翘嘴鲌生长激素基因侧翼2个微卫星位点与生长性状的关联分析
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摘要
通过120尾同塘养殖翘嘴鲌体长和体质量的测定,以及生长激素基因(GH)侧翼的2个微卫星位点Cal-GH01和Cal-GH02基因型检测,分析了翘嘴鲌GH基因侧翼的2个微卫星多态性与生长性能的相关性。结果表明:Cal-GH01共检测到3个等位基因(404、407和410 bp)和6种基因型,等位基因407 bp是优势等位基因,407/407 bp是优势基因型;该微卫星位点多态信息含量为0. 420,观测杂合度为0. 207 2,属中度多态性位点。Cal-GH02共检测到6个等位基因(366、375、378、381、384和387 bp)和11种基因型,等位基因381 bp是优势等位基因,381/381 bp是优势基因型;该微卫星座位多态信息含量为0. 450,观测杂合度为0. 466 1,属中度多态座位。微卫星位点Cal-GH01的分析结果表明,404/407型个体占样本数的13. 3%,其体长最长。407/410型个体占样本数的5%,其体质量最大。Cal-GH01不同基因型个体体长和体质量均有差异,但差异均不显著(P> 0. 05)。在微卫星位点Cal-GH02中,366/381型个体占样本数的3. 3%,其体长和体质量均最大,其体质量显著大于其他基因型个体(P <0. 05)。384/387型个体占样本数的2. 5%,其体长和体质量均最小,其体长显著小于其他基因型个体(P <0. 05)。翘嘴鲌GH基因多态性与生长性状存在关联,推测相关变异位点可作为翘嘴鲌生长的候选辅助标记。
The body length and weight of 120 topmouth culter (Culter alburnus Basilewsky) cultured in the same pond were measured. The polymorphism information and correlation with growth traits of two microsatellite loci,CalGH01 and Cal-GH02,located in the flanking regions of the growth hormone (GH) gene were analyzed. The results showed that three Cal-GH01 alleles (404,407 and 410 bp) and six genotypes could be detected. The 407 bp allele was the dominant allele,and 407/407 bp was the dominant genotype. The polymorphic information content of the microsatellite locus was 0. 420,and the heterozygosity was 0. 207 2. A total of six alleles (366,375,378,381,384 and 387 bp) and 11 genotypes were detected for Cal-GH02. The 381 bp allele was the dominant allele,and 381/381 bp was the dominant genotype. The polymorphic information content of the microsatellite locus was 0. 450,and the heterozygosity was 0. 466 1,which made it a moderately polymorphic locus. The associations between the polymorphisms of the two loci and growth traits were analyzed using general linear models. The analysis results for microsatellite locus Cal-GH01 showed that 404/407-type individuals accounted for 13. 3% of the sample size and had the longest body length. The 407/410-type individuals accounted for 5% of the sample weight and were the heaviest. There were some differences in body length and body weight among the Cal-GH01 genotypes; however,the differences were not significant (P > 0. 05). For microsatellite locus Cal-GH02,366/381-type individuals accounted for 3. 3% of the number of samples,and their body length and body weight were the highest. Their weight was significantly higher than that of the other genotypes (P < 0. 05). 384/387-type individuals accounted for 2. 5% of the sample size,and their body length and weight were the lowest. Their body length was significantly lower than that of other genotypes (P < 0. 05). In conclusion,we studied the genetic characteristics of two microsatellite loci in 120 fish sampled from the same pond. The correlation analysis for body length and body weight provided a scientific basis for marker-assisted selection for growth traits. Overall,there were associations between polymorphisms in the GH gene and the growth traits of C. alburnus. Therefore,the two microsatellite loci in GH gene could be used as markers for practical breeding programs for growth traits in C. alburnus.
