Use of a molecular marker for detecting a long hair trait in a cashmere goat

Abstract

The application belongs to the technical field of animal breeding, and particularly relates to application of a detection reagent of a molecular marker related to a long hair trait of a cashmere goat. The molecular marker is located at 155,344,701-155,346,600 on a goat chromosome 1, and a nucleotide sequence thereof is shown as SEQ ID NO. 1. It is found by the application that the length of wool of an individual with a normal type of the molecular marker region is greater than the length of wool of an individual with a deletion type of the molecular marker region, and thus a cashmere goat breed with long wool can be selected by detecting a deletion mutation of the molecular marker on the goat chromosome 1.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of animal breeding, and particularly relates to application of a detection reagent of a molecular marker related to a wool length trait of a cashmere goat. BACKGROUND

[0002] Inner Mongolia cashmere goat is an excellent local livestock breed formed through long-term natural selection and artificial systematic breeding. Copy number variation (CNV) is widely present in the genome, including deletion and duplication. With the deepening understanding of complex animal and plant genomes, more and more evidence shows that CNVs play an important role in determining breeding and agricultural important traits. Although CNV is an important source of animal genetic diversity, the influence of CNV on the genome of goats and the wool trait is still largely unknown. Genome-wide association study (GWAS) is a powerful tool for trait-related gene mapping and has been widely used to identify candidate genomic loci affecting phenotypic variation. However, the research of GWAS usually relies on the information of single nucleotide polymorphism (SNP) associated with the phenotype, while ignoring large fragment CNVs with larger phenotype effect size. Through CNV detection and CNV-GWAS analysis of Inner Mongolia cashmere goats, new insights can be provided for the genetic breeding of Inner Mongolia cashmere goats. It is found that with the increase of wool length, the wool yield and body weight will also increase. Selecting long wool type individuals for breeding can accelerate the genetic progress of wool yield and body weight of cashmere goats, realize the indirect selection of wool on wool yield and body weight, and facilitate processing in subsequent application. Therefore, mining molecular markers affecting the wool length of Inner Mongolia cashmere goats at the genome level is conducive to breeding cashmere goat breeds with larger wool yield and body weight. SUMMARY

[0003] In view of the above technical problems, the application provides application of a detection reagent of a molecular marker related to a wool length trait of a cashmere goat. The molecular marker can be used to breed cashmere goats with long wool length and indirectly improve the wool yield and body weight of cashmere goats.

[0004] In a first aspect of the application, application of a detection reagent of a molecular marker related to a wool length trait of a cashmere goat in identifying the wool length trait of the cashmere goat is provided. The nucleotide sequence of the molecular marker is shown in SEQ ID NO. 1.

[0005] As a preferred embodiment of the application, the method for identifying the wool length trait of the cashmere goat comprises the following steps:

[0006] Detecting deletion or insertion of the molecular marker on chromosome 1 of the genome of the goat to be tested. The wool length of an individual with insertion is greater than that of an individual with deletion.

[0007] In the second aspect of the present application, the application of a detection reagent of a molecular marker related to the hair length of a cashmere goat to genetic breeding of the cashmere goat is provided, and the nucleotide sequence of the molecular marker is shown in SEQ ID NO. 1.

[0008] As a preferred embodiment of the present application, the genetic breeding is breeding a cashmere goat with long hair length.

[0009] Further preferably, the method for breeding a cashmere goat with long hair length comprises the following steps:

[0010] The deletion or insertion of the molecular marker on the chromosome 1 of the goat genome is detected, and an individual with insertion is selected as a parent to increase the hair length of the offspring cashmere goat.

[0011] As a preferred embodiment of the present application, the genetic breeding is breeding a cashmere goat with large wool yield or large body weight.

[0012] Further preferably, the method for breeding a cashmere goat with large wool yield comprises the following steps:

[0013] The deletion or insertion of the molecular marker on the chromosome 1 of the goat genome is detected, and an individual with insertion is selected as a parent to increase the wool yield of the offspring cashmere goat.

[0014] Further preferably, the method for breeding a cashmere goat with large body weight comprises the following steps:

[0015] The deletion or insertion of the molecular marker on the chromosome 1 of the goat genome is detected, and an individual with insertion is selected as a parent to increase the body weight of the offspring cashmere goat.

[0016] The goat genome of the present application is the reference genome ARS1 of the goat.

[0017] The cashmere goat of the present application is Inner Mongolia cashmere goat.

