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The objective of this work was to sequence the MyoD1, MyoG, MyF5, MyF6, and MSTN genes and to identify polymorphisms in Santa Inês sheep (Ovis aries). A total of 192 lambs with 240 days of age were evaluated, and these genes were sequenced to be compared with the reference sequence in the Ovis aries genome. Genotype and allele frequencies were estimated, and the Hardy-Weinberg equilibrium was tested. Fragments containing 2,493 bp (MyoD1), 1,836 bp (MyoG), 2,813 bp (MyF5), 1,126 bp (MyF6), and 2,380 bp (MSTN) were obtained, and, in these sequences, 160 variants were identified. These polymorphisms were distributed as follows: 59 (MyoD1), 24 (MyoG), 63 (MyF5), 4 (MyF6), and 10 (MSTN). One hundred and four were novel polymorphisms, 45 in MyoD1, 2 in MyoG, 56 in MyF5, and 1 in MSTN. Regarding site, 61 were in intron (27 in MyoD1, 16 in MyoG, 5 in MyF5, 3 in MyF6, and 10 in MSTN), 87 in coding region (22 in MyoD1, 8 in MyoG, 56 in MyF5, and 1 in MyF6), and 12 on 3’UTR (10 in MyoD1 and 2 in MyF5). Therefore, the MyoD family and MSTN genes have several polymorphisms in Santa Inês sheep, which can be useful for association studies. Resumo: O objetivo deste trabalho foi sequenciar os genes MyoD1, MyoG, MyF5, MyF6 e MSTN e identificar polimorfismos em ovinos Santa Inês (Ovis aries). No total, 192 cordeiros com 240 dias de idade foram avaliados, e estes genes foram sequenciados para comparação com a sequência-referência no genoma de Ovis aries. As frequências genotípicas e alélicas foram estimadas, e o equilíbrio de Hardy-Weinberg, testado. Foram obtidos fragmentos contendo 2.493 pb (MyoDl), 1.836 pb (MyoG), 2.813 pb (MyF5), 1.126 pb (MyF6) e 2.380 pb (MSTN), e, nessas sequências, foram identificadas 160 variantes. Esses polimorfismos foram distribuídos da seguinte forma: 59 (MyoD1), 24 (MyoG), 63 (MyF5), 4 (MyF6) e 10 (MSTN). Foram encontrados 104 novos polimorfismos, sendo 45 no MyoD1, 2 no MyoG, 56 no MyF5 e 1 no MSTN. Com relação ao local, 61 variantes estavam em íntron (27 no MyoD1, 16 no MyoG, 5 no MyF5, 3 no MyF6 e 10 no MSTN), 87 em região codificante (22 no MyoD1, 8 no MyoG, 56 no MyF5 e 1 no MyF6) e 12 na região 3’UTR (10 no MyoD1 e 2 no MyF5). Portanto, os genes da família MyoD e o MSTN possuem vários polimorfismos em ovinos da raça Santa Inês, os quais podem ser úteis em estudos de associação.

Growth hormone (GH), insulin-like growth factor 1 (IGF1), and leptin (LEP) can be candidate genes for association studies because they play vital roles in the metabolism process. Thus, this study aimed to identify variants in these genes associated with body traits in Santa Inês sheep. The following were recorded: body weight at 100 (BW100) and 240 days (BW240), average daily gain (ADG), withers (WH) and croup (CH) heights, body length (BL), thoracic (TG) and leg (LG) girths, thoracic (TW) and croup (CW) widths, body depth (BD), rib eye area (REA), fat thickness (FT), and carcass finishing score (CFS). Single-locus association analysis was performed with 11 variants in IGF1, 18 in LEP, and 16 in GH. Moreover, two haplotypes in IGF1 and one haplotype in LEP were evaluated in haplotype association analysis. The single-locus analysis revealed 23 suggestive additive effects (p < 0.05), but no additive effect was found at the Bonferroni threshold. Haplotype association analysis revealed 19 additive effects, of which ten were at the Bonferroni threshold (p < 0.0074). In IGF1 gene, haplotype replacements were associated with ADG 20.51(7.37), CH 4.09(1.21), WH 3.52(1.20), BL 3.94(1.19), TG 3.88(1.30), TW 1.13(0.36), and LG 3.40(1.08); while in the LEP gene the haplotype replacement was associated with BW100 1.83(0.51), BD −2.51(0.56), and CFS −0.24(0.06). Therefore, there are haplotypes in IGF1 and LEP genes associated with body traits in Santa Inês sheep, which can be useful in marker-assisted selection.

µ-calpain (CAPN1) and calpastatin (CAST) genes play key roles in protein turnover. The present study aimed to identify the variants in these genes associated with growth and ultrasound carcass traits in Santa Inês sheep. A sample of 192 no full sibling Santa Inês lambs was used. Fragments of the CAST and CAPN1 genes were amplified and next-generation sequencing was performed in the MiSeq platform. Variants in the CAPN1 and CAST sequences were then detected using bioinformatic tools. Withers and croup heights, body length, thoracic and croup widths, thoracic and leg girths, body depth, carcass fat score, rib eye area, fat thickness, body weights were recorded at weaning and at 140 days post-weaning, and average daily gain post-weaning was calculated. Both single-locus and haplotype association analyses were performed with the model as follows: farm (2 levels), year (4 levels), the month of birth (12 levels), and the covariate age of the animal. The fragments amplified included 4,514 bp between the 20th and 23rd exons of CAST as well as 3,927 bp between the 12th and 21st exons of CAPN1. In these regions, 58 (CAST) and 45 (CAPN1) variants were identified. In the CAST gene, the single-locus analysis revealed 22 suggestive additive effects (PCAPN1 gene, the single-locus analysis identified seven suggestive additive effects, while only one haplotype replacement effect on fat thickness (–0.0143±0.0053) was detected. The results of the present study suggest that variants in the CAPN1 and CAST genes are associated with growth and ultrasound carcass traits in Santa Inês sheep, which may be a source of information to improve knowledge regarding the genetic control of these traits.

Myostatin and MyoD family genes play vital roles in myogenesis and this study aimed to identify association of variants in MyoD1, MyoG, MyF5, MyF6, and MSTN genes with meat traits in Santa Ines sheep. A dataset with 44 variants and records of seven meat traits in 192 lambs (pH0, pH24, a*, b*, L*, tenderness assessed by shear force, and water-holding capacity) was used. Single-locus and haplotype association analyses were performed, and the significance threshold was established according to Bonferroni's method. Single-locus analysis revealed two associations at a Bonferroni level, where the variant c.935-185C > G in MyoD1 had an additive effect (-4.31 ± 1.08 N) on tenderness, while the variant c.464 + 185G > A in MyoG had an additive effect (-2.86 ± 0.64) on a*. Additionally, the haplotype replacement GT>AC in MSTN was associated with pH0 (1.26 ± 0.31), pH24 (1.07 ± 0.27), a* (-1.40 ± 0.51), and tenderness (3.83 ± 1.22 N), while the replacement GT > AG in MyoD1 was associated with pH0 (1.43 ± 0.26), pH24 (1.25 ± 0.22), b* (-1.06 ± 0.39), and tenderness (-4.13 ± 1.16 N). Our results have demonstrated that some variants in MyoG, MyF6, MyoD1, and MSTN can be associated with physicochemical meat traits in Santa Ines sheep.