De Novo Assembly And Transcriptome Analysis Of Mycelium Formed Sclerotial And Sclerotial Development In Wolfiporia Cocos

Wolfiporia cocos,a well-known traditional Chinese medicine(TCM)in the Basidiomycetes,is widely distributed in East Asia.It is has a very high medicinal value and dietary nutritional value,are widely used in traditional Chinese medicine and health food.Polysaccharides and triterpenes are the main active chemical constituents of sclerotium of W.Cocos.Due to the growth of W.cocos mainly depends on the parasitic on pine,pine of scarcity limits the size of W.cocos production year by year,and the main researching contents of W.cocos has focused on the pharmacological active ingredients,such as polysaccharide and triterpene active ingredient research,research report at the molecular level is only triterpene synthesis genes,so molecular genetic breeding of W.cocos has become improve the planting scale,rich market resources important areas.With the rapid development of sequencing technology,The illumina deep RNA sequencing has laid a solid foundation to explore relevant genes of W.cocos sclerotium formation and development.This study is based on Illumina HiSeq 2000 high-throughput sequencing,won the mycelium growth of W.cocos 7 days,sclerotium growth minus 2 months(M,S1,S2,S3)transcriptome data,using the Trinity and TGICL software for Unigene De Novo,based on the transcriptome BLAST get comments,annotation of Unigene protein function,classification,GO functional annotation,KOG functional annotation,KEGG metabolic pathways and Heatmap analysis.Based on poria cocos transcriptome data,through gene expression abundance and real-time fluorescence quantitative(qRT-PCR)experimental results of a score,screening and sclerotium formation evolution and development of the related functional genes.The results of the study are as follows:1.Through poria cocos transcriptome data splicing,quality pretreatment and De Novo splicing comments received 36312 Unigene,17018 Unigene is hit to in NR about homologous protein information,Swissport database with 9932,the KOG database,there are 6780,GO in the data,there are 9073,KEGG database,there are 9033.However,there are still 19294 Unigene is unknown function.2.Four periods of W.cocos different transcriptome data are analysised for revealing the W.cocos sclerotium at the molecular level in the process of the formation and development,related functional genes play a role of regulation and control.3.The period of W.cocos M and S1,transcriptome data analysis show that there are 304 carbohydrate enzyme,and gene expression abundance(RPKM)contrast,there are 69 carbohydrate enzyme,found for the formation of W.cocos sclerotium continuous carbon source,the glycoside hydrolase(GHs)of 43,which is dominant the family,carbohydrate esterase(CEs)of 5,polysaccharide lyase(PLs)5,carbohydrates(CBMs)combined with area 0,glycosyl transferase(GTs)17,subsidiary vigor enzyme 2,degradation of dextran,respectively,xylan,chitin,pectin,hemicellulose.4.W.cocos S1,S2,S3 period the transcriptome data analysis reveal that:GO in development of cluster analysis showed that W.cocos into molecular metabolism in extremely strong,with 28 differentially expressed genes throughout the three times,including 25 in M times,KEGG Heatmap and DEG differentially expressed analysis poria cocos sclerotium in S3,limonene degradation enzyme,protein enzymes,MAPK signaling pathways such as highly expressed,show that sclerotium is ripe;S3 divinyl benzene polymers,triterpene substances synthetic S2 period,unsaturated fatty acid,fatty acid,respectively in S2,S3 period significantly synthesis,fungi to store a lot of nutrients by the formation of unsaturated fatty acid,at the same time,using C18 unsaturated fatty acid and its derivatives have proved to have a significant relationship on sclerotium development.S1 period peroxide dismutase(sod),superoxide dismutase(sod)is extremely active,also exists in M period at the same time,oxygen pressure and hydrogen peroxide in the metabolism of cells may be associated with sclerotium formation.5.In candidate genes selected 28 genes in RPKM value and qRT-PCR experiment data model,consistent with the experimental results prove that gene expression.Through the high throughput of W.Cocos transcriptome sequencing data analysis,we find out the related genes of W.Cocos sclerotium formation and development,reveals the W.Cocos sclerotium at the molecular level in the process of the formation and development,related functional genes play a role of regulation and control.Laid foundatation for further large-scale poria cocos herbs cultivation research work.