SNP molecular marker related to growth and development traits of Anhui-east cattle and application of SNP molecular marker

Abstract

The invention relates to the technical field of animal molecular breeding, in particular to an SNP (Single Nucleotide Polymorphism) molecular marker related to growth and development traits of Anhui-east cattle and application of the SNP molecular marker. The SNP molecular marker related to the growth and development traits of the Anhui-east cattle comprises at least one of a first SNP molecular marker, a second SNP molecular marker, a third SNP molecular marker, a fourth SNP molecular marker and a fifth SNP molecular marker, the first SNP molecular marker, the second SNP molecular marker, the third SNP molecular marker, the fourth SNP molecular marker and the fifth SNP molecular marker are all located in a TNNI3 gene of a No.18 chromosome of a bovine genome. A primer pair for detecting the SNP marker comprises a T3S2 primer pair or / and a T3S4 primer pair, the sequence of the T3S2 primer pair is as shown in SEQ ID NO: 1 and SEQ ID NO: 2, and the sequence of the T3S4 primer pair is as shown in SEQ ID NO: 3 and SEQ ID NO: 4.

Description

Technical Field

[0001] The present invention relates to the technical field of animal molecular breeding, and in particular to SNP molecular markers related to the growth and development traits of Wandong cattle and applications thereof. Background Art

[0002] As a breed of cattle with local characteristics and excellent genetic resources, Wandong cattle not only excel in meat performance, but also have significant advantages in environmental adaptation and genetic diversity. These characteristics make Wandong cattle a valuable livestock and poultry genetic resource in China, and have important development and utilization value for high-quality beef cattle production.

[0003] Therefore, exploring the growth-related genes of Wandong cattle will play a positive role in promoting the development of China's cattle industry. Studies have shown that the TNNI3 gene SNP locus provides important information on the muscle quality and myocardial tissue function of Tianfu mutton sheep, but no results show that muscle quality is correlated with growth traits. Summary of the invention

[0004] The purpose of the present invention is to solve the shortcomings of the prior art and to propose SNP molecular markers related to the growth and development traits of Wandong cattle and their applications.

[0005] One of the purposes of the present invention is to provide SNP molecular markers related to the growth and development traits of Wandong cattle, including: at least one of a first SNP molecular marker, a second SNP molecular marker, a third SNP molecular marker, a fourth SNP molecular marker, and a fifth SNP molecular marker; the first SNP molecular marker, the second SNP molecular marker, the third SNP molecular marker, the fourth SNP molecular marker, and the fifth SNP molecular marker are all located in the TNNI3 gene of chromosome 18 of the cattle genome.

[0006] The TNNI3 gene can be found in NCBI Reference Sequence: AC_000175.1.

[0007] Preferably, the first SNP molecular marker is located at the base at position 62720648 on chromosome 18 of the cattle genome, and the SNP molecular marker site has a C / T polymorphism; the second SNP molecular marker is located at the base at position 62720925 on chromosome 18 of the cattle genome, and the SNP molecular marker site has a C / G polymorphism; the third SNP molecular marker is located at the base at position 62719179 on chromosome 18 of the cattle genome, and the SNP molecular marker site has an A / G polymorphism; the fourth SNP molecular marker is located at the base at position 62719285 on chromosome 18 of the cattle genome, and the SNP molecular marker site has a G / C polymorphism; the fifth SNP molecular marker is located at the base at position 62719317 on chromosome 18 of the cattle genome, and the SNP molecular marker site has a G / A polymorphism.

[0008] Preferably, the body weight of 6-month-old Wandong cattle with CC genotype and CT genotype at the first SNP molecular marker polymorphism site is significantly higher than that with TT genotype.

[0009] Preferably, the body oblique length of 24-month-old Wandong cattle with CC genotype at the second SNP molecular marker polymorphic site is significantly higher than that with GG genotype.

