A method for detecting and / or evaluating quality traits of Chinese Simmental beef

Abstract

The present invention discloses a method for detecting and / or evaluating the quality traits of Chinese Simmental beef. Using genomic DNA from Chinese Simmental cattle blood as a template and primer pairs SEQ ID NOs: 1-2 as primers, PCR amplification identified six SNPs significantly associated with traits such as carcass fat coverage, dressing percentage, and liver weight in Chinese Simmental cattle. The method also establishes a method for detecting and / or evaluating the quality traits of Chinese Simmental beef. The present invention provides a method for early selection and breeding of Chinese Simmental cattle and for improving meat quality.

Description

Technical Field

[0001] The present invention relates to the technical field of breeding of Chinese Simmental cattle, and in particular to a method for detecting and / or evaluating quality traits of Chinese Simmental beef. Background Art

[0002] Since the Simmental cattle were introduced to my country in the 1950s, crossbreeding and breeding with Chinese yellow cattle has resulted in the genetically stable Chinese Simmental cattle, which boast superior production performance and increased adaptability. They are now widely distributed throughout my country. Chinese Simmental cattle boast excellent growth and meat production, making them a dual-purpose breed for both meat and milk production and the dominant breed in my country's beef cattle industry. Compared to other meats, beef is richer in protein and amino acids, providing essential nutrients for growth and development. Beef has a unique flavor and high nutritional value. With rising living standards, people's demand for both quantity and quality of beef is also increasing. Beef quality traits primarily include meat quality and carcass characteristics, and selecting individuals with superior meat quality and carcass traits is a key focus of beef cattle breeding.

[0003] Vanin (VNN) hydrolyzes pantetheine into pantothenic acid and cysteamine. Currently, three Vanin gene isoforms have been identified: Vanin-1, Vanin-2, and Vanin-3. Vanin-1, also known as vascular non-inflammatory molecule 1 (Vanin-1; VNN1), is the most extensively studied isoform of the Vanin family. The protein it encodes is pantetheine, a membrane-associated extracellular enzyme that belongs to the CN hydrolase family.

[0004] VNN1 is present in various mammalian tissues and plays a crucial role in various life processes, including tumorigenesis and immunity. In the liver, VNN1 is a target gene of peroxisome proliferator-activated receptor α (PPARα). Two PPRE binding sites are present in the VNN1 promoter, which bind to PPARα and promote VNN1 expression. Studies have also shown that hepatic miR-122 negatively regulates VNN1 gene expression, thereby affecting VNN1's function in regulating hepatic lipid metabolism. Van Diepen et al. found that VNN1 expression and enzyme activity are upregulated in humans after fasting or taking lipid-lowering drugs. VNN1 knockout mice and fasting mice showed increased triglyceride accumulation, but no significant changes in free fatty acids. However, treatment with inhibitors in rats exacerbated hepatic triglyceride accumulation. Li Jie's research found that VNN1 expression is highest in goose livers, and that overfeeding significantly downregulated VNN1 expression in adipose tissue, suggesting that VNN1 may participate in lipid metabolism pathways and regulate hepatic fat synthesis. Xing et al. found that in intrauterine growth retardation (IUGR) asthmatic mice, increased VNN1 expression can activate the PI3K / Akt / NFκB signaling pathway, leading to increased reactive oxygen species and inflammatory response. Summary of the Invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and to provide a method for detecting and / or evaluating the quality traits of Chinese Simmental beef.

[0006] The first object of the present invention is to provide a reagent for predicting and / or evaluating the quality traits of Chinese Simmental beef.

[0007] The second object of the present invention is to provide the use of the reagent in preparing a kit for detecting and / or evaluating the quality traits of Chinese Simmental beef, and / or detecting and / or evaluating the quality traits of Chinese Simmental beef.

[0008] The third object of the present invention is to provide a method for detecting and / or evaluating the quality traits of Chinese Simmental beef.

[0009] A fourth object of the present invention is to provide a kit for detecting and / or evaluating the quality traits of Chinese Simmental beef.

[0010] The fifth object of the present invention is to provide the use of the reagent, method, and / or kit described in 1 in molecular breeding of quality traits of Simmental beef in China.

[0011] In this study, genomic DNA from 129 Chinese Simmental cattle was used as a template to amplify the VNN1 gene. SNPs were screened and genotyped by sequencing. The genetic polymorphism of these SNPs was analyzed and their associations with beef quality and carcass traits were analyzed. Six SNPs were found in the VNN1 gene of Chinese Simmental cattle. SNP1 and SNP4 had only two genotypes, with the dominant genotypes being AA and CC, respectively, and the dominant alleles being A and C. The heterozygosity for these loci was less than 0.25, indicating low polymorphism at both loci. SNP2, SNP3, SNP5, and SNP6 each had three genotypes, with the dominant genotypes being TC, TC, AG, and GG, respectively. At SNP2, the frequencies of the T and C alleles were equal, while the dominant allele at SNP3, SNP5, and SNP6 was G. The heterozygosity for all four SNPs ranged from 0.25 to 0.50, indicating moderate polymorphism at all loci.

[0012] Chi-square tests demonstrated that all six SNPs were in Hardy-Weinberg equilibrium in the experimental population, indicating that the Chinese Simmental cattle population was in a state of random mating, had good heredity, and was rarely or not affected by artificial selection. SNP1 was significantly associated with dressing percentage (P<0.05); SNP2 and SNP3 were significantly associated with carcass fat coverage and marbling (P<0.05); SNP4 was significantly associated with liver weight (P<0.05); SNP5 was significantly associated with carcass fat coverage (P<0.05); and SNP6 was significantly associated with dressing percentage and whip weight (P<0.05).