[0018] Compared with the prior art, the present application has the following beneficial effects:

[0019] The present application provides a molecular marker related to the hair length of a cashmere goat, and it is found that by detecting the deletion mutation of the molecular marker on the chromosome 1 of the goat genome, a cashmere goat breed with long hair length can be bred, specifically: by selecting an individual with insertion of the molecular marker on the chromosome 1 of the goat genome as a father or a mother, the hair length of the offspring can be increased, and the wool yield and the body weight of the offspring can be indirectly increased. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1These are the characteristics of CNVs in IMCGs: A. Distribution of CNV numbers on the 29 autosomes; B. Correlation between chromosome length and CNV number; C. Distribution of CNV numbers within different length ranges.

[0021] Figure 2 These are descriptive statistics of CNVRs in IMCGs: A. The proportion of CNVRs in the three categories; B. The distribution of CNVRs on different chromosomes; C. The distribution of CNVRs in different length intervals; D. The distribution of CNVRs on autosomes based on a 1Mb window.

[0022] Figure 3 This is a PCA visualization; the X and Y axes represent the percentages of variance explained by the first two axes, respectively.

[0023] Figure 4 yes TDRD3 qPCR validation of key candidate CNVs. The x-axis represents 20 cashmere goat individuals, and the y-axis represents the relative copy number. A relative copy number < 2 indicates that the CNV is a deletion type.

[0024] Figure 5 yes KRT222 qPCR validation of key candidate CNVs. The x-axis represents 20 cashmere goat individuals, and the y-axis represents the relative copy number. A relative copy number < 2 indicates that the CNV is a deletion type.

[0025] Figure 6 yes MDGA1 qPCR validation of key candidate CNVs. The x-axis represents 20 cashmere goat individuals, and the y-axis represents the relative copy number. A relative copy number < 2 indicates that the CNV is a deletion type.

[0026] Figure 7 yes NOX4 qPCR validation of key candidate CNVs. The x-axis represents 20 cashmere goat individuals, and the y-axis represents the relative copy number. A relative copy number < 2 indicates that the CNV is a deletion type.

[0027] Figure 8 yes DYRK1A qPCR validation of key candidate CNVs. The x-axis represents 20 cashmere goat individuals, and the y-axis represents the relative copy number. A relative copy number < 2 indicates that the CNV is a deletion type.

[0028] Figure 9 yes SATB1 qPCR validation of key candidate CNVs. The x-axis represents 20 cashmere goat individuals, and the y-axis represents the relative copy number. A relative copy number < 2 indicates that the CNV is a deletion type.

[0029] Figure 10 yesSH2D5 qPCR validation of top important candidate CNVs. The horizontal axis represents 20 Cashmere goat individuals, and the vertical axis represents the relative copy number. Relative copy number < 2 represents the CNV is deletion type.

[0030] Figure 11 is MMADHC qPCR validation of top important candidate CNVs. The horizontal axis represents 20 Cashmere goat individuals, and the vertical axis represents the relative copy number. Relative copy number < 2 represents the CNV is deletion type.

[0031] Figure 12 is TPK1 qPCR validation of top important candidate CNVs. The horizontal axis represents 20 Cashmere goat individuals, and the vertical axis represents the relative copy number. Relative copy number < 2 represents the CNV is deletion type. DETAILED DESCRIPTION

[0032] It should be noted that the following detailed description is merely exemplary and is intended to provide further description of the application. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.

[0033] It is also important to note that the terms used herein are not intended to limit the exemplary embodiments to the specific embodiments which can be illustrated by the examples, and it is contemplated that other embodiments can fall within the scope of the exemplary embodiments according to the application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms "comprises" and / or "comprising", when used in this specification, specify the presence of stated features, steps, operations, devices, components and / or combinations thereof, but do not preclude the presence or addition of one or more other features, steps, operations, devices, components and / or combinations thereof.

[0034] The application will be further described with reference to the following examples, which are intended to be purely exemplary of the application. The following examples are presented in order to more fully illustrate the application and are not intended to limit the application. In the following examples, the test samples and test procedures used include the following (if the specific experimental conditions are not specified in the examples, the conditions are generally conventional or recommended by the reagent company; the reagents, consumables, etc. used in the following examples, if not specifically stated, can be obtained from commercial sources.

