基于转录组测序的卤虫卵生和卵胎生相关基因表达研究

TRANSCRIPTOME-BASED ANALYSIS OF OVIPAROUS AND OVOVIVIPAROUS-RELATED GENE EXPRESSION IN ARTEMIA

  • 摘要: 为揭示卤虫(Artemia)不同繁殖模式的发生机制, 文章通过构建孤雌生殖卤虫卵生和卵胎生差异转录组文库并结合生物信息学分析, 对两种繁殖模式间的差异表达基因进行查找筛选, 然后利用qRT-PCR对候选繁殖模式相关基因的表达进行分析。转录组测序显示有1452个差异表达基因, 包括601个上调基因和851个下调基因。根据差异表达基因GO功能分类结果可知, 注释到生物过程、细胞组成和分子功能的unigene分别有1243、306和530个。KEGG富集分析结果显示差异基因显著富集在抗原加工和核糖体通路中。结合转录组分析, 进一步筛选得到6个生殖相关基因, 并针对不同繁殖模式下的卤虫进行qRT-PCR, 结果表明, 6个生殖相关基因在卵生卤虫卵巢中的表达量均显著高于卵胎生卤虫。此外, 对6个候选生殖相关基因编码的蛋白质保守结构域进行预测, 发现均与之前报道的相应基因保守结构域一致。综上所述, 研究所选择的6个基因可能影响参与了卤虫的生殖过程。研究结果为孤雌卤虫繁殖模式分子机制调控的研究提供了基础信息, 有助于完善卤虫的生殖生物学理论。

     

    Abstract: Artemia is not only one of the most important live feed for larvi culture, but also an ideal experimental organism for scientific research. Female Artemia produce either nauplii via ovoviviparous pathway or diapause cyst via oviparous pathway. In order to reveal the mechanism of different reproductive modes of Artemia, the reproductive differential transcriptomes of parthenogenetic Artemia were constructed, bioinformatics analysis were performed to screen reproductive differential expression genes, and the gene expression patterns were studied by using qRT-PCR. Transcriptome analysis showed that there were 1452 differentially expressed genes, of which 601 genes were up-regulated and 851 down-regulated in the abscising carpopodium. According to GO function classification, 1243306 and 530 unigene were annotated into biological process, cell composition and molecular function respectively. KEGG enrichment analysis showed that differential genes were significantly enriched in antigen processing and ribosome pathways. Combined with transcriptome data and qRT-PCR analysis, six reproductive-related genes were screened and verified. The results showed that all the six reproductive-related genes had higher expression in oviparous Artemia than in ovoviviparous Artemia. In addition, the conserved domains of the proteins encoded by six candidate reproductive related genes were predicted and phylogenetic trees were constructed respectively. The results showed that the protein domains were consistent with the previously reported reproductive genes. Over all, our study indicated that the selected six genes may influence the reproductive process of Artemia. This study provides valuable information for dissecting the molecular mechanism of reproductive pattern in the parthenogenetic Artemia, and may also help to refine the reproductive biological theory of Artemia.

     

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