A SNP molecular marker related to foraging behavior traits on pig rbfox1 locus and application thereof

Abstract

The application belongs to the technical field of molecular biology and molecular marker, and particularly relates to a SNP molecular marker related to a foraging behavior trait on a pig RBFOX1 locus and application. The SNP site of the SNP molecular marker corresponds to a G>T mutation at position 35094559 on chromosome 3 of an international pig reference genome 11.1 version reference sequence. The application can increase the frequency of the dominant allele generation by generation by selecting the dominant allele of the molecular marker, reduce the average daily foraging times of pigs, increase the average foraging time of pigs, and improve the feed conversion efficiency of pigs. Selecting the superior pigs with the above traits can help to accelerate the genetic improvement of pigs and improve the economic benefits of pig breeding.

Description

Technical Field

[0001] This invention belongs to the fields of molecular biology and molecular marker technology, specifically relating to a SNP molecular marker located at the pig RBFOX1 locus that is associated with feeding behavior traits and its application. Background Technology

[0002] In pig production, feed costs account for 50% to 85% of total production costs, making it the largest expense. Therefore, feed conversion ratio (FCR) is an extremely important trait in pigs, closely related to the economic benefits of the pig industry. FCR is regulated by multiple genes with minor effects, and progress in improving FCR has been slow, with few major QTLs affecting FCR identified. Therefore, strengthening research on FCR is essential. Previous studies have shown a strong correlation between feeding behavior traits and feeding efficiency traits in pigs. Under the same conditions, individuals with fewer average daily feeding frequency and longer average feeding times have higher feed conversion ratios. Therefore, it is necessary to conduct in-depth research on feeding behavior traits in pigs, aiming to improve feed conversion ratio through screening of these traits.

[0003] Feeding behavior traits in pigs include mean daily feed intake (NVD), mean total feed volume (TPV), and mean feed rate (FR). NVD and TPV are often used as indicators to improve feed conversion ratio (FCR) in pigs. However, simply studying the correlation between feeding behavior traits and feed efficiency traits cannot directly reveal the genetic loci affecting feeding behavior traits and FCR. Therefore, using modern genetic methods to identify molecular genetic markers affecting feeding behavior traits and FCR is of great significance for improving feed conversion efficiency in pigs and promoting the development of the pig industry.

[0004] Genome-wide association studies (GWAS) have become one of the mainstream methods for identifying genetic markers of traits with low heritability. By analyzing the association between genome-wide genetic markers (SNPs, InDels, SVs, etc.) and phenotypes, GWAS can accurately locate the genetic factors affecting phenotypes and economic traits, showing unique advantages in the genetic improvement of complex traits. Compared with candidate gene methods and QTL mapping, GWAS has significant advantages in terms of mapping accuracy and discovery of new genes.

[0005] Duroc pigs are widely used in production due to their excellent growth performance; however, few molecular genetic markers associated with feed conversion efficiency have been identified in Duroc pig populations. Therefore, identifying candidate genes associated with feeding behavior and feed efficiency traits in Duroc pigs using GWAS is of great significance for improving feed conversion efficiency in pigs and promoting the development of the pig industry. Summary of the Invention

[0006] To overcome the shortcomings and disadvantages of the prior art, the primary objective of this invention is to provide a SNP molecular marker located at the pig RBFOX1 locus that is associated with feeding behavior traits.

[0007] Another object of the present invention is to provide a primer pair for detecting the above-mentioned SNP molecular markers.

[0008] Another object of the present invention is to provide a kit for detecting the above-mentioned SNP molecular markers, the kit comprising the above-mentioned primer pairs.

[0009] The fourth objective of this invention is to provide applications of the above-mentioned SNP molecular markers, primer pairs, and kits.

[0010] The fifth objective of this invention is to provide a method for genetic improvement of pigs.

