Polimorfismo no íntron 1-PstI do gene do hormônio do crescimento em linhagens de tilápia do Nilo (Oreochromis niloticus).

Molecular genetics studies are becoming more common in tilapicultura, because the Nile tilapia shows great potential in genetic engineering dedicated to the improvement stocks. With the development of different Nile tilapia (Oreochromis niloticus) strains have been increased the need to identify them, aimed the maintenance and the certification of genetic material quality of improved strains acquired by the producers. Improvement of fish natural growth rates in aquaculture has been widely explored, with gains resulting from improvements in livestock production, nutrition and genetic selection. Due to the high impact of GH in growth regulation and by being involved in several other metabolic functions, the GH gene is a potential target for genetic variation studies and its association with characteristics of fish growth. Thus, this study aimed to: describe the gene intron 1 variability of the growth hormone (GH1) of strains Chitralada and GIFT (Genetically Improvement Farmed Tilapia) from the Nile tilapia through molecular marker PCR-RFLP; study the association of this gene polymorphisms with strains performance characteristics; and seek a molecular marker that could be used in the strains identification and as a selection tool for growth characteristics in genetic improvement programs for the Nile tilapia. 200 animals were used of each strain from witch it was collected fragments of tail fin and the following measurements: total length (CT), standard length (CP), height (AL), width (LA), head length (CC), slaughter weight (PA), fillet weight (PF), fillet yield (RF) and the carcass weight (PC). From collected fin, it was done the genomic material extraction using saturated NaCl (5M). For PCR-RFLP analysis, it was necessary a specific pair of primer design based on the available sequence-reference in the GenBank (access M97766). The generated PCR products were digested with enzyme PstI, with restriction locus located in the intron of the gene GH1. Allelic frequencies, genotypic, rates of heterozygosity, test of the Hardy-Weinberg equilibrium and estatistic F were determined with Popgen 1.31 program. The analysis of the association between the polymorphisms and the performance characteristics were evaluated by the SAS GLM procedure. Duncan test (p <0.05) was used for the variables that showed effect of interaction between genotype and lineage. The three pairs of primers designed amplified satisfactorily in the two strains, producing fragments of 652, 441 and 396 bp. For 652 bp fragment, it was observed the three digestion standard possible for both strains, Chitralada and GIFT, corresponding to the genotypes PstI+/+, PstI+/- and PstI-/-. The genotypic frequencies found for the Chitralada strain were 0.707, 0.282 and 0.011 respectively for the genotypes PstI+/+, PstI+/- and PstI-/- and the GIFT strain, 0.930, 0.075 and 0.005 for the same genotypes. The allelic frequencies obtained in this study (GH1-PstI) show that the PstI+ is dominant in the Nile tilapia. The Chitralada strain (Ho = 0.282) showed greatest heterozygosity to GIFT strain (Ho = 0.065), having therefore a higher variability to the locus in question. The index of differentiation (FST) of the strains was 0.038. The allelic frequencies in the two strains were in Hardy-Weinberg equilibrium. About the association analysis of polymorphisms for production characteristics, there was no correlation only for CC and RF. So the major averages were recorded for animals that carriers the heterozygous genotype. Only to the RF characteristic there was an interaction effect between genotype and strain, to the point that the GIFT strain with homozygous genotype PstI+/+ was 2.72 percent higher than the genotype heterozygous PstI+/-. Within each genotype, the Chitralada strain was higher. With the results obtained here, it is suggested that the combination of performance characteristic of the Nile tilapia strains to the genetic polymorphisms of GH may be due to the effect of the gene regulation of growth hormone. Later detailed identifications of polymorphisms GH1-PstI structure may prove link between RFLPs described and the characteristics controlled by GH, which can contribute to the implementation of MAS in the tilapia genetic programs improvement.

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Bibliographic Details
Main Author: Blanck, D. V.
