Luxi Cattle Research Articles

FSHR genotype affects estrogen levels but not pregnancy rates in Luxi cattle subjected to embryo transfer

Follicle-stimulating hormone receptor (FSHR), which mediates the functioning of FSH, plays a central role in reproduction. We investigated bovine FSHR gene polymorphisms and analyzed their relationships with pregnancy rates after embryo transfer and with hormone concentrations on the day of embryo transfer. One reported SNP of FSHR, G-278A, located in the 5'-upstream region, was analyzed and three genotypes (GG, GA and AA) were detected in 132 Luxi cattle recipients. Statistical analysis revealed that recipients with the GG genotype had significantly higher estrogen levels on the day of embryo transfer than did GA and AA genotypes. There were no significant differences in pregnancy rates among genotypes, after embryo transfer. We conclude that variation at these loci of the FSHR gene has no significant effect on pregnancy rates in Luxi cattle.

Effects of PGR and ESRα genotypes on the pregnancy rates after embryo transfer in Luxi cattle

Progesterone receptor (PGR) and estrogen receptor alpha (ESRα), which mediate the biological effects of the steroid hormones progesterone and estrogen, play a central role in the establishment and maintenance of pregnancy. The objectives of this study were to detect bovine PGR and ESRα genes polymorphisms and analyze their relationships with the pregnancy rates after embryo transfer and the hormone concentrations at the day of embryo transfer. One reported SNP of PGR G59752C and a novel SNP of ESRα G75935C were analyzed in 132 recipients of Luxi cattle. For the PGR gene, recipients with g.59752 GG and g.59752 GC genotypes had obviously higher pregnancy rates than g.59752 CC genotype. For the ESRα gene, recipients with g.75935 GC and g.75935 CC genotypes had obviously higher pregnancy rates than g.75935 GG genotype. Furthermore, the same tendency was observed for these two genes that the same genotype groups with high pregnancy rates had high progesterone concentration and low estrogen concentration at the day of embryo transfer. These results showed for the first time that PGR G59752C and ESRα G75935C polymorphisms had obvious effects on the pregnancy rates after embryo transfer, and indicated that PGR G59752C and ESRα G75935C polymorphisms could be potential markers for recipient selection of embryo transfer.

Polymorphisms of the bovine WNT10B gene and their associations with growth traits

Wingless-type MMTV integration site family, member 10B (WNT10B) may play an important role in inhibiting differentiation of preadipocytes in vitro and impairing adipose development in vivo. In this study, based on DNA sequencing and PCR–RFLP methods, we attempted to characterize the associations between common genetic polymorphisms in WNT10B and growth traits of 435 female cattle from three breeds (Jiaxian, Qinchuan and Luxi cattle). The results indicated that g.220A>G was in the intron 1, and g.1617C>T, g.3980G>T, g.4711G>C were in the coding region. At the g.3980G>T locus, Jiaxian cattle individuals with genotype TT had greater body length than those with genotypes GG and GT (P<0.05). At the g.220A>G locus, Qinchuan cattle individuals with genotype GG had greater growth traits than those with genotype AA and AG (P<0.05 or P<0.01). These statistical results showed that the WNT10B gene might be a potential candidate gene for marker-assisted selection (MAS).

Association of polymorphisms in the Pit-1 intron 5 with body measurements in Chinese Cattle

The Pit-1 gene was studied as a candidate for genetic markers of growth traits in Chinese cattle in this study. The single-strand conformation polymorphism method was used to identify polymorphism in the Pit-1 intron 5. One single nucleotide polymorphism, Pit1I5, was found in intron 5. The frequencies of allele A of Nanyang cattle (NY), Qinchuan cattle (QC), Jiaxianhong cattle (JX), Luxi cattle (LX) and Holstein cow (H) populations were 0.444, 0.477, 0.538, 0.523 and 0.475, respectively. Associations of the polymorphisms with growth traits in Nanyang cattle were analyzed using a general linear model procedure. The following parameters were greater in individuals with AA genotype than with AB genotype: body weight, average day gain, body height, chest girth at six, 12, 18 and 24 months (P allele B in other age groups. Therefore, genotype AA maybe a dominant genotype and allele A may be a dominant allele. These results imply that allele A of Pit-1 gene may likely affect growth traits positively. Key words : Beef cattle, growth traits, Pit- 1 intron 5, polymorphism.