[0011] The objective of this invention is achieved through the following technical solution:

[0012] A SNP molecular marker located at the RBFOX1 locus in pigs and associated with feeding behavior traits is disclosed. The SNP site of the molecular marker corresponds to the G>T mutation at position 35094559 on chromosome 3 of the International Swine Reference Genome Version 11.1. Polymorphism of the base at this site affects feeding behavior in pigs. Among them, pigs with TG and GG genotypes have better feeding behavior or feed conversion rate than pigs with TT genotypes.

[0013] The feeding behavior traits mentioned include, but are not limited to, the average number of feed intakes per day (NVD) and the average time per feed (TPV); among them, the average number of feed intakes per day for TG and GG genotype pigs is lower than that for TT genotype pigs, and the average time per feed for TG and GG genotype pigs is higher than that for TT genotype pigs.

[0014] The pigs mentioned are Duroc and its synthetic lines;

[0015] The preferred pigs are the Canadian Duroc strain and its synthetic strains;

[0016] The preferred nucleotide sequence of the SNP molecular marker is shown in SEQ ID NO: 1, wherein M in the sequence is G or T, leading to differences in pig feeding behavior traits;

[0017] The SNP site of the SNP molecular marker is the G121-T121 nucleotide mutation at position 121 of the sequence marked in SEQ ID NO: 1 (corresponding to the G>T mutation at position 35094559 on chromosome 3 of the International Pig Reference Genome Version 11.1, named: g.121G>T);

[0018] A primer pair for detecting the above-mentioned SNP molecular markers comprises primers P001-F and P002-R, the nucleotide sequences of which are shown below:

[0019] P001-F: 5'-TGCCTTAGAGTGGGTCAT-3';

[0020] P002-R: 5'-AATAGGGCCTTCAACAGG-3';

[0021] A kit for detecting the above-mentioned SNP molecular markers, comprising the above-mentioned primer pairs;

[0022] The application of the SNP molecular markers, primer pairs or kits in identifying feeding behavior-related traits or feed conversion rate traits, screening pig breeds with low average daily feeding frequency, long average feeding time per session or high feed conversion rate, or in the genetic breeding of pig feeding behavior-related traits or feed conversion rate traits.

[0023] The genetic breeding method is preferably marker-assisted breeding;

[0024] The application of the aforementioned SNP molecular markers in gene editing;

[0025] The pigs mentioned are Duroc and its synthetic lines;

[0026] The preferred pigs are Canadian Duroc pigs and their synthetic lines;

[0027] A method for detecting traits related to pig feeding behavior or feed conversion ratio includes the following steps:

[0028] The above-mentioned SNP molecular markers on the RBFOX1 locus in pigs were detected. Based on whether the single nucleotide of the SNP site of the SNP molecular marker is G or T, the pig feeding behavior-related traits were determined. Among them, the feeding behavior or feed conversion rate of pigs with TG and GG genotypes was better than that of pigs with TT genotypes.

[0029] The feeding behavior traits mentioned include, but are not limited to, the average number of feed intakes per day (NVD) and the average time per feed (TPV); among them, the average number of feed intakes per day for TG and GG genotype pigs is lower than that for TT genotype pigs, and the average time per feed for TG and GG genotype pigs is higher than that for TT genotype pigs.

[0030] A method for screening pig breeds with low average daily feeding frequency, long average feeding time, or high feed conversion ratio using the above molecular markers includes the following steps:

[0031] The above-mentioned SNP molecular markers on the RBFOX1 locus in pigs were detected. Based on the SNP sites of the SNP molecular markers, individuals with the TT genotype were eliminated, while individuals with the TG or GG genotypes were retained. Among them, the feeding behavior or feed conversion rate of pigs with the TG and GG genotypes was better than that of pigs with the TT genotype.

[0032] The feeding behavior traits mentioned include, but are not limited to, the average number of feed intakes per day (NVD) and the average time per feed (TPV); among them, the average number of feed intakes per day for TG and GG genotype pigs is lower than that for TT genotype pigs, and the average time per feed for TG and GG genotype pigs is higher than that for TT genotype pigs.