Format: Thesis/Dissertation biblioteca
Language:Portuguese
Published: Universidade Estadual de Maringá. Centro de Ciências Agrárias. Programa de Pós-Graduação em Zootecnia 2008
Subjects:Growth hormone, Niloticus niloticus, Genetic variability, Gene, Restriction enzyme, Brazil, Variabilidade genética, Enzima de restrição, Genética molecular, Tilápia do Nilo, Hormônio do crescimento, Oreochromis niloticus, Nile tilapia, Brasil, Molecular genetics,
Online Access:http://hdl.handle.net/1834/9842
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id dig-aquadocs-1834-9842
record_format koha
institution UNESCO
collection DSpace
country Francia
countrycode FR
component Bibliográfico
access En linea
databasecode dig-aquadocs
tag biblioteca
region Europa del Oeste
libraryname Repositorio AQUADOCS
language Portuguese
topic Growth hormone
Niloticus niloticus
Genetic variability
Gene
Restriction enzyme
Brazil
Variabilidade genética
Gene
Enzima de restrição
Genética molecular
Tilápia do Nilo
Hormônio do crescimento
Oreochromis niloticus
Nile tilapia
Brasil
Molecular genetics
Growth hormone
Niloticus niloticus
Genetic variability
Gene
Restriction enzyme
Brazil
Variabilidade genética
Gene
Enzima de restrição
Genética molecular
Tilápia do Nilo
Hormônio do crescimento
Oreochromis niloticus
Nile tilapia
Brasil
Molecular genetics
spellingShingle Growth hormone
Niloticus niloticus
Genetic variability
Gene
Restriction enzyme
Brazil
Variabilidade genética
Gene
Enzima de restrição
Genética molecular
Tilápia do Nilo
Hormônio do crescimento
Oreochromis niloticus
Nile tilapia
Brasil
Molecular genetics
Growth hormone
Niloticus niloticus
Genetic variability
Gene
Restriction enzyme
Brazil
Variabilidade genética
Gene
Enzima de restrição
Genética molecular
Tilápia do Nilo
Hormônio do crescimento
Oreochromis niloticus
Nile tilapia
Brasil
Molecular genetics
Blanck, D. V.
Polimorfismo no íntron 1-PstI do gene do hormônio do crescimento em linhagens de tilápia do Nilo (Oreochromis niloticus).
description Molecular genetics studies are becoming more common in tilapicultura, because the Nile tilapia shows great potential in genetic engineering dedicated to the improvement stocks. With the development of different Nile tilapia (Oreochromis niloticus) strains have been increased the need to identify them, aimed the maintenance and the certification of genetic material quality of improved strains acquired by the producers. Improvement of fish natural growth rates in aquaculture has been widely explored, with gains resulting from improvements in livestock production, nutrition and genetic selection. Due to the high impact of GH in growth regulation and by being involved in several other metabolic functions, the GH gene is a potential target for genetic variation studies and its association with characteristics of fish growth. Thus, this study aimed to: describe the gene intron 1 variability of the growth hormone (GH1) of strains Chitralada and GIFT (Genetically Improvement Farmed Tilapia) from the Nile tilapia through molecular marker PCR-RFLP; study the association of this gene polymorphisms with strains performance characteristics; and seek a molecular marker that could be used in the strains identification and as a selection tool for growth characteristics in genetic improvement programs for the Nile tilapia. 200 animals were used of each strain from witch it was collected fragments of tail fin and the following measurements: total length (CT), standard length (CP), height (AL), width (LA), head length (CC), slaughter weight (PA), fillet weight (PF), fillet yield (RF) and the carcass weight (PC). From collected fin, it was done the genomic material extraction using saturated NaCl (5M). For PCR-RFLP analysis, it was necessary a specific pair of primer design based on the available sequence-reference in the GenBank (access M97766). The generated PCR products were digested with enzyme PstI, with restriction locus located in the intron of the gene GH1. Allelic frequencies, genotypic, rates of heterozygosity, test of the Hardy-Weinberg equilibrium and estatistic F were determined with Popgen 1.31 program. The analysis of the association between the polymorphisms and the performance characteristics were evaluated by the SAS GLM procedure. Duncan test (p <0.05) was used for the variables that showed effect of interaction between