The body length and weight of 120 topmouth culter (Culter alburnus Basilewsky) cultured in the same pond were measured. The polymorphism information and correlation with growth traits of two microsatellite loci,CalGH01 and Cal-GH02,located in the flanking regions of the growth hormone (GH) gene were analyzed. The results showed that three Cal-GH01 alleles (404,407 and 410 bp) and six genotypes could be detected. The 407 bp allele was the dominant allele,and 407/407 bp was the dominant genotype. The polymorphic information content of the microsatellite locus was 0. 420,and the heterozygosity was 0. 207 2. A total of six alleles (366,375,378,381,384 and 387 bp) and 11 genotypes were detected for Cal-GH02. The 381 bp allele was the dominant allele,and 381/381 bp was the dominant genotype. The polymorphic information content of the microsatellite locus was 0. 450,and the heterozygosity was 0. 466 1,which made it a moderately polymorphic locus. The associations between the polymorphisms of the two loci and growth traits were analyzed using general linear models. The analysis results for microsatellite locus Cal-GH01 showed that 404/407-type individuals accounted for 13. 3% of the sample size and had the longest body length. The 407/410-type individuals accounted for 5% of the sample weight and were the heaviest. There were some differences in body length and body weight among the Cal-GH01 genotypes; however,the differences were not significant (P > 0. 05). For microsatellite locus Cal-GH02,366/381-type individuals accounted for 3. 3% of the number of samples,and their body length and body weight were the highest. Their weight was significantly higher than that of the other genotypes (P < 0. 05). 384/387-type individuals accounted for 2. 5% of the sample size,and their body length and weight were the lowest. Their body length was significantly lower than that of other genotypes (P < 0. 05). In conclusion,we studied the genetic characteristics of two microsatellite loci in 120 fish sampled from the same pond. The correlation analysis for body length and body weight provided a scientific basis for marker-assisted selection for growth traits. Overall,there were associations between polymorphisms in the GH gene and the growth traits of C. alburnus. Therefore,the two microsatellite loci in GH gene could be used as markers for practical breeding programs for growth traits in C. alburnus.
引文
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[10]KANG J,LEE S,PARK S,et al.DNA polymorphism in the growth hormone gene and its association with weight in olive flounder Paralichthys olivaceus[J].Fisheries Science,2010,68(3):494-498.
[11]ALMULY R,CAVARI B,FERSTMAN H,et al.Genomic structure and sequence of the gilthead seabream(Sparus aurata)growth hormone-encoding gene:identification of minisatellite polymorphism in intron I[J].Génome,2000,43(5):836-845..
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[13]乐佩琦,罗云林.鲌亚科鱼类系统发育初探(鲤形目:鲤科)[J].水生生物学报,1996,20(2):182-185.YUE P Q,LUO Y L.Preliminary studies on phylogeny of subfamily Cultrinae(Cypriniformes:Cyprinidae)[J].Acta Hydrobiologica Sinica,1996,20(2):182-185.(in Chinese)
[14]LIU S L,GU Z M,JIA Y Y,et al.Isolation and characterization of 32 microsatellite loci for topmouth culter(Culter alburnus Basilewsky)[J].Genetics and Molecular Research,2014,13(3):7480-7483.
[15]LIU S L,GU Z M,ZHAO J L,et al.Characterization of microsatellites and repetitive flanking sequences(Re FS)from the topmouth culter(Culter alburnus Basilewsky)[J].Biochemical Systematics and Ecology,2015,63(5):127-135.
[16]刘士力,程顺,蒋文枰,等.翘嘴鲌胰岛素样生长因子-Ⅰ(IGF-I)基因的克隆及序列分析[EB/OL].(2018-12-24)[2019-01-04].http://ssswxb.ihb.ac.cn/article/id/fb324b22-122f-48a6-8c63-65fcbd8ccb3c.LIU S L,CHENG S,JIANG W P,et al.Cloning and sequence analysis of insulin-like growth factor I gene in topmouth culter(Culter alburnus Basilewsky)[EB/OL].(2018-12-24)[2019-01-4].http://ssswxb.ihb.ac.cn/article/id/fb324b22-122f-48a6-8c63-65fcbd8ccb3c.