[0033] The detection method includes the following steps:

[0034] (1) Extract genomic DNA from the pigs to be tested;

[0035] (2) Using the primer pairs mentioned above or the primer pairs in the kit mentioned above as amplification primers, and using the genomic DNA of the pig to be tested obtained in step (1) as template DNA, PCR amplification is performed to obtain PCR amplification products.

[0036] (3) Sequencing the PCR amplification products to obtain sequencing results;

[0037] (4) Based on the sequencing results, determine the genotype of the SNP molecular markers;

[0038] The pigs mentioned are Duroc and its synthetic lines;

[0039] The preferred pigs are Canadian Duroc pigs and their synthetic lines;

[0040] A method for genetic improvement of pigs, comprising the following steps:

[0041] Identify the SNP sites of the above-mentioned SNP molecular markers in the core breeding pig population, and make corresponding selections based on the molecular markers: select breeding pig individuals with TG and GG genotypes at locus 35094559 on chromosome 3 of the International Swine Reference Genome Version 11.1 in the core breeding pig population, and cull breeding pig individuals with TT genotype at this locus, so as to increase the frequency of allele G at this locus generation by generation, thereby improving the pig's feeding behavior or feed conversion rate;

[0042] The pigs mentioned are Duroc and its synthetic lines;

[0043] The preferred pigs are Canadian Duroc pigs and their synthetic lines;

[0044] The present invention has the following advantages and effects compared with the prior art:

[0045] (1) This invention selects NVD and TPV as indicators for improving the feed conversion rate of pigs. It studies and determines that the molecular markers related to pig feeding behavior traits are located on the nucleotide sequence of the RBFOX1 locus on chromosome 3 of pigs. Specifically, it verifies the effects of these markers on pig feeding behavior, such as the average number of feedings per day and the average time of each feeding. Finally, it establishes an efficient and accurate molecular marker-assisted breeding technology, which is applied to the genetic improvement of reducing the average number of feedings per day and increasing the average time of each feeding in breeding pigs. This improves the feed conversion efficiency of offspring pigs, reduces feed costs, increases the economic profit of enterprises, and enhances core competitiveness.

[0046] (2) The present invention provides a primer pair and kit for detecting the above-mentioned molecular markers. With the primer pair and kit, an efficient and accurate molecular marker-assisted breeding technology can be established, and the average daily feeding frequency and average feeding time can be selected quickly and accurately, thereby accelerating the breeding process.

[0047] (3) By selecting the superior alleles of the above molecular markers, the present invention can increase the frequency of superior alleles generation by generation, reduce the average number of times the breeding pigs eat per day and increase the average time of each feeding, select superior breeding pigs with fewer average number of times they eat per day and longer average time of each feeding, accelerate the progress of pig genetic improvement and thus effectively improve the economic benefits of pig breeding. Attached Figure Description

[0048] Figure 1 This is a Manhattan plot of the average daily feed intake of Duroc pigs on chromosome 3, generated using the mrMLM model in mrMLM software. The plot shows the x-axis representing the chromosome number of the pig and the principal y-axis representing -log[i, j]. 10 The P-value is represented by the secondary y-axis, which indicates the LOD value.

[0049] Figure 2 This is a Manhattan plot of multi-site GWAS on chromosome 3 of Canadian Duroc pigs, using the FASTmrMLM model in mrMLM software to represent the average daily feed intake. The x-axis represents the chromosome number of the pig, and the principal y-axis represents -log... 10 The P-value is represented by the secondary y-axis, which indicates the LOD value.

[0050] Figure 3 This is a Manhattan plot of multi-site GWAS on chromosome 3 of Canadian Duroc pigs using the FASTmrEMMA model in mrMLM software, showing the average daily feed intake. The x-axis represents the chromosome number of the pig, and the principal y-axis represents -log[i, j]. 10 The P-value is represented by the secondary y-axis, which indicates the LOD value.

[0051] Figure 4This is a box plot showing the phenotypic differences in the average daily feed intake of pigs with different genotypes.