[0010] Preferably, the cross section and body oblique length of 12-month-old Wandong cattle with GG genotype at the third SNP molecular marker polymorphic site are significantly higher than those with AG genotype.

[0011] Preferably, the body oblique length of 12-month-old Wandong cattle with CC genotype at the fourth SNP molecular marker polymorphic site is significantly higher than that with GG genotype and GC genotype.

[0012] Preferably, the height and weight of 18-month-old Wandong cattle with the GG genotype at the fifth SNP molecular marker polymorphic site are higher than those with the GA genotype, the height, oblique body length and weight of 24-month-old Wandong cattle with the GG genotype are significantly higher than those with the GA genotype, but the weight of 6-month-old Wandong cattle with the GA genotype is significantly higher than that with the GG genotype.

[0013] The second purpose of the present invention is to provide the application of the above-mentioned SNP molecular marker in the detection of growth and development traits of Wandong cattle or in the breeding of Wandong cattle.

[0014] The third object of the present invention is to provide a primer pair for detecting the above-mentioned SNP marker, including: a T3S2 primer pair and / or a T3S4 primer pair.

[0015] The upstream primer of the T3S2 primer pair is shown as SEQ ID NO: 1, and the downstream primer of the T3S2 primer pair is shown as SEQ ID NO: 2.

[0016] The upstream primer of the T3S4 primer pair is shown as SEQ ID NO:3, and the downstream primer of the T3S4 primer pair is shown as SEQ ID NO:4.

[0017] The fourth object of the present invention is to provide the application of the above primer pair in the detection of growth and development traits of Wandong cattle or in the breeding of Wandong cattle.

[0018] A fifth object of the present invention is to provide a kit comprising the above-mentioned primer pair.

[0019] The sixth object of the present invention is to provide a method for predicting the growth and development traits of Wandong cattle, and to detect the genotype of the polymorphic sites of the above-mentioned SNP molecular markers.

[0020] Preferably, the above primer pair or the above kit is used for detection.

[0021] The seventh object of the present invention is to provide a method for breeding Wandong cattle, extracting the genomic DNA of Wandong cattle, amplifying it using the primer pair as described in claim 5, and making corresponding selections with reference to the above-mentioned SNP molecular markers:

[0022] (1) Selecting the CC genotype and the CT genotype of the first SNP molecular marker polymorphic site, and eliminating the TT genotype of the first SNP molecular marker polymorphic site;

[0023] (2) selecting the CC genotype of the second SNP molecular marker polymorphic site and eliminating the GG genotype of the second SNP molecular marker polymorphic site;

[0024] (3) Selecting the GG genotype of the third SNP molecular marker polymorphic site and eliminating the AG genotype of the third SNP molecular marker;

[0025] (4) selecting the CC genotype of the fourth SNP molecular marker polymorphic site, and eliminating the GG genotype and GC genotype of the fourth SNP molecular marker polymorphic site;

[0026] (5) If 6-month-old Wandong cattle are needed, the GA genotype of the fifth SNP molecular marker polymorphic site is selected, and the GG genotype of the fifth SNP molecular marker polymorphic site is eliminated;

[0027] If 18-month-old or 24-month-old Wandong cattle are needed, the GG genotype of the fifth SNP molecular marker polymorphic site is selected, and the GA genotype of the fifth SNP molecular marker polymorphic site is eliminated.

[0028] Beneficial effects:

[0029] The SNP molecular marker provided by the present invention is significantly correlated with the growth and development traits of Wandong cattle. The SNP molecular marker can be used for auxiliary selection of the growth and development traits of Wandong cattle, and screen Wandong cattle with excellent growth and development potential. It has important practical application value for further improving the growth and development of Wandong cattle and using specific Wandong cattle as material for variety or strain selection, which facilitates breeders to breed and cultivate Wandong cattle. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 This is the result of electrophoresis test of genomic DNA in the blood of Wandong cattle.

[0031] Figure 2 This is the electrophoresis detection result of PCR amplification using T3S2 primer pair.

