Abstract
The blood acts as a transfer channel for a variety of factors in the whole body. The ricefield eel (Monopterus albus) is a protogynous hermaphrodite vertebrate. Until now, no research has reported an analysis of the blood transcriptome during the process of sexual development in the ricefield eel. In this study, the transcriptome sequencing of blood samples from male and female ricefield eels was completed with a total of 34.70 Gb clean data. The clean data of each sample all reached 5.23 GB, and the percent of the Q30 basic group was 88.62% and above. A total of 106,369 unigenes were obtained after assembly, including 13,296 unigenes with a length of more than 1 kb. Further functional annotation analysis showed that there are 28,522 unigenes that can be annotated. The annotations of genes with differential expression revealed that there were 563 genes with significant differential expression in the blood of male and female ricefield eels, including 91 upregulated genes and 472 downregulated genes. Among which, 14 genes may be closely related to sex differentiation, the qPCR was used to confirmed the expression pattern of those genes and result shown that 11 genes were downregulated and 3 genes were upregulated, consistent with the results of our RNA-Seq analysis. This blood transcript dataset will open future research avenues on ricefield eel sex development and differentiation.
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Data availability
Raw sequence data is available on the NCBI Sequence Read Archive under the BioProject Accession number PRJNA388266.
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Funding
This study was funded by the Central Public-Interest Scientific Institution Basal Research Fund (2015JBFM06).
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Contributions
WD performed most of the experimental studies and wrote this manuscript.
LC isolated DNA and mRNA and contributed to the PCR analysis.
ZC performed the sequencing and analyses.
XB designed part of the experiments and reviewed the paper.
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Ricefield eel (Monopterus albus) is not an endangered or protected species, and all experiments were approved by the Institutional Animal Care and Use Committee of the Ministry of Freshwater Fisheries Research Center of the Chinese Academy of Fishery Sciences and were undertaken in accordance with the national legislation for fish welfare established by the Ministry of Science and Technology of the People’s Republic of China.
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Significance statement
A total of 106,369 unigenes were obtained after transcriptome sequencing. The annotations of genes with differential expression revealed that there were 563 genes with significant differential expression in the blood of male and female ricefield eels, including 91 upregulated genes and 472 downregulated genes. Among which, 14 genes may be closely related to sex differentiation, but their specific functions need to be further studied.
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Fig. S1
The gonadal histology of ricefield eel. T01/T02/T03, Spermatophore could be observed in testicle. T04/T05/T06, Oocytes could be observed in ovary. SP:spermatophore; O:ooctytes. (JPG 3013 kb)
Fig. S2
The length distribution of all unigene (JPG 245 kb)
Fig. S3
Phylogenetic tree of nucleotide sequences of potential sex-related gene in ricefield fish along with several other vertebrate species. The tree was estimated by the maximum likelihood method. Each nucleotide sequence at the terminal is presented by the name of species and gene from which the nucleotide sequence originated. The statistical robustness of the tree was estimated by bootstrapping with 1000 replicates. Bar represents the p distance. Words with a yellow background indicate the queried genes related to sex development. a,piwi; b,DNAJB1; c,Uhrf1; d,plcx2; e,FSTL5; f,zmziz1; g,sox11a; h,eomes; i,NASP; j,oocyte zinc finger protein; k,hsp; l,sushi; m,vhsv; n,pfam (PDF 6595 kb)
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Ding, W., Cao, L., Cao, Z. et al. Transcriptome analysis of blood for the discovery of sex-related genes in ricefield eel Monopterus albus. Fish Physiol Biochem 46, 1507–1518 (2020). https://doi.org/10.1007/s10695-020-00809-5
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DOI: https://doi.org/10.1007/s10695-020-00809-5