[0052] Figure 5 This is a Manhattan plot of multi-site GWAS for Duroc pigs on chromosome 3, using the mrMLM model in mrMLM software, to show the average feeding time per session. The x-axis represents the chromosome number of the pig, and the principal y-axis represents -log... 10 The P-value is represented by the secondary y-axis, which indicates the LOD value.

[0053] Figure 6 This is a Manhattan plot of multi-site GWAS on chromosome 3 of Canadian Duroc pigs using the FASTmrMLM model in mrMLM software, showing the average feeding time per session. The x-axis represents the chromosome number of the pig, and the principal y-axis represents -log... 10 The P-value is represented by the secondary y-axis, which indicates the LOD value.

[0054] Figure 7 This is a Manhattan plot of multi-site GWAS on chromosome 3 of Canadian Duroc pigs using the FASTmrEMMA model in mrMLM software, showing the average feeding time per session. The x-axis represents the chromosome number of the pig, and the principal y-axis represents -log... 10 The P-value is represented by the secondary y-axis, which indicates the LOD value.

[0055] Figure 8 This is a box plot showing the phenotypic differences in the average feeding time per feeding session among pigs of different genotypes. Detailed Implementation

[0056] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0057] Example 1: Experimental Subjects, Phenotypic Determination, and DNA Sample Collection

[0058] (1) Experimental animals

[0059] The experimental pig herd used in this study was a core group of Canadian Duroc pigs from a company in Guangdong Province. A total of 1251 Canadian Duroc pigs were selected from this core group, and the herd's pedigree was recorded in detail. The pigs in this experiment had free access to feed and water, and the feeding methods and rearing conditions remained consistent throughout the experiment, following standard practices.

[0060] (2) Phenotypic determination

[0061] The average number of feed intakes per day and the average time of each feed intake were measured using standard methods at the testing station for 1251 Canadian Duroc pigs.

[0062] (3) Collection of pig tissue samples

[0063] To extract DNA, ear samples were collected from the aforementioned 1251 Canadian Duroc pigs and stored in a -80°C freezer. These samples were subsequently transferred to the company for SNP chip genotyping.

[0064] Example 2: SNP chip genotyping and genome-wide association analysis

[0065] (1) Genotyping using 50K SNP chip

[0066] ① The pig ear sample obtained in Example 1 was sent to Neogene Biotech (Shanghai) Co., Ltd. The pig whole genome 50K SNP chip (Illumina, USA) genotype was determined on the IlluminaBeadstration platform according to the company's standard procedure. The specific steps included: DNA extraction, filtering according to DNA quality, etc., and finally the original PLINK format (.map, .ped) file was obtained.

[0067] ② Use PLINK v2.0 to convert the original PLINK format (.map, .ped) files into binary PLINK format files.

[0068] ③ The 50K chip scan genotyping data of all the above samples were quality controlled using PLINK v2.0 software. Individuals with a detection rate below 90%, a family Mendelian error rate above 0.1, a minimum allele frequency below 0.05, and a Hardy-Weinberg equilibrium significance level above 10 were excluded. -6 The SNPs are then used to obtain PLINK format (.fam, .bim, .bed) files containing the variant sites.

[0069] (2) Multisite GWAS analysis

[0070] Multi-site GWAS analysis was performed using the mrMLM, FASTmrMLM, and FASTmrEMMA models from the mrMLM software package developed by Professor Yuan-Ming Zhang of Huazhong Agricultural University to analyze the average daily feeding frequency and average feeding time. The specific methods are as follows:

[0071] ① Use the PLINK file containing the final variant sites obtained in step (1) of PLINK2.0 to fill in the genotype file;

[0072] ② Extract the average daily feeding frequency and average feeding time per session from the original phenotypic file as the phenotypic file;

[0073] ③ Use GCTA software to convert the PLINK format file into a GRM format genotype file, select the --pca parameter, input the GRM format genotype file to calculate PCA and take the first three principal components; extract the sex, batch number, and the first three principal components of the PCA calculated by GCTA from the original records as covariance files;

[0074] ④ Prepare the genotype, phenotype, and covariance files according to the software's required format, input them into the software, select the corresponding model for calculation, and obtain the significant site results. The significance threshold is an LOD value of 3.

