Analysis of the genetic characteristics and variations in disease-resistant grass carp based on whole-genome resequencing and transcriptome sequencing

Huifang Tan , Yude Wang , Biao Hu , Yongjie Zhang , An'min Liao , Wuxia Liu , Chen Gen , Kaikun Luo , Min Tao , Chun Zhang , Qinbo Qin , Shaojun Liu
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Abstract

Disease-resistant grass carp (DR-GGC) was obtained by backcrossing gynogenetic grass carp (GGC, 2n = 48, female) with common grass carp (Ctenopharyngodon idellus [GC], 2n = 48, male). The genetic constitution and the potential mutation associated with growth dominance in DR-GGC were analyzed by genomic in situ hybridization (GISH), whole-genome resequencing (WGRS) and RNA-sequencing (RNA-seq). The GISH results showed partial translocations and deletions in the chromosomes of DR-GGC compared with GC. The WGRS results demonstrated that there were numerous genetic variations between DR-GGC and its parents, among which DR-GGC had the highest genomic heterozygosity and GGC had the highest genomic purity. The RNA-seq results revealed that the differentially expressed genes (DEGs) were significantly enriched in cellular processes and immune pathways in DR-GGC and GGC compared with GC. Meanwhile, the combined analysis of WGRS and RNA-seq identified DEGs with overlapping mutation loci in DR-GGC and GGC compared with GC. These DEGs were significantly enriched in the pathways of growth, metabolism, and immunity, including cellular metabolism, biosynthesis, glycolysis/gluconeogenesis, and extra cellular matrix (ECM) receptor interactions. These results provide a genetic basis for exploring the better growth characteristics and disease resistance of DR-GGC than GC, valuable information on the genetic breeding for grass carp, and a further understanding of the genetic characteristics of gynogenetic individuals.

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基于全基因组重测序和转录组测序的抗病草鱼遗传特征和变异分析
抗病草鱼(DR-GGC)是通过雌性草鱼(GGC,2n = 48,雌性)与普通草鱼(Ctenopharyngodon idellus [GC],2n = 48,雄性)回交获得的。通过基因组原位杂交(GISH)、全基因组重测序(WGRS)和 RNA 测序(RNA-seq)分析了 DR-GGC 的遗传结构和与生长优势相关的潜在突变。GISH 结果显示,与 GC 相比,DR-GGC 染色体中存在部分易位和缺失。WGRS结果表明,DR-GGC与其亲本之间存在大量遗传变异,其中DR-GGC的基因组杂合度最高,GGC的基因组纯度最高。RNA-seq结果显示,与GC相比,DR-GGC和GGC的差异表达基因(DEGs)在细胞过程和免疫通路中显著富集。同时,结合WGRS和RNA-seq分析发现,与GC相比,DR-GGC和GGC中的DEGs有重叠突变位点。这些DEGs在生长、代谢和免疫途径中明显富集,包括细胞代谢、生物合成、糖酵解/糖元生成和细胞外基质(ECM)受体相互作用。这些结果为探索 DR-GGC 比 GC 更好的生长特性和抗病性提供了遗传基础,为草鱼遗传育种提供了宝贵信息,并进一步了解了雌核发育个体的遗传特征。
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