genotype and lineage. The three pairs of primers designed amplified satisfactorily in the two strains, producing fragments of 652, 441 and 396 bp. For 652 bp fragment, it was observed the three digestion standard possible for both strains, Chitralada and GIFT, corresponding to the genotypes PstI+/+, PstI+/- and PstI-/-. The genotypic frequencies found for the Chitralada strain were 0.707, 0.282 and 0.011 respectively for the genotypes PstI+/+, PstI+/- and PstI-/- and the GIFT strain, 0.930, 0.075 and 0.005 for the same genotypes. The allelic frequencies obtained in this study (GH1-PstI) show that the PstI+ is dominant in the Nile tilapia. The Chitralada strain (Ho = 0.282) showed greatest heterozygosity to GIFT strain (Ho = 0.065), having therefore a higher variability to the locus in question. The index of differentiation (FST) of the strains was 0.038. The allelic frequencies in the two strains were in Hardy-Weinberg equilibrium. About the association analysis of polymorphisms for production characteristics, there was no correlation only for CC and RF. So the major averages were recorded for animals that carriers the heterozygous genotype. Only to the RF characteristic there was an interaction effect between genotype and strain, to the point that the GIFT strain with homozygous genotype PstI+/+ was 2.72 percent higher than the genotype heterozygous PstI+/-. Within each genotype, the Chitralada strain was higher. With the results obtained here, it is suggested that the combination of performance characteristic of the Nile tilapia strains to the genetic polymorphisms of GH may be due to the effect of the gene regulation of growth hormone. Later detailed identifications of polymorphisms GH1-PstI structure may prove link between RFLPs described and the characteristics controlled by GH, which can contribute to the implementation of MAS in the tilapia genetic programs improvement.
format Thesis/Dissertation
topic_facet Growth hormone
Niloticus niloticus
Genetic variability
Gene
Restriction enzyme
Brazil
Variabilidade genética
Gene
Enzima de restrição
Genética molecular
Tilápia do Nilo
Hormônio do crescimento
Oreochromis niloticus
Nile tilapia
Brasil
Molecular genetics
author Blanck, D. V.
author_facet Blanck, D. V.
author_sort Blanck, D. V.
title Polimorfismo no íntron 1-PstI do gene do hormônio do crescimento em linhagens de tilápia do Nilo (Oreochromis niloticus).
title_short Polimorfismo no íntron 1-PstI do gene do hormônio do crescimento em linhagens de tilápia do Nilo (Oreochromis niloticus).
title_full Polimorfismo no íntron 1-PstI do gene do hormônio do crescimento em linhagens de tilápia do Nilo (Oreochromis niloticus).
title_fullStr Polimorfismo no íntron 1-PstI do gene do hormônio do crescimento em linhagens de tilápia do Nilo (Oreochromis niloticus).
title_full_unstemmed Polimorfismo no íntron 1-PstI do gene do hormônio do crescimento em linhagens de tilápia do Nilo (Oreochromis niloticus).
title_sort polimorfismo no íntron 1-psti do gene do hormônio do crescimento em linhagens de tilápia do nilo (oreochromis niloticus).
publisher Universidade Estadual de Maringá. Centro de Ciências Agrárias. Programa de Pós-Graduação em Zootecnia
publishDate 2008
url http://hdl.handle.net/1834/9842
work_keys_str_mv AT blanckdv polimorfismonointron1pstidogenedohormoniodocrescimentoemlinhagensdetilapiadonilooreochromisniloticus
AT blanckdv polymorphisminintron1pstgrowthhormonegeneinniletilapiastrainsoreochromisniloticus
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spelling dig-aquadocs-1834-98422021-05-19T06:27:06Z Polimorfismo no íntron 1-PstI do gene do hormônio do crescimento em linhagens de tilápia do Nilo (Oreochromis niloticus). Polymorphism in intron 1- Pst growth hormone gene in Nile tilapia strains (Oreochromis niloticus) Blanck, D. V. Growth hormone Niloticus niloticus Genetic variability Gene Restriction enzyme Brazil Variabilidade genética Gene Enzima de restrição Genética molecular Tilápia do Nilo Hormônio do crescimento Oreochromis niloticus Nile tilapia Brasil Molecular genetics Molecular genetics studies are becoming more common in tilapicultura, because the Nile tilapia shows great potential in genetic engineering dedicated to the improvement stocks. With the development of different Nile tilapia (Oreochromis niloticus) strains have been increased the need to identify them, aimed