,nitrogen,pH and organic additives were preliminary studied by single factor experiment.The results shows that suitable carbon source for the growth of mycelia and primordia both were lactose,the pH were 5,the organic additive were bran,and followed by rice,as to nitrogen sources,were respectively NH4NO3 and beef extract with some diffeences.Using]H-NMR technology to detect the water extraction of three stage of mycelia,primordia and fruit bodies of X.spadiceus,by the Topspin software carrying through fourier transform,phase adjustment,baseline correction and calibration,referring to Chenomx nuclear magnetic resonance platform and reference literature 38 metabolit(17 kinds of amino acids,7 kinds of organic acids,4 kinds of sugar,4 kinds of alcohols,3 types of organic alkali,2 kinds of nucleoside and 1 kind of salts)were identified totally.Respectively utilizing MestRenova and SIMCA analysis software,to spectrum peak conducting segmentation integral and date export,differential metabolites of three periods above were analysed comparatively in pairs,the results show that between mycelia and fruit bodies,significant differential metabolites were 16 species(valine,isoleucine,threonine,glucose,trehalose,ethanolamine,alanine,glutamine,succinic acid,acetic acid,ethanol,4-aminobutyric acid,choline,tyrosine,arginine,phenylalanine);between primordia and fruit bodies,13 species(valine,isoleucine,threonine,glucose,trehalose,ethanolamine,alanine,phenylalanine,trehalose,acetic acid,ethanol,4-aminobutyric acid,choline);and 10 species between mycelia and primordia(valine,isoleucine,threonine,trehalose,ethanolamine,glutamine,phenylalanine,choline,tyrosine and methanol).There are 7 metabolites all with significant differential:valine,isoleucine,threonine,phenylalanine,biliary and trehalose between the three of mycelia,primordia and fruit bodies,their relative content(relative peak area)of the first six of them were successively:mycelia<primordia<fruit bodies,but to the trahalose,its relative content:Mycelia>primordia>fruit bodies.Using 1H-NMR technology to shudy metabolomics,there are still have some difficulties in metabolite peak assignments and accurate attribution,and there haven't find report about the use of this technology for macro fungi research.In this paper,some attempts in this respcct were finished and related establish a extraction and analysis method were preliminarily established,the above were the main innovation of this paper. |
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