[0013] Furthermore, it was found that the four SNPs sites, SNP2, SNP3, SNP5 and SNP6, formed a haplotype domain, with a total of 8 different haplotypes with biological statistical significance (number of individuals n≥3). They are H1H1 (the genotypes of the four SNPs sites are CC, GG, GG, GG), H1H2 (the genotypes of the four SNPs sites are CT, GA, GA, GG), H1H3 (the genotypes of the four SNPs sites are CT, GA, GA, GA), H1H4 (the genotypes of the four SNPs sites are CT, GA, GG, GG), H2H2 (the genotypes of the four SNPs sites are TT, AA, AA, GG), H2H3 (the genotypes of the four SNPs sites are TT, AA, AA, GA), H2H4 (the genotypes of the four SNPs sites are TT, AA, AG, GG), and H3H3 (the genotypes of the four SNPs sites are TT, AA, AA, AA).

[0014] The slaughter rate of individuals with the H1H4 haplotype was significantly lower than that of individuals with the H1H1, H1H2, and H2H2 haplotypes (P<0.05); the carcass fat coverage of individuals with the H1H1 haplotype was significantly higher than that of individuals with the H2H2 haplotype (P<0.05); the bullwhip weight of individuals with the H1H2 haplotype was significantly higher than that of individuals with the H1H3 haplotype (P<0.05); the testicle weight of individuals with the H1H2 haplotype was significantly higher than that of individuals with the H2H3 haplotype (P<0.05). The meat color score of individuals with H1H2 haplotype was significantly higher than that of individuals with H1H1 and H1H3 haplotypes (P<0.05); the marbling grade of individuals with H3H3 haplotype was significantly lower than that of individuals with H1H2, H2H2 and H2H3 haplotypes (P<0.05), and the marbling grade of individuals with H1H2 haplotype was significantly higher than that of individuals with H1H1 haplotype (P<0.05).

[0015] In order to achieve the above object, the present invention is implemented through the following technical solutions:

[0016] The present invention claims a reagent for predicting and / or evaluating the quality traits of Chinese Simmental beef, wherein the reagent is used to detect one or more of the genotypes of the following SNP sites:

[0017] SNP site 1 is located at NC_037336.1:70835469, which is the 70835469 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate of individuals with the AA genotype is significantly higher than that of individuals with the GA genotype.

[0018] SNP site 2 is located at NC_037336.1:70835516, which is the 70835516 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with CC genotype is significantly higher than that of individuals with TT genotype, and the marbling grade of individuals with TT and TC genotypes is significantly higher than that of individuals with CC genotype.

[0019] SNP site 3 is located at NC_037336.1:70835542, which is the 70835542 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype. The marbling grade of individuals with the AA and AG genotypes is significantly higher than that of individuals with the GG genotype.

[0020] SNP site 4 is located at NC_037336.1:70835571, which is the 70835571 position on chromosome 9 of the ARS-UCD1.2 version of the bovine genome. The liver weight of individuals with CC genotype is significantly higher than that of individuals with TC genotype.

[0021] SNP site 5 is located at NC_037336.1:70835623, which is the 70835623 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype.

[0022] SNP site 6 is located at NC_037336.1:70835638, which is the 70835638th position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate and bullwhip weight of GG genotype individuals are significantly higher than those of AG genotype individuals.

[0023] Preferably, the slaughter rate of individuals whose genotype at SNP site 2 is TC, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG is significantly lower than the slaughter rate of individuals with the following genotypes:

[0024] Individuals with a genotype of CC at SNP site 2, a genotype of GG at SNP site 3, a genotype of GG at SNP site 5, and a genotype of GG at SNP site 6;

[0025] Individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG;

[0026] An individual whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG.

[0027] Preferably, the carcass fat coverage of individuals whose genotype at SNP site 2 is CC, whose genotype at SNP site 3 is GG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG is significantly higher than the carcass fat coverage of individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG.

[0028] Preferably, the bullwhip weight of an individual whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than the bullwhip weight of an individual whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is AG.

[0029] Preferably, the testicular weight of an individual whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than the testicular weight of an individual whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AG.

[0030] Preferably, the meat color score of an individual whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than the meat color scores of individuals with the following genotypes:

[0031] The genotype of SNP site 2 is CC, the genotype of SNP site 3 is GG, the genotype of SNP site 5 is GG, and the genotype of SNP site 6 is GG;

[0032] The genotype of SNP site 2 is CT, the genotype of SNP site 3 is AG, the genotype of SNP site 5 is AG, and the genotype of SNP site 6 is AG.

[0033] Preferably, the marbling grade of an individual whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AA is significantly lower than the marbling grade of an individual with the following genotypes:

[0034] Individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG;

[0035] Individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG;

[0036] An individual whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AG.

[0037] Preferably, the marbling grade of an individual whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is GA, whose genotype at SNP site 5 is GA, and whose genotype at SNP site 6 is GG is significantly higher than that of an individual whose genotype at SNP site 2 is CC, whose genotype at SNP site 3 is GG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG.

[0038] Preferably, the reagent is a primer having a nucleotide sequence as shown in SEQ ID NO: 1-2.

[0039] The invention also claims the use of the reagent in preparing a kit for detecting and / or evaluating the quality traits of Chinese Simmental beef, and / or detecting and / or evaluating the quality traits of Chinese Simmental beef.

[0040] And a method for detecting and / or evaluating the quality traits of Chinese Simmental beef, using the reagent to detect Chinese Simmental cattle.

[0041] Preferably, the reagent is a primer having a nucleotide sequence as shown in SEQ ID NO: 1-2.

[0042] The upstream primer sequence was: 5′-GGCTAATATCCCAAACAAGTGA-3′ (SEQ ID NO: 1);

[0043] The downstream primer sequence is: 5′-CAGAAGAACTCAGACCTCCAA-3′ (SEQ ID NO: 2).