[0035] 1. Materials and methods

[0036] 1.1. Experimental animals and samples

[0037] The Inner Mongolian cashmere goats used in the present application are all from Erlangshan Ranch of Inner Mongolia Beiping Textile Co., Ltd. The phenotype data collection method used is consistent with the previous research published by the present application team, see "Rong Y, Wang X, Na Q, et al. Genome-wide association study for cashmere traits in Inner Mongolian cashmere goat population reveals new candidate genes and haplotypes. BMC genomics 2024; 25:658." Specifically, the cashmere trait phenotype data of 3842 individuals were collected between 2020 and 2023, including 1401 males and 2441 females, and the pedigree can be traced back to two generations. The phenotype records include 6966 cashmere yield, 6814 cashmere fineness, 5830 cashmere thickness and 5831 cashmere length. Among the 404 individuals used for resequencing, 84 were rams, of which 75 were one year old, 4 were two years old and 5 were three years old, and 320 were ewes, of which 92 were one year old, 54 were two years old and 174 were three years old. For the correction of the phenotype value, the present application uses the method of "breeding value + residual", and the descriptive statistics of the corrected phenotype value are shown in Table 1.

[0038] Table 1 Descriptive statistics of four cashmere traits

[0039]

[0040] 1.2, Quality control of whole genome sequencing data and sequence alignment

[0041] Genomic DNA was extracted from ear tissue samples using the phenol-chloroform method, and the concentration and purity of the DNA sample were determined by ultraviolet spectrophotometry, A 260nm / A 280nmThe ratio was 1.8-2.0. Subsequently, high-quality DNA samples were submitted to Shijiazhuang Boruitai Biotechnology Co., Ltd. for 25 X deep whole genome resequencing using the DNBSEQ-T7 platform. The original reads were filtered using V0.20.0 fastp software, the adapter sequence was removed, paired reads with N content exceeding 10 bases were removed, and paired reads with Q≤20 low-quality base number exceeding 40% of the length of the read were removed. The reference genome was indexed, and the quality-controlled clean reads were aligned with the goat reference genome ARS1, GCF_001704415.1 using V0.7.17 Burrows-Wheeler Aligner software. The aligned sam file was converted into a bam file using V1.20 Samtools software, and the bam file was sorted. The sorted bam file was removed from the duplicate data to obtain the final bam file using the MarkDuplicates program in V3.8 Genome Analysis Toolkit software.

[0042] 1.3, Whole genome CNV detection

[0043] CNV calling of Inner Mongolia Cashmere Goat population by V1.3.2 CNVpytor, which is a read-depth-based CNV detection method. Referring to the literature record "Abyzov A, Urban AE, Snyder M, Gerstein M. CNVnator: an approach to discover, genotype, and characterize typical and atypical CNVs from family and population genome sequencing. Genome research 2011;21:974-84.", the bin size of all individuals is set to 100bp. In order to improve the accuracy of CNV detection, the following standards are used to filter false positive CNV calls: P-value <0.01, fraction of reads mapped with q0 quality in call segments >0.5, size of CNV >1 kb, fraction of reference genome gaps within call region >5%, calls close to gaps in reference genome <100 kb. This produces high-quality CNVs suitable for subsequent analysis. Among them, CNVs of the duplication type refer to regions that include more than two copy numbers, and CNVs of the deletion type represent regions that only contain 0 or 1 copy number. According to the following principles, a Python script is written: the calculation method of CNV length is termination position-starting position+1, and the length interval is set to: 1 Kb~2 Kb, 2 Kb~5 Kb, 5 Kb~10 Kb, 10 Kb~20 Kb, 20 Kb~50 Kb, 50 Kb~100 Kb, 100 Kb~300 Kb. The present application uses a self-programmed Python script to count the length and number of all CNVs on each chromosome. In addition, the present application respectively counts the length and number of deletion and duplication types. The Graphpad Prism 8.0.2 software is used to draw the distribution diagram of CNVs in different chromosomes and different length intervals, and the fitting diagram of the number of CNVs and the length of chromosomes.