[0032] Figure 3 This is the electrophoresis detection result of PCR amplification using T3S4 primer pair.

[0033] Figure 4 This is the sequencing peak diagram of different genotypes of the g.62720648C>T mutation site.

[0034] Figure 5 This is the sequencing peak diagram of different genotypes of the g.62720753A>C mutation site.

[0035] Figure 6 This is the sequencing peak diagram of different genotypes at the g.62720859A>G mutation site.

[0036] Figure 7 This is the sequencing peak diagram of different genotypes of the g.62720911T>C mutation site.

[0037] Figure 8 This is the sequencing peak diagram of different genotypes at the g.62720925C>G mutation site.

[0038] Fig. 9 This is the sequencing peak diagram of different genotypes at the g.62719179A>G mutation site.

[0039] Fig.10 This is the sequencing peak diagram of different genotypes of the g.62719285G>C mutation site.

[0040] Fig.11 This is the sequencing peak diagram of different genotypes of the g.62719317G>A mutation site. DETAILED DESCRIPTION

[0041] The present invention will be further explained below in conjunction with specific embodiments.

[0042] Example 1

[0043] At the Anhui Wandong cattle breeding farm, Chuzhou City (Fengyang County) Daming Agriculture and Animal Husbandry Development Co., Ltd. collected body size and length data of 220 6-month-old, 12-month-old, 18-month-old and 24-month-old Wandong cattle from multiple cow sheds, and drew 5 mL of blood from the jugular vein or tail vein of all cattle, placed the blood in a blood vessel with an anticoagulant, and stored it in a refrigerator.

[0044] Example 2

[0045] 2.1 Extraction of genomic DNA from blood samples

[0046] Thaw the frozen blood sample anticoagulant tube in a refrigerator at 4°C in advance to prevent the blood anticoagulant tube from bursting due to the large temperature difference and causing the loss of blood sample. Extract the sample DNA with the purchased blood DNA extraction kit according to the instructions. Seal the extracted 220 blood DNA samples and freeze them at -20°C for later use.

[0047] 2.2 Detection of blood DNA samples

[0048] 2.2.1 Agarose gel electrophoresis detection

[0049] After adding a small amount of 1×TAE buffer to the cooled and completely solidified agarose gel plate, slowly pull out the electrophoresis comb, put the electrophoresis gel plate into the electrophoresis tank and pour in 1×TAE electrophoresis buffer until the electrophoresis gel plate is submerged. First extract 5μL DL 2000DNA Marker and add it to the first spotting well of the gel plate, then extract blood DNA samples from different individuals and mix them with loading buffer at a ratio of 1μL:5μL and inject them into the corresponding spotting wells, perform electrophoresis at 120V for 40min, place the gel on a UV imager to analyze the electrophoresis results, and take photos for preservation.

[0050] 2.2.2 Ultra-micro UV spectrophotometer detection

[0051] The ultra-micro-volume UV spectrophotometer was set to DNA determination mode and ddH 2 Wash the sample wells of the ultra-micro UV spectrophotometer several times and perform blank correction. After the instrument calibration is completed, take 1 μL of DNA sample from each sample for spot detection. After each detection, wash with ddH 2 O is used to clean and calibrate the sample wells. 260 / OD 280 The two values ​​are used to determine the concentration and purity of each sample.

[0052] The Conce value represents the DNA concentration of the measured sample, and its unit is ng / mol.

[0053] OD 260 / OD 280The theoretical value is between 1.7 and 2.2, indicating that the purity of each sample DNA is high; the sample OD 260 / OD 280 The value is less than 1.7, the sample DNA may be contaminated by incomplete digestion of proteins and other substances during the extraction process; if the OD 260 / OD 280 If the value is greater than 2.2, it may be due to contamination such as RNA during the extraction process.