the maintenance and the certification of genetic material quality of improved strains acquired by the producers. Improvement of fish natural growth rates in aquaculture has been widely explored, with gains resulting from improvements in livestock production, nutrition and genetic selection. Due to the high impact of GH in growth regulation and by being involved in several other metabolic functions, the GH gene is a potential target for genetic variation studies and its association with characteristics of fish growth. Thus, this study aimed to: describe the gene intron 1 variability of the growth hormone (GH1) of strains Chitralada and GIFT (Genetically Improvement Farmed Tilapia) from the Nile tilapia through molecular marker PCR-RFLP; study the association of this gene polymorphisms with strains performance characteristics; and seek a molecular marker that could be used in the strains identification and as a selection tool for growth characteristics in genetic improvement programs for the Nile tilapia. 200 animals were used of each strain from witch it was collected fragments of tail fin and the following measurements: total length (CT), standard length (CP), height (AL), width (LA), head length (CC), slaughter weight (PA), fillet weight (PF), fillet yield (RF) and the carcass weight (PC). From collected fin, it was done the genomic material extraction using saturated NaCl (5M). For PCR-RFLP analysis, it was necessary a specific pair of primer design based on the available sequence-reference in the GenBank (access M97766). The generated PCR products were digested with enzyme PstI, with restriction locus located in the intron of the gene GH1. Allelic frequencies, genotypic, rates of heterozygosity, test of the Hardy-Weinberg equilibrium and estatistic F were determined with Popgen 1.31 program. The analysis of the association between the polymorphisms and the performance characteristics were evaluated by the SAS GLM procedure. Duncan test (p <0.05) was used for the variables that showed effect of interaction between genotype and lineage. The three pairs of primers designed amplified satisfactorily in the two strains, producing fragments of 652, 441 and 396 bp. For 652 bp fragment, it was observed the three digestion standard possible for both strains, Chitralada and GIFT, corresponding to the genotypes PstI+/+, PstI+/- and PstI-/-. The genotypic frequencies found for the Chitralada strain were 0.707, 0.282 and 0.011 respectively for the genotypes PstI+/+, PstI+/- and PstI-/- and the GIFT strain, 0.930, 0.075 and 0.005 for the same genotypes. The allelic frequencies obtained in this study (GH1-PstI) show that the PstI+ is dominant in the Nile tilapia. The Chitralada strain (Ho = 0.282) showed greatest heterozygosity to GIFT strain (Ho = 0.065), having therefore a higher variability to the locus in question. The index of differentiation (FST) of the strains was 0.038. The allelic frequencies in the two strains were in Hardy-Weinberg equilibrium. About the association analysis of polymorphisms for production characteristics, there was no correlation only for CC and RF. So the major averages were recorded for animals that carriers the heterozygous genotype. Only to the RF characteristic there was an interaction effect between genotype and strain, to the point that the GIFT strain with homozygous genotype PstI+/+ was 2.72 percent higher than the genotype heterozygous PstI+/-. Within each genotype, the Chitralada strain was higher. With the results obtained here, it is suggested that the combination of performance characteristic of the Nile tilapia strains to the genetic polymorphisms of GH may be due to the effect of the gene regulation of growth hormone. Later detailed identifications of polymorphisms GH1-PstI structure may prove link between RFLPs described and the characteristics controlled by GH, which can contribute to the implementation of MAS in the tilapia genetic programs improvement. Os estudos em genética molecular estão se tornando mais comuns na tilapicultura, pois a tilápia do Nilo apresenta grande potencial em termos de engenharia genética voltada ao melhoramento dos estoques. Com o desenvolvimento de diferentes linhagens de tilápia do Nilo (Oreochromis niloticus) têm aumentado a necessidade de identificá-las, visando a manutenção e certificação da qualidade do material genético de