Construction of recombinant proteins for reprogramming of endangered Luxi cattle fibroblast cells

Generation of induced pluripotent stem cells from endangered cattle breeds makes a significant contribution to the establishment of embryonic stem cell line, conservation and utilization of genetic resources in endangered cattle. In our study, we are trying to construct recombinant proteins of transcription factors Oct-4, Nanog, Sox2 and Lin28 for reprogramming of endangered Luxi cattle fibroblast cells in induced pluripotent stem cells. To test the cell permeability and stability of the recombinant proteins, we designed and fused a pep-1 protein transduction domain to the C-terminus of enhanced green fluorescent protein, we treated Luxi cattle fibroblast cells with the purified proteins at various concentrations by adding them to the cell culture media for 2-48 h and assayed cell morphology and protein presence. We found that the purified EGFP-tagged recombinant proteins readily entered cells at a concentration of 0.12 mg/ml within 2 h and some of them could translocate into nucleus. In addition, we found that the transduced proteins appeared to be stable inside cells for up to 48 h. Then codons of Oct-4, Nanog, Sox2 and Lin28 were optimized for high level of expression in E. coli., they were synthesized using DNA oligo based, PCR gene assembling method, the reprogramming factors were designed with optimized transcription factors fused a pep-1 protein transduction domain to the C-terminus, for high level protein expression of the reprogramming factors, inducer concentration, and induction time were tested, the optimum inducer concentration for the expression of reprogramming factors Oct-4, Nanog, Sox2 and Lin28 were 0.01 mM, the optimum induction time were 10, 8, 2 and 12 h, respectively.

Effect of cattle breed on finishing performance, carcass characteristics and economic benefits under typical beef production system in China

This study compared the finishing performance carcass characteristics and economic benefits of two imported (Limousin and Simmental) and three local (Luxi, Jinnan and Qinchuan) cattle breeds slaughtered at 18.5 months of age under thetypical Chinese beef production system. All cattle (n=71) were reared under the same production system and fed the same finishing diet for 105 days. Eight bulls from each breed were randomly selected for slaughtering. Compared with the three local breeds, the two imported breeds had higher average daily gain, dry matter intake and gain efficiency. Regarding carcass characteristics, the two imported breeds had higher carcass weight, bone weight, net meat weight, and ribeye area (P<0.001). However, the local breeds had higher (P<0.01) marbling scores than the imported breeds. The imported breeds showed higher economic benefits (P<0.001) than the local breeds. In conclusion, the imported cattle breeds had better growth performance, carcass traits and economic benefits compared with the local cattle breeds at 18.5 months old under the typical Chinese feeding conditions whereas, in this study, the local breeds may have some advantage in terms of meat quality.

Association of SCD1 and DGAT1 SNPs with the intramuscular fat traits in Chinese Simmental cattle and their distribution in eight Chinese cattle breeds

Intramuscular fat (IMF) is a key parameter for evaluation of nutritional quality of beef, with its endogenous synthesis regulated by stearoyl CoA desaturase (SCD1) and diacylglycerol-acyl transferase 1 (DGAT1) genes in cattle. The object of this research was to evaluate the effect of SCD1 and DGAT1 polymorphisms on IMF trait in beef cattle and to estimate the frequency distribution of SNPs in the two genes in Chinese cattle populations. The SCD1 and DGAT1 polymorphisms were detected by PCR-single strand conformation polymorphism (PCR-SSCP) method in Chinese Simmental cattle and their associations with IMF traits were analyzed using the general linear model (GLM). The frequency distribution of SNPs in SCD1 and DGAT1 genes were detected by PCR-SSCP method and analyzed in seven other cattle populations. The results showed significant associations of SNPs SCD1-878, SCD1-762, and DGAT1 10433 and 10434 with IMF (%) and shearing force values (SFV; kg) in Chinese Simmental cattle. A haplotype combining SCD1-878C, SCD1-762T, and DGAT1 10433 and 10434-GC had the highest IMF, marbling score and shearing force. The polymorphic investigation indicated that the frequency of SCD1-878C or SCD1-762T was significantly higher in Chinese southern cattle (Leiqiong, Yunnan High pump, BMY or Minnan Cattle) than in Chinese northern cattle (Chinese Simmental, Luxi Cattle, Bohai Black or Chinese Holstein), while the frequency of DGAT1 10433 and 10434-GC in Chinese indigenous breed (Leiqiong, Yunnan High pump, BMY, Luxi Cattle, Bohai Black, or Minnan Cattle) was significantly lower than breeds with imported blood (Chinese Simmental or Chinese Holstein). These findings demonstrated that both the SCD1 and DGAT1 SNPs were prospect genetic markers for IMF traits, and the SCD1 SNPs could be used as a genetic marker for southern or northern blood in Chinese cattle.

