Transcriptome Analysis Of Purpureocillium Lilacinum E22 And Key Lethal Gene Families Analysis Of Bursaphelenchus Xylophilus

Pine wilt disease(PWD)is caused by Bursaphelenchus xylophilus,which is extremely harmful to Chinese pines,causing inestimable loss of forestry resources.With the gradual elimination of chemical agents and the inability of biological agents to completely cure pine wilt,the research of genetically engineered disease-resistant plants will become more and more popular.Purpureocillium lilacinum has significant effects on controlling plant nematodes,and it can control them by producing chitinase and serine protease as important virulence factors.In this test,P.lilacinum E22 was used to test the B.xylophilus,and P.lilacinum E22 was used for parameter-free transcriptome analysis.and the differential gene screening,GO enrichment analysis and pathway enrichment analysis in KEGG were used to excavate chitinase gene family and serine protease gene family that are toxic to B.xylophilus,and use bioinformatics to analyze all genes for subsequent transgenes research on disease-resistant pine plants paves the way.1.Through transcriptome sequencing,a total of 73.91 Gb of valid data was obtained,and a total of 167,018 transcript sequences were assembled.A total of 107,830 non-redundant single genes were assembled and compared to Nr,Nt,Pfam,KOG,and Swiss-prot,KEGG,GO on seven public databases.51,067 differential genes were obtained,of which 49,265 genes were up-regulated and 1,802 genes were down-regulated.2.By increasing the expression of differential genes by 6 times and screening through the Pfam database annotation,eight GH19 family chitinase class I genes were obtained,which are Cluster-7269.0,Cluster-11977.0,Cluster-34695.5883,and Cluster-16977.0,Cluster-19046,0,Cluster-31483.0,Cluster-12366.0,Cluster-34695.18565.Seven serine protease S10 family genes,they are Cluster-45122.0,Cluster-34695.26144,Cluster-39710.0,Cluster-42428.0,Cluster-21153.0,Cluster-38672.0,Cluster-7477.0.3.In the GO enrichment analysis,the eight GH19 family chitinase class I genes are involved in three processes,namely the macromolecular decomposition process,the cell decomposition process and the catabolic process,which are related to the decomposition of the B.xylophilus body wall.Related to cells.The seven serine protease S10 family genes are involved in three processes,which are protein metabolism,proteolysis and peptidase activity,which are related to the degradation of proteins in B.xylophilus.Due to the limited number of KEGG annotated genes,only the ribosome metabolic pathway and the endocytosis metabolic pathway are enriched.These two important metabolic pathways play an important role in responding to the stress of B.xylophilus.4.Bioinformatics analysis of eight GH19 family chitinase class I genes.Their open reading frames(ORF),amino acid lengths,and protein molecular weights were roughly the same.The isoelectric points were very close.They were all stable hydrophilic proteins.Cell localization shows extracellular or membrane-bound extracellular proteins;8 chitinases are generally 100 on the phylogenetic tree,and the evolutionary relationship is highly consistent,all with only one exon;Multiple amino acid sequence alignments revealed that multiple conserved binding sites constitute a conserved domain,and the conserved motifs are also highly similar;8 chitinases are conserved domains rich in ?-helix through secondary and tertiary structure prediction analysis This lays the foundation for destroying the macromolecules of the B.xylophilus body wall.5.Bioinformatics analysis of seven serine protease S10 family genes.The open reading frame(ORF),amino acid length,and protein molecular weight were basically the same.All were hydrophilic proteins,most of which were stable proteins,and the subcellular localization showed that most of them were Extracellular or membrane-bound extracellular proteins;phylogenetic tree shows that the total of 7 serine proteases is 99,all with only one exon;in the amino acid multiple sequence alignment,there are many conservative binding sites together to form a conserved domain,conserved The motifs are completely consistent,indicating that the evolution process is highly unified;the seven serine proteases are analyzed by the prediction of secondary and tertiary structures,and the function of cleaving the peptide bonds of B.xylophilus is mainly based on the conserved domain of a combination of ?-helix and random coils.