CN116574816A - Boar sperm deformity rate-related molecular genetic marker and obtaining method and application thereof - Google Patents
Boar sperm deformity rate-related molecular genetic marker and obtaining method and application thereof Download PDFInfo
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Abstract
The invention discloses a molecular genetic marker related to boar sperm malformation rate and an obtaining method and application thereof. The molecular genetic marker is a SNP locus of TMCO5A gene, the SNP locus is positioned at 133586233 th position of chromosome 1 of pig genome, and the base of the locus is A or G. Among the three genotypes corresponding to the boar sperm malformation rate, the sperm malformation rate of the individual boars with the GG genotype is low, the malformation rate is obvious from the individual boars with the AA genotype, the boar sperm quality can be evaluated by detecting the molecular genetic markers related to the boar sperm malformation rate, the boar with low sperm malformation rate is bred, the boar sperm malformation rate is reduced, and the utilization rate of the boar is improved. The invention is beneficial to the genetic improvement of reproductive performance of the boars in China and solves the problem that the boars produced in large-scale boar stations are eliminated in large quantity due to poor semen quality.
Description
Technical Field
The invention belongs to the technical field of molecular genetic markers. More particularly, relates to a molecular genetic marker related to boar sperm malformation rate, and an obtaining method and application thereof.
Background
In a modern live pig production system, genes of excellent boars can be rapidly diffused into a group through an artificial insemination technology, so that the production performance of commercial pigs is improved efficiently, and a great effect is exerted on a pork protection area. Duroc boars are used as terminal male parents in the currently widely used Duroc production system of Duroc-Chang-Dada or Duroc-Chang ternary hybrid commercial pigs, and the commercial pigs contain Duroc blood system proportion of 50%, so Duroc pigs have a vital role in the pig industry. The conventional breeding scheme is to select and breed the characters of the Duroc boar such as growth speed, backfat thickness, feed efficiency and the like, and select and breed the boar reproductive performance in a neglected way. Therefore, in the process of continuously improving the conventional characters such as the growth rate of the Duroc boar, the semen quality is necessary to be bred simultaneously. Especially in the 'post-non-pestilence' age, the socialized sperm supply model is gradually raised and widely adopted by most large-scale breeding enterprises, and the rise of large-scale boar stations has become the necessary trend of the pig industry. The improvement of boar semen quality through breeding technology is an important guarantee for social semen supply mode and economic benefit of large boar stations.
Researches show that the boar semen belongs to the medium and low genetic trait and has the potential of genetic improvement. However, the number of boars in a single population is small, and semen quality detection data is small. So far, genetic improvement plans for boar semen traits are fresh worldwide. Boar semen trait genetic analysis is an important basis for implementing genetic improvement. By analyzing the boar semen character genetic mechanism, the molecular genetic marker for improving the boar semen character is excavated, which is expected to provide a basis for the genetic improvement of reproductive performance of the boar in China, and solves the key problem that the production of the boar in large-scale boar stations is largely eliminated (the ratio is 40% -61%) due to poor semen quality. In addition, the raising amount of the boars can be effectively reduced by improving the sperm producing capacity of the boars, and the production cost of the boar station is greatly reduced.
In recent years, the commercial application of high-throughput sequencing technology and high-density SNP (Single Nucleotide Porlymorphism) chips has enabled researchers to quickly and cheaply obtain high-density SNP and even DNA sequence data covering the entire genome. Based on high-density SNP data and phenotype records of important characters of livestock and poultry, gene localization is carried out on complex characters, and the application of the information to genetic improvement of livestock and poultry is tried to become a new research hotspot in the field of livestock and poultry breeding. The whole genome association analysis becomes a conventional technology for QTL (Quantitative Trait Loci) localization and is widely applied to localization of important economic character key genes of human complex diseases and livestock and poultry.
