尼罗罗非鱼补体C9基因单核苷酸多态性及其与抗无乳链球菌感染的关联分析

发布时间：2019-01-19 16:54

【摘要】：为获得与尼罗罗非鱼(Oreochromis niloticus)链球菌病抗性相关的单核苷酸多态性(single nucleotide polymophisms,SNPs)标记,本研究克隆了尼罗罗非鱼补体C9基因(On C9)的基因组DNA序列,通过直接测序法从20个家系的39条尼罗罗非鱼On C9的测序序列中分析得到22个SNPs位点,其中S1(C483T)、S3(G954A)、S4(T1180C)、S5(G1539A)、S6(G1627T)、S7(A2590T)、S10(T2876G)、S11(C2915G)、S12(T2939G)、S13(A2942C)、S14(A3544C)、S15(T3604C)、S16(T3670C)、S17(T3841C)、S18(T4339C)、S19(G4494C)、S20(A4678G)、S21(T5790A)、S22(T5816G)等19个SNPs位点在内含子中,S2(C644T)、S8(C2660T)和S9(A2751T)等3个SNPs位点位于外显子中,其中S2、S9属于非同义突变,S8为同义突变。此外,本研究获得的On C9 SNPs位点中有一半为碱基转换产生,另一半为碱基颠换产生。通过snapshot分型法对易感群体(susceptible group,SG)(84尾)和抗病群体(resistance group,RG)(81尾)进行了分型,并利用Popgen32软件统计分析了尼罗罗非鱼On C9各个SNPs位点在两个群体的多态性和遗传参数,结果显示多态信息含量(polymorphism information content,PIC)值为0.09~0.37,表明所有SNPs位点均为中度或低度多态。采用SPSS 20软件分析了22个SNPs位点在两个群体中的基因型频率、等位基因频率,并采用卡方检验其显著性,分析其与抗病性状之间的相关性,发现S8和S21两个位点与抗病性状显著相关。通过Haploview 4.2软件分析了On C9中的22个SNPs位点所形成的单倍块和连锁不平衡状态。结果表明,22个位点可构成4个单倍块和11个单倍型。其中GTTAG和ATGA两个单倍型与尼罗罗非鱼链球菌病易感性显著相关(P0.05)。本研究显示,筛选到的尼罗罗非鱼On C9的两个SNPs位点和两个单倍型具有用于尼罗罗非鱼抗链球菌(Streptococcus agalactiae)病性状的分子辅助育种的潜力。
[Abstract]:In order to obtain a single nucleotide polymorphism (single nucleotide polymophisms,SNPs) marker associated with (Oreochromis niloticus) streptococcus disease resistance in tilapia niloticus, the genomic DNA sequence of complement C9 gene (On C9) in Tilapia niloticus was cloned. By direct sequencing, 22 SNPs loci were obtained from 39 On C9 sequences from 20 families, including S1 (C483T), S3 (G954A), S4 (T1180C), S5 (G1539A), S6 (G1627T), S7 (A2590T), S10 (T2876G), S5 (G1539A), S6 (G1627T), S7 (A2590T), S10 (T2876G), S4 (T1180C), S5 (G1539A), S6 (G1627T), S7 (A2590T), S10 (T2876G). S11 (C2915G), S12 (T2939G), S13 (A2942C), S14 (A3544C), S15 (T3604C), S16 (T3670C), S17 (T3841C), S18 (T4339C), S19 (G4494C), S20 (A4678G), S21 (T5790A), S22 (T5816G) in the intron, S2 (C644T), Three SNPs loci, S8 (C2660T) and S9 (A2751T), were located in exons, among which S2 + S9 was nonsynonymous and S8 was synonymous. In addition, half of the On C9 SNPs sites were produced by base transversion and the other half by base transversion. The susceptible population (susceptible group,SG) (84) and disease-resistant population (resistance group,RG) (81) were classified by snapshot typing. The polymorphism and genetic parameters of On C9 SNPs loci in two populations were analyzed by Popgen32 software. The results showed that the polymorphic information content (polymorphism information content,PIC) value was 0.09 ~ 0.37. All SNPs loci were moderately or lowly polymorphic. The genotype frequency and allele frequency of 22 SNPs loci in two populations were analyzed by SPSS 20 software. S8 and S21 loci were found to be significantly correlated with disease resistance traits. Haplotypes and linkage disequilibrium of 22 SNPs loci in On C9 were analyzed by Haploview 4.2 software. The results showed that 22 loci could form 4 haplotypes and 11 haplotypes. GTTAG and ATGA haplotypes were significantly associated with susceptibility to Streptococcus niloticus (P0.05). The results showed that the two SNPs loci and two haplotypes of On C9 of tilapia niloticus had the potential to be used in molecular assisted breeding for resistance to streptococcus (Streptococcus agalactiae) disease in Tilapia niloticus.
【作者单位】：中国水产科学研究院珠江水产研究所/农业部热带亚热带水产资源利用与养殖重点实验室;上海海洋大学水产与生命学院;
【基金】：国家自然科学基金(No.31502205) 现代农业产业技术体系专项资金(CARS-49) 广东省自然科学基金(No.2014A030310337)
【分类号】：S917.4

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