[0054] 2.3 Primer design and synthesis

[0055] Aiming at the variant site of the third exon of the TNNI3 gene in the experimental Wandong cattle population, the Primer3Plus-PickPrimers software was used to design amplification primers for different SNP sites in the TNNI3 gene and named them T3S2 and T3S4. The primer information is as follows:

[0056]

[0057]

[0058] 2.4 PCR amplification

[0059] The above primers were amplified using 2×Taq PCR Master mix, and multiple gradient tests were performed to explore the optimal annealing temperature, and ultimately the optimal reaction conditions and reaction system for each primer were determined.

[0060] (1) The PCR reaction system is as follows:

[0061]

[0062] (2) Different primers were designed according to different SNP sites. The PCR reaction conditions are shown in the following table. After each pair of primers was amplified according to the reaction system, 5 μL of the obtained PCR product was extracted for agarose gel (3%) electrophoresis detection. The electrophoresis results of the PCR product were observed using a UV imager. The successfully amplified TNNI3 primers will be used for large-scale verification experiments in the later stage.

[0063]

[0064]

[0065] 2.5 Association analysis

[0066] The GLM process of SPSS27.0 software was used to conduct association analysis between the growth and development traits of Wandong cattle and the SNPs of candidate genes. The association analysis model was:

[0067] Y=μ+G+m+e

[0068] In the formula, Y represents the measured value of the phenotypic trait, μ represents the population mean, G represents the genotype effect, m represents the sex effect, and e represents the residual effect.

[0069] Example 3

[0070] 3.1 Results of extraction of Wandong cattle genomic DNA

[0071] Blood sample DNA was tested using agarose gel electrophoresis. Figure 1 As shown, it can be observed that the bands are clear and bright without any mixed bands. The concentration and OD value of the sample DNA are tested using an ultra-micro spectrophotometer and both meet the test requirements and can be used for subsequent tests.

[0072] 3.2 PCR amplification with different primers

[0073] The designed primers were used for PCR amplification. The gel electrophoresis of the two primer pairs T3S2 and T3S4 showed clear bands without any other bands. The electrophoresis results were as follows: Figure 2 and Figure 3 shown.

[0074] in, Figure 2 This is the PCR result of the mutation site with primer pair T3S2, and the length of the amplified band is 630 bp; Figure 3 This is the PCR result of the mutation site using the T3S4 primer pair. The length of the amplified band is 584bp, which is consistent with the fragment length of the designed primers. The PCR amplification is normal, so these two pairs of primers can continue to the next step of the experimental operation.

[0075] 3.3 Mutation site sequencing

[0076] Sanger sequencing showed that there were mutations at 8 sites in the TNNI3 gene. GeneBank search revealed that 3 SNP sites were located in the 5th intron region and 5 SNP sites were located in the 7th intron region. The specific information is shown in the table below.

[0077]

[0078] Afterwards, peak sequencing was performed on 8 sites, among which:

[0079] g.62720648C>T mutation Figure 4 As shown, there are three genotypes, namely CC, CT, and TT genotypes. CC is the wild homozygous type, TT is the mutant homozygous type, and CT is the mutant heterozygous type. Therefore, the mutant heterozygous type will be displayed as an overlapping peak state in the sequencing peak graph;

[0080] g.62720753A>C mutation Figure 5As shown, there are three genotypes, namely AA, AC, and CC genotypes. AA is the wild homozygous type, GG is the mutant homozygous type, and AG is the mutant heterozygous type. Therefore, the mutant heterozygous type will be displayed as an overlapping peak state in the sequencing peak graph.