Based on high-density SNP data covering the whole genome and trait phenotype recordings of large populations, candidate genes can be pinpointed by whole genome association analysis techniques (Genome wide association study, GWAS), but this technique has some drawbacks. Classical GWAS generally performs single-marker regression analysis on all markers one by one based on software such as Plink, and then sets a significant threshold to screen significant sites, so that the problems of high calculation intensity, overestimation of the marker effect, certain randomness in the significance threshold setting and the like exist. To further increase the efficiency of GWAS, new methods and software are continually being developed. The one-step whole genome association analysis (weighted single step genome-wide association study, wsGWAS) simultaneously utilizes pedigree, historical individual phenotype records and genotype data for association analysis, is suitable for the situation that a large number of individuals possess phenotype records and only a small number of individuals possess genotype data, and is particularly suitable for whole genome association analysis of important economic traits of livestock and poultry. In addition, through sequence filling, SNP chip data can be filled to a sequence level, ultra-high density SNP variation data of the sequence level is obtained, and correlation analysis of the sequence level is carried out through GCTA software.
Spermatogenesis involves three important steps, mitosis, meiosis and spermatogenesis, involving the coordinated process of many genes and cells in the seminiferous tubules. Mutations in the relevant genes and changes in the state of the corresponding cells may affect sperm quality and male reproductive capacity. From the standpoint of genetic mechanism analysis and animal breeding, locating candidate genes related to sperm quality and reproductive ability is a crucial step. Although researchers have located candidate genes or genome regions which partially influence boar semen traits, the genetic background of research groups, the size of boar groups, the marker density and the like have great differences, and the gene location results reported by different researches are different, so that the molecular genetic markers which can be used for improving boar semen traits are lacking. The candidate genes related to boar semen characters have the problems of insufficient marker density, single QTL positioning method and the like.
Disclosure of Invention
The invention aims to overcome the defects and the shortcomings of the prior art and provide a molecular genetic marker related to boar sperm malformation rate, and an obtaining method and application thereof.
The first object of the invention is to provide a molecular genetic marker related to boar sperm malformation rate.
It is a second object of the present invention to provide a method for obtaining said molecular genetic markers.
The third object of the invention is to provide the application of the molecular genetic marker in the aspect of evaluating the semen quality of boars or breeding boars with low sperm malformation rate.
It is a fourth object of the present invention to provide a reagent for detecting the molecular genetic marker.
It is a fifth object of the present invention to provide the use of said agent for assessing boar semen quality.
It is a sixth object of the present invention to provide the use of said agent for the preparation of a product for assessing boar semen quality.
The seventh object of the invention is to provide the application of the reagent in breeding boars with low sperm malformation rate.
An eighth object of the invention is to provide the application of the reagent in the preparation of products for breeding boars with low sperm malformation rate.
The ninth object of the invention is to provide a method for breeding boars with low sperm malformation rate.
The above object of the present invention is achieved by the following technical scheme:
according to the invention, key individual re-sequenced data genetic variation is detected by collecting the data of the semen character phenotype of the Duroc boar, data filling is carried out based on the SNP chip sequence, key genes of the semen character of the Duroc boar are mined, key genes affecting the sperm malformation rate of the boar are mined by adopting GCTA software based on the data of the semen character phenotype-genome sequence, and TMCO5A genes in the range of No. 1 chromosome 133.1-134.2 Mb (NC_010443.5:c133383696-133291660Sus scrofa isolate TJ Tabasco breed Duroc chromosome 1,Sscrofa11.1) are positioned, and the total length of the genes is 92kb, so that the genes are candidate genes for the sperm malformation rate of the boar. On the basis of the TMCO5A gene, the invention also obtains a molecular genetic marker related to the boar sperm abnormality rate. The sperm malformation rates of the boars with different genotypes based on the molecular genetic markers are extremely obviously different, and the sperm malformation rates of the boars can be used for evaluating the semen quality of the boars to select and breed the boars with low sperm malformation rates of the boars. Thus, the invention claims said molecular genetic markers, methods for obtaining them and uses thereof.
The invention provides a molecular genetic marker related to boar sperm malformation rate, which is a SNP locus of TMCO5A gene, wherein the SNP locus is positioned at 133586233 th position of chromosome 1 of pig genome, and the base of the locus is A or G.
Specifically, the pig is a Duroc boar, the reference pig genome is Srcrofa 11.1, and the sequence information of the TMCO5A gene is: NC-010443.5:c133383696-133291660Sus scrofa isolate TJ Tabasco breed Duroc chromosome 1,Sscrofa11.1.
