C/EBPα gene as a genetic marker for beef quality improvement.

Intramuscular fat (IMF) or intramuscular triglycerides are interspersed throughout the skeletal muscles. The IMF, also called marbling, imparts meat with flavor and juiciness and is one of the core criteria for judging carcass value. The quantity of IMF is influenced entirely by genetics. Recently, understanding the underlying genetic bases of IMF has been a focus particularly in the beef industry. In this study, with the deep insights of ameliorating the beef quality by genetic means, the role of the CCAAT/enhancer binding protein alpha (C/EBPα) gene was investigated by over-expressing C/EBPα in bovine muscle stem cells (MSCs) to initiate the adipogenic program. Prior to this, bovine MSCs were isolated and induced to differentiate into adipocytes from cells that were exposed to dexamethasone isobutylmethylxanthine and indomethacin; the presence of insulin and fetal bovine serum was examined. Either ectopic expression of C/EBPα or treatment with dexamethasone and insulin induced the accumulation of fat droplets and the expression of adipogenic induction genes (LPL, PPARγ, C/EBPß, and C/EBPδ). The expression levels of myoblast-related genes (MyoD, Myf5, and Pax7) were also measured to assess the accuracy of the differentiation process. This study provides evidence that the C/EBPα gene is essential for cattle adipose tissue growth and development. Hence, this finding can contribute to improving beef carcass quality.

Production of transgenic beef cattle rich in n-3 PUFAs by somatic cell nuclear transfer.

OBJECTIVES: Omega-3 polyunsaturated fatty acids (n-3 PUFAs) are beneficial to human health. However, the n-3 PUFAs contents of the livestock meat that we consume daily are relatively low. Utilization of transgenic technology to increase n-3 PUFAs contents in livestock may solve this problem. RESULTS: The omega-3 fatty acid desaturase (FAD3), encoded by fat1 gene derived from Caenorhabditis elegans (C. elegans), converts omega-6 polyunsaturated fatty acids (n-6 PUFAs) to n-3 PUFAs. In the study, a plasmid containing the codon-optimized C. elegans fat1 gene (mfat1) was constructed and used to produce transgenic beef cattle by somatic cell nuclear transfer. Fourteen transgenic calves were obtained, and the ratio of n-6 to n-3 PUFAs in the transgenic calves decreased from 5.33: 1 to 0.95: 1 compared with negative controls. CONCLUSIONS: Our results demonstrated that the codon-optimized C. elegans mfat1 gene can be functionally expressed in the beef cattle and converts n-6 PUFAs to n-3 PUFAs.

Polymorphisms of the Osteocrin gene and its association with meat quality traits in Qinchuan cattle.

Here, we detected 2 SNPs, A85C and T335C, that were located on the 3rd exon and the 3 untranslated regions of the bovine Osteocrin gene, respectively, using 413 Qinchuan cattle DNA samples. PCR-SSCP and DNA sequencing methods were specifically used. Three genotypes (AA, AC, and CC) were found at A85C; yet, only 2 genotypes (TC and CC) were found at T335C. Association analysis showed that both loci were associated with certain meat quality traits, including back fat thickness and loin muscle area. At the A85C locus, individuals with the CC genotype had greater back fat thickness. In comparison, at the T335C locus, individuals with the TC genotype had greater back fat thickness and a larger loin muscle area. Therefore, these 2 SNPs could be used as genetic markers to enhance Qinchuan cattle breeding programs.

Polymorphisms in the delta-like 2 homolog gene and their association with growth and meat-quality traits in Qinchuan cattle.

The delta-like 2 homolog (DLK2) modulates adipogenesis, hematopoiesis, osteogenesis, and other cell-differentiation processes. In the present study, we detected potential polymorphisms in the DLK2 gene in 604 individuals of Qinchuan cattle by using PCR-RFLP and DNA-sequencing methods. Herein, we identified five novel single-nucleotide polymorphisms (SNPs) (g.888G>A, g.910A>G, g.995G>A, g.4321A>G, g.4850A>G) and analyzed their association with measured traits. Four of the five analyzed polymorphisms were associated with at least one of the following traits: body weight (BW), chest depth (CD), chest circumference (CC), back fat thickness (BT), and rib-eye area (REA). To the best of our knowledge, our research is the first to report the association of DLK2 gene polymorphisms with growth and meat quality traits in Qinchuan cattle. In summary, the results of our study suggest that the DLK2 gene can be used as a candidate gene in beef cattle breeding.