The polymorphisms of bovine PCSK1 gene and their associations with growth traits

Proprotein convertase 1 (PCSK1) is an endopeptidase involved in proteolytic processing of peptide hormone precursors in granules of the regulated secretory pathway of endocrine cells and mutations in PCSK1 gene are thought to cause obesity. In the present study, based on PCR-SSCP and DNA sequencing methods, polymorphisms of the PCSK1 gene were detected in 858 individuals from five breeds (Nanyang cattle, Qinchuan cattle, Jiaxian cattle, Luxi cattle and Chinese Holstein). The results showed that only P8 locus showed polymorphisms and 3 synonymous SNPs of PCSK1 gene were identified. Additionally, significant statistical difference was found in bovine birth weight and diplotype MM were 7.35% higher than diplotype XY.

Polymorphism of bovine Y-STR UMN0929 and its correlation with carcass traits in five Chinese beef cattle populations

The correlations between Y chromosome polymorphisms and the carcass traits were studied in five Chinese beef cattle populations by PCR, single strand conformation polymorphism and Y-STR sequence analysis. Nine alleles and their frequencies were identified on Y-STR UMN0929 region in Qinchuan (n=116), Luxi (n=112), Jinnan (n=104) pure breeds, Simmental×Qinchuan crossbred (n=80) and Angus×Qinchuan crossbred (n=96). The most popular A-176 and B-178 alleles were presented in all 5 cattle populations in the range of 12% (Jinnan) to 66% (Simmental×Qinchuan). The allele I-194 presented Luxi and Angus×Qinchuan. In Qinchun cattle, G-190 and E-186 alleles had bigger effect on BPI (4.23±0.32 and 4.22±0.48 kg/cm, P<0.01) and CW (325.40±49.42 and 316.73±45.29 kg, P<0.01), respectively. In Luxi cattle, I-194 allele affected higher BPI (4.08±0.35 kg/cm, P<0.01) and CW (302.07±17.55 kg, P<0.01), respectively. In Jinnan cattle breed, H-192 had higher BPI (4.32±0.50 kg/cm, P<0.05) and CW (327.87±59.37 kg, P<0.05), respectively. In Simmental×Qinchuan cross breed, C-180 allele affected largely on BPI (5.16±0.25 kg/cm, P<0.05) and CW (393.16±25.92 kg, P<0.05). In Angus×Qinchuan cross breed, I-194 had higher BPI (4.43±0.33 kg, P<0.05) and CW (346.63±29.77 kg, P<0.05). Correlations between alleles and other carcass traits (net meat weight, top grade weight, slaughter rate, net meat rate, loin-eye muscle area, carcass length, meet tenderness and shear force) were also analyzed using mixed-effect model. Cattle Y-STR UMN0929 loci alleles and its correlation with carcass traits in beef cattle populations could be implemented into the cattle breeding program for choosing beef cattle with better carcass traits.

Detecting a deletion in the coding region of the bovine bone morphogenetic protein 15 gene (BMP15)

The aim of this study was to detect polymorphism in the bovine bone morphogenetic protein 15 (BMP15) gene. On the basis of PCR-SSCP and DNA sequencing, a 4-bp deletion was identified in the coding region of the gene. Sequence analysis revealed that the deletion altered the reading frame and introduced a stop codon at position 264. Eight breeds (Luxi, Qinchuan, Nanyang, Jinnan, Bohai Black, Menggolian, Holstein, and Simmental) were genotyped by PCR-SSCP. No cows homozygous for this mutation were observed in these breeds. Heterozygous cows were detected in Luxi, Qinchuan, Nanyang, Jinnan and Bohai Black cattle. Fecundity was not increased in heterozygous individuals.

