AGPAT6 polymorphism and its association with milk traits of dairy goats

As one of the eight members in the 1-acylglycerol-3-phosphate-O-acyltransferase (AGPATs) family, AGPAT6 is a crucial enzyme for the biosynthesis of glycerolipids and triacylglycerol in eukaryotes, as well as catalyzing the conversion from lysophosphatidic acid to phosphatidic acid. AGPAT6 can be considered as a candidate gene for regulating milk composition. DNA sequencing and PCR-RFLP methods were applied to detect genetic variation in the AGPAT6 gene in 549 Chinese dairy goats. Four polymorphisms (NC_007328.3:g.152G>C, 8124G>A, 9263C>G, 16436G>A) were detected in 5'UTR, intron 2, exon 4, and 3'UTR, respectively. For the KpnΙ locus, the frequencies of the AGPAT6-G allele were 0.955 and 0.936 for SN (Xinong Sannen) and GZ (Guanzhong) dairy goat breeds, respectively. In the PCR-RFLP analysis for KpnΙ, EcoRII, NcoΙ, and BglΙ, the frequencies of the G allele of AGPAT6 were 0.955 and 0.936, 0.694 and 0.819, 0.206 and 0.254, 0.729 and 0.623 for SN and GZ dairy goat breeds, respectively. The 9263C>G mutation revealed a synonymous genetic code of Thr (threonine). Associations between the four mutations and milk traits were analyzed in two dairy goat breeds. At the 9263C>G locus, genotype GG and CG individuals showed significantly better milk performance than genotype CC individuals (P < 0.05). Therefore, the G allele is suggested to be a molecular marker for milk production in dairy goats.     

5个山羊品种CSN1S2基因的Alw26I酶切

利用PCR-RFLP技术对西农萨能奶山羊、关中奶山羊、陕南白山羊、安哥拉山羊和波尔山羊5个山羊品种的170个个体的αs 2酪蛋白(CSN1S2)基因进行多态性分析,结果表明：扩增大小为310 bp的片段经限制性内切酶Alw26I酶切后表现多态,且5个山羊品种该基因座位均处于Hardy-Weinberg平衡状态。西农萨能奶山羊、关中奶山羊、陕南白山羊、安哥拉山羊和波尔山羊的基因杂合度/有效等位基因数/Shaanon信息熵/PIC值分别为0.1589/1.1889/0.2955/0.1463, 0.4114/1.6981/0.6017/0.5171,0.1653/1.1980/0.3046/0.1516,0.0646/1.0691/0.1463/0.0625,0.0541/1.0572/ 0.1270/ 0.0526。分析结果显示,关中奶山羊的遗传多样性最丰富,表现为高度多态;其次是西农萨能奶山羊和陕南白山羊,而安哥拉山羊和波尔山羊的遗传变异程度最低。     

A novel SNP of α-lactalbumin gene in Chinese dairy goats

The α-lactalbumin (α-LA) plays a key role in lactose synthesis in mammary glands of domestic animals. Mutations in the α-LA gene are associated with the milk traits in dairy cattle. In our study, a novel SNP: NO_X06366: g.875 C > T was detected in 708 dairy goat individuals—268 of the Xinong Saanen breed and 440 of Guanzhong breed, which revealed a synonymous mutation in the exon 1 of α-LA gene. The Polymerase Chain Reaction-Single Strand Conformation Polymorphism (PCR-SSCP) and sequencing techniques showed that there were three genotypes: CC, CT and TT. Moreover, the χ²-test showed that the genotype frequencies of the two breeds were in good agreement with the Hardy-Weinberg equilibrium (P > 0.05). The relationship of the polymorphism of dairy goat α-LA gene with the milk trait and the body size trait was revealed. Individuals with the CC genotype were significantly smaller at chest circumference than those with CT (P < 0.05) in both breeds. But the milk trait and other body size traits of the two dairy goat breeds had no significant association with genotypes studied.     