The invention also provides a method for obtaining the molecular genetic marker, which is characterized by comprising the following steps:
s1, collecting boar semen character phenotype data; counting phenotype data of 4 characters of semen volume, density, vitality and sperm malformation rate of the boar collected each time of the complete pedigree breeding pig, and calculating to obtain the effective sperm number of the boar obtained by single semen collection according to the phenotype data of the 4 characters;
s2, detecting genetic variation of the key individual resequencing data; comparing the data obtained by removing low-quality sequences from the whole genome resequencing original data to a reference genome by using BWA software based on the whole genome resequencing original data of the breeding pigs to obtain a sam file; converting the.samtools file into a.bam file using SAMtools; removing repeated sequences appearing in the bam file through the marked repeated sequences, and finally obtaining the bam file for carrying out SNP detection;
s3, filling SNP chip-sequence data; filling large-population gene chip data to a sequence level by taking SNP detected by re-sequencing data as a reference, and obtaining genome sequence data of the breeding boar in ultra-high density through individual weight sequencing, wherein the genome sequence data is used for mining key genes of boar semen characters;
s4, boar semen character association analysis; digging key genes affecting the sperm abnormality rate of the boars based on semen character phenotype-genome sequence data; estimating individual breeding values based on a mixed linear model equation set, converting the breeding values into inverse regression breeding values DRP, performing whole genome association analysis by taking the DRP as an independent variable, and positioning molecular genetic markers affecting the sperm abnormality rate of the boars.
The invention also claims the application of the molecular genetic marker in evaluating the semen quality of boars or breeding boars with low sperm malformation rate.
Specifically, the quality of boar semen can be evaluated by detecting the genotype of the molecular genetic marker; when the genotype is AA or AG, the abnormal rate of boar sperm is high, and the quality of boar semen is low; when the genotype is GG, the sperm malformation rate of the boar is low, and the sperm quality of the boar is high.
The invention also provides a reagent for detecting the molecular genetic marker.
Alternatively, the reagents are primers or primer and probe combinations, and the sequences containing the molecular genetic marker nucleotide sites can be obtained through primer amplification, and the genotypes of the sequences are obtained through common sequencing.
The invention also claims the application of the reagent for detecting the molecular genetic marker in evaluating the quality of boar semen.
The invention also claims the application of the reagent for detecting the molecular genetic marker in the preparation of products for evaluating the quality of boar semen.
The invention also claims the application of the reagent for detecting the molecular genetic marker in breeding boars with low sperm malformation rate.
The invention also claims the application of the reagent for detecting the molecular genetic marker in the preparation of the products for breeding the boars with low sperm malformation rate.
The invention also provides a method for breeding boars with low sperm malformation rate, which comprises the following steps: extracting the genome DNA of the boar to be detected, detecting the 133586233 nucleic acid locus of the chromosome 1, and determining the genotype of the boar; and selecting the boar with the genotype GG for further seed selection or breeding.
Based on the molecular genetic marker, the difference of the individual malformation rate of the AA and GG genotype boars is 1.64%, and the G allele obviously reduces the sperm malformation rate of the boars. Therefore, the molecular UAN marker genotype of the invention can be detected to assist the breeding of the boar, and the boar sperm malformation rate is reduced and the utilization efficiency of the boar is improved by selecting and reserving GG homozygous boar to enter a boar station.
The invention has the following beneficial effects:
the invention provides a boar sperm malformation rate candidate gene: the invention also provides a molecular genetic marker related to the boar sperm abnormality rate, wherein the molecular genetic marker is a SNP locus of the TMCO5A gene, the SNP locus is positioned at 133586233 th site of chromosome 1 of a pig genome, and the base of the locus is A or G. Among the three genotypes corresponding to the boar sperm malformation rate, the sperm malformation rate of the individual boars with the GG genotype is low, the malformation rate of the individual boars with the AA genotype is different by 1.64%, the sperm malformation rate of the boars is obviously reduced by the G allele, the sperm quality of the boars can be evaluated by detecting the molecular genetic markers related to the sperm malformation rate of the boars, the boars with low sperm malformation rate are bred, the sperm malformation rate of the boars is reduced, and the utilization efficiency of the boars is improved.