[0044] 1.4, CNVR determination

[0045] To obtain CNVRs, the present invention wrote a Python script according to the following principle: Considering two CNVs, CNV1 starts at position A and ends at position C, CNV2 ranges from B to D, where A < B < C < D. If the overlap between the two CNVs is at least 1 bp, they are merged into a CNVR on the genome that ranges from X to W. Using this script, two or more CNVs with an overlap of at least 1 bp are merged into one CNVR. Moreover, to reduce the false positive rate, only CNVRs found in three or more samples are considered, thus generating a final list of CNVRs. This list contains the following columns: CNVR_ID, Chr, Start, End, n_Sample, Type, Length, n_CNV. CNVR_ID is the generated CNVR number; Chr is the chromosome number; Start is the starting position of the generated CNVR; End is the ending position of the generated CNVR; n_Sample is the number of samples involved in this CNVR; Type is the type of CNVR. CNVRs are divided into two categories: "Deletion" and "Duplication". Overlapping "Deletion" and "Duplication" type CNVRs are merged into a single region, called a "Mixed" CNVR; Length is the length of the generated CNVR; n_CNV is the number of CNVs contained in this CNVR. In addition, the present invention used a self-written Python script to count the lengths and quantities of all CNVRs on each chromosome, as well as the respective lengths and quantities of the three types. The length intervals of CNVRs are set as: 1 Kb - 2 Kb, 2 Kb - 5 Kb, 5 Kb - 10 Kb, 10 Kb - 20 Kb, 20 Kb - 50 Kb, 50 Kb - 2000 Kb. The total length of the autosomes of the goat ARS1 reference genome is approximately 2,466,191,353 bp. The present invention calculated the proportion of all CNVRs in the whole genome. Statistical analysis was performed on the three types of CNVRs. The number of CNVRs in different chromosomes and different length intervals was visualized using Graphpad Prism 8.0.2 software. The distribution of CNVRs in 29 pairs of autosomes was visualized using the R package CMplot.

[0046] 1.5, PCA Analysis

[0047] Based on the obtained whole-genome CNVs data, the present invention used the "--pca 3" parameter of Vial.90 Plink software to calculate the first three principal components PC, and used V4.2.2 R to draw the principal component analysis PCA graph.

[0048] 1.6, Genome-wide Association Analysis

[0049] CNV association analysis with the traits of fleece yield CY, fine count CD, fine thickness CT, and fleece length FL was performed using the fastGWA-mlm model in the V1.94.1 GCTA software. The software will automatically filter CNVs with MAF < 0.0001 and missingness rate > 10%. As shown below:

[0050] ;

[0051] where y is the n x 1 phenotype vector; X cnv is the genotype vector, whose effect is β cnv ; g is the vector of total genetic effects captured by the CNV-derived genetic relationship matrix GRM, g ~ N(0, ); π is the CNV-derived GRM vector, where all off-diagonal elements are set to 0; e is the residual vector, e ~ N(0, ).

[0052] Since the Bonferroni correction method is too strict to determine the significance threshold of GWAS, the present application adjusts the threshold of genome-wide significant association to P = 1 x 10 -3 The genome inflation factor λ of the test statistic is calculated by the slope of the linear regression between the observed quantiles and the theoretical quantiles in V4.2.2 R. Significant SNPs are displayed with the threshold line in the Manhattan plot. The Manhattan plot and quantile-quantile QQ plot are generated by the CMplot package in R.

[0053] 1.7, Functional enrichment analysis of candidate genes overlapping with CNVs

[0054] To determine the candidate genes associated with the traits of fleece, the present application uses the V2.30.0 Bedtools software to annotate the significantly associated CNVs based on the ARS1 version of the goat genome. The CNVs are positioned on the identified genes, and only those genes located within 1 Mb ± 500 kb around each CNV are considered. In addition, the candidate genes are subjected to GO functional annotation and KEGG enrichment analysis using the clusterProfiler package in R. The KEGG pathways and GO biological processes of the candidate genes are manually searched to infer the potential gene functions.

[0055] 1.8, Real-time fluorescent quantitative PCR verification

[0056] To verify the accuracy of CNVs significantly associated with wool traits, the present application used real-time quantitative PCR to verify 10 randomly selected deletion-type CNVs. These CNVs were verified by qPCR on 20 Inner Mongolia Cashmere Goat samples. After comparing the reference genome ARS1 and determining the sequences of the 10 CNVs, primers were designed using Primer premier 5.0 software, synthesized by Beijing Sinonow Gold Biotech Co., Ltd., and qPCR reactions were performed using TB Green ® Premix Ex Taq™ II kit. See Table 2 for detailed information on all CNVs and primers. The genomic DNA of the sample to be tested was used as the template for qPCR, and β-actin, which did not undergo copy number variation, was selected as the internal reference gene. The CNVs to be verified and the internal reference gene were each repeated three times. The formula for calculating the relative copy number is:

[0057] ;

[0058] where 2 indicates that a normal individual has 2 copies of a certain gene; 2 -ΔΔCt represents the fold change of a certain DNA fragment between a normal individual and a variant individual, and the base number 2 refers to the fold change in product amount during PCR amplification, ΔCt = Ct value of the target fragment - Ct value of the corresponding internal reference gene, and ΔΔCt = ΔCt value of the sample with copy number variation - ΔCt value of the control sample. Finally, the calculated results are compared with the normal value 2, and values close to 2 are considered normal, while values of 3 or more indicate copy number duplication, and values of 1 or less indicate copy number deletion.