Identification of a functional SNP in the 3′‐UTR of caprine MTHFR gene that is associated with milk protein levels

Xinong Saanen (n = 305) and Guanzhong (n = 317) dairy goats were used to detect SNPs in the caprine MTHFR 3′‐UTR by DNA sequencing. One novel SNP (c.*2494G>A) was identified in the said region. Individuals with the AA genotype had greater milk protein levels than did those with the GG genotype at the c.*2494 G>A locus in both dairy goat breeds (P < 0.05). Functional assays indicated that the MTHFR:c.2494G>A substitution could increase the binding activity of bta‐miR‐370 with the MTHFR 3′‐UTR. In addition, we observed a significant increase in the MTHFR protein level of AA carriers relative to that of GG carriers. These altered levels of MTHFR protein may account for the association of the SNP with milk protein level.     

Effects of genetic variability of the dairy goat growth hormone releasing hormone receptor (<Emphasis Type="Italic">GHRHR</Emphasis>) gene on growth traits

Growth hormone-releasing hormone receptor (GHRHR) plays a critical role in growth hormone (GH) synthesis, release and regulation of pituitary somatotroph expansion in vertebrates. The objective of this study was to investigate variations in goat GHRHR gene and their associations with growth traits in 668 dairy goats. The results showed four novel single nucleotide polymorphisms (SNPs): NC_007302:g.5203C>T, 7307C>G, 9583G>A and 9668A>C. In detail, the novel SNP C>T in the 5203rd nucleotide identified a missense mutation: CCC (Pro)>TCC (Phe) at position 116aa of the goat GHRHR (423aa). Besides, 9583G>A and 9668A>C polymorphism were in complete linkage disequilibrium. The genetic diversity analysis revealed that the Guanzhong dairy goat possessed intermediate genetic diversity in P3 and P7 loci, and the Xinong Sannen dairy goat belonged to poor genetic diversity in P4 locus. Significant associations between the genotypes of P3 locus and body length, body height and chest circumference was observed in Guanzhong goat (P < 0.05). However, in Xinong saanen population, significant statistical difference was only found in body height and body length (P < 0.05). In P4 and P7 loci, no significant associations were detected between any variant sites and body length, body height and chest circumference, as well as for the milk traits (P > 0.05). These results strongly suggested that the goat GHRHR gene is a candidate gene that influences growth traits in dairy goat.     

Polymorphism identification in goat GNRH1 and GDF9 genes and their association analysis with litter size

This study investigated the polymorphisms of GNRH1 and GDF9 genes in 641 goats of three breeds: Xinong Saanen, Guanzhong and Boer. Two allelic variants were identified in the GNRH1 gene (JN645280:g.3548A>G and JN645281:g.3699G>A) and one allelic variant was found in the GDF9 gene (JN655693:g.4093G>A). Furthermore, g.4093G>A was a missense mutation (p.Val397Ile of GDF9). Results of an association analysis indicated that SNPs g.3548A>G and g.4093G>A had significant effects on litter size (P < 0.05). The combination genotypes of SNPs g.3548A>G, g.3699G>A and g.4093G>A also affected litter size with the C₅ genotype having the highest litter size in the first, third, fourth and average parity. Hence, the biochemical and physiological functions, together with the results obtained in our investigation, suggest that the GNRH1 and GDF9 genes could serve as genetic markers for litter size in goat breeding.     

The CSN1S1?N and F alleles identified by PCR-SSCP and their associations with milk yield and composition in Chinese dairy goats

A method was depicted to identify null allele CSN1S1 N and low allele CSN1S1 F of the CSN1S1 gene of goat using PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism). First, primer A was designed to amplify the exon 9 of CSN1S1 gene which produced three genotypes AA, AB, and BB. Among these three genotypes, only AA and AB individuals had a cytosine deletion at exon 9 after DNA sequencing, which cannot be used to identify the N and F alleles. Therefore, primer B was used to amplify intron 14 of CSN1S1 of described AA and AB individuals. Genotypes FF, FN and NN were detected within AA individuals and genotypes FO and NO were detected in the above AB individuals. The frequencies of F and N alleles in 708 samples from Xinong Saanen (XS) and Guanzhong (GZ) dairy goat breeds were 0.1139, 0.0927, and 0.2376, 0.1193, respectively. In 268 XS samples, the individuals with NN genotype contained a significant lower protein content than that of other genotypes (P<0.01). Individuals of FF genotype had significant higher milk yield than that of NO genotype in the first milk lactation of 202 XS individuals (P<0.05). Therefore, the variability at CSN1S1 locus contains enough genetic diversity to be potentially useful in improving the quality and production of milk in Chinese dairy goat breeds.     

