银鲫不同克隆系疱疹病毒抗性差异及全基因组关联分析

RESISTANCE DIFFERENCES TO CARASSIUS AURSTUS HERPESVIRUS AND THE GENOME-WIDE ASSOCIATION STUDY AMONG DIFFERENT CLONES OF GIBEL CARP

  • 摘要: 为了探究银鲫不同克隆系在鲫疱疹病毒抗性上的差异及鉴定潜在抗病相关基因, 研究首先构建了7个雌核生殖克隆系, 并对其母本进行了微卫星标记鉴定。这7个克隆系包括方正银鲫克隆系1 (FZ1)、方正银鲫克隆系2 (FZ2)、方正银鲫克隆系3 (FZ3)、鄱阳银鲫克隆系(PY)和彭泽鲫克隆系(PZ), 以及作为对照的高抗品系H系和易感品种“中科3号”A+系。随后, 对这些银鲫克隆系进行鲫疱疹病毒人工感染试验, 存活率由高到低依次为H (100%) > FZ2 (95%) > PY (40%) > FZ1 (20%) = FZ3 (20%) = PZ (20%) > A+ (10%)。人工感染后48h, 高抗克隆系FZ2和H的肝脏细胞排列更紧密, 且鲫疱疹病毒基因表达水平更低, 暗示其肝脏病毒载量低; 而易感克隆系FZ1、FZ3、PZ和A+的肝脏细胞更肿大, 且鲫疱疹病毒基因表达水平更高, 暗示肝脏病毒载量高。此外, 研究对每个克隆系的10尾子代进行了基因组重测序, 鉴定到35,278,852个SNP (Single Nucleotide Polymorphisms)位点。基于全基因组关联分析, 研究筛选到63个与抗病性状相关的基因组区域, 这些区域总共含有213个注释基因。通过与转录组数据联合分析及荧光定量PCR验证, 最终筛选到11个潜在抗病相关基因。研究不仅鉴定出具有较高鲫疱疹病毒抗性的优良种质, 还筛选出银鲫潜在抗病相关基因, 为银鲫抗病育种工作奠定了基础。

     

    Abstract: To investigate the resistance differences against Carassius auratus herpesvirus (CaHV) among different gynogenetic clones of gibel carp (Carassius gibelio) and identify potential antiviral resistance-related genes, seven gynogenetic clones and performed microsatellite identification on their maternal individuals. These seven clones were established, including three Fangzheng clones (FZ1, FZ2, and FZ3), one Poyang clone (PY), one Pengze clone (PZ), along with two control clones: the highly resistant clone H and the susceptible variety allogynogenetic gibel carp “CAS III” (clone A+). Subsequently, artificial infection experiments with CaHV were conducted on these clones. The survival rates, ranked from highest to lowest, were: H (100%)>FZ2 (95%)>PY (40%)>FZ1 (20%)=FZ3 (20%)=PZ (20%)>A+ (10%). At 48h post-infection, histological observations revealed that the liver cells of the highly resistant clones FZ2 and H exhibited tighter cellular arrangements and significantly lower expression levels of CaHV gene orf72, indicating low viral load. In contrast, susceptible clones FZ1, FZ3, PZ, and A+ displayed swollen liver cells and elevated orf72 expression, suggesting high viral load. Additionally, whole-genome resequencing of 10 offspring from each clone identified 35278852single nucleotide polymorphism loci. Genome-wide association study mapped 63 genomic regions associated with disease resistance traits, encompassing 213 annotated genes. Integrative analysis with transcriptomic data and qPCR validation identified 11 potential antiviral resistance-related genes. This study not only identifies superior germplasm with high resistance to CaHV but also screens potential genes for disease resistance, laying a foundation for antiviral disease-resistant breeding in gibel carp.

     

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