The application of asymmetric PCR-SSCP in gene mutation detecting

The advantages and disadvantages of asymmetric PCR-SSCP and the traditional PCR-SSCP were compared in this study. The mutations in 3′UTR of Smad4 gene of Luxi cattle and the Holstein cow were analyzed by asymmetric PCR-SSCP and one insert “T” mutation and one G/A mutation in this region were found. The G/A mutation created a HhaI restriction enzyme digestion position and the frequencies studied by asymmetric PCR-SSCP and HhaI-RFLP in 116 Luxi cattle and 75 Holstein cows were all the same. The asymmetric PCR-SSCP had fewer, clearer and more stabile bands than traditional PCR-SSCP. This indicates that the asymmetric PCR-SSCP is suited for mutation detection.

Establishment and biological characteristics of Luxi cattle fibroblast bank

A fibroblast line from ear marginal tissue of Luxi cattle (LXCEM2/2) was successfully established by direct culturing of explants. Biological analysis showed that the population doubling time (PDT) for reviving cells was approximately 24 h. Measurement of lactic dehydrogenase (LDH) and malic dehydrogenase (MDH) isoenzymes showed no cross-contamination among the cells. Karyotyping showed that the frequency of cells with chromosome number 2n = 60 was 90.7–92.2%. Tests for bacteria, fungi, viruses and mycoplasma were negative. The efficiencies of expression of pEGFP-N3, pEYFP-N1 and pDsRed1-N1 were between 6.3% and 31.6% at 24 h, 48 h and 72 h after transfer; at 24 h, fluorescence was well distributed in the cytoplasm and nucleus except for some cryptomeric vesicles. Every index of the Luxi cattle cell line meets the quality control standards of the American Type Culture Collection (ATCC). Not only has the germline of this important cattle breed been preserved at the cell level, but also valuable material had been provided for genome, postgenome and somacloning research. Moreover, the establishment of this technical platform may provide both technical and theoretical support for storing the genetic resources of other animals and poultry at the cell level.

소 Y 염색체 특이 Microsatellite를 이용한 품종별 대립유전자 빈도 분석

Bovine microsatellite 마커인 INRA124는 소에 있어 Y 염색체 특이 마커로서 130 및 132 bp의 두 개 대립유전자를 가지고 있는데, Bos taurus 특이 대립유전자는 132 bp이고, Bos indicus 특이대립유전자는 130 bp이다. 이번 연구는 이 유전좌위를 이용하여 한우집단에 대한 Bos taurus 및 Bos indicus 유래의 대립유전자 분석을 통하여 한우의 유전적 특성을 이해하고자 하였다. 동북아시아, 중국내륙, 유럽, 인도 및 아프리카 유래의 20개 소 품종 822두의 수컷을 공시하여 분석해 본 결과, 유럽계통, 일본 화우 및 한우에서는 Bos indicus 유래의 대립유전자는 전혀 검출되지 않았으며, 인도 및 아프리카 유래의 Bos indicus 품종에서는 132 bp의 대립유전자가 전혀 검출되지 않았다. 한우, 브라만 및 샤롤레의 교잡우 집단(CBK)에서는 Bos indicus 특이 대립유전자인 130 bp가 0.19의 빈도로 검출되었고, 중국내륙 품종인 노서우 및 난양우는 각각 0.46 및 0.29의 빈도로 검출되었다. 이로서 한우의 기원은 기존 혼합설과 달리 유럽계통의 Bos taurus만으로 기원하였을 가능성을 제기한다. The INRA124 is a bovine Y-chromosomal specific microsatellite locus that has been revealed a polymorphism. This locus has two alleles. The 132 bp allele is specific to cattle (humpless) of taurine origin and the 130 bp allele is specific to cattle (humped) of indicine origin. A total 822 males of 20 breeds or populations; North Eastern Asian breeds (Hanwoo, Korean Black cattle, Chik-so, CBK, Japanese Black cattle, Japanese Brown cattle, Yanbian cattle), Chinese yellow cattle (Luxi cattle, Nanyang cattle), European origin (Angus, Hereford, Charolais, Simmental, Brown swiss, Holstein, Limousin), African origin (Kavirondo zebu, White Fulani, crossbreed of N'Dama and Boran), Indian origin (Sahiwal) were characterized the distribution of alleles using INRA124 locus. Any individuals of European, Japanese origins and Hanwoo were not detected 130 bp allele, Bos indicus specific allele. Bos indicus breeds of Indian and African origins were not detected 132 bp allele, Bos taurus specific allele. CBK population that the crossbreed of Hanwoo, Brahman and Charolais showed the frequency of 0.19 in indicine specific allele. The breeds of Chinese mainland, Luxi and Nanyang cattle were detected 0.46 and 0.29 frequencies in indicine specific allele, respectively. These results suggest that Korean cattle, Hanwoo, had not been originated from a crossbred between Bos primigenius in Europe and Bos indicus in India.

