石斑鱼PPAR-δ基因SNP位点和单倍型与抗虹彩病毒和神经坏死病毒抗性的关联

    Association between PPAR-δ gene SNPs, haplotypes and resistances to SGIV and RGNNV in the orange-spotted grouper, Epinephelus coioides

    • 摘要:
      目的  获得石斑鱼Epinephelus spp.抗病分子标记,选育石斑鱼抗病品系以解决石斑鱼病害频发的问题。
      方法  基于免疫基因PPAR-δ的基因组DNA序列开展单核苷酸多态性(Single nucleotide polymorphism,SNP)位点筛选,并对这些位点分别进行石斑鱼虹彩病毒(Singapore grouper iridovirus, SGIV)和神经坏死病毒(Red-spotted grouper nervous necrosis virus, RGNNV)抗性的关联分析。
      结果  在SGIV感染的易感组和抗感组样品中共筛查到9个SNP位点,均位于内含子中;多态信息含量(Polymorphism information content, PIC)的范围为0.177~0.375,其中,SNP-S1(g.940T>A)属于低度多态(PIC<0.25),其余SNP位点属于中度多态(0.25≤PIC<0.50);关联分析结果显示,SNP-S7(g.4595T>A)基因型频率在SGIV易感组和抗感组中存在显著差异分布(P<0.05),SNP-S7的TT和AA这2种纯合子基因型与SGIV抗感性状相关,而AT杂合子基因型与SGIV易感性相关。在RGNNV感染的易感组和抗感组样品中共筛查到8个SNP位点,其中,SNP-N1位于外显子中,属于同义突变,其余SNP均位于内含子中;PIC的范围为0.106~0.317,其中,SNP-N1(g.324G>A)和SNP- N2(g.883A>G)属于低度多态(PIC<0.25),其余SNP属于中度多态(0.25≤PIC<0.50);关联分析表明,SNP-N5(g.2510C>T)基因型频率在RGNNV易感组和抗感组中存在显著差异分布(P<0.05),SNP-N5的CT基因型与RGNNV抗感性状相关, CC基因型与RGNNV易感性状相关。
      结论  本研究在PPAR-δ的基因组DNA序列中分别筛选到与SGIV和RGNNV抗性相关的SNP标记各1个,可以为石斑鱼抗病育种提供技术支持和理论依据。

       

      Abstract:
      Objective  To obtain the disease-resistant molecular markers in grouper (Epinephelus spp.), and serve for the selective breeding program of disease-resistant grouper strains, so as to solve the problem of frequent occurrence of grouper disease.
      Method  Single nucleotide polymorphisms (SNPs) were screened based on PPAR-δ genomic DNA sequence, and association analysis of resistance to Singapore grouper iridovirus (SGIV) and red-spotted grouper nervous necrosis virus (RGNNV) was performed on these SNPs.
      Result  A total of nine SNPs were detected in the susceptible and resistant groups against SGIV infection, all of which were located in the introns, with the polymorphism information contents (PICs) ranging from 0.177−0.375. Among the SNPs, SNP-S1(g.940T>A) showed low degree polymorphism (PIC<0.25), while the rest SNPs showed moderate degree polymorphism(0.25≤PIC<0.50). The association analysis showed that the genotype frequencies of SNP-S7 (g.4595T>A) were significantly different between SGIV susceptible and resistant groups (P<0.05), the TT and AA homozygous genotypes of SNP-S7 were correlated with SGIV resistance traits, while the AT heterozygous genotypes were correlated with SGIV susceptibility traits. In addition, a total of eight SNPs were detected in the susceptible and resistant groups agasinst RGNNV infection, among which SNP-N1 was located in the exon, with a synonymous mutation, and the rest SNPs were located in the introns, with the PICs ranging from 0.106−0.317. Among the SNPs, SNP-N1 (g.324G>A) and SNP-N2 (g.883A>G) showed low degree polymorphism (PIC<0.25), while the rest SNPs showed moderate degree polymorphism (0.25≤PIC<0.50). The association analysis showed that SNP-N5 (g.2510C>T) genotype frequencies were significantly different between RGNNV susceptible and resistant groups (P<0.05), the CT genotype of SNP-N5 was correlated with RGNNV resistance traits and the CC genotype was correlated with RGNNV susceptibility traits.
      Conclusion  In this study, we successfully screened one SNP marker related to SGIV resistance and one SNP marker related to RGNNV resistance from PPAR-δ genomic DNA sequence. This finding can offer a technical support and a theoretical basis for resistance breeding of grouper.

       

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