[0044] PCR amplification system (total volume 20 μL): Taq PCR Mix 10 μL, upstream and downstream primers 0.5 μL each, DNA template 2 μL, ddH2O 7 μL.

[0045] PCR amplification program: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 56°C for 30 s, extension at 72°C for 1 min, for a total of 35 cycles; and extension at 72°C for another 5 min.

[0046] The invention also claims a kit for detecting and / or evaluating the quality traits of Chinese Simmental beef, comprising the reagent.

[0047] Preferably, it also contains PCR reagents.

[0048] More preferably, the PCR reagents are ddH2O and Taq PCR Mix.

[0049] Preferably, it also contains CutSmart Buffer and NsiI enzyme.

[0050] The use of the reagent, the method, and / or the kit in molecular breeding of quality traits of Simmental beef in China also falls within the scope of protection of the present invention.

[0051] Preferably, the beef quality traits are one or more of slaughter rate, carcass fat coverage, liver weight, bullwhip weight, testicle weight, meat color score, and marbling.

[0052] Compared with the prior art, the present invention has the following beneficial effects:

[0053] This invention discloses a method for detecting and / or evaluating meat quality traits in Chinese Simmental cattle. Using genomic DNA from Chinese Simmental cattle blood as a template and primer pairs corresponding to SEQ ID NOs: 1-2, PCR amplification revealed six SNPs significantly associated with traits such as carcass fat coverage, dressing percentage, and liver weight. This method provides a method for early-stage selection and breeding of Chinese Simmental cattle and for improving meat quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 PCR amplification results of the Chinese Simmental VNN1 gene; M: DL-2 000 Marker; 1-5: PCR amplification products.

[0055] Figure 2 This is the sequencing map of the two genotypes at SNP site 1 of the VNN1 gene; Figure I is an individual with genotype AG; Figure II It is an individual with genotype AA.

[0056] Figure 3 This is the sequencing map of the three genotypes at SNP site 2 of the VNN1 gene; Figure I is an individual with genotype TT; Figure II is an individual with genotype CT; Figure III is an individual with genotype CC.

[0057] Figure 4 This is the sequencing map of the three genotypes at SNP site 3 of the VNN1 gene; Figure I is an individual with genotype AA; Figure II is an individual with genotype AG; Figure III It is an individual with genotype GG.

[0058] Figure 5 This is the sequencing map of the two genotypes at SNP site 4 of the VNN1 gene; Figure I is an individual with CC genotype; Figure II It is an individual with genotype CT.

[0059] Figure 6 Electrophoresis patterns of three genotypes identified by PCR-RFLP at SNP site 5 of the VNN1 gene; M: DL-2 000 Marker; lanes 1, 2, and 6 are AG genotypes, lanes 3 and 5 are AA genotypes, and lane 4 is GG genotypes.

[0060] Figure 7 This is the sequencing map of the three genotypes at SNP site 6 of the VNN1 gene; Figure I is an individual with genotype GG; Figure II is an individual with genotype AG; Figure III It is an individual with genotype AA.

[0061] Figure 8 This is the SNPs linkage reaction of the VNN1 gene in the Chinese Simmental cattle population. DETAILED DESCRIPTION

[0062] The present invention is further described in detail below with reference to the accompanying drawings and specific examples. The examples are intended only to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods used in the following examples are conventional methods unless otherwise specified; the materials and reagents used are commercially available unless otherwise specified.

[0063] Example 1 PCR amplification and SNPs genotyping of Chinese Simmental cattle

[0064] 1. Experimental Sample

[0065] Blood samples were collected from 129 34-month-old male Chinese Simmental cattle from the Baolongshan Beef Cattle Fattening Farm in Tongliao, Inner Mongolia. Genomic DNA was extracted from these blood samples and stored at -80°C.

[0066] 2. Experimental Methods

[0067] 1. Primer design

[0068] The bovine VNN1 gene sequence (SEQ ID NO: ENSBTAT00000020086.6) was searched on the Ensembl website, and a pair of primers was designed using Primer Premier 6.0 software and synthesized by Sangon Biotech (Shanghai) Co., Ltd. The upstream primer sequence was: 5′-GGCTAATATCCCAAACAAGTGA-3′ (SEQ ID NO: 1); the downstream primer sequence was: 5′-CAGAAGAACTCAGACCTCCAA-3′ (SEQ ID NO: 2).

[0069] 2. PCR amplification and sequencing to identify individual genotypes

[0070] PCR amplification was performed using genomic DNA from the blood of 129 Chinese Simmental cattle as templates.

[0071] PCR amplification system (total volume 20 μL): Taq PCR Mix 10 μL, upstream and downstream primers 0.5 μL each, DNA template 2 μL, ddH2O 7 μL.

[0072] PCR amplification program: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 30 s, annealing at 56°C for 30 s, extension at 72°C for 1 min, for a total of 35 cycles; and extension at 72°C for another 5 min.

[0073] The PCR amplification products were detected by electrophoresis on 1.5% agarose gel at 120V for 18 min. After the target band was confirmed to be correct, they were sent to Sangon Biotech (Shanghai) Co., Ltd. for sequencing.

[0074] SeqMan software was used to observe the sequencing results and confirm the genotype of each sample individual based on the peak plot and sequence.

[0075] 3. PCR-RFLP identification of individual genotypes

[0076] PCR amplification was performed using genomic DNA from the blood of 129 Chinese Simmental cattle as template.

[0077] PCR amplification system (total volume 20 μL): Taq PCR Mix 10 μL, 0.5 μL each of upstream and downstream primers (SEQ ID NOs: 1-2), 2 μL DNA template, 7 μL ddH2O. PCR amplification program: 95°C initial denaturation for 5 min; 35 cycles of denaturation at 95°C for 30 s, annealing at 56°C for 30 s, and extension at 72°C for 1 min; followed by an additional extension at 72°C for 5 min.

