Based On Transcriptome Sequencing To Mine Of Autolysis-related Genes In Coprinus Comatus

Coprinus comatus is a kind of edible mushroom with high nutritional value,which has the characteristics of reducing blood sugar and blood fat,anti-tumor and anti-oxidation.However,autolysis always happened with age,which seriously affects the commercial value of C.comatus.In this study,transcriptome sequencing of different phases fruiting bodies of C.comatus based on high-throughput sequencing technology.The genes related to autolysis were excavated,in order to inhibite the occurrence of autolysis at the gene level and provide theoretical basis for the healthy development of the cultivation industry of the C.comatus.The main results of this study are as follows:1.Observation of the micromorphological characteristics of C.comatus.On the 27 th day,the fruit body of C.comatus has no longer continued to grow.The lamellae were whitish.The microscopic morphology showed that basidia stema appeared on the basidia,the basidiospore were colorless,and the cell walls of all the tissues were smooth and complete.This period was defined the mature fruiting phase(M).On the 30 th day,the color of the lamellae began to turn pink,and the micromorphology found that some basidiospores appeared black,while the cell walls of all tissues were still smooth and complete,which was defined as the discoloration phase(D).From this we speculate that the color change of the lamellae in the discoloration phase is the key period in the autolysis process.2.Transcriptome sequencing.Using the fruiting bodies of C.comatus mature fruiting phase and discoloration phase as the research material,total RNA was extracted and transcriptome sequenced.After sequence quality control filtering,29,281 transcripts and 12,941 uingenes were obtained,with a total length of 49,031,913 bp,the longest transcript length is 16,420 bp,the shortest transcript length is 201 bp,the average length of all assembled transcripts is 1,674.53 bp,the N50 value is 2,606 bp,the GC content is 44.1%.There were 10,178 unigene expressions in the M phase and 9,722 unigene expressions in the D phase,and the correlation coefficients of both time periods were greater than 0.85,indicating that the transcriptome sequencing data were good,and the mature fruiting phase and the discoloration phase had high repeatability,which could be used for subsequent analysis.3.Excavating of autolysis-related genes.Analysis of differentially expressed genes between M and D phases revealed a total of 4,516 differentially expressed genes,of which 2,201 were up-regulated and 2,315 were down-regulated.Functional annotation and enrichment analysis of these differentially expressed genes revealed that the ribosome pathway,DNA replication pathway,and cell cycle pathway may be related to autolysis.In addition,glucanase,chitinase,pigment synthesis-related enzymes,autophagy and programmed death-related genes,F-box and HMG domains,and transcription factor Rlm1 genes may be related to autolysis.4.Verification of partial autolysis related genes.From ribosome pathway,cell cycle-yeast,DNA replication pathway and autologous glucanase,chitinase,pigment synthesis related enzymes,autophagy and programmed death related genes,F-box domain,HMG domain and transcription factor Rlm1,20 genes were randomly selected for verification,and the data of 12 genes were consistent with the transcriptome data.