| With the continuous development of DNA sequencing technology,the research on metabolism and phylogeny based on transcriptome assembly analysis has shown its unique advantages.Transcriptome sequencing is an important means to obtain gene expression information of non-model organisms without reference genome.In this paper,transcriptome data of decapoda Palaemon gravieri,Ibacus novemdentatus and Cancer pagurus were sequenced and assembled,and functional annotation was performed.The results of related studies can provide basic data for functional gene research and phylogenetic research of the three species.The main achievements of the paper are as follows:After transcriptome sequencing of the gill tissue and muscle tissue of Palaemon gravieri,404,65,590 original sequences were obtained and 40,282,258 high-quality sequences(Clean Data)were obtained after removing impurities,with a data volume of 6.04 G.A large number of transcripts were obtained by assembling the obtained high-quality sequences,and 15,089 non-redundant genes with the N50 value of 1909 bp were obtained by further de-redundant assembly.By blastx comparison,gene functional annotation of Nr,Nt,Pfam,KOG/COG,swis-prot,KEGG and GO databases was performed for the non-redundant genes obtained by redundant assembly.8,593 Unigene genes were annotated,and the annotation rate was 56.95%.GO function was annotated into three categories: biological process,cell component and molecular function.Unigene in cell transformation,metabolic process and single biological process in biological process was annotated the most,with 3488,3083 and 2704 respectively.Among the cell components,most Unigene were annotated into cell parts,cells and organelles,which were 2116,2116 and 1491 respectively.The most annotated Unigene molecular functions are binding function and catalytic activity,which are respectively 3555 and 2562.Meanwhile,we found that the endocrine system,immune system,metabolic pathway and other pathways accounted for the most Unigene in the KEGG metabolic pathway analysis.In addition,we found 6008 microsatellite markers.Transcriptome sequencing was performed on the liver and muscle tissues of Ibacus novemdentatus,and 7.22 Gb of high-quality Clean Data was obtained,with a GC content of 44.81%.The obtained high-quality sequences were assembled,and the obtained transcripts were further disassembled to obtain 100,014 Unigenes with N50 of 980 bp.By blastx comparison of non-redundant genes,gene functional annotation was performed for Nr,Nt,Pfam,KOG/COG,swis-prot,KEGG and GO databases,and 24,561 Unigene genes were annotated,with an annotation rate of 24.56%.Among the three categories annotated by GO,the cell component obtained the most annotated genes,with 19,174 Unigene,accounting for 38.69% of the total annotated information.In the biological process,the number of Unigene notes to cell transformation,metabolism and single biological process was the most,which were 4914,4546 and 2971 respectively.The most annotated molecular functions were binding function and catalytic activity,and the number of Unigene annotated were 4710 and 4699,respectively.The analysis of KEGG pathway showed that most Unigene was annotated into ribosome,oxidative phosphorylation,endoplasmic reticulum protein processing,RNA transport,carbon metabolism and other pathways.At the same time,we found 10,114 microsatellite markers in the transcriptome.Transcriptome sequencing was performed on the liver,gonad and gill tissues of Cancer pagurus,and 6.77 Gb of high-quality sequence(Clean Data)was obtained,with a GC content of 40.47%.A large number of transcripts were obtained by assembling the obtained high-quality sequences,and 65,725 Unigenes(N50 is equal to 980 bp)were obtained after further de-redundant assembly.By blastx comparison of non-redundant genes,gene functional annotation was performed for Nr,Nt,Pfam,KOG/COG,swiss-prot,KEGG and GO databases,and 19,216 Unigene genes were annotated,with an annotation rate of 29.24%.GO was annotated into three categories of biological processes,cell components and molecular functions.Among them,cell transformation,metabolic process and single biological process of biological processes were annotated with a large number of Unigene,including 3295,2928 and 1947 respectively.Among the cell components,most Unigene were annotated into cell part,cell and membrane,respectively 2541,2529 and 2032.The most annotated Unigene molecular functions are binding function and catalytic activity,which are 3368 and 2562 respectively.In the KEGG pathway,the most Unigene was noted in ribosomeoxidative phosphorylation,endoplasmic reticulum protein processing,RNA transport,carbon metabolism and other pathways.In addition,7808 microsatellite markers were found in the transcriptome of the species.Finally,this study of the above three kinds of shrimp or crab and Seven species of prawns of system evolutionary relationships are studied,after the transcriptome assembly,ORF prediction,gene families to make use of OrthoMCL clustering,to obtain 160790 gene clustering,eventually selected 83 single copy orthologous genes,molecular phylogenetic tree was constructed,and the phylogenetic relationships among 10 species were discussed. |