Two mutations in the 5′‐flanking region of the KITLG gene are associated with litter size of dairy goats

In this study, Xinong Saanen (SN) and Guanzhong (GZ) dairy goat breeds were used to detect single nucleotide polymorphisms (SNPs) in the 5′‐flanking region of the KITLG gene by DNA sequencing and primer‐introduced restriction analysis–polymerase chain reaction. Two novel SNPs (g.13090G>T and g.13664C>A) were identified (GenBank Accession no. KM658964). Furthermore, g.13090G>T and g.13664C>A loci were closely linked in SN and GZ breeds (r² > 0.33). Association analysis results showed that g.13090G>T and g.13664C>A SNPs significantly affected litter size (P < 0.05). The litter size of individuals with the combined genotype GG/CC from both dairy goat breeds was greater than that of individuals with TT/AA in average parity (P < 0.05). Known biochemical and physiological functions, along with our results, indicated that GG/CC could be used in marker‐assisted selection to choose individuals with greater litter size from both breeds. These results extend the spectrum of genetic variation in the caprine KITLG gene and may contribute to genetic resources and breeding of goats.     

Pseudomonas aeruginosa in Dairy Goats: Genotypic and Phenotypic Comparison of Intramammary and Environmental Isolates

Following the identification of a case of severe clinical mastitis in a Saanen dairy goat (goat A), an average of 26 lactating goats in the herd was monitored over a period of 11 months. Milk microbiological analysis revealed the presence of Pseudomonas aeruginosa in 7 of the goats. Among these 7 does, only goat A showed clinical signs of mastitis. The 7 P. aeruginosa isolates from the goat milk and 26 P. aeruginosa isolates from environmental samples were clustered by RAPD-PCR and PFGE analyses in 3 genotypes (G1, G2, G3) and 4 clusters (A, B, C, D), respectively. PFGE clusters A and B correlated with the G1 genotype and included the 7 milk isolates. Although it was not possible to identify the infection source, these results strongly suggest a spreading of the infection from goat A. Clusters C and D overlapped with genotypes G2 and G3, respectively, and included only environmental isolates. The outcome of the antimicrobial susceptibility test performed on the isolates revealed 2 main patterns of multiple resistance to beta-lactam antibiotics and macrolides. Virulence related phenotypes were analyzed, such as swarming and swimming motility, production of biofilm and production of secreted virulence factors. The isolates had distinct phenotypic profiles, corresponding to genotypes G1, G2 and G3. Overall, correlation analysis showed a strong correlation between sampling source, RAPD genotype, PFGE clusters, and phenotypic clusters. The comparison of the levels of virulence related phenotypes did not indicate a higher pathogenic potential in the milk isolates as compared to the environmental isolates.     

A novel polymorphism of the lactoferrin gene and its association with milk composition and body traits in dairy goats

Milk composition and body measurement traits, influenced by genes and environmental factors, play important roles in value assessments of efficiency and productivity in dairy goats. Lactoferrin (LF), involved in the efficient expression of protein in milk, is also an anabolic factor in skeletal tissue and a potent osteoblast survival factor. Therefore, it is an important candidate gene for milk composition and body measurement trait selection in marker-assisted selection. We employed PCR-SSCP and DNA sequencing to screen the genetic variations of the LF gene in 549 Chinese dairy goats. A novel single-nucleotide polymorphism (SNP) (G198A in exon II) of the LF gene was detected. The frequencies of the AA genotype were 0.0285 and 0.0261 in GZ and SN populations, respectively. Both populations were found to have low levels of polymorphism and were in Hardy-Weinberg disequilibrium (P < 0.05). We found significant (P < 0.05) associations of the SNP marker with milk protein and acidity in the total population; animals with the AA genotype had higher mean values for milk protein than those with the GA genotype. Animals with genotype AA had higher mean values for withers height than those with genotype GG (P < 0.05). We concluded that this SNP of the LF gene has potential as a genetic marker for milk composition and body traits in dairy goat breeding.     