[0059] Table 2 List of qPCR verification primer information for CNVs

[0060]

[0061] 2. Data analysis

[0062] The association between the genotypes of 31 SNPs and wool traits was evaluated using independent sample t-test in V25.0 SPSS statistical software. When the variance analysis results of a certain group of genotypes showed significant differences, the statistical differences between two genotypes were evaluated using Bonferroni correction, and the data were expressed as "mean ± standard error".

[0063] 3. Results

[0064] 3.1. Identification of copy number variation

[0065] The application obtains 1,451,950 CNVs from 404 Inner Mongolia Cashmere Goats, and then retains 400,578 high-quality CNVs from the CNVs, with an average of 991.5 per animal. The CNVs include 193,741 deletions, accounting for 48.37%, and 206,837 duplications, accounting for 51.63%. The length of all CNVs ranges from 1000 bp to 255,600 bp, and the average length of CNVs is 6,351.66 bp, wherein the average length of deletions is 4,718.59 bp, and the average length of duplications is 7,881.34 bp, as shown in Table 3. Among the 29 pairs of autosomes, the most deletions are on chromosome 12, which is 11,905; the most duplications are on chromosome 3, which is 12,678, as shown in Figure 1 Table 4. The number of CNVs is significantly positively correlated with the length of the chromosome, R 2 = 0.49, P = 0.0001, as shown in Figure 1 Table 3 and Table B in the middle. Among all the CNVs of deletion type, the CNVs with a length of 2Kb-5Kb account for the highest proportion, which is 47.16%. Among all the CNVs of duplication type, the CNVs with a length of 5Kb-10Kb account for the highest proportion, which is 41.62%. When the length is 50Kb-300Kb, the number of CNVs decreases significantly, and the CNVs of deletion type in this length interval only account for 0.23%, and the CNVs of duplication type only account for 0.63%, as shown in Figure 1 Table 5.

[0066] Table 3 Statistics of CNVs and CNVRs

[0067]

[0068] Table 4 Distribution of CNVs of 29 pairs of autosomes of Inner Mongolia Cashmere Goat

[0069]

[0070] Table 5 Distribution of CNVs in different length intervals

[0071]

[0072] 3.2, Distribution of copy number variation region

[0073] Combining high-quality CNVs between all samples, a total of 12,609 CNVRs were obtained, with an average length of 12,258.59 bp, of which the average length of deletion was 6,211.79 bp, the average length of duplication was 17,534.39 bp, and the average length of mixed was 26,248.65 bp. These CNVRs included 8,131 deletions, accounting for 64.49%, 1,547 duplications, accounting for 12.27%, and 2,931 mixed, accounting for 23.24%, as shown in Table 3, and Figure 2 Table 3, and Figure 2 Table 3, and Figure 2 Table 3, and Figure 2 Table 6 and Supplementary Table 6. The coverage of CNVRs on chromosome 18 was the highest, which was 14.26%. Compared with other chromosomes, the coverage of CNVRs on chromosome 18 was the highest for all three types of deletions, duplications, and mixed, which were 4.00%, 2.35%, and 7.91%, respectively. The coverage of CNVRs on chromosome 20 was the lowest, which was 3.80%; compared with other chromosomes, the coverage of CNVRs on chromosome 10 was the lowest for deletions, which was 1.26%; the coverage of CNVRs on chromosome 12 was the lowest for duplications, which was 0.64%; and the coverage of CNVRs on chromosome 20 was the lowest for mixed, which was 1.38%, as shown in Table 6 and Supplementary Table 6.

[0074] All CNVRs on 29 autosomes covered 154,568,600 bp, which was equivalent to 6.27% of the reference genome. The length of 77.64% of CNVRs was 2 Kb~20 Kb, as shown in Figure 2 Table 7. It can be seen from Figure 2 Table 7 that these CNVRs had good uniformity on 29 autosomes.

