Transcriptomic Insights and the Development of Microsatellite Markers to Assess Genetic Diversity in the Broodstock Management of Litopenaeus stylirostris

Ya-Chi Yang, Pei-Yun Chu, Che-Chun Chen, Wen-Chin Yang, Te-Hua Hsu, Hong-Yi Gong, I-Chiu Liao, Chang‐Wen Huang
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Abstract

Simple Summary The Pacific blue shrimp (Litopenaeus stylirostris) is essential to the global seafood market, but intensive farming has increased disease and reduced genetic diversity. This study aimed to develop microsatellite markers for broodstock management using high-throughput transcriptome sequencing from various tissues. We identified 40 robust markers, revealing significant genetic diversity crucial for sustainable shrimp farming. These markers are valuable for breeding programs and help in the conservation and improvement of shrimp populations. Abstract The Pacific blue shrimp (Litopenaeus stylirostris) is a premium product in the international seafood market. However, intensified farming has increased disease incidence and reduced genetic diversity. In this study, we developed a transcriptome database for L. stylirostris and mined microsatellite markers to analyze their genetic diversity. Using the Illumina HiSeq 4000 platform, we identified 53,263 unigenes from muscle, hepatopancreas, the intestine, and lymphoid tissues. Microsatellite analysis identified 36,415 markers from 18,657 unigenes, predominantly dinucleotide repeats. Functional annotation highlighted key disease resistance pathways and enriched categories. The screening and PCR testing of 42 transcriptome-based and 58 literature-based markers identified 40 with successful amplification. The genotyping of 200 broodstock samples revealed that Na, Ho, He, PIC, and FIS values were 3, 0.54 ± 0.05, 0.43 ± 0.09, 0.41 ± 0.22, and 0.17 ± 0.27, respectively, indicating moderate genetic variability and significant inbreeding. Four universal microsatellite markers (CL1472.Contig13, CL517.Contig2, Unigene5692, and Unigene7147) were identified for precise diversity analysis in Pacific blue, Pacific white (Litopenaeus vannamei), and black tiger shrimps (Penaeus monodon). The transcriptome database supports the development of markers and functional gene analysis for selective breeding programs. Our findings underscore the need for an appropriate genetic management system to mitigate inbreeding depression, reduce disease susceptibility, and preserve genetic diversity in farmed shrimp populations.
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转录组学洞察力和微卫星标记的开发,用于评估石斑裸鲤鱼苗种管理中的遗传多样性
简单摘要 太平洋青虾(Litopenaeus stylirostris)是全球海鲜市场的必需品,但集约化养殖增加了疾病,降低了遗传多样性。本研究旨在利用各种组织的高通量转录组测序技术开发用于种群管理的微卫星标记。我们确定了 40 个稳健的标记,揭示了对可持续对虾养殖至关重要的重要遗传多样性。这些标记对育种计划很有价值,有助于对虾种群的保护和改良。摘要 太平洋青虾(Litopenaeus stylirostris)是国际海产品市场上的优质产品。然而,集约化养殖增加了疾病发生率,降低了遗传多样性。在这项研究中,我们开发了一个L. stylirostris的转录组数据库,并挖掘微卫星标记来分析其遗传多样性。利用 Illumina HiSeq 4000 平台,我们从肌肉、肝胰腺、肠道和淋巴组织中鉴定出了 53,263 个单基因。微卫星分析从 18,657 个单位基因中鉴定出 36,415 个标记,主要是二核苷酸重复序列。功能注释突出了关键的抗病途径和富集类别。对 42 个基于转录组的标记和 58 个基于文献的标记进行了筛选和 PCR 测试,确定了 40 个扩增成功的标记。对 200 份鱼种样本进行基因分型后发现,Na、Ho、He、PIC 和 FIS 值分别为 3、0.54 ± 0.05、0.43 ± 0.09、0.41 ± 0.22 和 0.17 ± 0.27,表明遗传变异性适中,近亲繁殖显著。确定了四个通用微卫星标记(CL1472.Contig13、CL517.Contig2、Unigene5692 和 Unigene7147),用于精确分析太平洋青虾、太平洋白对虾和黑虎虾的多样性。转录组数据库可为选择性育种计划的标记开发和功能基因分析提供支持。我们的研究结果表明,有必要建立适当的遗传管理系统,以减轻近亲繁殖抑制,降低疾病易感性,保护养殖对虾种群的遗传多样性。
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