Application Of Transcriptome And Proteomics To Excavate The Key Points Of The Formation Of Ophiocordyceps Sinensis And The Differences Between The Protein Groups Of Wild Ophiocordyceps Sinensis And Artificial Ophiocordyceps Sinensis

Ophiocordyceps sinensis is a dry complex of stroma,which is Ophiocordyceps sinensis(Beck.)Sacc parasitic on larvae of genus Hepiaus and corpses of larval.Ophiocordyceps sinensis has the effect of “nourish lungs and kidneys and relieving cough and reducing sputum”,and has long been regarded as a valuable Chinese medicines.However,due to the particularity and complexity of its physiological and biochemical characteristics,the research on the formation mechanism of its fruiting body is not clear at present.In this study,Illumina/Solexa HiSeq 2500 reverse transcriptional sequencing technology was used for sequencing,data assembly and analysis of the larvae transcriptome of the mycelium and fruiting body of Ophiocordyceps sinensis and larva of Hepialus xiaojinensis,and it has found that high identities were shared among mycelium and fruiting body,while larva of Hepialus xiaojinensis assembly shared very low identities with the others;The differential expression analysis showed that 9238 genes were expressed in all four samples,corresponding to a percentage of 72.57 %.It has been identified that 1003 genes are DEGs,and KEGG enrichment analysis showed that these DEGs were enriched in the pathway of“Ribosome”;A total of 161 differentially expressed genes involved in enriched GO terms.Only58 of them were down-regulated in fruiting-body,while the other 103 DEGs were up-regulated,and these results indicated that the protein synthesis basic metabolism was altered in Ophiocordyceps sinensis after infecting Hepialus xiaojinensis.Forty-seven specifically expressed genes were also found.And we also have been found that the alteration of actin and tubulin related genes expression may play roles in the initiation of fruiting-body development.These results are all related to the formation of fruiting body of Ophiocordyceps sinensis.In the meantime,we used proteomic techniques to analyze the proteome of three stages of the development(mycotic complex,fruiting body,complete body)of Ophiocordyceps sinensis(mycotic complex,fruiting body,complete body),and validate the indicative proteins of Ophiocordyceps sinensis among quality goods and counterfeit drugs in the early stage of our research group.And in the study,we selected Five stable proteins and three proteins with significant differences.After further identifying the eight proteins,the functional annotationanalysis(GO,KEGG)was performed,and the results were found that the three proteins(spots 1,5and 6)with significant differences were all related to cell composition.Spot 1 is involved in a large number of nucleotide binding and enzymatic activities.Spot 5 is involved in a large number of metabolic processes and compounds.Spot 6 is involved in a large number of metabolic processes and cell growth processes.The spots 2 and 3 of the 5 stable proteins were correlated with the stimulation response and nucleoside triphosphatase.Spot 4 is related to organic nitrogen compound metabolism and ion binding.Spot 7 was involved in biological regulation and hydrolytic enzyme activity,and spot 8 was associated with transcriptional synthesis.Spot 1 was identified as actin,GO annotation found that the protein is involved in cell,cell part,cellular component,intracellular,intracellular part,KEGG Pathway enrichment analysis found that its active metabolic pathway is Phosphatidylinositol signaling system,phagosome.It is suggested that spot 1 may play an important role in the growth and development of Ophiocordyceps sinensis.This study also used two-dimensional electrophoresis(2-DE)on protein groups of mycotic complex,fruiting body,complete body of wild Ophiocordyceps sinensis and artificial Ophiocordyceps sinensis.The specific protein of wild Ophiocordyceps sinensis was selected.We also validates the indicative proteins of Ophiocordyceps sinensis among quality goods and counterfeit drugs in the early stage of our research group.The differences between wild Ophiocordyceps sinensis and it laid the foundation for the identification of wild Ophiocordyceps sinensis and artificial Ophiocordyceps sinensis.