Genetic variation of LEI0258 locus at major histocompatibility complex (MHC) region in Ethiopian indigenous village chicken

A. Kebede, K. Tesfaye, G. Belay, M. Kyallo, D. Githae, T. Dessie, N. Sparks, O. Hanotte, R. Pelle
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Abstract

  Indigenous chickens are locally adapted to environmental challenges and provide subsistence to millions of farmers in Africa.  However, their productivity remains low compared to exotic strains. Efforts are being made to combine the local adaptation of indigenous chicken with productive traits of exotic chicken. Understanding the link between genetic diversity and environmental challenges leads to marker-assisted breed improvement programs for sustainable chicken production at smallholder level. Genetic variation at LEI0258 VNTR locus located within the MHC region has been linked to infectious disease resistance/susceptibility in commercial breeds. The aim of this study was to investigate allelic variability, genetic diversity and genetic relationships of 24 chicken populations in Ethiopia. Here, the diversity at LEI0258 in 236 chickens from 24 Ethiopian indigenous chicken populations using the major Histo-compatibility Complex linked LEI0258 marker is reported. A total of 236 DNA samples were genotyped by capillary electrophoresis from 24 chicken populations. The number of alleles, allele frequency, and heterozygosity levels were used to measure genetic variation at LEI0258 locus in Ethiopian indigenous village chicken. Twenty-nine LEI0258 alleles were observed using capillary electrophoresis that ranged from 185 to 569 bp with no significant difference in allele frequencies between populations.  The number of alleles ranged from 179 (Meseret) to 569 (Batambie), with an average of 9.6 alleles per population. Allelic polymorphism was further evaluated through genotyping by Sanger sequencing. Twenty-three DNA samples with different fragment sizes were re-amplified and their alleles sequenced to depict polympormisms based on the combination of two repeat regions at 12 bp and 13 bp, respectively, and flanking regions with SNP and indels. The repeat region at 12 bp appeared 2 to 18 times, whereas the region at 13 bp appeared invariant in all populations. Sequence relationships revealed two distinct groups of alleles. The number of indels and mutations were 33 and 17, respectively. From capillary electrophoresis, the fixation coefficient of the sub-population within the total population (FST), inbreeding/fixation/ coefficient of an individual in a sub population (FIS) and total inbreeding /heterozygosity deficit/ coefficient of an individual within the total population (FIT) in the locus was 0.03, 0.08 and 0.11, respectively. Three percent of the genetic diversity was due to differences among populations, where as 8% and 89% were variations among individuals and variations within individuals, respectively. Despite the overall low genetic differentiation, both fragment and sequencing analysis revealed high allelic and genetic variability across the 24 populations. The high diversity at LEI0258 in Ethiopian indigenous village chicken populations supports the importance of the MHC region in relation to the disease challenges faced by smallholder poultry farmers across Ethiopia. We recommend that breed improvement programs ensure the maintenance of this diversity by selecting breeding stock as diverse as possible at the LEI0258 locus. 
埃塞俄比亚地方鸡主要组织相容性复合体(MHC)区LEI0258位点的遗传变异
本土鸡在当地适应了环境挑战,为非洲数百万农民提供了生计。然而,与外来菌株相比,它们的生产力仍然很低。正在努力将本地鸡的地方适应性与外来鸡的生产特性结合起来。了解遗传多样性和环境挑战之间的联系,有助于在小农水平上进行标记辅助的品种改良计划,以实现可持续的鸡肉生产。位于MHC区域的LEI0258 VNTR位点的遗传变异与商品品种的传染病抗性/易感性有关。本研究的目的是调查埃塞俄比亚24个鸡种群的等位基因变异、遗传多样性和遗传关系。本文报道了来自24个埃塞俄比亚地方鸡群体的236只鸡的LEI0258的多样性,这些鸡使用主要的组织相容性复合体连接LEI0258标记。对24个鸡群体的236份DNA进行了毛细管电泳分型。采用等位基因数量、等位基因频率和杂合度测定了埃塞俄比亚土着村鸡LEI0258位点的遗传变异。毛细管电泳检测到29个LEI0258等位基因,等位基因频率在185 ~ 569 bp之间,不同群体间无显著差异。等位基因数量从179个(Meseret)到569个(Batambie)不等,平均每个群体有9.6个等位基因。通过Sanger测序进行基因分型进一步评估等位基因多态性。对23份不同片段大小的DNA样本进行了重新扩增,并对其等位基因进行了测序,以分别在12 bp和13 bp处的两个重复区域以及具有SNP和indels的侧翼区域的组合为基础,描绘了多态性。12bp的重复序列出现2 ~ 18次,而13bp的重复序列在所有群体中没有变化。序列关系揭示了两组不同的等位基因。索引数33个,突变数17个。毛细管电泳结果表明,该位点亚种群内固定系数(FST)、亚种群内个体近交/固定/系数(FIS)和种群内个体总近交/杂合缺陷/系数(FIT)分别为0.03、0.08和0.11。3%的遗传多样性是由于群体之间的差异,其中8%和89%分别是个体之间和个体内部的差异。尽管总体上遗传分化程度较低,但片段和测序分析显示,24个群体的等位基因和遗传变异都很高。埃塞俄比亚土著村庄鸡群LEI0258的高度多样性支持了MHC区域在埃塞俄比亚各地小农家禽养殖户面临的疾病挑战方面的重要性。我们建议通过在LEI0258位点选择尽可能多样化的种畜,以确保维持这种多样性。
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