[0078] The PCR amplification products were detected by specific enzyme digestion using the restriction endonuclease NsiI.

[0079] Enzyme digestion system (total volume 10 μL): CutSmart Buffer 1 μL, NsiI enzyme 0.2 μL, PCR amplification product 5 μL, ddH2O 3.8 μL. Enzyme digestion conditions: Incubate at 37°C for 30 min.

[0080] The enzyme digestion products were detected by electrophoresis on 2% agarose gel at 120V for 20min to confirm the individual genotype.

[0081] 3. Experimental Results

[0082] The electrophoresis results of PCR amplification products are as follows Figure 1 As shown, a total of 129 Chinese Simmental cattle blood genomic DNAs were amplified, and the amplified product size was 976 bp. The electrophoresis bands were bright and clear, and there were no non-specific amplification bands. The length was consistent with expectations and could be used for the next experiment.

[0083] The genotyping results of SNPs sites are as follows Figures 2 to 7 As shown, the results showed that 6 SNP sites were found in the VNN1 gene:

[0084] SNP site 1 has two genotypes, AA and AG, and is located at NC_037336.1:70835469, which is position 70835469 on chromosome 9 of the ARS-UCD version 1.2 bovine genome;

[0085] SNP site 2 has three genotypes: TT, TC, and CC, and is located at NC_037336.1:70835516, which is position 70835516 on chromosome 9 of the ARS-UCD1.2 version of the bovine genome;

[0086] SNP site 3 has three genotypes: AA, AG, and GG, and is located at NC_037336.1:70835542, which is position 70835542 on chromosome 9 of the ARS-UCD version 1.2 bovine genome;

[0087] SNP site 4 has two genotypes, TC and CC, and is located at NC_037336.1:70835571, which is position 70835571 on chromosome 9 of the ARS-UCD version 1.2 bovine genome;

[0088] SNP site 5 has three genotypes: AA, AG, and GG, and is located at NC_037336.1:70835623, which is position 70835623 on chromosome 9 of the ARS-UCD version 1.2 bovine genome;

[0089] SNP site 6 has three genotypes: GG, AG and AA, and is located at NC_037336.1:70835638, which is the 70835638th position of chromosome 9 in the ARS-UCD1.2 version of the bovine genome.

[0090] Example 2 Population genetic characteristics of SNPs sites of the VNN1 gene

[0091] 1. Experimental Methods

[0092] Chromas software was used to observe overlapping peaks in sequencing results; Popgene 32 software was used to calculate gene frequency, genotype frequency, genetic heterozygosity (H), and effective number of alleles (Ne); PIC 0.6 software was used to calculate polymorphism information content (PIC); and the formula X was used to calculate 2 The values ​​were tested for Hardy-Weinberg equilibrium;

[0093] 2. Experimental Results

[0094] The population genetic characteristics of SNPs are shown in Table 1.

[0095] Table 1 Population genetic characteristics analysis results of 6 SNPs in the VNN1 gene

[0096]

[0097] The results showed that SNP1 and SNP4 sites had only two genotypes, the dominant genotypes were AA and CC, the genotype frequencies were 0.9670 and 0.9612, respectively, the dominant alleles were A and C, the genotype frequencies were 0.9535 and 0.9806, respectively, and the heterozygosity of the two loci was less than 0.25, indicating that SNP1 and SNP4 sites were both low polymorphic in VNN1; SNP2, SNP3, SNP5 and SNP6 sites had three genotypes, the dominant genotypes were T The genotype frequencies of C, TC, AG, and GG were 0.4728, 0.4729, 0.4574, and 0.7287, respectively. The frequencies of the alleles T and C at the SNP2 site were the same, with gene frequencies of 0.5000. The dominant alleles at SNP3, SNP5, and SNP6 were all G, with gene frequencies of 0.5078, 0.5388, and 0.8450, respectively. The genetic heterozygosity of the four SNPs were all in the range of 0.25 to 0.50, indicating that the four SNPs were moderately polymorphic.

[0098] The chi-square test showed that the above six SNPs were in Hardy-Weinberg equilibrium in the experimental population, indicating that the Chinese Simmental cattle population was in a state of random mating, had good population heredity, and was rarely or not affected by artificial selection.

[0099] Example 3 Correlation Analysis between SNPs of the VNN1 Gene and Quality Traits of Chinese Simmental Beef

[0100] 1. Experimental Methods

[0101] According to the method in the national standard "Fresh and Frozen Cut Beef" (GB / T17238-2008), the meat quality traits of Chinese Simmental cattle in Example 1, such as slaughter rate, carcass fat coverage, liver weight, bullwhip weight, meat color score, and testicle weight, were measured and counted.

[0102] SPSS 23.0 software was used to analyze the correlations between different SNP genotypes and beef quality and carcass traits of Chinese Simmental cattle. P < 0.05 was considered significant. Data are expressed as "mean ± standard error."

[0103] 2. Experimental Results

[0104] The results of correlation analysis between SNPs and meat quality and carcass traits are shown in Table 2.

[0105] Table 2 Associations between VNN1 gene SNPs and meat quality and carcass traits

[0106]

[0107] The results showed that the SNP1 locus was significantly correlated with the dressing rate trait (P<0.05): the dressing rate of the AA genotype individuals was significantly higher than that of the GA genotype individuals;

[0108] Both SNP2 and SNP3 were significantly correlated with carcass fat coverage and marbling traits (P<0.05): the carcass fat coverage of individuals with CC genotype at SNP2 was significantly higher than that of individuals with TT genotype, and the marbling grade of individuals with TT and TC genotypes was significantly higher than that of individuals with CC genotype; the carcass fat coverage of individuals with GG genotype at SNP3 was significantly higher than that of individuals with AA genotype, and the marbling grade of individuals with AA and AG genotypes was significantly higher than that of individuals with GG genotype.

