凡纳滨对虾α-淀粉酶基因PCR-RFLP多态性与生长性状的相关分析
PCR-RFLP polymorphism of alpha-amylase gene and its association with growth traits of Litopenaeus vannamei
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摘要: 采用PCR-RFLP法对379尾凡纳滨对虾α-淀粉酶基因(AMY)多态性进行检测,并分析AMY基因多态性与生长性状的相关性。结果表明:引物AMY-5与AMY-6扩增片段均具多态性,引物AMY-5扩增产物有CC, CT和TT 3种基因型,AMY-6扩增产物有AA, AB和BB 3种基因型。AMY基因AMY-5引物位点TT基因型与CC基因型相比在外显子5的123 bp处发生C→T突变,AMY-6引物位点BB基因型与AA基因型相比在外显子6的81 bp处发生C→T突变,2处突变均未引起氨基酸变化,表现为沉默突变。最小二乘法分析结果表明,CC, CT基因型在体重、体长、头胸甲长、头胸甲宽、第一腹节长及尾节长6个生长指标上均显著高于TT基因型(P<0.05)。AA, BB基因型在体长、第一腹节长及尾节长3个指标上显著高于AB基因型(P<0.05),在体重、头胸甲长及头胸甲宽指标上AA基因型显著高于AB基因型(P<0.05)。结论:AMY基因多态性对凡纳滨对虾生长性状有显著影响,可以作为影响凡纳滨对虾生长性状的候选基因,为凡纳滨对虾选育提供分子遗传标记。Abstract: The polymorphism of the alpha-amylase (AMY) gene was analyzed by PCR-RFLP method in 379 Litopenaeus vannamei, and association analyses were carried out to evaluate the effects of genotypes of the candidate genes on growth traits in Litopenaeus vannamei. The results showed that both the products amplified by primer AMY-5 and AMY-6 displayed polymorphisms, and there were three genotypes (CC, TT and CT) for AMY-5 and three genotypes (AA, AB and BB) for AMY-6 respectively. The results indicated that there was one single nucleotide mutation (C→T) in the exon5 (123 bp) and one single nucleotide mutation (C→T) in the exon6 (81 bp) of the alpha-amylase gene. The two mutations did not cause any amino acid changes and were silence mutations. Least square was applied to analyze the correlation of different genotypes and the growth traits. The CC and CT genotypes showed significant differences in body mass, body length, carapace length, carapace width, first abdominal segment length and the last abdominal segment length compared with the genotype TT(P<0.05). The AA and BB genotypes showed significant differences in body length, first abdominal segment length and the last abdominal segment length compared with the genotype AB (P<0.05). The AA genotype showed significant differences in body mass, carapace length, carapace width compared with the genotype AB (P<0.05). It implied that the AMY gene could have a major effect on the growth traits of Litopenaeus vannamei, and could be used as the gene that significantly affect the growth traits in Litopenaeus vannamei, and these molecular genetic markers may be applied for Litopenaeus vannamei breeding.
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Key words:
- Litopenaeus vannamei /
- alpha-amylase gene /
- PCR-RFLP /
- growth traits
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