许尤厚,李鑫,彭金霞,李蔚,张兴志,官俊良,何庆松,陈泳先,张立,张彬,韦嫔媛,何苹萍

• 1:北部湾大学海洋学院
• 2:广西水产科学研究院广西水产遗传育种与健康养殖重点实验室

摘要(Abstract)：

【目的】鉴定筛选出与凡纳滨对虾（itopenaeus vannmei）性腺发育相关的miRNA及靶基因，为揭示其性腺分化的分子调控机制提供基础。【方法】通过高通量测序平台对凡纳滨对虾性腺进行miRNA测序分析。利用DESeq 2筛选出差异表达miRNA，并使用MiRanda对其进行靶基因预测，对靶基因进行GO功能注释和KEGG富集分析；运用实时荧光定量PCR(RT-qPCR)验证4个显著差异表达miRNAs及6个候选靶基因的表达模式。【结果】分别从精巢和卵巢小RNA文库获得18～30 nt的高质量序列27 665 604和28 431 611条，其中精巢中鉴定出117个已知的成熟miRNAs和214个未知miRNAs；卵巢中鉴定出106个已知的成熟miRNAs和159个未知miRNAs。分析获得153个精巢和卵巢显著差异表达的miRNAs，共预测得到57 412个差异表达miRNAs的靶基因。KEGG分析表明，这些靶基因显著富集的前20条KEGG通路包括白细胞介素-17信号通路、磷脂酶D信号通路、ECM-受体相互作用和Hedgehog信号通路等。RT-qPCR结果显示，miR-92b-3p＿3、miR-263a-5p＿1、novel＿mir23及novel＿mir67等miRNAs及其靶基因的表达模式与高通量测序结果基本一致，表明测序数据准确可靠。FoxL2、Dsx、NR1D1以及Octβ2R为精巢上调基因，在精巢中高表达；Vitellogenin为卵巢上调基因，在卵巢中特异性表达；而Huntingtin在精巢和卵巢中均高表达。【结论】mi R-92b-3p＿3和miR-263a-5p＿1等4个差异表达miRNAs以及FoxL2、Dsx、NR1D1和Vitellogenin等6个靶基因与凡纳滨对虾性腺发育密切相关，共同调节性腺的成熟过程。

关键词(KeyWords)： 凡纳滨对虾;性腺;miRNA;靶基因;RT-qPCR

Abstract:

Keywords:

基金项目(Foundation): 广西自然科学基金青年基金项目(2019GXNSFBA185022）;; 国家现代农业产业技术体系建设专项资助资金（CARS-48）

作者(Author): 许尤厚,李鑫,彭金霞,李蔚,张兴志,官俊良,何庆松,陈泳先,张立,张彬,韦嫔媛,何苹萍

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