[17]刘士力,贾永义,蒋文枰,等.翘嘴鲌生长激素(GH)基因与侧翼区的克隆及分析[J].浙江农业学报,2017,29(8):1281-1289.LIU S L,JIA Y Y,JIANG W P,et al.Cloning and sequence analysis of growth hormone gene and its flanking region in topmouth culter(Culter alburnus Basilewsky)[J].Acta Agriculturae Zhejiangensis,2017,29(8):1281-1289.(in Chinese with English abstract)
[18]BLACKET M J,ROBIN C,GOOD R T,et al.Universal primers for fluorescent labelling of PCR fragments:an efficient and cost-effective approach to genotyping by fluorescence[J].Molecular Ecology Resources,2012,12(3):456-463.
[19]STEFFENS D L,SUTTER S L,ROEMER S C.An alternate universal forward primer for improved automated DNA sequencing of M13[J].Bio Techniques,1993,15(4):580,582.
[20]SCHUELKE M.An economic method for the fluorescent labeling of PCR fragments[J].Nature Biotechnology,2000,18(2):233-234.
[21]ALMULY R,SKOPAL T,FUNKENSTEIN B.Regulatory regions in the promoter and first intron of Sparus aurata growth hormone gene:repression of gene activity by a polymorphic minisatellite[J].Comparative Biochemistry and PhysiologyPart D:Genomics and Proteomics,2008,3(1):43-50.
[22]鲁双庆,刘峰,刘臻,等.三种鳜鱼(Siniperca)生长激素基因的克隆及序列比较[J].海洋与湖沼,2008,39(4):354-361.LU S Q,LIU F,LIU Z,et al.Comparison in cloning and sequence of growth hormone gene in three species of genus Siniperca[J].Oceanologia et Limnologia Sinica,2008,39(4):354-361.(in Chinese with English abstract)
[2]CAVARI B,FUNKENSTEIN B,CHEN T T,et al.Effect of growth hormone on the growth rate of the gilthead seabream(Sparus aurata),and use of different constructs for the production of transgenic fish[J].Aquaculture,1993,111(1/4):189-197.
[3]SAKAMOTO T,SHEPHERD B S,MADSEN S S,et al.Osmoregulatory actions of growth hormone and prolactin in an advanced teleost[J].General and Comparative Endocrinology,1997,106(1):95-101.
[4]FORSYTH I A,WALLIS M.Growth hormone and prolactin:molecular and functional evolution[J].Journal of Mammary Gland Biology&Neoplasia,2002,7(3):291-312.
[5]李美娟.黄颡鱼生长激素基因多态性与生长性状的关联性研究[D].广州:华南农业大学,2016.LI M J.Correlation between the polymorphism of growth hormone gene with the growth traits in the yellow catfish(Pelteobagrus Fulvidraco)[D].Guangzhou:South China Agricultural University,2016.(in Chinese with English abstract)
[6]王文君,陈克飞,任军,等.中外不同猪品种生长激素基因遗传多态性检测[J].农业生物技术学报,2003,11(1):103-104.WANG W J,CHEN K F,REN J,et al.Detection of the polymorphisms of growth hormone gene in Chinese and European pig breeds[J].Journal of Agricultural Biotechnology,2003,11(1):103-104.(in Chinese with English abstract)
[7]CHUNG E R,LEE C S.A previously unreported Dra I polymorphism within the regulatory region of the bovine growth hormone gene and its association with growth traits in Korean Hanwoo cattle[J].Animal Genetics,2004,35(2):152-158.
[8]聂庆华,张细权,杨关福,等.鸡生长激素基因的PCR-RFLPs分析[J].农业生物技术学报,2001,9(3):282-285.NIE Q H,ZHANG X Q,YANG G F,et al.PCR-RFLPs analysis of chicken growth hormone gene[J].Journal of Agricultural Biotechnology,2001,9(3):282-285.(in Chinese with English abstract)
[9]YOWE D L,EPPING R J.A minisatellite polymorphism in intron III of the barramundi(Lates calcarifer)growth hormone gene[J].Genome,1996,39(5):934-940.