Genetic Structure and Differentiation of Three Chinese Indigenous Cattle Populations

Levels of genetic differentiation, gene flow, and genetic structure of three indigenous cattle populations (Luxi, Bohai, and Minnan) and two reference cattle populations (Chinese Holstein and Qinhai yak) in China were estimated using the information from 12 microsatellites, and 141 microsatellite alleles were identified. The mean number of alleles per locus ranged from 2.9005 in yak to 4.9722 in Holstein. The observed heterozygosity ranged from 0.5325 (yak) to 0.7719 (Holstein); 29 private alleles were detected. The global heterozygote deficit across all populations amounted to 58.5% (p < 0.001). The overall significant (p < 0.001) deficit of heterozygotes because of inbreeding within breeds amounted to 43.2%. The five cattle populations were highly differentiated (F (st) = 26.9%, p < 0.001) at all loci. The heterozygote deficit within the population was highest in Luxi cattle and lowest in yak. The average number of effective migrants exchanged per generation was highest (1.149) between Luxi and Holstein, and lowest (0.509) between Luxi and yak. With the application of prior population information, cluster analysis achieved posterior probabilities from 91% to 98% of correctly assigning individuals to populations. Combining the information of cluster analysis, gene flow, and Structure analysis, the five cattle populations belong to three genetic clusters, a taurine (Luxi and Chinese Holstein), a zebu (Bohai and Minnan), and a yak cluster. This indicates that Bohai black is closer to Bos indicus than Luxi cattle. The evolution and development of three indigenous cattle populations are discussed.

Relationship between genotypes at MyoD locus and carcass traits in cattle

A 261 bp sequence of the bovine MyoD gene intron 2 was cloned and was found to bear no similarities to the human MyoD gene sequence. Polymorphisms of the Myod gene in cattle including three Chinese breeds (Luxi cattle, Jinnan cattle and Qinchuan cattle) and four cross-breeding populations (Limousin x Luxi cattle, Simmental x Luxi cattle, Charolais x Luxi cattle and Angus x Luxi cattle) were detected by PCR-SSCP. Two SSCP alleles (A and B) were detected, which caused by a C-->T at 39 bp and a C-->G transition at 112 bp. Only two genotypes AA and AB occurred in the population. In Chinese local cattle, B allele was dominant, and this locus was at Hardy-Weinberg equilibrium except for the Charolais x Luxi cattle and Angus x Luxi cattle. The association of these polymorphisms with cattle carcass traits was analyzed using the general linear model (GLM). Statistical analysis revealed a higher value of living weight, carcass weight and loin eye area for individuals with genotype AA than AB (P < 0.05). Further studies on a bigger population size are needed to confirm the observed effect of MyoD genotypes on carcass quality traits.

The Genetic Diversity and Phylogenetic Status of Luxi Cattle

A total of 87 individuals of Luxi cattle from Juanchen and Liangshan counties, Shangdong Province, China, were sampled by simple random sampling in typical colony. Twenty-one blood proteins and enzymes loci were detected by polyacrylamide gel electrophoresis (PAGE) and starch gel electrophoresis (SGE). In the meantime, the data of 7 loci of 13 cattle populations in China and other countries were collected and phylogeny relationships were studied. The results indicated that 9 out of 21 loci showed polymorphism (42.86%); the level of genetic variation in Luxi cattle population was relatively high, the mean heterozygosity was 0.1416. The Luxi cattle have a close phylogenetic relationship with the cattle populations of east and south of Asia, and this further confirmed the fact that Luxi cattle were the cross-breed between the Bos taurus and Bos indicus in China, but it is impossible that yellow cattle contained the blood of of Bali.