Physiological and production response of dairy goats bred in a tropical climate

The aim of this work was to determine the adaptability of Saanen and ½Saanen?×?½Anglo-Nubian (½S½AN) goats bred in tropical climates. The study included 30 goats, 15 Saanen and 15 ½S½AN. The data was collected during the rainy and dry seasons. During the whole experimental period, the environment variables were recorded, as well as rectal temperature (RT), superficial temperature (ST), respiratory rate (RR) and heart rate (HR) and milk production (MP). The adaptability coefficient (AC) was calculated for both genotypes. The averages were evaluated by ANOVA at 5 % probability. There was a genotype and period of year effect, as well as the interaction genotype?×?period of year. Pearson's simple correlation analysis was then carried out between milk production and physiological and environment variables. There was a statistical difference (p?p?p?相似文献   

Combined effects of four SNPs within goat PRLR gene on milk production traits

Xinong Saanen (SN, n = 323) and Guanzhong (GZ, n = 197) goat breeds were used to detect single nucleotide polymorphisms (SNPs) in the coding regions with their intron–exon boundaries of prolactin receptor (PRLR) gene by DNA sequencing, primer-introduced restriction analysis-polymerase chain reaction (PIRA-PCR) and PCR-restriction fragment length polymorphism (PCR-RFLP). Four novel SNPs (g.40452T > C, g.40471G > A, g.61677G > A and g.61865G > A) were identified. The g.61677G > A and g.61865G > A SNPs caused amino acid variations p.Ser485Asn and p.Val548Met, respectively. Both g.40452T>C and g.40471G>A loci were closely linked in SN and GZ goat breeds (r² > 0.33). In addition, there was also a close linkage between g.61677G>A and g.61865G>A loci in both goat breeds. Statistical results indicated that the g.40452T > C, g.61677G > A and g.61865G > A SNPs were significantly associated with milk production traits in SN and GZ breeds. Further analysis revealed that combinative genotype C1 (TTAAGGGG) was better than the others for milk yield in SN and GZ goat breeds. These results are consistent with the regulatory function of PRLR in mammary gland development, milk secretion, and expression of milk protein genes, and extend the spectrum of genetic variation of the caprine PRLR gene, which might contribute to goat genetic resources and breeding.     

Combined effects of three novel SNPs within goat LHX3 gene on milk performance

In this study, single nucleotide polymorphisms (SNPs) of LHX3 gene were detected by DNA sequencing based on DNA pools and PCR-RFLP method in 792 goats belonging to three Chinese indigenous breeds (Guanzhong dairy, Xinong Sannen dairy, Inner Mongolia white cashmere). The results revealed three novel mutations (AY923832:g.7778A>T; g.8035T>C; g.10592C>T), which were named as P2-DraI, P3-HinfI and P4-MspI loci, respectively. The linkage disequilibrium analysis demonstrated P3-HinfI and P4-MspI loci were strongly linked (r ²>0.33) in all the analyzed populations. Each SNP produced no significant (p>0.05) effects on milk performance. However, the two-loci and three-loci combined genotypes had more profound impacts on milk yield than in separation. The individuals with diplotype AATT (P2-DraI/P3-HinfI) showed significantly (p<0.05) higher milk yield than those with diplotypes ATTT, TTTT, ATTC, and AACC. Diplotype TTCCTT (P2-DraI/P3-HinfI/P4-MspI) carriers had significantly (p< 0.05) higher milk yield than those with diplotypes ATTTCC and AACCTC. These combined effects of LHX3 gene polymorphisms indicated that this gene had significant effect on milk performance and its corresponding combined diplotypes could be regarded as potential genetic markers of milk performance.     

