Scientia Agricultura Sinica ›› 2017, Vol. 50 ›› Issue (23): 4644-4655.doi: 10.3864/j.issn.0578-1752.2017.23.017

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

The Comparative Transcriptome Analysis of Parabronema skrjabini at Different Developmental Stages

WANG WenLong1, FENG ChenChen1, HONG Mei1, YUE JianWei1, Huhebateer1, LIU ChunXia2   

  1. 1College of Veterinary Medicine, Inner Mongolia Agricultural University/Key Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease, Ministry of Agriculture, Hohhot 010018 ; 2College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018
  • Received:2016-06-20 Online:2017-12-01 Published:2017-12-01

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Abstract: 【Objective】 The objective of this study was to identify the differentially expressed genes(DEG) and describe biological characteristics involved in functional classifications and metabolic pathways at different developmental stages of Parabronema skrjabini infecting camel, which is necessary to better understand functional genes involved in growth and development and enrich the transcriptome data of parasitical nematodes.【Method】Eggs, the third-stage larvae(L3s) and females of Parabronema skrjabini were sequenced by Illumina HiSeq2000TM sequencing platform and constructed their cDNA libraries after quality filtering. De novo assembling and assembly efficiency assessment were carried out using Trinity, a short-read assembly program. Then all the effective sequential data obtained were assigned to the relevant databases to perform functional annotation and bioinformatic analysis.【Result】The results showed that 47 717, 76 342 and 54 624 unigenes were obtained respectively in eggs，the third-stage larvae and female stages. 33 579 differentially expressed genes(DEGs) were identified by comparing the unigenes obtained from eggs and L3s stages, of which 20 477 were up-regulated and 13 102 were down-regulated. There were 32 199 differentially expressed genes between L3s and female stages, of these genes, 9 293 were up-regulated genes and 22 906 were down-regulated genes. The differentially expressed genes of two pairwise comparisons were respectively enriched in Gene Ontology. 6 617, 3 891 and 8 755 differentially expressed genes comparing eggs and L3s stages were annotated into database of biological process, cellular component and molecular function respectively, while the number by comparing L3s and female stages were 7 043, 3 686 and 10 177 respectively. In KEGG pathways identification, 6 521 differentially expressed genes comparing eggs and L3s stages were assigned to 251 KEGG pathways, and clustered significantly in MARK, Wnt signaling pathways and oxidative phosphorylation. In comparison of L3s and female stages, 6 528 differentially expressed genes were enriched in metabolic pathways, DNA replication and cell cycle. Functional cluster analysis indicated that the regulation related differential genes of growth rate and the reproductive and genital development were highly expressed in eggs and females stages, then the genes of the defense and carbohydrate metabolism were enriched in exclusive L3s stage. Differential genes of embryonic and post-embryonic development were highly expressed in all three stages, and 196 function genes from embryonic development and 166 function genes from post-embryonic development were co-expressed in all three stages, implied that these genes played a crucial role in (post-)embryonic development. Moreover, we identified 9 heterochronic genes, such as LIN-28, LIN-14 and RHEB-1, 48 nuclear hormone receptors (NHRs) genes, including NHR-49, NHR-48, NHR-40 and NHR-1, and 36 zinc metalloproteinase(NAS) genes, including NAS-36, NAS-33 and NAS-14. Then the analysis of enrichment capacity in three stages showed these genes were necessary to regulate the different development stages of Parabronema skrjabini. 【Conclusion】The transcriptomic research of Parabronema skrjabini at three developmental stages using RNA-seq revealed biological characteristics of differentially expressed genes involved in development- related GO functional classification, KEGG pathway and functional cluster and identified many kinds of heterochronic genes and developmental genes, which provides a foundation and reference for further investigation of the whole genome sequence analysis, interaction between Parabronema skrjabini and host, pathogenic mechanism and immune evasion.

Key words: Parabronema skrjabini, transcriptome, differentially expressed genes, development-related genes

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