Novel SNPs in the exon region of bovine DKK4 gene and their association with body measurement traits in Qinchuan cattle.

The aim of this study was to determine whether single nucleotide polymorphisms (SNPs) of bovine Dickkopf homolog 4 (DKK4) are associated with body measurement traits in Qinchuan cattle. By using PCR-SSCP technology and DNA sequencing, we discovered 5 DKK4 SNPs in Qingchuan cattle, including -65G>A and -77G>T in the 5'-untranslated region, 1532C>G and 1533T>C in exon 2, and 2088C>T in exon 3. The sequencing map showed that 1532C>G and 1533T>C were in close linkage disequilibrium and were treated as 1532C>G-1533T>C in this study. Allele frequencies were calculated and analyzed by the chi-square test, which showed that -65G>A and 1532C>G-1533T>C were in Hardy-Weinberg equilibrium (P > 0.05), whereas -77G>T and 2088C>T were not in all 633 tested Qinchuan cattle individuals (P < 0.01). Gene heterozygosity (HE), effective allele number (NE), and polymorphism information content (PIC) were 0.407, 1.686, and 0.324 at -65G>A; 0.472, 1.894, and 0.361 at -77G>T; 0.476, 1.908, and 0.363 at 1532C>G-1533T>C; and 0.218, 1.279, and 0.195 at 2088C>T. We also evaluated the potential association of these SNPs with body measurement traits in all 633 individuals; the results suggest that several SNPs in Qinchuan cattle DKK4 were significantly associated with body length, hip height, rump length, hip width, heart girth, and pin bone width (P < 0.05 and P < 0.01). These results suggest that bovine DKK4 could be used as candidate gene for Qinchuan cattle breeding.

Bovine GDF10 gene polymorphism analysis and its association with body measurement traits in Chinese indigenous cattle.

The objective of this research was to detect bovine GDF10 gene polymorphism and analyze its association with body measurement traits (BMT) of animals sampled from 6 different Chinese indigenous cattle populations. The populations included Xuelong (Xl), Luxi (Lx), Qinchuan (Qc), Jiaxian red (Jx), Xianang (Xn) and Nanyang (Ny). Blood samples were taken from a total of 417 female animals stratified into age categories of 12-36 months. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) was employed to find out GDF10 single polymorphism nucleotide (SNPs) and explore their possible association with BMT. Sequence analysis of GDF10 gene revealed 3 SNPs in total: 1 in exon1 (G142A) and 2 in exon3 (A11471G, and T12495C). G142A and T12495C SNPs are both synonymous mutation. They showed 2 genotypes namely respectively (GG, GA) and (PP and PB). A11471G SNP is a missense mutation leading to the change of Alanine to Threonine amino acid. It showed three genotypes namely AA, BB and AB. Analysis of association of polymorphism with body measurement traits at the three locus showed that there were significant effects on BMT in Qc, Jx and Ny cattle population. These results suggest that the GDF10 gene might have potential effects on body measurement traits in the above mentioned cattle populations and could be used for marker-assisted selection.

Back fat thickness and meat tenderness are associated with a 526 T→A mutation in the exon 1 promoter region of the MyF-5 gene in Chinese Bos taurus.

