ABSTRACT: Here single nucleotide polymorphisms (SNPs) were associated with white spot syndrome virus (WSSV) resistance in black tiger shrimp Penaeus monodon. SNPs were identified by single-strand conformation polymorphism (SSCP) screening and DNA sequencing of shrimp sampled from 3 families (100 shrimp per family) challenged with WSSV. Shrimp that died over the 14 d challenge trial were designated susceptible, with those remaining alive on Day 14 designated resistant. To compare SNPs, 10 samples from the susceptible and resistant groups, each comprising DNA pooled from 3 shrimp, were amplified by polymerase chain reaction (PCR) using primers to 12 selected genes and screened by SSCP. SNPs were only identified in the anti-lipopolysaccharide factor 3 (ALFPm3) gene product. Analysis of complete ALFPm3 gene sequences confirmed the existence of 3 SNPs (g.934C>G, g.1186A>G, and g.1898C>G) that were polymorphic between the susceptible and resistant groups. Further analyses using specific tetra-primer amplification refractory mutation system PCR primer sets associated these 3 SNPS, and particularly the g.1186A>G SNP, with WSSV resistance. This SNP thus has potential for use as a DNA marker to select for WSSV resistance in P. monodon breeding programs.

中文翻译：

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黑虎虾斑节对虾ALFPm3单核苷酸多态性与白斑综合症病毒抗性的关系

摘要：在这里，单核苷酸多态性（SNPs）与黑虎虾对虾对虾白斑综合症病毒（WSSV）抗性有关。通过单链构象多态性（SSCP）筛选和对来自WSSV攻击的3个家庭（每个家庭100只虾）的虾样品进行DNA测序，鉴定出SNP。在14天挑战试验中死亡的虾被指定为易感虾，而在第14天仍存活的虾被指定为抗性虾。为了比较SNP，使用来自12个选定基因的引物，通过聚合酶链反应（PCR）扩增了来自易感和抗性组的10个样品，每个样品都包含从3个虾中收集的DNA，并通过SSCP进行了筛选。仅在抗脂多糖因子3（ALFPm3）基因产物中鉴定出SNP。对完整的ALFPm3基因序列的分析证实了存在3个SNP（g.934C> G，g.1186A> G和g.1898C> G），这些SNP在易感和抗性基团之间具有多态性。使用特异性四引物扩增难治性突变系统PCR引物的进一步分析将这3个SNPS，特别是g.1186A> G SNP与WSSV抗性相关。因此，该SNP有潜力用作DNA标记，以选择WSSV抗性。体育斑节对虾繁殖计划。