Archivos de Zootecnia ISSN: 0004-0592 [email protected] Universidad de Córdoba España Serrano, G. M.; Egito, A. A.; McManus, C.; Mariante, A. da S. Genetic population structure of brazilian bovine breeds inferred by rapd markers Archivos de Zootecnia, vol. 54, núm. 206-207, 2005, pp. 409-414 Universidad de Córdoba Córdoba, España Available in: http://www.redalyc.org/articulo.oa?id=49520745 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative GENETIC POPULATION STRUCTURE OF BRAZILIAN BOVINE BREEDS INFERRED BY RAPD MARKERS ESTRUTURA GENÉTICA DE POPULAÇÕES DE RAÇAS BOVINAS BRASILEIRAS INFERI- DA POR MARCADORES RAPD Serrano, G.M. 2, A.A. Egito 1, C. McManus 2 and A. da S. Mariante 1 1Embrapa Recursos Genéticos e Biotecnologia. Brasília-DF. Brasil. E-mail: [email protected] 2Universidade de Brasília. Brasília-DF. Brasil. E-mail: [email protected] ADDITIONAL KEYWORDS PALAVRAS CHAVE ADICIONAIS Bovine native breeds. Populations structure. Raças bovinas nativas. Estrutura de populações. Conservation genetics. Conservação genética. SUMMARY Conservation and improvement strategies two populations have high genetic similarity. In should be based on the association between the dendrogram generated by UPGMA method all genetic and phenotypic characteristics. In this groups clustered in their respective breed. In this study 10 different populations of five native study we could demonstrate the existence of Brazilian cattle breeds (Caracu, Mocho Nacional, regional genetic differences between the Crioulo Lageano, Curraleira and Pantaneira) were populations of native breeds. These estimates characterized using RAPD techniques to estimate could be useful in crossbreeding and genetic their genetic relationships and to verify the sample exchange between different nuclei. existence of possible regional differences within each breed. Two commercial breeds were used as control and outgroup respectively (Holstein RESUMO and Nellore). 120 arbitrarily primers were screened, of which 22 were selected, and Estratégias para a conservação e mel- generated 122 polymorphic bands. When all horamento animal devem ser baseadas na groups were analysed in pairs, non-significant associação de características fenotípicas e genetic variability (p>0.05) observed only genéticas. Neste estudo realizou-se a carac- between two populations of Curraleiro breed terização genética de 10 diferentes populações located in States of Piaui (CUPI) and Minas Gerais de cinco raças bovinas nativas brasileiras (CUMG). Using Popgene program Nei's 1978 (Caracu, Mocho Nacional, Crioulo Lageano, genetic distance was estimated. The smallest Curraleira e Pantaneira) através da técnica de divergences were observed between groups of RAPD, visando estimar a relação genética das the same breed and the least distance between populações e verificar a possibilidade de existir the two groups of Caracu breed (0.0159). In the diferenças regionais dentro de cada raça. Duas four populations of Curraleiro the groups CUPI raças comerciais foram utilizadas como controle and CUMG presented the smallest difference e outgroup respectivamente (Holandesa e Nelore). (0.0285). These results could indicate that these Uma triagem com 120 primers foi realizada dos Arch. Zootec. 54: 409-414. 2005. GeneticSerrano.p65 409 07/12/2005, 14:53 SERRANO, EGITO, MCMANUS AND MARIANTE quais foram selecionados 22, que geraram 122 conservation and improvement, that bandas polimórficas. Quando todos os grupos these must be based on the combination foram confrontados em pares, a variabilidade of the phenotypic and genetic data. genética não foi significativa (p>0,05) apenas Studies related to the genetic para dois grupos da raça Curraleira localizados characterization of the native Brazilian no estado do Piauí (CUPI) e no de Minas Gerais bovine breeds are being carried out in (CUMG). Utilizando o programa Popgene, foram the Laboratory of Animal Genetics - estimadas as distâncias genéticas, através do Brasilia, DF – using DNA molecular método de Nei (1978). As menores divergências markers. One of those molecular foram observadas entre os grupos da mesma markers used is the RAPD (random raça e a menor delas foi encontrada entre os dois amplified polymorphic DNA). The grupos da raça Caracu (0,0159). Entre as quatro RAPD metodology is being widely used populações da raça Curraleira, os grupos CUPI in studies on conservation, as it is e CUMG apresentaram a menor diferença realively simple, low cost and that does (0,0285). Este resultado indica que esses dois grupos possuem grande similaridade genética. not require specific knowledge on the Os grupos de cada raça se agruparam em clusters population to be studied, helping in the diferentes, pertencentes às respectivas raças choice of strategies to preserve no dendrograma gerado pelo método de UPGMA. threatened species and breeds. Here Neste estudo pode-se demonstrar a existência these markers were used to study 10 de diferenças