Transcriptome Sequencing And Comparative Transcriptome Analysis Of The Dikaryotic Mycelium And Monocaryon Mycelium Of Pleurotus Ostreatus

In the study, combined with the isozyme method and the traditional biology method(biology characteristic and the fruiting experiment) and RAPD method which was adopted to identify the genetic relationship of the monocaryon mycelium and dikaryotic mycelium of Pleurotus ostreatus. Then, the monocaryon mycelium and dikaryotic mycelium of Pleurotus ostreatus were analysed by Transcriptome sequencing, which revealed the biological function of genes. The differentially expressed genes was analysed and the fruiting mechanism of Pleurotus ostreatus was explored from the results of sequencing. Then, we confirmed the sequencing results by RT-PCR, The main results and conclusion were as follows:(1) Combined with three method to indentify the genetic relationship of the monocaryon mycelium and dikaryotic mycelium of Pleurotus ostreatus, we confirmed that the genetic relationship of dikaryotic mycelium13×3 and the monocaryon mycelium 3 were close to Tian Da, even the monocaryon mycelium 13.The results of fruiting indentified that there were no significant differences on growth rate and the product from zoogamy. It also provided some significance for preventing strains degradation and hybridization breeding.(2) From the analysis of transcriptome sequencing of Pleurotus ostreatus, We argued that factors such as length distribution, sequence errors, and read length impact on the prediction capacity of sequence census experiments.37145 All-Unigene were obtained and a third of the gene function was unknown. Unigene sequences were firstly annotated and calculated by blastx to protein databases like nr, Swiss-Prot, and COG, we got the functional classification of COG,GO and KEGG by software. From the functional classification we obtained the systematized biological function of gene and found the function of some new genes and alternative splicing. After using a greater statistically significant value FDR≤0.001 and |log2Ratio|≥1, we found most of differentially expressed genes participate in different pathway like cell signaling, starch and sucrose metabolism and biosynthesis of secondary metabolites. In the analysis of transcriptome, the dikaryotic mycelium 13x3 and the monocaryon mycelium 3 had less differentially expressed genes than the dikaryotic mycelium 13x3 and the monocaryon mycelium 13, also proved the genetic relationship between the three strains, which showed agreement with the result of genetic relationship. It can become a new method to indentify the genetic relationship.(3) We confirmed the sequencing result by detecting relative expression level of laccase genes using RT-PCR. The expression patterns of these genes showed agreement with the solexa data, which basically confirmed the reliability of our transcrpitome analysis. There are some differences between cultivation material mycelium and fermentation broth mycelium and it may be that the induction of some ion or cellulose etc that conduce the change of gene expression.