[0109] SNP4 locus was significantly correlated with liver weight (P<0.05): the liver weight of individuals with CC genotype was significantly higher than that of individuals with TC genotype;

[0110] SNP5 locus was significantly correlated with carcass fat coverage (P<0.05): the carcass fat coverage of GG genotype individuals was significantly higher than that of AA genotype individuals;

[0111] SNP6 locus was significantly correlated with the traits of slaughter rate and bullwhip weight (P<0.05): the slaughter rate and bullwhip weight of individuals with GG genotype were significantly higher than those with AG genotype.

[0112] Example 4 Correlation Analysis between SNPs Haplotypes of VNN1 Gene and Quality Traits of Chinese Simmental Beef

[0113] 1. Experimental Methods

[0114] Haploview 4.2 software was used to analyze the linkage disequilibrium relationship of the six SNPs in Examples 2 and 3. The results showed that four SNPs, SNP2, SNP3, SNP5, and SNP6, were strongly linked. A one-way analysis of variance in SPSS 23.0 software was further used to analyze the correlation between the eight haplotypes at these SNPs and beef quality and carcass traits of Chinese Simmental beef. P < 0.05 was used as the criterion for significant differences. Experimental data are expressed as "mean ± standard error."

[0115] 2. Experimental Results

[0116] The results of linkage disequilibrium analysis are shown in Tables 3 and Figure 8It was found that the four SNPs sites, SNP2, SNP3, SNP5 and SNP6, formed a haplotype block, with a total of 8 different haplotypes with biological statistical significance (number of individuals n≥3). They are H1H1 (the genotypes of the four SNPs sites are CC, GG, GG, GG), H1H2 (the genotypes of the four SNPs sites are CT, GA, GA, GG), H1H3 (the genotypes of the four SNPs sites are CT, GA, GA, GA), H1H4 (the genotypes of the four SNPs sites are CT, GA, GG, GG), H2H2 (the genotypes of the four SNPs sites are TT, AA, AA, GG), H2H3 (the genotypes of the four SNPs sites are TT, AA, AA, GA), H2H4 (the genotypes of the four SNPs sites are TT, AA, AG, GG), and H3H3 (the genotypes of the four SNPs sites are TT, AA, AA, AA).

[0117] Table 3 Haplotype frequencies of SNPs in the VNN1 gene

[0118]

[0119] The correlation analysis results of 8 haplotype combinations with biological statistical significance (n≥3 individuals) and traits are shown in Table 4.

[0120] The slaughter rate of individuals with the H1H4 haplotype was significantly lower than that of individuals with the H1H1, H1H2, and H2H2 haplotypes (P<0.05); the carcass fat coverage of individuals with the H1H1 haplotype was significantly higher than that of individuals with the H2H2 haplotype (P<0.05); the bullwhip weight of individuals with the H1H2 haplotype was significantly higher than that of individuals with the H1H3 haplotype (P<0.05); the testicle weight of individuals with the H1H2 haplotype was significantly higher than that of individuals with the H2H3 haplotype (P<0.05). The meat color score of individuals with H1H2 haplotype was significantly higher than that of individuals with H1H1 and H1H3 haplotypes (P<0.05); the marbling value of individuals with H3H3 haplotype was significantly lower than that of individuals with H1H2, H2H2 and H2H3 haplotypes (P<0.05), and the marbling value of individuals with H1H2 haplotype was significantly higher than that of individuals with H1H1 haplotype (P<0.05).

[0121] Table 4 Correlation analysis between VNN1 gene SNPs haplotypes and meat quality and carcass traits

[0122]

[0123] Example 5 A method for detecting and / or evaluating the quality traits of Chinese Simmental beef

[0124] 1. PCR Amplification and Genotyping

[0125] 1. PCR amplification and sequencing to identify individual genotypes

[0126] PCR amplification was performed using genomic DNA from the blood of Chinese Simmental cattle as a template.

[0127] The upstream primer sequence is: 5′-GGCTAATATCCCAAACAAGTGA-3′ (SEQ ID NO: 1), and the downstream primer sequence is: 5′-CAGAAGAACTCAGACCTCCAA-3′ (SEQ ID NO: 2).

[0128] PCR amplification system (total volume 20 μL): Taq PCR Mix 10 μL, 0.5 μL each of upstream and downstream primers, 2 μL DNA template, 7 μL ddH2O. PCR amplification program: 95°C initial denaturation for 5 min; 35 cycles of denaturation at 95°C for 30 s, annealing at 56°C for 30 s, and extension at 72°C for 1 min; followed by an additional extension at 72°C for 5 min.

[0129] The PCR amplification products were detected by electrophoresis on 1.5% agarose gel at 120V for 18 min. After the target band was confirmed to be correct, they were sent to Sangon Biotech (Shanghai) Co., Ltd. for sequencing.

[0130] SeqMan software was used to observe the sequencing results and confirm the genotype of each sample individual based on the peak plot and sequence.

[0131] 2. Interpretation of results

[0132] SNP site 1 is located at NC_037336.1:70835469, which is the 70835469 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate of individuals with the AA genotype is significantly higher than that of individuals with the GA genotype.

[0133] SNP site 2 is located at NC_037336.1:70835516, which is the 70835516 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with CC genotype is significantly higher than that of individuals with TT genotype, and the marbling grade of individuals with TT and TC genotypes is significantly higher than that of individuals with CC genotype.

