Scientific evidence of African origin Breed Brazilian Curraleiro

MATERNAL ORIGIN AND GENETIC DIVERSITY OF POPULATIONS OF CATTLE

Curraleiro FROM DIFFERENT REGIONS OF BRAZIL

Summary

Analyses of mitochondrial DNA (mtDNA) have demonstrated their potential for understanding the origin and

nature of the processes of domestication, dispersion analysis and gene flow analysis, expansion

demographic, genetic drift and mixing of populations. The breed originated in bovine Curraleiro

animals introduced by settlers who adapted health conditions, climate and management in Brazil.

The objective of this study was to assess the genetic diversity of this breed female mediated by

sequencing and analysis of the mtDNA control region. Twelve animals were analyzed from five

distinct regions. Three haplogroups were observed: two of taurine African origin (YY and T1) and a

European origin taurine (T3). The nucleotide diversity observed was 0.013. The highest diversity

haplotype was observed in (0.012) and not between populations (0.001). The rate of differentiation observed

was 0.074. There were no significant genetic differences between populations of different

regions. For the analysis it can be concluded that the populations of Curraleiro have the same

maternal origin and influenced animal origin African taurine.

Introduction

Analysis of genetic diversity in the sequence of mtDNA in cattle have demonstrated the potential

this tool for understanding the origin and nature of the processes of domestication (Bradley et al.

1998) as well as studies on the diversification of current populations of cattle (Carvajal-Carmona

et al. 2003). Essentially uniparentalmente haploid and transmitted via maternal mtDNA opened a new

perspective in the study of population genetics. Since the molecular marker used in studies of

domestication, mtDNA is used to identify the probable wild ancestor, the number of lines

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mothers in the study population and geographical origin. With the observed data can trace a pattern

geographic diversity and evolution of a species, dispersal and gene flow, check the expansions

demographic, genetic drift and miscegenation (Bruford et al., 2003).

By analyzing the mtDNA control region of the bovine species was separated into two major clusters (Afroeuropeu

and Asia), whose divergence occurred ~ 12,000 years ago, coinciding with the events independent of

domestication of taurine and zebu subspecies (Loftus et al., 1994a). Subsequent studies showed that

There was a wide divergence between bullfighting originated in Africa and Europe, showing the origin and

Expansion of the two populations from two distinct and separate ancestral sources (Bradley et al. 1996).

Haplotypes unique and highly frequent in both groups represented the African taurine breeds (Afcons -

African consensus, also called T1) and the European taurine (Eucons - European consencus,

referred to as T3), being the difference between the two strains related mitochondrial three

substitutions within a region of 240bp in the control region (d-loop) and its variants.

The breed originated in Curraleiro bovine animals introduced by settlers for more than five centuries in

Brazil. These animals have adapted to the sanitary conditions of climate and management and after the introduction of the races

Zebu suffered a great risk of extinction. The preservation, maintenance and genetic characterization of this

population is working object of Animal Genetic Resources Network Platform for Genetic Resources

EMBRAPA and its partners. The objective of this study was to determine the nucleotide diversity of this breed,

mediated by the female, by sequencing and analysis of the mtDNA control region.

Material and methods

We analyzed 12 animals of Curraleiro from 5 regions: Distrito Federal (DF-CU, n = 3),

Goi‡s (GO-CU, n = 3), Maranh‹o (MA-CU, n = 2), Piau’ (PI-CU, n = 2) and Minas Gerais (MG-CU, n = 2). A

sequence of 375bp located in the control region (d-loop) used in studies involving different races

Iberia (Loftus et al., 1994b; Bradley et al., 1996; Cymbron et al., 1999) and Latin America (Miretti

et al. 2002; Carvajal-Carmona et al. 2003; Mirol et al., 2003) was amplified with the primers described by

Cymbron et al. (1999). Amplification was performed in a final volume of 20ul containing 1.5 mM MgCl2,

0.25 mM of each primer, 200μM of each dNTP, 10-25ng of DNA and Taq DNA polymerase 1UI. The

Programming of PCRs had the following configuration: 94 ¼ C / 5 ', 30x (94 ¼ C / 1', 56oC / 1 'and 72 ¡ C / 1') and final extension

of 72oC/30 '. After purification of the sequencing reaction was performed by the chain termination method with

dideoxinucleot’deos marked with fluorochromes. Electrophoresis was performed on an automatic sequencer

ABI PRISM 3100.

