Changes In Proteomic Profile At Different Development Stages Of Volvariella Volvacea By Comparative Analysis

Volvariella volvacea (Bull. ex Fr.) Sing., which is commonly known as the paddystraw mushroom, is the fifth most popular edible mushroom in the global market. Atpresent, China is the leading producer and exporter of V. volvacea in the world,accounting for about60%of its total production.In this study, isobaric tags for relative and absolute quantification(iTRAQ)-coupled two-dimensional liquid chromatography tandem mass spectrometry(2D LC-MS/MS) proteomics approach was first applied to investigate changes ofproteomic profiles during different development stages of V. volvacea. After that,bioinformatics was used to analyze the detected results. The chief results are showedas follows:(1) One pairs of non-fruiting homokaryotic strains, PYD15and PYD21, andheterokaryotic strain PYD1521obtained from mon-mon mating between PYA15andPYD21were firstly identified by observations of mycelium colony shapes andstructure variation (SV) molecular markers. Then three sort mycelia of V. volvacea,PYD15, PYD21and PYD1521, were respectively collected by stationary cultivation.Five fruit-body samples of primordium stage, button stage, egg stage, elongation stageand mature stage were determined and collected by fruit-body shapes and innermicroscopic examination in fruit-bodies of V. volvacea.(2)1390protein groups, included a total of2335unique peptides, were identifiedby using iTRAQ)-coupled2D LC–MS/MS. There are667protein groups matched bytwo or more of unique peptide fragments among1030protein groups providedquantitative information. The data of identified proteins results were combined bycontrol data of heterokaryon strain PYD1521. Differentially expressed proteins wereanalyzed for significant down-or up-regulation, which was calculated by twice folds,a total of385unique proteins were identified at different development stages of V. volvacea, of which there were83up-regulated and97down-regulated significantlydifferential proteins.(3) Results of the experiment and analysis of principal component, hierarchicalclustering, correlation clustering, correlation coefficient thermogram and K-meansshowed that biologically relevant changes during different development stages of V.volvacea could be displayed by the data of identified proteins. The results revealedthat iTRAQ-coupled2D LC-MS/MS was an excellent method to isolated andidentified protein groups of V. volvacea at different development stages.(4) Analysis of Gene Ontology (GO), KEGG pathways and International ProteinIndex (IPI) showed that163different expression proteins at different developmentstages of V. volvacea were involved in80unique metabolism pathways from fourfungi of Agaricales, Coprinopsis cinerea, Laccaria bicolor, Moniliophthoraperniciosa and Schizophyllum commune. This work displayed the whole picture ofmetabolic networks of mycelium growth and fruit body development and identifiedproteins mapped in regulation nodes of metabolic networks. Twenty-eight differentexpression proteins, which are involved in Glycosylphosphatidylinositol(GPI)-anchorbiosynthesis, N-glycan biosynthesis, biosynthesis of secondary metabolites, terpenoidbackbone biosynthesis, cell cycle-yeast, meiosis-yeast, ubiquitin mediated proteolysis,proteasome, fructose and mannose metabolism, tryptophan metabolism andsphingolipid metabolism, respectively might play some important roles inphysiological regulation of mycelium growth and fruit body formation, developmentand mature of V. volvacea.Furthermore, the results of this work might be helpful to insight into the molecularmechanism of biological characteristics of mycelia and morphogenesis of fruit body atdifferent development stages of V. volvacea, even other macro-basidiomycetes, in thefuture.