[0134] SNP site 3 is located at NC_037336.1:70835542, which is the 70835542 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype. The marbling grade of individuals with the AA and AG genotypes is significantly higher than that of individuals with the GG genotype.

[0135] SNP site 4 is located at NC_037336.1:70835571, which is the 70835571 position on chromosome 9 of the ARS-UCD1.2 version of the bovine genome. The liver weight of individuals with CC genotype is significantly higher than that of individuals with TC genotype.

[0136] SNP site 5 is located at NC_037336.1:70835623, which is the 70835623 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype.

[0137] SNP site 6 is located at NC_037336.1:70835638, which is the 70835638th position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate and bullwhip weight of GG genotype individuals are significantly higher than those of AG genotype individuals.

[0138] The carcass fat coverage of individuals whose genotype at SNP site 2 is CC, whose genotype at SNP site 3 is GG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG.

[0139] The bullwhip weight of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is AG.

[0140] The testicular weight of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AG.

[0141] The meat color score of individuals with the genotype of SNP site 2 being CT, the genotype of SNP site 3 being AG, the genotype of SNP site 5 being AG, and the genotype of SNP site 6 being GG was significantly higher than that of individuals with the following genotypes:

[0142] The genotype of SNP site 2 is CC, the genotype of SNP site 3 is GG, the genotype of SNP site 5 is GG, and the genotype of SNP site 6 is GG;

[0143] The genotype of SNP site 2 is CT, the genotype of SNP site 3 is AG, the genotype of SNP site 5 is AG, and the genotype of SNP site 6 is AG.

[0144] Individuals with a genotype of TT at SNP 2, AA at SNP 3, AA at SNP 5, and AA at SNP 6 had significantly lower marbling grades than individuals with the following genotypes:

[0145] Individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG;

[0146] Individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG;

[0147] An individual whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AG.

[0148] The marbling grade of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is GA, whose genotype at SNP site 5 is GA, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is CC, whose genotype at SNP site 3 is GG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG.

[0149] Example 6 A kit for detecting and / or evaluating quality traits of Chinese Simmental beef

[0150] 1. Composition

[0151] The nucleotide sequences are shown in SEQ ID NO: 1-2, primers, ddH2O, Taq PCR Mix, and NsiI.

[0152] 2. Usage

[0153] Same as Example 5.

[0154] Example 7 A method for detecting and / or evaluating the quality traits of Chinese Simmental beef

[0155] 1. PCR amplification was performed using genomic DNA from Chinese Simmental cattle blood as a template.

[0156] The upstream primer sequence is: 5′-GGCTAATATCCCAAACAAGTGA-3′ (SEQ ID NO: 1), and the downstream primer sequence is: 5′-CAGAAGAACTCAGACCTCCAA-3′ (SEQ ID NO: 2).

[0157] PCR amplification system (total volume 20 μL): Taq PCR Mix 10 μL, 0.5 μL each of upstream and downstream primers (shown in SEQ ID NOs: 1 to 2), 2 μL DNA template, 7 μL ddH2O. PCR amplification program: 95°C initial denaturation for 5 min; 35 cycles of denaturation at 95°C for 30 s, annealing at 56°C for 30 s, and extension at 72°C for 1 min; followed by an additional extension at 72°C for 5 min.

[0158] The PCR amplification products were detected by electrophoresis on 1.5% agarose gel at 120V for 18 min. After the target band was confirmed to be correct, they were sent to Sangon Biotech (Shanghai) Co., Ltd. for sequencing.

[0159] 2. The PCR amplification products were detected by specific enzyme digestion using restriction endonuclease NsiI.

[0160] Enzyme digestion system (total volume 10 μL): CutSmart Buffer 1 μL, NsiI enzyme 0.2 μL, PCR amplification product 5 μL, ddH2O 3.8 μL. Enzyme digestion conditions: Incubate at 37°C for 30 min.

[0161] The enzyme digestion products were detected by electrophoresis on 2% agarose gel at 120V for 20min to confirm the individual genotype.

[0162] 3. Genotype determination

[0163] When the base at NC_037336.1:70835623 (SNP site 5) is an A, this position can be cut by the restriction endonuclease NsiI, meaning that the 976bp amplified product can be cut into two fragments of different sizes, 605bp and 371bp. When the base at this position is a G, it cannot be cut. Therefore, the electrophoresis results for the GG genotype show a single band of 976bp in length; the electrophoresis results for the AG genotype show three bands of 976bp, 605bp, and 371bp in length; and the electrophoresis results for the AA genotype show two bands of 605bp and 371bp in length.

[0164] The genotypes of other sites (SNP site 1 is located at NC_037336.1: 70835469, SNP site 2 is located at NC_037336.1: 70835516, SNP site 3 is located at NC_037336.1: 70835542 and SNP site 4 is located at NC_037336.1: 70835571) are determined based on the sequencing results.

[0165] SNP site 1 is located at NC_037336.1:70835469, which is the 70835469 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate of individuals with the AA genotype is significantly higher than that of individuals with the GA genotype.

[0166] SNP site 2 is located at NC_037336.1:70835516, which is the 70835516 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with CC genotype is significantly higher than that of individuals with TT genotype, and the marbling grade of individuals with TT and TC genotypes is significantly higher than that of individuals with CC genotype.

[0167] SNP site 3 is located at NC_037336.1:70835542, which is the 70835542 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype. The marbling grade of individuals with the AA and AG genotypes is significantly higher than that of individuals with the GG genotype.

[0168] SNP site 4 is located at NC_037336.1:70835571, which is the 70835571 position on chromosome 9 of the ARS-UCD1.2 version of the bovine genome. The liver weight of individuals with CC genotype is significantly higher than that of individuals with TC genotype.

[0169] SNP site 5 is located at NC_037336.1:70835623, which is the 70835623 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype.