The invention not only provides a molecular genetic marker for boars with low sperm abnormality rate, but also provides an obtaining method of the molecular genetic marker and a method for breeding boars with low sperm abnormality rate, which are beneficial to genetic improvement of reproductive performance of boars in China and solve the problem that large-scale boar stations for producing boars are eliminated in large quantity due to poor semen quality.
Drawings
FIG. 1 shows the results of a sperm cell rate high density chip and resequencing whole genome correlation analysis.
FIG. 2 shows the results of TMCO5A gene expression pattern analysis.
FIG. 3 shows the expression level of TMCO5A gene in 3 developmental stages of spermatogonia, pachytic sperm cells and round sperm.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
Example 1 Duroc boar semen trait Key Gene excavation and semen trait association analysis
The invention provides a molecular genetic marker related to boar sperm malformation rate, which is obtained by the following steps:
(1) Boar semen trait phenotype data collection
Based on daily semen collection data of a large-scale live pig genetic improvement boar station, collecting and sorting 28 ten thousand semen collection data of 2253 Duroc boars, wherein the 28 ten thousand semen collection data mainly comprises phenotype data of 4 characters of semen volume, density, vitality and deformity rate, and calculating to obtain the sperm effective number of single semen collection of the boars according to the phenotype data of the 4 characters to obtain a Duroc boar semen character phenotype data set. The calculation formula of the sperm effective number is as follows: sperm availability = volume x density x motility x (1-rate of malformation); the deformity rate was obtained by analysis of fresh semen using the UltiMateTM CASA system.
(2) Key individual body weight sequencing data genetic variation detection
Comparing the whole genome resequencing original data (clean reads) obtained by removing low-quality sequences with BWA software to a reference genome based on the whole genome resequencing original data (short reads) of the breeding pigs to obtain a sam file; converting the.samtools file into a.bam file using SAMtools; the repeated sequences possibly occurring due to PCR reactions in the bam file are removed using markups (marker repeats) in Picard, and finally the bam file is obtained for SNP detection by GATK software.
(3) SNP chip-sequence data filling
Based on GGP 50k SNP (GeneSeek, US) chip data of 2000 Duroc pigs and ultra-high-density SNP data of 95 pigs, filling large-population gene chip data (5 ten thousand gene loci) into sequence level (ten million gene loci) by taking SNP detected by resequencing data as a reference, and obtaining ultra-high-density genotype data of more than 2000 Duroc pigs by resequencing a small amount of key individuals, wherein the ultra-high-density genotype data is used for mining semen character key genes of Duroc boars.
(4) GCTA software genome-wide association analysis
Calculating a breeding value of a large number of sperm collecting data of the boar with repeated observation values, converting the breeding value into an inverse regression breeding value, carrying out whole genome association analysis by adopting a linear model of GCTA (general purpose chemical amplification) by taking the inverse regression breeding value as an dependent variable, obtaining p values of the association significance of all genetic markers and sperm malformation rate of the boar, screening genetic variation with the strongest association from the p values, and identifying major genes in the significant association region through functional annotation, linkage disequilibrium analysis and large data expression analysis.
Example 2 verification of molecular markers
(1) Boar semen trait association analysis
Based on the boar semen character phenotype data and the ultra-high density genome sequence data of more than 2000 boars obtained in the example 1, semen character phenotype-genome sequence data is formed, and key genes affecting the sperm abnormality rate of the boars are mined by adopting GCTA software. Firstly, estimating individual breeding values based on a mixed linear model equation set, then converting the breeding values into inverse regression breeding values (DRP), carrying out whole genome association analysis by taking the DRP as an dependent variable, and positioning a molecular genetic marker affecting the sperm abnormality rate of the boar. The breeding value calculation hybrid linear model adopted by the invention is as follows:
y＝Xb+Za+Wp+Age+Intv+e
wherein y is a character observation value vector; x, Z and W are design matrices; b is a fixed effector vector (ensemble mean, year-season effect, and semen collector effect);is a breeding value vector; />Permanent environmental effects for the individual; age and Intv are month-old and semen collection frequency (day) of semen collection of boars respectively, and are covariates; />Is the residual.