Qualitative trait loci (QTL) for growth and meat quality traits in cattle (Bos taurus) have been previously mapped to three chromosome regions, 0 to 30, 55 to 70, and 70 to 80 cM on chromosome 5. We evaluated the allele frequencies and gene-specific single nucleotide polymorphisms (SNPs) of bovine myogenic factor 5 (MyF-5) in the QTL regions and their associations with live weight and meat characteristics in indigenous Chinese cattle breeds. PCR-SSCP methodology showed a T>A mutation at 526 bp. Least square analysis revealed a significant association of this SNP with backfat thickness and meat tenderness (P < 0.05), while no significant association was found with live weight, loin eye height, loin eye area, rib area, or water holding capacity. Allele frequencies of MyF-5-A/B in the five breeds were 0.760/0.239, 0.752/0.247, 0.629/0.370, 0.715/0.284, and 0.750/0.250, for JiaXian red, Luxi, Nanyang, Qinchuan, and XiaNan crossbreed, respectively. The genotype distributions for these alleles in two of the Chinese cattle breeds (Luxi and Qinchuan) were not in Hardy-Weinberg equilibrium (P < 0.05); while those for the other three breeds (JiaXian red, Nanyang, and XiaNan) were in agreement with Hardy-Weinberg equilibrium (P > 0.05). The genotypic frequencies among all five cattle breeds showed moderate diversity (0.25 < polymorphism information content < 0.5). Based on our findings, we suggest that the MyF-5 gene influences back fat thickness and meat tenderness in Chinese Bos taurus. This SNP could be useful for marker-assisted selection for meat quality traits in these cattle.

Lack of an association between a single nucleotide polymorphism in the bovine myogenic determination 1 (MyoD1) gene and meat quality traits in indigenous Chinese cattle breeds.

The myogenic determination 1 (MyoD1) gene is a member of the MyoD gene family. It encodes for skeletal muscle-specific transcription factors containing highly conserved basic helix-loop-helix regions that perform important roles in the initiation, maintenance, and regulation of phenotypic traits. We investigated a new single nucleotide polymorphism (SNP) in the MyoD1 gene to evaluate whether this polymorphism affects meat quality traits in five Chinese indigenous cattle breeds, namely Qinchuan (QC), Xia-Nan (XN), Nan-yang (NY), Luxi (LX), and Jia-xian red (JXR). A C→G transversion at position 624 was detected in exon 1 of the MyoD1 gene; it causes an amino acid substitution ((624)serine/(624)cysteine). Least squares analysis showed that this SNP is not significantly associated with back fat thickness, eye muscle area, intramuscular fat, or marbling. The A/B allelic frequencies in the five breeds were 0.810/0.189, 0.779/0.220, 0.768/0.231, 0.820/0.180, and 0.801/0.198, respectively. Based on the χ(2) test, the genotype distributions of four cattle breeds (LX, NY, QC, and XN) did not agree with Hardy-Weinberg equilibrium (P < 0.05); one breed (JXR) did not deviate significantly from Hardy-Weinberg equilibrium (P > 0.05). The genotypic frequencies among all five cattle breeds showed moderate diversity (0.25 < polymorphism information content < 0.5). We concluded that the C624G SNP of the MyoD1 gene does not influence meat quality traits in indigenous Chinese cattle breeds; however, this SNP could be included in breed composition and population admixture analyses due to the marked differences in allelic frequencies among these five breeds.

A novel polymorphism of the GDF10 gene and its association with body measurement traits in Chinese indigenous cattle.

Body measurement traits are known to play numerous important roles in the assessment of productivity and economic value. They are influenced by several factors, among which genetic factors are predominant. The gene GDF10 is involved in skeletal morphogenesis and is associated with body measurement traits. It may be an important candidate gene for marker-assisted selection. We used the PCR-SSCP technology to examine a possible association of the single nucleotide polymorphism (SNP) (G142A) of the bovine GDF10 gene with body measurement traits in 417 animals belonging to six different Chinese cattle populations: Xue long (Xl), Luxi (Lx), Qinchuan (Qc), Jiaxian red (Jx), Xianang (Xn), and Nanyang (Ny). In the Jx population, least squares analysis revealed significant effects on hip width, chest depth and chest circumference. The animals with the GG genotype had higher mean values than those with the GA genotype for all three traits. We conclude that the SNP of the GDF10 gene could be a very useful genetic marker for body traits in Jx cattle reproduction and breeding.