Identification of allelic vatriants in melanocortin-1 receptorgene in llama (Lama glama)

DAVERIO MARIA S, DI ROCCO F, ARBELETCHE VIDAL RIOJA, L

Cairns

Congreso; 33rd Conference of the ISAG; 2012

International Society for Animal Genetics

Poster Nº3016 - ISAG 2012Identification of allelic variants in Melanocortin-1 receptor gene in llama (Lama glama)MARIA .S. DAVERIO1; FLORENCIA. DI ROCCO1 and LIDIA VIDAL RIOJA11Laboratorio de Genética Molecular, Instituto Multidisciplinario de Biología Celular (IMBICE), CIC-PBA. CCT-CONICET, Calle 526 e/ 10 y 11, CP(1900), La Plata, Buenos Aires, Argentina. m.sil.daverio@hotmail.comINTRODUCTIONThe llama (Lama glama) is a South American camelid widely distribuited in Argentina where an interesting regional variation of coat color phenotype is shown.The coat color in mammals is regulated by several genes among wich, MC1R (melanocortin-1 receptor), plays an essential role in pigmentation development. In llamas, segregation of coat color have been assessed by classical genetics (Frank et al.,2006), but molecular bases of this trait remain unknown. The objectives of this research were: 1) characterize and locate single nucleotide polymorphisms (SNPs) in MC1R gene of llama 2) identify allelic variants 3) determine the association between allelic variants and coat color patterns.MATERIALS & METHODSDNA was obtained from 48 llamas located in Villaguay (Entre Ríos), Laguna Blanca (Catamarca) and Abra Pampa (Jujuy) and from 10 guanacos (Lama guanicoe) from Argentinean Patagonia. Four groups of coat color pattern (red-brown with black face and trim; dark brown, light brown and white no albine) were analized (Fig.1). PCR amplification of the complete MC1R gene was performed in all animals using 2 pairs of primers designed on the mRNA sequence of alpaca (GenBank: EU135880.1). Amplicons were visualised in 2% agarose electrophoresis, purified and automatically sequenced. Sequences were aligned by Clustal W software and SNPs identified with Geneious v5.6. Arlequin v3.5 was used for allelic variants determination. Then, association of alleles with the coat color was analysed by comparing each allele with the phenotype of each llamaRESULTSThe llama MC1R gene comprised 1173bp including 130bp of 5´UTR, a single exon of 954bp and 89bp of 3´UTR. Twelve out of thirteen SNPs were located in coding region nine of wich showed amino acid substitutions. C205A and A638G are novel SNPs for llamas not reported previously in alpacas (Table1).To construct the alleles 3 SNPs (A259G/A376G/T383C) seen in more than five animals were selected (Table 1,)The llama MC1R gene three alleles are shown in Table 2.The wild-type allele (MC1R*3) was determined in guanaco and observed in all colour groups. MC1R*1 was found in red-brown with black face and trim phenotype and other colored patterns but it was absent in white animals. MC1R*2 in homozygote condition was found, in high frequency in white animals but not in red-brown with black face and trim phenotype. (Table 3)CONCLUSIONSOur results suggest a relationship between the white and red-brown with black face and trim phenotypes with and allele variants of the MC1R.MC1R*2 was found in white llamas but was absent in those red-brown with black face and trim. Direct correlation between MC1R alleles and eumelanic/pheomelanic patterns could not be established.