genéticas regionais nas popu- populations of five native Brazilian lações das raças nativas. Estas estimativas bovine breeds in order to investigate podem ser utilizadas em cruzamentos e trocas the existence of regional genetic de amostras genéticas entre os diferentes nú- differences. cleos. MATERIAL AND METHODS INTRODUCTION In these study we analised possible Brazilian native bovine breeds have regional diferences between Curraleiro developed from those brought by the (CU) from the Northeast and Central Portuguese soon after the discovery. West States; Caracu (CA) from Cen- With importation of some exotic breeds, tral West and Southern Brazil and mainly zebus, at the beginning of the National Polled breed (MN) from Mi- past century, the native animals have nas Gerais State, South-eastern Brazil. gradually been substituted making most Two other naturalized Brazilian cattle of them threatened with extinction. breeds and two commercial breeds Although the exotic breeds are were also studied. These included the considered more productive, they do Pantaneiro (PAN, n=48) from the not necessarily possess the adaptation, Pantanal region, South Western Brazil resistance to illnesses and parasites and Crioulo Lageano (CL, n=48) from found in the native breeds. Santa Catarina State, Southern Brazil. The techniques for the analysis of As all native cattle breeds are of taurine the genetic variability are essential origin the Holstein-Friesian (HOL, ingredients for programs of rational n=48) was included as a control group Archivos de zootecnia vol. 54, núm. 206-207, p. 410. GeneticSerrano.p65 410 07/12/2005, 14:53 GENETIC STRUCTURE OF BRAZILIAN BOVINE BREEDS and Nellore (NEL, n=48) ( Bos indicus) 1 min, 72oC for 2 min. A final extension as an outgroup. at 72oC was carried out for 7 min The Caracu breed was divided in followed by a cooling at 4oC. PCR two groups, in accordance with the products were separated by electro- aptitude of the populations, the Caracu phoresis in 1.4 percent agarose gels in Caldeano (CA1, n=17) selected for 1 x TBE, and were stained with ethidium milk and the other collected in the bromide (0,6 mg/ml) and observed under remaining portion of Brazil (CA2, n= UV light. 31), usually selected for meat pro- The fragment data were entered in duction. The Curraleiro was divided in a computer file as a binary matrix: 0 four groups of origin and/or region of coded for absence and 1 for presence the herd. The first group (CUPI, n=12) of a band. The genetic distances was composed of animals from Piauí, between populations were calculated the II group (CUMG, n=7) of Minas using the POPGENE program (Popu- Gerais, and the other two were from lation Genetic Analysis) version 1.31 Goiás, group III (CUGO1, n=9) of (Yeh et al., 1999). This program Maranhão origin and the IV group establishes standardized genetic (CUGO2, n=2) from Natividade-GO. distance matrices (Nei, 1972) and The National Mocho was divided in matrices of genetic distances corrected two groups, in the first group (MN1, for small samples (Nei, 1978). All clus- n=13) from the Três Barras Farm - ter analyses done in this work used MG and the other (MN2, n=34) animals UPGMA and the resulting clusters from the Bank of Animal Germplasm were expressed as dendrograms. The located in Embrapa Recursos Gené- bootstrap analysis was carried out using ticos e Biotecnoloiga, Brasília-DF. the TFPGA program (Tools for DNA extraction was based on a Population Genetics Analyses, version non-organic protocol described on 1.3) (Miller, 1997). The analysis of Miller et al. (1988) with slight molecular variance (AMOVA) (Exco- modifications. A total of 120 arbitrary ffier et al., 1992) was used to analyze primers (Operon Technologies Inc., the variance between the populations. Alameda, Calif.: A, B, E, J, K and AB) were screened using one sample of each breed. RESULTS AND DISCUSSION Amplification reactions were performed in 50mM KCl, 20mM Tris- Of the 120 primers investigated, 72 were polymorphic (60 percent), 23 HCl (pH 8,4), 2,5mM MgCl2, 200mM of each dNTP, 0,4mM primer, 9ng monomorphic (19,2 percent) and 25 template DNA, 8 percent of BSA (2,5 did not amplify or had poor amplification mg/ml) and 1.5UI Taq DNA polimerase (20.8 percent). Only those that in a final volume of 13ml. DNA presented at least 4 polymorphic bands amplification was performed in a were used, thus reducing, the time and thermal cycler programmed as follows: the cost of the experiments. Using 94oC for 5 min followed by 40 cycles these criteria, 22 primers had been consisting of 94oC for 1 min, 36oC for selected and generated a total of 122 Archivos de zootecnia vol. 54, núm. 206-207, p. 411. GeneticSerrano.p65 411 07/12/2005, 14:53 SERRANO, EGITO, MCMANUS AND MARIANTE polymorfics bands (5.5 bands/primer). between them. These measures also In relation to the genetic variability express the degree of genetic diver- between groups, when CUPI and gence between the groups.