H is an affinity matrix for integrating the pedigree and the SNP marker simultaneously, and the calculation formula of the inverse matrix is as follows:
wherein A is a family-based affinity matrix; a is that 22 The method comprises the steps of A, dividing a block matrix corresponding to genotype individuals; g w ＝0.9G+0.1A 22 ，For the relative moment based on whole genome sequence data, Z is a genotype matrix corrected for minor allele frequencies (minor allele frequency, MAF), where 0-2p,1-2p and 2-2p represent the three genotypes AA, AA and AA, respectively, and p is the minor allele frequency; d is a diagonal matrix, representing the weight of the SNP; p (P) i Minor allele frequencies for the ith marker; m is the number of marks.
And (3) for the mixed linear model, estimating a variance component by adopting an AI-REML (average information restricted maximum likelihood) method, and obtaining a breeding value by solving a mixed linear model equation set. Calculating a reverse breeding value (DRP), carrying out sequence-level whole genome association analysis by using the DRP as an dependent variable through GCTA software, and positioning a molecular genetic marker affecting the sperm abnormality rate of the boar. After obtaining the p value of correlation between sequence level SNP variation and boar sperm abnormal rate, drawing a Manhattan diagram to display a significant correlation region, and carrying out linkage disequilibrium analysis on all SNP markers within the range of 1Mb of the significant correlation region to obtain the haplotype of the region. Key candidate genes for the significantly associated regions were obtained by comparison with the NCBI database. The effect of the most obvious mutation site on the sperm malformation rate of the boar and the expression pattern of the key candidate gene are further analyzed, and the candidate gene is verified.
Sequence level whole genome association analysis manhattan plots, significant association region haplotypes, and association region major genes are shown in figure 1. As can be seen from FIG. 1, the TMCO5A gene in the range of chromosome 1 133.1 to 134.2Mb (NC-010443.5: c 13338396-133291660Sus scrofa isolate TJ Tabasco breed Duroc chromosome 1,Sscrofa11.1) was located at a position 202kb upstream of the center position (SNP with the smallest p-value) of the genome segment over the entire length of 92kb by sequence-level genome-wide association analysis.
EXAMPLE 3TMCO5A Gene expression Pattern analysis
The invention uses the pigGTEx project (http:// piggytex. Farm ex /) data to carry out mapping analysis on the tissue expression mode of the pig TMCO5A gene.
The results of analysis of the expression pattern of the porcine TMCO5A gene are shown in FIG. 2. The analysis result of the large sample RNAseq data of the pig GTEx based on the public database shows that the TMCO5A gene is highly specifically expressed in the testis tissue of the pig (figure 2) and hardly expressed in the rest tissues, which shows that the gene is very likely to play a unique function in the spermatogenesis process of the boar.
In addition, the invention uses Single cell transcriptome sequencing disclosed by Zhang et al (Single-cell RNA-sequencing reveals the dynamic process and novel markers in porcine spermatogenesis, journal of Animal Science and Biotechnology, 2021) to sequence the sperm cell transcriptome data of 3 development stages of spermatogonia, pachytic sperm cells and round sperm to count the expression quantity of TMCO5A gene in the sperm cells of the 3 development stages, and the result is shown in figure 3, and the result shows that the TMCO5A gene is low-expressed in the spermatogonia, the expression quantity is increased in the pachytic sperm cells of the pachytic sperm, and the expression quantity in the round sperm cells is obviously increased, namely, the gene is specifically and highly expressed when sperm allergic development is started, so that the gene possibly plays an important role in the sperm allergic development process.
Example 4 individual sperm malformations differ from genotype to genotype
Based on the above results, further analysis of the TMCO5A gene revealed a molecular genetic marker related to boar sperm abnormality rate, which was a SNP site of the TMCO5A gene located at the 133586233 nucleic acid site of chromosome 1 of the Duroc boar genome (Srcrofa 11.1), the base of which was A or G.
Specifically, the influence of 133586233-base on the semen index of the Duroc boar on chromosome 1 is analyzed through analysis of variance, F test and multiple comparison, all boar individuals are grouped according to the genotype of the locus, and then, whether the genetic locus is obviously related to the sperm abnormality rate of the boar is tested by adopting single-factor analysis of variance and F test. Analysis results show that the genotype of the 133586233 th site on the Duroc boar chromosome 1 significantly affects the sperm malformation rate of the Duroc boar (p= 0.0000218). The sperm malformation rate of the boars with different genotypes and the multiple comparison results are shown in the following table 1, and as can be seen from the table 1, the G allele can significantly reduce the sperm malformation rate of the boars, and the sperm malformation rate of the boars with the AA genotype is 1.64 percent higher than that of the boars with the GG genotype (p= 0.0000023).