[0075] The results of multi-site GWAS analysis of average daily feeding frequency are as follows: Figure 1-3 As shown in the figure, there is a significant SNP site on Duroc pig chromosome 3 that affects the average daily feed intake, corresponding to the G>T mutation at position 35094559 on chromosome 3 of the International Swine Reference Genome Version 11.1 (nucleotide g.121G>T in SEQ ID NO: 1). The LOD value of the mrMLM model is 4.2723, the LOD value of the FASTmrMLM model is 4.7951, and the LOD value of the FASTmrEMMA model is 3.3233.

[0076] The results of multi-site GWAS analysis of average feeding time per feeding are as follows: Figure 5-7 As shown in the figure, there is a significant SNP site on Duroc pig chromosome 3 that affects the average feeding time per session. This SNP corresponds to the G>T mutation at position 35094559 on chromosome 3 of the International Swine Reference Genome Version 11.1 (nucleotide g.121G>T in SEQ ID NO: 1). The LOD value of the mrMLM model is 5.4296, the LOD value of the FASTmrMLM model is 6.129, and the LOD value of the FASTmrEMMA model is 7.1194.

[0077] (3) Association analysis between different genotypes and average daily feeding frequency and average feeding time patterns

[0078] Comparing the analysis results of different multi-locus GWAS models, we retained the SNP loci that significantly affected the average daily feed intake or the average feeding time in the analysis results of the mrMLM, FASTmrMLM, and FASTmrEMMA models of multi-locus GWAS. The results showed that there is a SNP locus on chromosome 3 located at the RBFOX1 locus that significantly affects both the average daily feed intake and the average feeding time, and this locus deserves special attention. According to Table 1, the SNP locus g.121G>T at the RBFOX1 locus is significantly correlated with both the average daily feed intake and the average feeding time (P<0.05), indicating that this molecular marker significantly affects the average daily feed intake and the average feeding time in pigs. Assisted selection at this molecular marker locus in pigs could reduce the average daily feed intake and increase the average feeding time, thereby improving feed conversion ratio.

[0079] Furthermore, Table 1 shows that the TT type has a higher average daily feeding frequency and a shorter average feeding time per session than the TG and GG types, indicating that homozygous TT and heterozygous TG have a higher average daily feeding frequency and a shorter average feeding time per session. Figure 4 and Figure 8 Further investigation revealed highly significant differences between homozygous TT and heterozygous TG genotypes and the GG genotype in average daily feed intake and average feed duration. This further indicates that homozygous TT and heterozygous TG genotypes have a higher average daily feed intake and a shorter average feed duration. Average daily feed intake and average feed duration are important traits affecting pig feed conversion ratio (FCR). Higher average daily feed intake and shorter average feed duration mean poorer FCR and higher feeding costs. Therefore, pigs with the TT genotype have poorer production performance. In breeding, it is necessary to cull TT genotype breeding pigs and retain TG and GG genotype breeding pigs to gradually increase the frequency of the G allele at this locus. Currently, the frequency of the inferior allele in this population is still as high as 34.05%, indicating significant potential for genetic improvement.

[0080] Table 1. Correlation analysis of the RFBOX1 gene locus of molecular markers with the average number of feedings per day.

[0081]

[0082] Note: Average daily feeding frequency is expressed as mean ± standard deviation (SD).

[0083] Table 2. Correlation analysis of RFBOX1 gene loci of molecular markers with average feeding time per session.

[0084]

[0085] Note: The average feeding time per session is expressed as mean ± standard deviation (SD).