[0170] SNP site 6 is located at NC_037336.1:70835638, which is the 70835638th position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate and bullwhip weight of GG genotype individuals are significantly higher than those of AG genotype individuals.

[0171] The carcass fat coverage of individuals whose genotype at SNP site 2 is CC, whose genotype at SNP site 3 is GG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG.

[0172] The bullwhip weight of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is AG.

[0173] The testicular weight of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AG.

[0174] The meat color score of individuals with the genotype of SNP site 2 being CT, the genotype of SNP site 3 being AG, the genotype of SNP site 5 being AG, and the genotype of SNP site 6 being GG was significantly higher than that of individuals with the following genotypes:

[0175] The genotype of SNP site 2 is CC, the genotype of SNP site 3 is GG, the genotype of SNP site 5 is GG, and the genotype of SNP site 6 is GG;

[0176] The genotype of SNP site 2 is CT, the genotype of SNP site 3 is AG, the genotype of SNP site 5 is AG, and the genotype of SNP site 6 is AG.

[0177] Individuals with a genotype of TT at SNP 2, AA at SNP 3, AA at SNP 5, and AA at SNP 6 had significantly lower marbling grades than individuals with the following genotypes:

[0178] Individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG;

[0179] Individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG;

[0180] An individual whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AG.

[0181] The marbling grade of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is GA, whose genotype at SNP site 5 is GA, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is CC, whose genotype at SNP site 3 is GG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG.

[0182] Example 8 A kit for detecting and / or evaluating quality traits of Chinese Simmental beef

[0183] 1. Composition

[0184] Primers with nucleotide sequences as shown in SEQ ID NO: 1-2, ddH2O, Taq PCR Mix, CutSmart Buffer and NsiI enzyme.

[0185] 2. Usage

[0186] Same as Example 7. Sequence Listing <120> A method for detecting and / or evaluating quality traits of Chinese Simmental beef <160> 2 <170> SIPOSequenceListing 1.0 <210> 1 <211> twenty two <212> DNA <213> Artificial Sequence <400> 1 ggctaatatc ccaaacaagt ga 22 <210> 2 <211> twenty one <212> DNA <213> Artificial Sequence <400> 2 cagaagaact cagacctcca a 21

Claims

1. An application of a reagent for predicting and / or evaluating the quality traits of Chinese Simmental cattle beef, characterized in that: The application is one or more of the following: Application in the preparation of a kit for detecting and / or evaluating quality traits of Chinese Simmental cattle beef, Application of the detection and / or evaluation of beef quality traits of Simmental cattle in China, Application of molecular breeding for beef quality traits in Simmental cattle in China, The beef quality traits are dressing percentage, carcass fat coverage, marbling grade, bullwhip weight, liver weight, testicle weight or meat color score; The reagent is used to detect one or more of the genotypes of the following SNP sites: SNP site 1 is located at NC_037336.1:70835469, which is the 70835469 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate of individuals with the AA genotype is significantly higher than that of individuals with the GA genotype. SNP site 2 is located at NC_037336.1:70835516, which is the 70835516 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with CC genotype is significantly higher than that of individuals with TT genotype, and the marbling grade of individuals with TT and TC genotypes is significantly higher than that of individuals with CC genotype. SNP site 3 is located at NC_037336.1:70835542, which is the 70835542 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype. The marbling grade of individuals with the AA and AG genotypes is significantly higher than that of individuals with the GG genotype. SNP site 4 is located at NC_037336.1:70835571, which is the 70835571 position on chromosome 9 of the ARS-UCD1.2 version of the bovine genome. The liver weight of individuals with CC genotype is significantly higher than that of individuals with TC genotype. SNP site 5 is located at NC_037336.1:70835623, which is the 70835623 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype. SNP site 6 is located at NC_037336.1:70835638, which is the 70835638th position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate and bullwhip weight of GG genotype individuals are significantly higher than those of AG genotype individuals.

2. The use according to claim 1, characterized in that The slaughter rate of individuals with the genotype of SNP site 2 being TC, the genotype of SNP site 3 being AG, the genotype of SNP site 5 being GG, and the genotype of SNP site 6 being GG was significantly lower than the slaughter rate of individuals with the following genotypes: Individuals with a genotype of CC at SNP site 2, a genotype of GG at SNP site 3, a genotype of GG at SNP site 5, and a genotype of GG at SNP site 6; Individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG; An individual whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG.

3. The use according to claim 1, characterized in that The carcass fat coverage of individuals whose genotype at SNP site 2 is CC, whose genotype at SNP site 3 is GG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG.

4. The use according to claim 1, characterized in that The bullwhip weight of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is AG.

5. The use according to claim 1, characterized in that The testicular weight of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AG.

6. The use according to claim 1, characterized in that The meat color score of individuals with the genotype of SNP site 2 being CT, the genotype of SNP site 3 being AG, the genotype of SNP site 5 being AG, and the genotype of SNP site 6 being GG was significantly higher than the meat color score of individuals with the following genotypes: The genotype of SNP site 2 is CC, the genotype of SNP site 3 is GG, the genotype of SNP site 5 is GG, and the genotype of SNP site 6 is GG; The genotype of SNP site 2 is CT, the genotype of SNP site 3 is AG, the genotype of SNP site 5 is AG, and the genotype of SNP site 6 is AG.

7. The use according to claim 1, characterized in that The marbling grade of individuals with the genotype of SNP site 2 being TT, the genotype of SNP site 3 being AA, the genotype of SNP site 5 being AA, and the genotype of SNP site 6 being AA was significantly lower than the marbling grade of individuals with the following genotypes: Individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG; Individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG; An individual whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AG.

