三疣梭子蟹低密度SNP芯片的设计及在家系鉴定中的应用评估

Development of a low-density SNP chip for pedigree identification in Portunus trituberculatus

  • 摘要:
    目的 本研究旨在设计一款三疣梭子蟹低密度SNP芯片并应用其建立高精度、低成本的家系鉴定技术。
    方法 基于298份重测序数据,根据测序深度、MAF和缺失率等指标,从参考基因组中筛选高质量SNP位点,进一步通过对来自8个群体((家系)的79个样本群体分析以检验其在家系鉴定中的潜力。根据SNP位点信息设计cGPS芯片,利用芯片对家系个体进行分型并结合KING-robust算法分析亲缘关系。
    结果 筛选到977个分布于53条染色体上的高质量SNP位点,平均间距935 512.44 bp,位点平均测序深度达13.57×,次等位基因频率(MAF)平均值为0.21,平均缺失率为0.11。群体分析结果显示,筛选的SNP位点能够清晰区分野生群体、人工群体及全同胞家系;根据SNP位点信息设计了低密度cGPS芯片,评估显示其位点检出率为99.48%,重复样本一致率为99.55%;利用芯片对8个家系的45个个体进行亲缘关系分析,结果显示子代检出率为91%,家系判别准确率100%。
    结论 本研究设计了一款低密度SNP芯片,经评估其分型性能准确、稳定。该芯片具有较高的家系辨别能力,能够满足育种研究中的家系鉴定及亲本溯源。本研究设计的低密度SNP芯片及家系鉴定方法为三疣梭子蟹育种提供了重要工具及技术支撑,有助于促进优良品种的选育。

     

    Abstract: Portunus trituberculatus is an important marine aquaculture crab species in China, widely distributed in the Bohai Sea, Yellow Sea, East China Sea, and South China Sea. Although the aquaculture production and area have been increasing year by year, the coverage of improved breeds remains low, and there is an urgent need to develop new varieties that are adapted to different farming regions. In aquaculture breeding, family information is crucial for accurately estimating genetic parameters, breeding values, and genomic selection. However, due to the large number of individuals and limited marker techniques, family management and information collection face significant challenges. SNP chips, as a high-throughput and low-cost genotyping platform, have been successfully applied in family identification in other species; however, no such application has been reported in P. trituberculatus. This study aimed to design a low-density SNP chip for P. trituberculatus and establish a high-accuracy, cost-effective method for pedigree identification. Based on 298 whole-genome resequencing datasets, high-quality SNP loci were screened from the reference genome with filtering criteria on sequencing depth, minor allele frequency (MAF), and missing rate. Their potential utility for parentage analysis was further evaluated through population genetic approaches. A cGPS chip was then designed using the selected SNP loci, and it was applied for family samples genotyping and kinship analysis combined with the KING-robust algorithm. In total, 977 high-quality SNPs distributed across 53 chromosomes were selected, with an average inter-marker distance of 935 512.44 bp. The mean sequencing of these loci was 13.57×, and the average MAF was 0.21, as well as the average missing rate was 0.11. Population structure analysis demonstrated that these selected SNPs clearly distinguished wild populations, cultured populations, and full-sib families. The designed low-density cGPS chip achieved a locus call rate of 99.48% and a concordance rate of 99.55% across duplicate samples. Genotyping of 45 individuals from eight families revealed an offspring detection rate of 91% and a family assignment accuracy of 100%. Overall, this study successfully developed low-density SNP chip with accurate and stable genotyping performance. It could meets the requirements for pedigree identification and parentage tracing in breeding programs. This chip, along with the associated pedigree analysis framework, provide an important molecular tool and technical foundation for the selective breeding of P. trituberculatus, thereby facilitating the development of enhanced genetic strains.

     

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