TABLE 1 individuals with different genotypes of sperm malformations at significantly related loci of pig No. 1 chromosome
The results show that the locus can be used as a molecular genetic marker, and can be used for evaluating the semen quality of the boars and breeding the boars with low sperm abnormality rate of the boars.
Example 5 method for Breeding boars with Low sperm Rate of malformation
Based on the molecular genetic markers disclosed in the present invention, one skilled in the art can easily design primers for amplifying the molecular markers or probes for identifying the molecular markers, thereby being used for detection of the genetic markers, for example, by PCR amplification to obtain the molecular genetic markers, clone sequencing to obtain corresponding sequences, or Bsm-RFLP polymorphism. Thus, the invention also includes primers for amplifying the molecular genetic markers or probes for identifying the molecular genetic markers, and kits containing the primers or probes.
The genetic molecular marker or the kit for detecting the genetic molecular marker can be used for evaluating the semen quality of the boar or for breeding the boar with low sperm abnormality rate.
The invention also provides a method for breeding boars with low sperm malformation rate, which comprises the following steps: extracting the genome DNA of the boar to be detected, detecting the 133586233 nucleic acid locus of the chromosome 1, and determining the genotype of the boar; and selecting the boar with the genotype GG for further seed selection or breeding.
The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.
Claims (10)
1. The molecular genetic marker is characterized in that the molecular genetic marker is a SNP locus of TMCO5A gene, the SNP locus is positioned at 133586233 th position of chromosome 1 of pig genome, and the base of the locus is A or G.
2. The molecular genetic marker of claim 1, wherein the pig is a doloque boar and the reference pig genome is sscroffa 11.1.
3. The method for obtaining a molecular genetic marker as claimed in claim 1, comprising the steps of:
s1, collecting boar semen character phenotype data;
counting phenotype data of 4 characters of semen volume, density, vitality and sperm malformation rate of the boar collected each time of the complete pedigree breeding pig, and calculating to obtain the effective sperm number of the boar obtained by single semen collection according to the phenotype data of the 4 characters;
s2, detecting genetic variation of the key individual resequencing data;
comparing the data obtained by removing low-quality sequences from the whole genome resequencing original data to a reference genome by using BWA software based on the whole genome resequencing original data of the breeding pigs to obtain a sam file; converting the.samtools file into a.bam file using SAMtools; removing repeated sequences appearing in the bam file through the marked repeated sequences, and finally obtaining the bam file for carrying out SNP detection;
s3, filling SNP chip-sequence data;
filling large-population gene chip data to a sequence level by taking SNP detected by re-sequencing data as a reference, and obtaining genome sequence data of the breeding boar in ultra-high density through individual weight sequencing, wherein the genome sequence data is used for mining key genes of boar semen characters;
s4, boar semen character association analysis;
digging key genes affecting the sperm abnormality rate of the boars based on semen character phenotype-genome sequence data; estimating individual breeding values based on a mixed linear model equation set, converting the breeding values into inverse regression breeding values DRP, performing whole genome association analysis by taking the DRP as an independent variable, and positioning molecular genetic markers affecting the sperm abnormality rate of the boars.
4. The use of the molecular genetic marker of claim 1 for evaluating boar semen quality or for breeding boars with low sperm malformation rate.
5. A reagent for detecting the molecular genetic marker of claim 1.
6. Use of the reagent according to claim 5 for assessing boar semen quality.
7. Use of the reagent according to claim 5 for the preparation of a product for assessing boar semen quality.
8. The use of the reagent of claim 5 in breeding boars with low sperm malformation rate.
9. Use of the agent of claim 5 for the preparation of a product for breeding boars with low sperm cell malformation.
10. A method for breeding a boar with low sperm malformation rate is characterized by comprising the following steps: extracting the genome DNA of the boar to be detected, detecting the molecular genetic marker in claim 1, and determining the genotype of the boar; and selecting the boar with the genotype GG for further seed selection or breeding.
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