Polymorphisms of the Toll-Like Receptor 2 of Goats (Capra hircus) may be Associated with Somatic Cell Count in Milk

Toll-like receptor 2 (TLR2) plays an important role in recognition by the innate immune system of Gram-positive bacteria. As Gram-positive bacteria cause mastitis, we examined variations in the region of the TLR2 gene that codes for the extracellular domain. Samples of forty goats from a single dairy herd were collected, half with low SCC (≤200,000 cells/mL), and half with higher SCC. Two synonymous single nucleotide polymorphisms (SNPs) were identified: 840G?>?A and 1083A?>?G. One nonsynonymous SNP 739G?>?A was identified. This coded for valine or isoleucine, which have similar physiochemical properties, and was not in a region coding for a known functional domain. Surprisingly, the least square mean SCC of the heterozygous goats (146,220) was significantly lower than the SCC of homozygous GG goats (537,700; p?=?0.004), although these two groups were similar in days in milk (p?=?0.984), and there was no significant difference by breed (p?=?0.941). Because factors other than mastitis can affect SCC and our sample sizes were limited, additional studies are needed to corroborate an association between TLR2 genotype and SCC or mastitis in goats.     

Insulin-like growth factor I (somatomedin C) in goats during normal lactation and in response to somatotropin treatment

1. Two experiments were performed. In Experiment 1, IGF-I concentrations in serum were followed in four lactating dairy goats during normal lactation and before, during and after 14 days of daily administration of 10 IU recombinant bovine somatotropin, starting 10 weeks post-partum. In Experiment 2, IGF-I was measured in single blood samples obtained from 40 dairy goats in the first part of lactation. 2. Measured IGF-I concentrations ranged from 4.4-9.0 nM. Concentrations varied during the day with coefficients of variation from 8.9 to 18.4% in the four goats in Experiment 1. There were no systematic trends in the diurnal variations (P greater than 0.75). 3. Concentrations of IGF-I remained fairly constant in all goats in Experiment 1 during the lactational period, and were not related to the changes in milk yield (P greater than 0.65). Similarly, no relationship was found in Experiment 2 between level of milk production and IGF-I concentration in serum. 4. Somatotropin treatment stimulated milk yields by 14 +/- 2.9% (P less than 0.05) and IGF-I concentration by 27.9 +/- 6.7% (P less than 0.05). 5. IGF-I is the most likely factor which increases mammary gland synthetic capacity in response to somatotropin treatment, and thus is responsible for the galactopoietic effect of this hormone being exerted. 6. However, serum IGF-I may primarily be involved in regulation of nutrient partitioning during normal lactation, but a paracrine synthesis with local effect on the mammary gland cannot be ruled out.     

Study on polymerization effect of polyembryony genes by SSCP marker and family trees in Chinese goats

The aim of experiment was to analyze the polymerization effect of genotypes and genotype combinations of PRLR and LH?? gene loci in Xinong Saanen goat by SSCP marker and family trees. The relationships of genotype combinations and litter size were compared in Xinong Saanen goat. The results indicate that there are genetic polymorphisms at the PRLR and LH?? gene loci, and there are 4 positive genotypes (GG, CC, PP and LL) and 4 negative genotypes (HH, DD, QQ and MM) effects on litter size, respectively in Xinong Saanen goat. Compared with the other genotype combinations, the polymerization effect of GGCCPPLL markedly improved (P?<?0.05). The polymerization effect value of CC genotype was higher than that of CD genotype by 14.12%, and MM genotype was higher than that of LM genotype by 3.80%, and PP genotype was higher than that of QQ genotype by 15.67%, and LL genotype was higher than that of LM genotype by 11.48%, and PQ genotype was higher than that of QQ genotype by 11.02%, and CD genotype was higher than that of DD genotype by 10.69%, and PQ genotype was higher than that of PP genotype by 6.09%. There was significantly polymerization effect in the course of reproduction from parental generation (F0) to F1 generation. However, there was significantly gene isolation effect in the course of reproduction from F1 generation to F2 generation. This result can be used to guide the goat breeding in polyembryony trait.     

Polymorphism identification in the goat KITLG gene and association analysis with litter size

This study reported the analysis of KIT ligand (KITLG) gene polymorphisms in 681 goats of three breeds: Xinong Saanen (SN), Guanzhong (GZ), and Boer (BG). In addition, the study identified three allelic variants: g.769T>C and g.817G>T in SN and GZ breeds, and g.9760G>C in the three goat breeds. The g.769T>C and g.817G>T loci were closely linked (r² > 0.33). All the single nucleotide polymorphism loci were in Hardy–Weinberg disequilibrium (P < 0.05). Significant associations were found for litter size with all three loci. Therefore, these results suggest that the KITLG gene is a strong candidate gene affecting litter size in goats.