[0086] Example 3: Target DNA Sequence Amplification and Sequencing

[0087] (1) Primer design

[0088] The DNA sequence of SEQ ID NO:1 on pig chromosome 3 was downloaded from the Ensembl website (http: / / asia.ensembl.org / index.html). Primers were designed using the primer design software Primer Premier 6.0 and synthesized by Sangon Biotech (Shanghai) Co., Ltd. The DNA sequences of the designed primers are shown below:

[0089] P001-F: 5'-TGCCTAGAGTGGGTCAT-3' (SEQ ID NO: 2);

[0090] P002-R: 5'-AATAGGGCCTTCAACAGG-3' (SEQ ID NO: 3);

[0091] (2) PCR amplification

[0092] To a 10 μL reaction mixture, add 1 μL of DNA template, 3.4 μL of double-distilled water, 5 μL of 2×TaqPCR StanMix with Loading Dye, and 0.3 μL each of primers P001-F and P002-R. The PCR reaction conditions were: 94℃ pre-denaturation for 5 min, followed by 35 cycles of 94℃ denaturation for 30 s, 64.5℃ annealing for 30 s, and 72℃ extension for 45 s, with a final extension at 72℃ for 5 min.

[0093] (3) DNA sequencing

[0094] DNA sequencing identification: Performed at BGI Genomics Co., Ltd. in Shenzhen, the gene fragments were sequenced using both forward and reverse reactions. The obtained sequences were compared with the NCBI genome sequence to identify the corresponding RBFOX1 gene locus mutation. The sequencing results are shown below:

[0095]

[0096] Note: M marked in the sequence listing is the mutation site, indicated by an underline (the mutated base in parentheses represents the allele mutation). The positions of the primer sequences are indicated by bolding at the beginning and end of the sequence.

[0097] Example 4: Analysis of the g.121G>T effect of the RBFOX1 gene locus using molecular markers

[0098] Table 1 shows that, for the average daily feed intake frequency, the dominant allele genotype g.121G>T (GG) at the RBFOX1 gene locus significantly reduced the frequency by 0.76 times / day compared to the TT phenotype. For the average feeding time per session, the dominant allele genotype g.121G>T (GG) at the RBFOX1 gene locus significantly increased the feeding time per session by 0.97 minutes compared to the TT phenotype. Fewer average daily feed intake frequencies and longer average feeding times in pigs result in higher feed conversion rates. This will significantly reduce feed costs in pig farming, creating greater profit margins for businesses. By selecting the dominant allele (G) at this gene locus in Duroc pigs, it is possible to ultimately improve the economic efficiency of commercial pigs, thereby increasing the profits of enterprises.

[0099] This invention provides a novel molecular marker for marker-assisted selection of pigs by detecting the mutation site at position 121 of the SEQ ID NO:1 sequence and conducting preliminary association analysis between its genotype and the average daily feeding frequency and average feeding time of pigs.

[0100] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. The application of a SNP molecular marker located at the pig RFBOX1 locus that is associated with feeding behavior traits in identifying feeding behavior traits, screening pigs with low average daily feeding frequency and long average feeding time, or in the genetic breeding of pig feeding behavior traits, characterized in that... The SNP molecular markers mentioned above correspond to the G>T mutation at position 35094559 on chromosome 3 of the International Swine Reference Genome Version 11.

1. The polymorphism of the bases at this site affects the feeding behavior of pigs. The feeding behavior traits mentioned above are the average number of feedings per day or the average time of each feeding. The average number of feedings per day for TG and GG genotype pigs is lower than that for TT genotype pigs, and the average time of each feeding for TG and GG genotype pigs is higher than that for TT genotype pigs. The pigs mentioned are Canadian Duroc pigs and their synthetic lines.

2. The application according to claim 1, characterized in that: The nucleotide sequence of the SNP molecular marker is shown in SEQ ID NO: 1, where M in the sequence is G or T.