8. The use according to claim 1, characterized in that The marbling grade of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is GA, whose genotype at SNP site 5 is GA, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is CC, whose genotype at SNP site 3 is GG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG.

9. The use according to claim 1, characterized in that The reagent is a primer with a nucleotide sequence as shown in SEQ ID NO: 1-2.

10. A method for detecting and / or evaluating the quality traits of Chinese Simmental beef, characterized in that: Detect one or more of the following SNP genotypes: SNP site 1 is located at NC_037336.1:70835469, which is the 70835469 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate of individuals with the AA genotype is significantly higher than that of individuals with the GA genotype. SNP site 2 is located at NC_037336.1:70835516, which is the 70835516 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with CC genotype is significantly higher than that of individuals with TT genotype, and the marbling grade of individuals with TT and TC genotypes is significantly higher than that of individuals with CC genotype. SNP site 3 is located at NC_037336.1:70835542, which is the 70835542 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype. The marbling grade of individuals with the AA and AG genotypes is significantly higher than that of individuals with the GG genotype. SNP site 4 is located at NC_037336.1:70835571, which is the 70835571 position on chromosome 9 of the ARS-UCD1.2 version of the bovine genome. The liver weight of individuals with CC genotype is significantly higher than that of individuals with TC genotype. SNP site 5 is located at NC_037336.1:70835623, which is the 70835623 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype. SNP site 6 is located at NC_037336.1:70835638, which is the 70835638th position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate and bullwhip weight of GG genotype individuals are significantly higher than those of AG genotype individuals.

11. The method according to claim 10, characterized in that The slaughter rate of individuals with the genotype of SNP site 2 being TC, the genotype of SNP site 3 being AG, the genotype of SNP site 5 being GG, and the genotype of SNP site 6 being GG was significantly lower than the slaughter rate of individuals with the following genotypes: Individuals with a genotype of CC at SNP site 2, a genotype of GG at SNP site 3, a genotype of GG at SNP site 5, and a genotype of GG at SNP site 6; Individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG; An individual whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG.

12. The method according to claim 10, characterized in that The carcass fat coverage of individuals whose genotype at SNP site 2 is CC, whose genotype at SNP site 3 is GG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG.

13. The method according to claim 10, characterized in that The bullwhip weight of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is AG.

14. The method according to claim 10, characterized in that The testicular weight of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AG.

15. The method according to claim 10, characterized in that The meat color score of individuals with the genotype of SNP site 2 being CT, the genotype of SNP site 3 being AG, the genotype of SNP site 5 being AG, and the genotype of SNP site 6 being GG was significantly higher than the meat color score of individuals with the following genotypes: The genotype of SNP site 2 is CC, the genotype of SNP site 3 is GG, the genotype of SNP site 5 is GG, and the genotype of SNP site 6 is GG; The genotype of SNP site 2 is CT, the genotype of SNP site 3 is AG, the genotype of SNP site 5 is AG, and the genotype of SNP site 6 is AG.

16. The method according to claim 10, characterized in that The marbling grade of individuals with the genotype of SNP site 2 being TT, the genotype of SNP site 3 being AA, the genotype of SNP site 5 being AA, and the genotype of SNP site 6 being AA was significantly lower than the marbling grade of individuals with the following genotypes: Individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is AG, whose genotype at SNP site 5 is AG, and whose genotype at SNP site 6 is GG; Individuals whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is GG; An individual whose genotype at SNP site 2 is TT, whose genotype at SNP site 3 is AA, whose genotype at SNP site 5 is AA, and whose genotype at SNP site 6 is AG.

17. The method according to claim 10, wherein: The marbling grade of individuals whose genotype at SNP site 2 is CT, whose genotype at SNP site 3 is GA, whose genotype at SNP site 5 is GA, and whose genotype at SNP site 6 is GG is significantly higher than that of individuals whose genotype at SNP site 2 is CC, whose genotype at SNP site 3 is GG, whose genotype at SNP site 5 is GG, and whose genotype at SNP site 6 is GG.

18. The method according to claim 10, wherein: Detection was performed using primers with nucleotide sequences as shown in SEQ ID NOs: 1-2.

19. A kit or method for detecting and / or evaluating beef quality traits of Chinese Simmental cattle, and its use in molecular breeding of beef quality traits of Chinese Simmental cattle, characterized in that: The beef quality traits are dressing percentage, carcass fat coverage, marbling grade, bullwhip weight, liver weight, testicle weight or meat color score; The method is the method according to any one of claims 10 to 18; The kit contains one or more reagents for detecting the genotype of the following SNP sites; SNP site 1 is located at NC_037336.1:70835469, which is the 70835469 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate of individuals with the AA genotype is significantly higher than that of individuals with the GA genotype. SNP site 2 is located at NC_037336.1:70835516, which is the 70835516 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with CC genotype is significantly higher than that of individuals with TT genotype, and the marbling grade of individuals with TT and TC genotypes is significantly higher than that of individuals with CC genotype. SNP site 3 is located at NC_037336.1:70835542, which is the 70835542 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype. The marbling grade of individuals with the AA and AG genotypes is significantly higher than that of individuals with the GG genotype. SNP site 4 is located at NC_037336.1:70835571, which is the 70835571 position on chromosome 9 of the ARS-UCD1.2 version of the bovine genome. The liver weight of individuals with CC genotype is significantly higher than that of individuals with TC genotype. SNP site 5 is located at NC_037336.1:70835623, which is the 70835623 position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The carcass fat coverage of individuals with the GG genotype is significantly higher than that of individuals with the AA genotype. SNP site 6 is located at NC_037336.1:70835638, which is the 70835638th position of chromosome 9 in the ARS-UCD1.2 version of the cattle genome. The slaughter rate and bullwhip weight of GG genotype individuals are significantly higher than those of AG genotype individuals.

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