Study On Optimization Of Culture Medium And Medicinal Components Of Dictyophora Rubrovolvata

Dictyophora rubrovolvata is a precious edible and medicinal fungus.It’s fruiting body is rich in proteins,amino acids,polysaccharides and vitamins.It has the effects of anti-tumor,antioxidation,antibacterial,anti-fatigue and improving immunity,and has high nutritional and medicinal value.At present,the production mod e of Dictyophora rubrovolvata is mainly solid strain technology,but the slow growth of solid bacteria,high pollution rate,low biotransformation,long cultivation cycle and other problems restrict the development of Dictyophora rubrovolvata industry.There are few reports on the liquid strain of Dictyophora rubrovolvata,and the molecular mechanism in the process of growth and development and the synthesis of related active components are not deep enough.In this study,according to the biological charact eristics of Dictyophora rubrovolvata,the development of liquid strain,selection of nutrients,optimization of culture medium composition and culture conditions were carried out.Different main materials and excipients were used to make Dictyophora rubrovolvata stick spawn,and the nutrients suitable for making Dictyophora rubrovolvata stick spawn were screened.The third generation sequencing technique Naopore was used to analyze the transcriptome sequencing and bioinformatics of Dictyophora rubrovolvata at different growth and development stages,to reveal and explore the key genes and pathways in the process of growth and development,and to regulate the biosynthesis pathway of active components of Dictyophora rubrovolvata.It is report the transcriptome analysis of different growth stages of Dictyophora rubrovolvata,which provides an important resource for further study on the biosynthesis regulation of active compounds of Dictyophora rubrovolvata.1.The components of liquid medium and culture conditions were optimized by single factor and orthogonal experiments.The best carbon and nitrogen sources were glucose and peptone.The best formula for first-class liquid strain is glucose 30g/L,peptone 1.5g/L,KH₂PO₄ 2g/L,Mg SO₄ 1g/L,（NH₄）₂SO₄ 1g/L,Vitamin C 0.016g/L,Indole acetic acid 0.0016g/L.The optimum growth conditions are 25℃,150r/min culture for 20 days,the best liquid volume of secondary liquid strain is 500ml in1000mL triangle bottle,the best inoculation amount is 15%,and the best culture days is 15 days.2.By comparing the mycelial growth inside and outside of the Dictyophora rubrovolvata stick spawn and measuring the growth rate of mycelium,and the main materials and excipients suitable for the mycelial growth of Dictyophora rubrovolvata were screened.The results showed that when alder,birch and cherry were used as main materials,soybean meal and wheat bran as auxiliary materials,the mycelium was dense and sturdy,the mycelium grew fast,and the quality of the Dictyophora rubrovolvata stick spawn was the best.3.The transcriptome analysis of Dictyophora rubrovolvata at different growth stages showed that the Clean reads was 630885562,and 87253 Unigene,were compared to 7 major databases,Uniprot,KEGG,NR,GO,KEGG Pathway,egg NOG and Pfam,respectively.Finally,235148 effectively annotated Unigenes,6245 SSR markers were obtained.The differentially expressed genes in adjacent two periods were analyzed by GO functional annotation and KEGG enrichment analysis,a total of458 significantly enriched gene entries and 35 significantly enriched metabolic pathways were obtained.4.Monooxygenase activity,oxidoreductase,lignin decomposition,cell division,ATP dehydrogenase activity and phosphorylation mechanism,translation extension factor activity gene and lysosome pathway,cGMP-PKG signal pathway,sphingolipid signal pathway,PPAR signal pathway,histidine metabolism pathway are related to the growth,development and maturation of Dictyophora rubrovolvata fruit body.5.The biosynthetic pathways of histidine,phenylpropanes and steroid active substances were expressed in large quantities in the fruiting body of Dictyophora rubrovolvata.Aldehyde dehydrogenase,Phenylalanine lyase andβ-glucosidase and Cholesterol 7α-hydroxylase were genes significantly expressed in these three pathways,respectively,may be the key genes expressed in these pathways.