[0081] The mutation of g.62720859A>G is as follows Figure 6 As shown, there are three genotypes, namely, AA, AG, and GG genotypes. AA is the wild homozygous type, GG is the mutant homozygous type, and AG is the mutant heterozygous type. Therefore, the mutant heterozygous type will be displayed as an overlapping peak state in the sequencing peak graph;

[0082] g.62720911T>C mutation Figure 7 As shown, there are three genotypes, namely, TT, TC, and CC genotypes. TT is the wild homozygous type, CC is the mutant homozygous type, and TC is the mutant heterozygous type. Therefore, the mutant heterozygous type will be displayed as an overlapping peak state in the sequencing peak graph;

[0083] The mutation of g.62720925C>G is as follows Figure 8 As shown, there are three genotypes, namely GG, GC, and CC genotypes. GG is the wild homozygous type, CC is the mutant homozygous type, and GC is the mutant heterozygous type. Therefore, the mutant heterozygous type will be displayed as an overlapping peak state in the sequencing peak graph;

[0084] The mutation of g.62719179A>G is as follows Fig. 9 As shown, there are three genotypes, namely, AA, AG, and GG genotypes. AA is the wild homozygous type, GG is the mutant homozygous type, and AG is the mutant heterozygous type. Therefore, the mutant heterozygous type will be displayed as an overlapping peak state in the sequencing peak graph;

[0085] g.62719285G>C mutation Fig.10 As shown, there are three genotypes, namely GG, GC, and CC genotypes. GG is the wild homozygous type, CC is the mutant homozygous type, and GC is the mutant heterozygous type. Therefore, the mutant heterozygous type will be displayed as an overlapping peak state in the sequencing peak graph;

[0086] The mutation of g.62719317G>A is as follows Fig.11 As shown, there are two genotypes, namely GG and GA genotypes, GG is the wild homozygous type, and GA is the mutant heterozygous type. Therefore, the mutant heterozygous type will be displayed as an overlapping peak state in the sequencing peak graph.

[0087] 3.4 Statistical analysis of sequencing results of experimental samples

[0088] The genotype frequencies and gene frequencies of the eight mutation sites of the TNNI3 gene are shown in the following table. 2 Value analysis showed that all eight sites were in Hardy-Weinberg equilibrium.

[0089]

[0090]

[0091] 3.5 Statistical analysis of polymorphic information content of eight mutation sites in the TNNI3 gene of the sample population

[0092] By counting the polymorphic information content of the eight mutation sites of the TNNI3 gene in the sample population, the statistical results shown in the following table were obtained.

[0093]

[0094]

[0095] Through analysis, we can get:

[0096] (1) The polymorphic information content (PIC) of the two loci g.62720648C>T and g.62719317G>A is less than 0.25, so these three loci are low polymorphic. Therefore, the two loci g.62720648C>T and g.62719317G>A on the TNNI3 gene in the sample population belong to a low level of genetic variation.

[0097] (2) The polymorphic information content of the six polymorphic loci g.62720753A>C, g.62720859A>G, g.62720911T>C, g.62720925C>G, g.62719179A>G, and g.62719285G>C is greater than 0.25 and less than 0.5, so these six loci are moderately polymorphic. Therefore, the six loci g.62720753A>C, g.62720859A>G, g.62720911T>C, g.62720925C>G, g.62719179A>G, and g.62719285G>C on the TNNI3 gene in the sample population all belong to a medium level of genetic variation.

[0098] 3.6 Association analysis between SNP loci and growth and development traits of Wandong cattle

[0099] SPSS27.0 software was used to analyze the correlation between the TNNI3 gene SNP sites and growth and development traits of the Wandong cattle experimental population.

[0100] The results of the correlation analysis between the g.62720648C>T locus and the growth traits of Wandong cattle at different ages are shown in the following table:

[0101]

[0102] The results of the correlation analysis between the g.62720753A>C locus and the growth traits of Wandong cattle at different ages are shown in the following table:

[0103]

[0104]

[0105] The results of the correlation analysis between the g.62720859A>G locus and the growth traits of Wandong cattle at different ages are shown in the following table:

[0106]

[0107]

[0108] The results of the correlation analysis between the g.62720911T>C locus and the growth traits of Wandong cattle at different ages are shown in the following table:

[0109]

[0110] The results of the correlation analysis between the g.62720925C>G locus and the growth traits of Wandong cattle at different ages are shown in the following table:

[0111]

[0112] The results of the correlation analysis between the g.62719179A>G locus and the growth traits of Wandong cattle at different ages are shown in the following table:

[0113]

[0114]

[0115] The results of the correlation analysis between the g.62719285G>C locus and the growth traits of Wandong cattle at different ages are shown in the following table:

[0116]

[0117]

[0118] The results of the correlation analysis between the g.62719317G>A locus and the growth traits of Wandong cattle at different ages are shown in the following table:

[0119]

[0120] In the above tables, two groups of data with the same letter on their shoulders indicate no significant difference, and two groups of data with different letters on their shoulders indicate significant difference. Lowercase letters indicate P<0.05, i.e. significant difference; uppercase letters indicate P<0.01, i.e. extremely significant difference.

[0121] The analysis results show that among the 8 SNP sites of TNNI3 gene, g.62720648C>T, g.62720925C>G, g.62719179A>G, g.62719285G>C, and g.62719317G>A are correlated with the growth and development traits of Wandong cattle.

[0122] In summary, the present invention screens mutation sites on the TNNI3 gene, analyzes the effects of these mutation sites on the growth and development traits of Wandong cattle, and then selects and breeds the growth and development traits of Wandong cattle from a molecular level.

[0123] The present invention has been found through experiments and the chi-square test shows that the eight sites g.62720648C>T, g.62720753A>C, g.62720859A>G, g.62720911T>C, g.62720925C>G, g.62719179A>G, g.62719285G>C, and g.62719317G>A in the TNNI3 gene are all in a Hardy-Weinberg equilibrium state (P>0.05), indicating that these eight sites have good adaptability after long-term selection evolution. The polymorphic information content showed that the six loci g.62720753A>C, g.62720859A>G, g.62720911T>C, g.62720925C>G, g.62719179A>G, and g.62719285G>C were in a moderate state (0.25≤PIC<0.5), indicating that these six loci had strong selection potential in Wandong cattle. The polymorphic information content PIC of the two loci g.62720648C>T and g.62719317G>A was less than 0.25, so these three loci were low polymorphic, so the two loci g.62720648C>T and g.62719317G>A on the TNNI3 gene in the sample population belonged to a low level of genetic variation.

[0124] Association analysis showed that among the 8 SNP sites of TNNI3 gene, g.62720648C>T, g.62720925C>G, g.62719179A>G, g.62719285G>C, and g.62719317G>A were correlated with the growth and development traits of Wandong cattle. Molecular selection of Wandong cattle was carried out through these 5 mutation sites to cultivate excellent growth and development traits that can be stably inherited to offspring; the SNP molecular marker provided by the present invention is significantly correlated with the growth and development traits of Wandong cattle, and the SNP marker can be used for auxiliary selection of growth and development of Wandong cattle, and Wandong cattle with excellent growth and development trait potential can be screened, which has important practical application value for further improving the growth and development of Wandong cattle and using specific Wandong cattle as materials for variety or strain selection, and is convenient for breeders to breed and cultivate Wandong cattle.

[0125] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. SNP molecular markers related to the growth and development traits of Wandong cattle, characterized in that: include: At least one of the first SNP molecular marker, the second SNP molecular marker, the third SNP molecular marker, the fourth SNP molecular marker, and the fifth SNP molecular marker; The first SNP molecular marker, the second SNP molecular marker, the third SNP molecular marker, the fourth SNP molecular marker, and the fifth SNP molecular marker are all located in the TNNI3 gene on chromosome 18 of the cattle genome.

