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对光棘球海胆卵巢成熟相关基因进行从头转录组测序及比较分析。

De novo transcriptome sequencing and comparative analysis to discover genes involved in ovarian maturity in Strongylocentrotus nudus.

作者信息

Jia Zhiying, Wang Qiai, Wu Kaikai, Wei Zhenlin, Zhou Zunchun, Liu Xiaolin

机构信息

College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.

College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China.

出版信息

Comp Biochem Physiol Part D Genomics Proteomics. 2017 Sep;23:27-38. doi: 10.1016/j.cbd.2017.05.002. Epub 2017 May 30.

Abstract

Strongylocentrotus nudus is an edible sea urchin, mainly harvested in China. Correlation studies indicated that S. nudus with larger diameter have a prolonged marketing time and better palatability owing to their precocious gonads and extended maturation process. However, the molecular mechanism underlying this phenomenon is still unknown. Here, transcriptome sequencing was applied to study the ovaries of adult S. nudus with different shell diameters to explore the possible mechanism. In this study, four independent cDNA libraries were constructed, including two from the big size urchins and two from the small ones using a HiSeq™2500 platform. A total of 88,581 unigenes were acquired with a mean length of 1354bp, of which 66,331 (74.88%) unigenes could be annotated using six major publicly available databases. Comparative analysis revealed that 353 unigenes were differentially expressed (with log2(ratio)≥1, FDR≤0.001) between the two groups. Of these, 20 differentially expressed genes (DEGs) were selected to confirm the accuracy of RNA-seq data by quantitative real-time RT-PCR. Furthermore, gene ontology and KEGG pathway enrichment analyses were performed to find the putative genes and pathways related to ovarian maturity. Eight unigenes were identified as significant DEGs involved in reproduction related pathways; these included Mos, Cdc20, Rec8, YP30, cytochrome P450 2U1, ovoperoxidase, proteoliaisin, and rendezvin. Our research fills the gap in the studies on the S. nudus ovaries using transcriptome analysis.

摘要

虾夷马粪海胆是一种可食用的海胆,主要在中国捕捞。相关性研究表明,直径较大的虾夷马粪海胆由于性腺早熟和成熟过程延长,其销售期延长且适口性更好。然而,这一现象背后的分子机制仍然未知。在此,应用转录组测序来研究不同壳径的成年虾夷马粪海胆的卵巢,以探索可能的机制。在本研究中,使用HiSeq™2500平台构建了四个独立的cDNA文库,其中两个来自大个体海胆,两个来自小个体海胆。共获得88,581个单基因,平均长度为1354bp,其中66,331个(74.88%)单基因可使用六个主要的公开可用数据库进行注释。比较分析显示,两组之间有353个单基因差异表达(log2(比值)≥1,FDR≤0.001)。其中,选择了20个差异表达基因(DEGs)通过定量实时RT-PCR来确认RNA-seq数据的准确性。此外,进行了基因本体和KEGG通路富集分析,以找到与卵巢成熟相关的假定基因和通路。八个单基因被鉴定为参与生殖相关通路的显著DEGs;这些包括Mos、Cdc20、Rec8、YP30、细胞色素P450 2U1、卵过氧化物酶、蛋白酶抑制剂和rendezvin。我们的研究填补了利用转录组分析对虾夷马粪海胆卵巢研究的空白。

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