發(fā)布時間：2019-01-19 16:54

【摘要】：為獲得與尼羅羅非魚(Oreochromis niloticus)鏈球菌病抗性相關(guān)的單核苷酸多態(tài)性(single nucleotide polymophisms,SNPs)標(biāo)記,本研究克隆了尼羅羅非魚補(bǔ)體C9基因(On C9)的基因組DNA序列,通過直接測序法從20個家系的39條尼羅羅非魚On C9的測序序列中分析得到22個SNPs位點,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個SNPs位點在內(nèi)含子中,S2(C644T)、S8(C2660T)和S9(A2751T)等3個SNPs位點位于外顯子中,其中S2、S9屬于非同義突變,S8為同義突變。此外,本研究獲得的On C9 SNPs位點中有一半為堿基轉(zhuǎn)換產(chǎn)生,另一半為堿基顛換產(chǎn)生。通過snapshot分型法對易感群體(susceptible group,SG)(84尾)和抗病群體(resistance group,RG)(81尾)進(jìn)行了分型,并利用Popgen32軟件統(tǒng)計分析了尼羅羅非魚On C9各個SNPs位點在兩個群體的多態(tài)性和遺傳參數(shù),結(jié)果顯示多態(tài)信息含量(polymorphism information content,PIC)值為0.09~0.37,表明所有SNPs位點均為中度或低度多態(tài)。采用SPSS 20軟件分析了22個SNPs位點在兩個群體中的基因型頻率、等位基因頻率,并采用卡方檢驗其顯著性,分析其與抗病性狀之間的相關(guān)性,發(fā)現(xiàn)S8和S21兩個位點與抗病性狀顯著相關(guān)。通過Haploview 4.2軟件分析了On C9中的22個SNPs位點所形成的單倍塊和連鎖不平衡狀態(tài)。結(jié)果表明,22個位點可構(gòu)成4個單倍塊和11個單倍型。其中GTTAG和ATGA兩個單倍型與尼羅羅非魚鏈球菌病易感性顯著相關(guān)(P0.05)。本研究顯示,篩選到的尼羅羅非魚On C9的兩個SNPs位點和兩個單倍型具有用于尼羅羅非魚抗鏈球菌(Streptococcus agalactiae)病性狀的分子輔助育種的潛力。
[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.
【作者單位】： 中國水產(chǎn)科學(xué)研究院珠江水產(chǎn)研究所/農(nóng)業(yè)部熱帶亞熱帶水產(chǎn)資源利用與養(yǎng)殖重點實驗室;上海海洋大學(xué)水產(chǎn)與生命學(xué)院;
【基金】：國家自然科學(xué)基金(No.31502205) 現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)體系專項資金(CARS-49) 廣東省自然科學(xué)基金(No.2014A030310337)
【分類號】：S917.4

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