2. The SNP molecular marker according to claim 1, characterized in that: The first SNP molecular marker is located at the base of position 62720648 on chromosome 18 of the cattle genome, and the SNP molecular marker site has a C / T polymorphism; the second SNP molecular marker is located at the base of position 62720925 on chromosome 18 of the cattle genome, and the SNP molecular marker site has a C / G polymorphism; the third SNP molecular marker is located at the base of position 62719179 on chromosome 18 of the cattle genome, and the SNP molecular marker site has an A / G polymorphism; the fourth SNP molecular marker is located at the base of position 62719285 on chromosome 18 of the cattle genome, and the SNP molecular marker site has a G / C polymorphism; The fifth SNP molecular marker is located at the base of position 62719317 on chromosome 18 of the cattle genome, and the SNP molecular marker site has a G / A polymorphism.

3. The SNP molecular marker according to claim 1, characterized in that: The body weight of 6-month-old Wandong cattle with CC genotype and CT genotype of the first SNP molecular marker polymorphism site was significantly higher than that with TT genotype; The CC genotype of the second SNP molecular marker polymorphism site had a significantly higher body length than the GG genotype in 24-month-old Wandong cattle. The cross section and body oblique length of the 12-month-old Wandong cattle with the GG genotype at the third SNP molecular marker polymorphism site were significantly higher than those with the AG genotype; The CC genotype of the fourth SNP molecular marker polymorphism site had a significantly higher body length than the GG and GC genotypes in 12-month-old Wandong cattle. The height and weight of 18-month-old Wandong cattle with GG genotype at the fifth SNP molecular marker polymorphism site were higher than those with GA genotype. The height, oblique body length and weight of 24-month-old Wandong cattle with GG genotype were significantly higher than those with GA genotype. However, the weight of 6-month-old Wandong cattle with GA genotype was significantly higher than that with GG genotype.

4. Application of the SNP molecular marker as described in any one of claims 1 to 3 in the detection of growth and development traits of Wandong cattle or in the breeding of Wandong cattle.

5. A primer pair for detecting the SNP marker according to any one of claims 1 to 3, characterized in that: include: T3S2 primer pair and / or T3S4 primer pair, the upstream primer of the T3S2 primer pair is shown as SEQ ID NO: 1, the downstream primer of the T3S2 primer pair is shown as SEQ ID NO: 2, the upstream primer of the T3S4 primer pair is shown as SEQ ID NO: 3, and the downstream primer of the T3S4 primer pair is shown as SEQ ID NO:

4.

6. Use of the primer pair as claimed in claim 5 in the detection of growth and development traits of Wandong cattle or in the breeding of Wandong cattle.

7. A kit, characterized in that: Containing the primer pair according to claim 5.

8. A method for predicting the growth and development traits of Wandong cattle, characterized in that: Detect the genotype of the polymorphic site of the SNP molecular marker as described in any one of claims 1-3.

9. The method according to claim 8, characterized in that The detection is performed using the primer pair described in claim 5 or the kit described in claim 7.

10. A method for breeding Wandong cattle, characterized in that: Extract the genomic DNA of Wandong cattle, amplify it using the primer pair described in claim 5, and make corresponding selections with reference to the SNP molecular markers described in any one of claims 1 to 3: (1) Selecting the CC genotype and the CT genotype of the first SNP molecular marker polymorphic site, and eliminating the TT genotype of the first SNP molecular marker polymorphic site; (2) selecting the CC genotype of the second SNP molecular marker polymorphic site and eliminating the GG genotype of the second SNP molecular marker polymorphic site; (3) Selecting the GG genotype of the third SNP molecular marker polymorphic site and eliminating the AG genotype of the third SNP molecular marker; (4) selecting the CC genotype of the fourth SNP molecular marker polymorphic site, and eliminating the GG genotype and GC genotype of the fourth SNP molecular marker polymorphic site; (5) If 6-month-old Wandong cattle are needed, the GA genotype of the fifth SNP molecular marker polymorphic site is selected, and the GG genotype of the fifth SNP molecular marker polymorphic site is eliminated; If 18-month-old or 24-month-old Wandong cattle are needed, the GG genotype of the fifth SNP molecular marker polymorphic site is selected, and the GA genotype of the fifth SNP molecular marker polymorphic site is eliminated.

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