Sequences were aligned and edited the program SeqScape v 2.1 (Applied Biosystems) and the

V00654 the reference sequence (Anderson et al. 1982). The nucleotide diversity and distances

within, between and total were obtained by using the program Mega 3. The matrices of distances generated

were based on the substitution pattern Kimura 2-parameter. From the distance matrix generated to

pairs a dendrogram was constructed by grouping neighbor-joining (Consensus Network), taking into

consideration the possibility of interbreeding between populations (Hybridization Network). This action was

SliptTree4 implemented by the program (http://www.Splitstree.org/).

Results and Discussion

Based on the observed haplotype animals were included into three groups predefined by others

(Troy et al. 2001; Miretti et al., 2002): 3 haplogroup AA animals showed that although share

positions that define the haplogroup T1 differs at positions 16053, 16122 and 16196 (Miretti et al., 2002) and

corresponding to one tap of the line of African origin taurine found in higher proportion in

Creole breeds in South America, 5 animals showed haplogroup T1 (defined by 16,050 positions,

16,113 and 16,255), corresponding to taurine lineage of African origin (Afcons) and 4 showed the strain

T3 (Eucons), which corresponds to European reference sequence (Anderson et al., 1982).

Given the colonization of the Americas and the introduction of cattle on our continent, it was assumed that

there should be a reduced number of mitochondrial haplotypes shared by native breeds of

South America Contradicting this hypothesis, Miretti et al. (2002) and Egypt (2007) observed the existence of

several distinct haplotypes source taurine breeds in Brazil, one of the AA haplogroup. This

haplogroup may reflect the evolutionary process of its own local Creole breeds (Miretti et al., 2002) or

may have been widespread in the Americas due to processes of genetic drift (Egypt, 2007). The latter

hypothesis is supported by the existence of taurine haplotypes of African origin have been observed in races

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Iberian, showing the introgression of African taurine breeds in Iberia occurred probably

moor the time domain in this region (Cymbron et al. 1999). Beja-Pereira et al. (2006) also suggest that

This interbreeding may have occurred due to demographic expansion observed through the Strait of Gibraltar, in

time of the Bronze Age (~ 3,000 B.C.).

The index of differentiation observed between populations of different origins of Curraleiro was 0.074 and

the nucleotide diversity of the breed was 0.013. When individuals of Curraleiro were sub-grouped

depending on its origin it was found that the nucleotide diversity within this population is 0.012

while among populations there is a small difference of 0,001.

There were no significant genetic differences in subpopulations of Curraleiro. Differences

observed at the level of sub-populations curraleiras, although showing some closer

populations (Figure 1), as in the case of animals from herds of Maranh‹o and Piau’ and animals

Herd Animal Germplasm Bank of Embrapa Genetic Resources and Biotechnology and the population

collected in Minas Gerais (Motto of jump), reflect a common origin or next of herds and analyzed

no differences in the structure and origin of Curraleiro. This fact can be better visualized when a

dendrogram is generated for the 12 individuals of Curraleiro that compose the sample (Figure 2).

Figure 1. Circular dendrogram based on genetic distance of Kimura 2p for the set of sub-populations

of Curraleiro by grouping Neighbor-joining (Unrooted dendrogram based on Kimura's genetic 2p

distances for all subpopulations Curraleiro breed made by the Neighbor-joining clustering method)

Figure 2. Dendrogram of individuals based on genetic distance of Kimura 2p done by grouping

Neighbor-joining (Kimura's Dendrogram based on genetic distances between 2p Individuals made by the

Neighbor-joining clustering method)

Conclusions

For the analysis we concluded that the sub-populations of Curraleiro have the same

maternal origin with no statistically significant differences between them. Moreover, there is a great

influence of animal origin in African taurine populations.

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Financing

EMBRAPA

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