晒田环境胁迫下克氏原螯虾卵巢的转录组学分析

Transcriptomic analysis of ovary of Procambarus clarkii under sun-drying environmental stress

  • 摘要: 【目的】运用转录组测序技术探究不同晒田环境胁迫下克氏原螯虾卵巢的基因表达变化,筛选出与其卵巢发育相关的候选基因及信号通路,为研究晒田胁迫方式促进克氏原螯虾卵巢发育的作用机制提供理论依据。【方法】以雌性克氏原螯虾为研究对象,分别在晒田环境胁迫0 d (CK)、3 d (T1)、7 d (T2)和10 d (T3)时采样,构建卵巢cDNA文库进行转录组测序,对差异表达基因(DEGs)进行GO功能注释及KEGG信号通路富集分析。【结果】共获得86377个Unigenes,在Nr、KOG/COG、Swiss-Prot和KEGG库中成功注释到22712个Unigenes;获得1115个胁迫组(T1、T2和T3)与CK的DEGs。KEGG信号通路富集分析结果表明,DEGs富集的PI3K-Akt信号通路、GnRH信号通路和雌激素信号通路与卵巢发育相关。实时荧光定量PCR验证结果表明,3个DEGs的表达模式与转录组分析结果基本一致,证明转录组数据准确可靠。筛选得到B2M、TUBA1B、MT-CYB、MT-ND1、MT-ND2、MT-ND4、DSX、Pck2、PIM3serine protease nudel-like等10个与卵巢发育相关的DEGs,并对其表达量进行分析,结果表明各基因在晒田胁迫期间均可调控卵巢发育。【结论】 B2M、TUBA1B、MT-CYB、MT-ND1、MT-ND2、MT-ND4、DSX、Pck2、PIM3serine protease nudel-like等10个DEGs及PI3K-Akt信号通路、GnRH信号通路、雌激素信号通路等3个富集通路在晒田胁迫期间参与卵巢发育调控。

     

    Abstract: 【Objective】The purpose of the study was to explore the gene expression changesin ovaries of Procambarus clarkii under different sun-drying stresses by transcriptomic sequencing technology, and to screen outthe candidate genes and signaling pathways related to ovarian development,so as toprovide a theoretical basis for studying the mechanism of sun-drying stress in promoting the ovarian development of P. clarkii.【Method】The female P. clarkii were sampled at 0 d(CK),3 d(T1),7 d(T2)and 10 d(T3)of sun-drying environmental stress,and ovary cDNA libraries were constructed for transcriptome sequencing,and the differentially expressed genes(DEGs)were subjected to GO functional annotation and KEGG signaling pathway enrichment analysis.【Result】A total of 86377 Unigenes were obtained. Of these, 22712 Unigenes were successfully annotated in Nr,KOG/COG, Swiss-Prot and KEGG databases. A total of 1115 DEGs were obtained between the stress group(T1,T2,T3)and the CK. The results of KEGG signaling pathway enrichment analysis showed that DEGs enriched on the pathways associated with ovarian development, including PI3K-Akt signaling pathway,GnRH signaling pathway and estrogen signaling pathway. The real-time fluorescence quantitative PCR verification results showed that the expression patterns of the three DEGs was basically consistent with the results of transcriptome analysis,proving that the transcriptome data were accurate and reliable. The ten DEGs related to ovarian development were screened out, including B2M, TUBA1B, MT-CYB, MT-ND1, MT-ND2, MT-ND4, DSX, Pck2, PIM3, and serine protease nudel-like. The expressions of these DEGs were analyzed, and the results showed that these genes regulated ovarian development during the period of sun-drying environmental stress.【Conclusion】 In this study,ten DEGs including B2M, TUBA1B,MT-CYB,MT-ND1,MT-ND2,MT-ND4, DSX,Pck2,PIM3 and serine protease nudel-like, and three enriched pathway sincluding PI3K-Akt signaling pathway, GnRH signaling pathway, and estrogen signaling pathway are involved in the regulation of ovarian development during the period of sun-drying environmental stress.

     

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