Polymorphisms in MC1R and ASIP genes associated with color phenotypes in alpaca Huacaya

PINARES, RUBÉN; CRUZ, ALAN; DAVERIO MARIA SILVANA; DI ROCCO FLORENCIA; PONCE DE LEÓN, FEDERICO ABEL; WURZINGER, MARÍA; GUTIÉRREZ, GUSTAVO AUGUSTO

Simposio; 8th European Symposium on South American Camelids 4th European Meeting on Fibre Animals; 2022

The wide phenotypic diversity in alpacas results from human selection by color phenotypes thatfavored the fixation of main alleles in MC1R and ASIP genes. In this context, the objectives ofthis study were: to characterize the fiber color by colorimetry and identify the mainpolymorphisms in MC1R and ASIP genes associated with black and brown alpacas. Fiber andblood samples of 98 alpacas were obtained from Pacomarca Research Station; and 9 vicuñasfrom Abancay province were considered as reference for this study (Peru). Fleece colorphenotypes were determined by colorimetry using Chroma Meter CR-210. DNA was extractedfrom 200 μL of EDTA anticoagulated blood using the commercial kit (Quick-DNA™ MiniprepPlus Kit). Polymerase chain reaction (PCR) primers were designed to amplify MC1R and ASIPgenes following Daverio et al. (2016). For all PCR products purification we used exo-sapmethod, with enzyme ExoASP-IT® (usb). The purified amplicons were sequenced by theSanger method at Macrogen Inc using the original PCR forward and reverse primers. CompleteMC1R and ASIP coding sequences for each animal were obtained and analyzed using GeneiousSoftware Version 11.1.5. The CIE L*a*b* system, L* = lightness showed low values ineumelanic alpacas (black and black-brown) and high values in white, pheomelanic brownalpacas and vicuñas. Coding sequence of MC1R (CDS) consisted of 954bp and encoded a 317amino acid protein. Inside of CDS a heterozygous deletion c.224_227del and nine SNPs wereobserved; 5 non synonymous SNPs: c.82A>G, c.259G>A, c.376G>A, c.587T>C, c.901C>T(p.T28A, p.M87V, p.G126S, p.F196S, p.R301C, respectively) and 4 synonymous SNPs:c.126C>T, c.354C>T, c.618G>A, c.933A>G. Two non-synonymous polymorphisms(c.292C>T and c.353G>A) and a 57bp deletion (c.325_381del) were identified within exon 4of ASIP gene. The five non synonymous SNPs at MC1R and the mutations at ASIP define therecessive genotypes (ASIP) together with the dominant genotypes at MC1R (EEaa) in blackand black-brown alpacas, heterozygote genotypes for both genes (EeAa) was observed inbrown, dark brown and black alpacas. The wild genotype (E+E+A+A+) was observed in white,brown alpaca and vicuña. The nine vicuñas have the wild allele without deletion in MC1R andASIP genes. In sum, there is more than one genotype for black and brown phenotypes. Likewise,for each genotype described we also observed black and brown phenotypes; then it would beexplained by the epistatic interaction between MC1R and ASIP genes.