3. A primer pair for detecting SNP molecular markers associated with feeding behavior traits located at the pig RFBOX1 locus in the application of identifying feeding behavior traits, screening pigs with low average daily feeding frequency and long average feeding time, or in the genetic breeding of pig feeding behavior traits, characterized in that... The primer pair comprises primer P001-F and primer P002-R, and their nucleotide sequences are shown below: P001-F: 5'-TGCCTTAGAGTGGGTCAT-3', P002-R: 5'-AATAGGGCCTTCAACAGG-3'; The SNP molecular marker associated with feeding behavior traits located at the RBFOX1 locus in pigs is the SNP molecular marker described in claim 1; the feeding behavior traits are the average number of feedings per day or the average time of each feeding; the average number of feedings per day for TG and GG genotype pigs is lower than that for TT genotype pigs, and the average time of each feeding for TG and GG genotype pigs is higher than that for TT genotype pigs; the pigs are Canadian Duroc pigs and their synthetic lines.

4. A kit for detecting SNP molecular markers associated with feeding behavior traits located at the pig RFBOX1 locus, used in identifying feeding behavior traits, screening pigs with low average daily feeding frequency and long average feeding time, or in the genetic breeding of pig feeding behavior traits, characterized in that... The kit comprises the primer pair as described in claim 3; The SNP molecular marker associated with feeding behavior traits located at the RBFOX1 locus in pigs is the SNP molecular marker described in claim 1; the feeding behavior traits are the average number of feedings per day or the average time of each feeding; the average number of feedings per day for TG and GG genotype pigs is lower than that for TT genotype pigs, and the average time of each feeding for TG and GG genotype pigs is higher than that for TT genotype pigs; the pigs are Canadian Duroc pigs and their synthetic lines.

5. A method for detecting traits related to pig feeding behavior, comprising the following steps: The SNP molecular markers described in claim 1 or 2 at the porcine RBFOX1 gene locus are detected, and the feeding behavior-related traits in pigs are determined based on whether the single nucleotide of the SNP site of the SNP molecular marker is G or T; wherein... The feeding behavior traits mentioned are the average number of feedings per day or the average time of each feeding. The average number of feedings per day for TG and GG genotype pigs is lower than that for TT genotype pigs, and the average time of each feeding for TG and GG genotype pigs is higher than that for TT genotype pigs. The pigs mentioned are Canadian Duroc pigs and their synthetic lines.

6. A method for screening pigs with low average daily feeding frequency and long average feeding time using SNP molecular markers associated with feeding behavior traits located at the pig RFBOX1 locus, characterized in that... It includes the following steps: The SNP molecular markers described in claim 1 or 2 are detected on the RBFOX1 locus in pigs. Based on the SNP sites of the SNP molecular markers, individuals with the TT genotype are culled, while individuals with the TG or GG genotypes are retained. Among these individuals, the average daily feed intake of pigs with the TG and GG genotypes is lower than that of pigs with the TT genotype, and the average feeding time per session of pigs with the TG and GG genotypes is higher than that of pigs with the TT genotype. The pigs mentioned are Canadian Duroc pigs and their synthetic lines.

7. The method according to claim 5 or 6, characterized in that: The detection method includes the following steps: (1) Extract genomic DNA from the pigs to be tested; (2) Using the primer pair described in claim 3 or the primer pair in the kit described in claim 4 as amplification primers, and using the genomic DNA of the pig to be tested obtained in step (1) as template DNA, PCR amplification is performed to obtain PCR amplification products; (3) Sequencing the PCR amplification products to obtain sequencing results; (4) Based on the sequencing results, determine the genotype of the SNP molecular marker.

8. A method for genetic improvement of pigs, characterized in that... It includes the following steps: Identify the SNP sites of the SNP molecular markers described in claim 1 or 2 in the core breeding pig population, and make corresponding selections based on the SNP molecular markers: select breeding pig individuals with TG and GG genotypes at locus 35094559 on chromosome 3 of the International Swine Reference Genome Version 11.1 in the core breeding pig population, and cull breeding pig individuals with TT genotype at this locus, so as to increase the frequency of the G allele at this locus generation by generation, thereby improving the pig's feeding behavior ability; The pigs mentioned are Canadian Duroc pigs and their synthetic lines.

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