linhagens melhoradas adquiridas pelos produtores. Melhoria das taxas de crescimento natural de peixe na aqüicultura tem sido amplamente explorada, com os ganhos decorrentes de melhorias na produção animal, nutrição e seleção genética. Em função do alto impacto do hormônio do crescimento (GH) na regulação do crescimento e por estar envolvido em diversas outras funções metabólicas, o gene GH é um potencial alvo para estudos de variação genética e sua associação com características de crescimento de peixes. Dessa maneira, com este estudo objetivou-se: descrever a variabilidade do íntron 1 do gene do hormônio do crescimento (GH1) das linhagens Chitralada e GIFT (Genetically Improvement Farmed Tilapia) de tilápia do Nilo através do marcador molecular PCR-RFLP; estudar a associação de polimorfismos deste gene as características de desempenho das linhagens; e buscar um marcador molecular que possa ser utilizado na identificação das linhagens e como ferramenta de seleção para características de crescimento em programas de melhoramento genético de tilápias do Nilo. Foram utilizados 200 animais de cada linhagem, dos quais foram coletados fragmentos de nadadeira caudal e as seguintes mensurações: comprimento total (CT), comprimento padrão (CP), altura (AL), largura (LA), comprimento da cabeça (CC), peso de abate (PA), peso do filé (PF), rendimento de filé (RF) e peso da carcaça (PC). Das nadadeiras coletadas, procedeu-se a extração do material genômico utilizando NaCl saturado (5M). Para as análises PCR-RFLP, foi necessário o desenho de pares de primers específicos com base na seqüência-referência disponível do GenBank (acesso M97766). Os produtos de PCR gerados foram digeridos com a enzima PstI, com sítio de restrição localizado no íntron 1 do gene GH1. As freqüências alélicas, genotípicas, índices de heterozigosidade, teste de equilíbrio de Hardy-Weinberg e estatítica F foram determinadas com o auxílio do programa Popgen 1.31. A análise de associação entre o polimorfismo e as características de desempenho foi avaliada pelo procedimento GLM do SAS. Teste de Duncan (p<0,05) foi utilizado para as variáveis que apresentaram efeito de interação entre linhagem e genótipo. Os três pares de primers desenhados amplificaram satisfatoriamente nas duas linhagens, produzindo fragmentos de 652, 441 e 396 pb. Para o fragmento de 652 pb, observou-se os três padrões de digestão possíveis tanto para a linhagem Chitralada como para a GIFT, correspondendo aos genótipos PstI+/+, PstI+/- e PstI-/-. As freqüências genotípicas encontradas para a linhagem Chitralada foram 0,707, 0,282 e 0,011, respectivamente para os genótipos PstI+/+, PstI+/- e PstI-/- e para a linhagem GIFT, 0,930, 0,075 e 0,005 para os mesmos genótipos. As freqüências alélicas obtidas neste estudo (GH1-PstI) mostram que o PstI+ é dominante na tilápia do Nilo. A linhagem Chitralada (Ho=0,282) apresentou maior heterozigosidade que a linhagem GIFT (Ho=0,065), tendo, portanto maior variabilidade para o loco em questão. O índice de diferenciação (FST) das linhagens foi de 0,038. As freqüências alélicas nas duas linhagens se mostraram em equilíbrio de Hardy-Weinberg. Quanto a análise de associação dos polimorfismos as características de produção, não houve correlação somente para CC e RF. Sendo que as maiores médias foram verificadas para os animais portadores do genótipo heterozigoto. Somente para a característica RF houve efeito de interação entre genótipo e linhagem, ao ponto que a linhagem GIFT com genótipo homozigoto PstI+/+ foi 2,72% superior em relação ao genótipo heterozigoto PstI+/-. Dentro de cada genótipo, a linhagem Chitralada foi superior. Com os resultados aqui obtidos, sugere-se que a associação das características de desempenho de linhagens de tilápia do Nilo ao polimorfismo genético do GH1 pode ser devido ao efeito da regulação do próprio gene do hormônio do crescimento. Posteriores identificações detalhadas da estrutura dos polimorfismos GH1-PstI podem revelar ligação entre os RFLPs descritos e as características controladas pelo GH, a qual pode contribuir para a aplicação de MAS no programa de melhoramento genético de tilápias. Masters 2017-08-24T09:05:23Z 2017-08-24T09:05:23Z 2008 Thesis/Dissertation http://hdl.handle.net/1834/9842 pt 53pp. Universidade Estadual de Maringá. Centro de Ciências Agrárias. Programa de Pós-Graduação em Zootecnia