Analysis of Microsatellite DNA Polymorphisms in Five China Native Cattle Breeds and Application to Population Genetics Studies

Five China native cattle breeds have been characterized by using 10 microsatellite DNA markers. The studied populations can be divided into five groups: Luxi cattle, Nanyang cattle, Jinnan cattle, Qinchuan cattle and Yanbian cattle. Allele frequencies were calculated and used for the characterization of the breeds and the study of their genetic relationships. Heterozygosity, polymorphism information content, the effective number of alleles was calculated. Nei's standard genetic distance (1978) was calculated and used for a neighbor-joining tree construction. NJ tree showed that Luxi cattle, Nanyang cattle, Jinnan cattle and Qinchuan cattle are closely related, whereas Yanbian cattle are clearly distinct from other four populations. The genetic relationship of five breeds corresponds to their history and geographic origins. This work analyzes the recent origin of these populations and contributes to the knowledge and genetic characterization of China native breeds. (Asian-Aust. J. Anim. Sci. 2005. Vol 18, No. 12 : 1696-1700)

Microsatellite loci 분석에 의한 한우와 타 품종간의 유전적 유연관계

For the genetic assessment of the cattle breeds including Hanwoo, eleven microsatellite markers on ten bovine autosomes were genetically characterized for 618 individuals of nineteen cattle breeds; North Eastern Asian breeds (Korean cattle, Korean Black cattle, Japanese Black cattle, Japanese Brown cattle, Yanbian cattle), Chinese yellow cattle (Luxi cattle, Nanyang cattle), European Bas taurus (Angus, Hereford, Charolais, Holstein, Limousin), African Bas taurus (N'Dama, Baoule), African Bas indicus (Kavirondo Zebu, White Fulani), Asian Bas indicus (Sahiwal, Nelore) and one Bali cattle, Bas banteng as an outbreed-reference population. Allele frequencies derived from the genotyping data were used in estimating heterozygosities, gene diversities and genetic distances. The microsatellite loci were highly polymorphic, with a total of 162 different alleles observed across all loci. Variability in allele numbers and frequencies was observed among the breeds. The average expected heterozygosity of North Eastern Asian breeds was higher than those of European and African taurines, but lower than those of Asian and African indicines. Genetic distances were estimated using Nei's DA genetic distance and the resultant DA matrix was used in the construction of the phylogenetic trees. The genetic distances between North Eastern Asian cattle breeds and Bas indicus were similar with those between European Bas taurus and Bas indicus, and African Bas taurus and Bas indicus, respectively. The clusters were clearly classified into North Eastern Asian, European and African taurines groups as well as different cluster with Chinese mainland breeds, firstly out-grouping with Bas indicus. These results suggest that Korean cattle, Hanwoo, had not been originated from a crossbred between Bas primigenius in Europe and Bas indicus in India and North Eastern Asian Bas taurus may be have separate domestication from European and African Bas taurus.

Titanium dioxide

Wingless-type MMTV integration site family, member 10B (WNT10B) may play an important role in inhibiting differentiation of preadipocytes in vitro and impairing adipose development in vivo. In this study, based on DNA sequencing and PCR–RFLP methods, we attempted to characterize the associations between common genetic polymorphisms in WNT10B and growth traits of 435 female cattle from three breeds (Jiaxian, Qinchuan and Luxi cattle). The results indicated that g.220A>G was in the intron 1, and g.1617C>T, g.3980G>T, g.4711G>C were in the coding region. At the g.3980G>T locus, Jiaxian cattle individuals with genotype TT had greater body length than those with genotypes GG and GT (P < 0.05). At the g.220A>G locus, Qinchuan cattle individuals with genotype GG had greater growth traits than those with genotype AA and AG (P < 0.05 or P < 0.01). These statistical results showed that the WNT10B gene might be a potential candidate gene for marker-assisted selection (MAS).