[10]KANG J,LEE S,PARK S,et al.DNA polymorphism in the growth hormone gene and its association with weight in olive flounder Paralichthys olivaceus[J].Fisheries Science,2010,68(3):494-498.
[11]ALMULY R,CAVARI B,FERSTMAN H,et al.Genomic structure and sequence of the gilthead seabream(Sparus aurata)growth hormone-encoding gene:identification of minisatellite polymorphism in intron I[J].Génome,2000,43(5):836-845..
[12]KASHI Y,KING D,SOLLER M.Simple sequence repeats as a source of quantitative genetic variation[J].Trends in Genetics,1997,13(2):74-78.
[13]乐佩琦,罗云林.鲌亚科鱼类系统发育初探(鲤形目:鲤科)[J].水生生物学报,1996,20(2):182-185.YUE P Q,LUO Y L.Preliminary studies on phylogeny of subfamily Cultrinae(Cypriniformes:Cyprinidae)[J].Acta Hydrobiologica Sinica,1996,20(2):182-185.(in Chinese)
[14]LIU S L,GU Z M,JIA Y Y,et al.Isolation and characterization of 32 microsatellite loci for topmouth culter(Culter alburnus Basilewsky)[J].Genetics and Molecular Research,2014,13(3):7480-7483.
[15]LIU S L,GU Z M,ZHAO J L,et al.Characterization of microsatellites and repetitive flanking sequences(Re FS)from the topmouth culter(Culter alburnus Basilewsky)[J].Biochemical Systematics and Ecology,2015,63(5):127-135.
[16]刘士力,程顺,蒋文枰,等.翘嘴鲌胰岛素样生长因子-Ⅰ(IGF-I)基因的克隆及序列分析[EB/OL].(2018-12-24)[2019-01-04].http://ssswxb.ihb.ac.cn/article/id/fb324b22-122f-48a6-8c63-65fcbd8ccb3c.LIU S L,CHENG S,JIANG W P,et al.Cloning and sequence analysis of insulin-like growth factor I gene in topmouth culter(Culter alburnus Basilewsky)[EB/OL].(2018-12-24)[2019-01-4].http://ssswxb.ihb.ac.cn/article/id/fb324b22-122f-48a6-8c63-65fcbd8ccb3c.
[17]刘士力,贾永义,蒋文枰,等.翘嘴鲌生长激素(GH)基因与侧翼区的克隆及分析[J].浙江农业学报,2017,29(8):1281-1289.LIU S L,JIA Y Y,JIANG W P,et al.Cloning and sequence analysis of growth hormone gene and its flanking region in topmouth culter(Culter alburnus Basilewsky)[J].Acta Agriculturae Zhejiangensis,2017,29(8):1281-1289.(in Chinese with English abstract)
[18]BLACKET M J,ROBIN C,GOOD R T,et al.Universal primers for fluorescent labelling of PCR fragments:an efficient and cost-effective approach to genotyping by fluorescence[J].Molecular Ecology Resources,2012,12(3):456-463.
[19]STEFFENS D L,SUTTER S L,ROEMER S C.An alternate universal forward primer for improved automated DNA sequencing of M13[J].Bio Techniques,1993,15(4):580,582.
[20]SCHUELKE M.An economic method for the fluorescent labeling of PCR fragments[J].Nature Biotechnology,2000,18(2):233-234.
[21]ALMULY R,SKOPAL T,FUNKENSTEIN B.Regulatory regions in the promoter and first intron of Sparus aurata growth hormone gene:repression of gene activity by a polymorphic minisatellite[J].Comparative Biochemistry and PhysiologyPart D:Genomics and Proteomics,2008,3(1):43-50.
[22]鲁双庆,刘峰,刘臻,等.三种鳜鱼(Siniperca)生长激素基因的克隆及序列比较[J].海洋与湖沼,2008,39(4):354-361.LU S Q,LIU F,LIU Z,et al.Comparison in cloning and sequence of growth hormone gene in three species of genus Siniperca[J].Oceanologia et Limnologia Sinica,2008,39(4):354-361.(in Chinese with English abstract)