[0075] Table 6 Distribution of CNVRs length and number on 29 autosomes of Inner Mongolia Cashmere Goat

[0076]

[0077] Supplementary Table 6 Distribution of CNVRs length and number on 29 autosomes of Inner Mongolia Cashmere Goat

[0078]

[0079] Table 7 Distribution of CNVRs in different length intervals

[0080]

[0081] 3.3 PCA Analysis

[0082] The first three principal components were calculated using Plink software. The population structure clustered by the first and second principal components is as follows: Figure 3 As shown, the individuals in the population roughly clustered into three categories, indicating a certain degree of population stratification. To correct for this stratification, the published method of "breeding value + residual" was used as the corrected phenotypic value to ensure the accuracy of the association analysis results. See "Rong Y, Wang X, Na Q, et al. Genome-wide association study for cashmere traits in Inner Mongolia cashmere goat population reveals new candidate genes and haplotypes. BMC genomics 2024;25:658."

[0083] 3.4 CNVs affecting hair length traits

[0084] Further research on CNVs that reached genome-wide significance revealed that deletion mutations at positions 155,344,701–155,346,600 on chromosome 1 of the goat genome can significantly affect the hair length trait of cashmere goats.

[0085]

[0086] The association analysis of the deletion mutation at 155,344,701~155,346,600 of goat chromosome 1 with the wool length trait is shown in Table 8:

[0087] Table 8 Polymorphism at 155,344,701~155,346,600 of goat chromosome 1

[0088]

[0089] Note: Different letters represent significant differences (P<0.05), and the same letter represents no significant difference (P>0.05) p p

[0090] As shown in Table 8, for individuals with the normal type in the region, the wool length is the largest; for individuals with the deletion type in the region, the wool length is the smallest. It should be noted that the normal type individual is an individual with an insertion at 155,344,701~155,346,600 of goat chromosome 1.

[0091] In the whole genome association analysis of CNV by using a mixed linear model, the CNV marker at 155,344,701~155,346,600 of goat chromosome 1 reaches the whole genome significant level, indicating that the marker is significantly related to the wool length trait of cashmere goats, and when the marker is normal, it is beneficial for cashmere goats to have a larger wool length.

[0092] Therefore, the existence of the deletion mutation at 155,344,701~155,346,600 of goat chromosome 1 can be used to select a cashmere goat breed with a large wool length, and by selecting an individual with a normal type as a sire or dam, the wool length of offspring can be improved. At the same time, according to the record in the prior art "Genetic Regularity of Cashmere Goat Coat Type and Its Indirect Selection on Important Economic Traits[D]. Li Xuewu, Inner Mongolia Agricultural University" that "selecting long wool type can reduce cashmere fineness and increase cashmere length and body weight", the molecular marker related to the cashmere length trait provided in the present application can also indirectly achieve the goal of increasing the cashmere yield and body weight of offspring.

[0093] 3.6, verification of the detected CNV by qPCR technology

[0094] In order to verify the association analysis results in the present application, 10 important candidate CNVs were randomly selected, which are located in 10 co-localization genes DYRK1A, MMADHC, TPK1, TDRD3, KRT222, MDGA1, NOX4, SH2D5, MIR17 and SATB1 , for qPCR verification.

[0095] The results are shown in Table 9:​​Figure 4~Figure 12 As shown, 10 CNVs are all deletion types in all individuals, proving that the CNVs obtained by the present application and significantly associated with the villus traits are accurate.

[0096] The above only describes the preferred embodiments of the present application, and it should be noted that those skilled in the art can make several improvements and refinements without departing from the principles of the present application, and these improvements and refinements should also be considered within the protection scope of the present application.

Claims

1. The use of a detection reagent of a molecular marker associated with the long hair trait of cashmere goats in identifying the long hair trait of cashmere goats or genetic breeding of cashmere goats, characterized in that, The nucleotide sequence of the molecular marker is shown as SEQ ID NO. 1, the cashmere goat is Inner Mongolia cashmere goat, and the genetic breeding is breeding cashmere goats with long hair length. The method for identifying the hair length of the cashmere goat comprises the following steps: Detecting the deletion or insertion of the molecular marker on the chromosome 1 of the genome of the to-be-tested goat, and the hair length of the insertion type individual is greater than that of the deletion type individual.

2. The application of the molecular marker for the long hair trait of cashmere goats according to claim 1 in identifying the long hair trait of cashmere goats or genetic breeding of cashmere goats, characterized in that, The method for breeding the cashmere goat with long hair length comprises the following steps: Detecting the deletion or insertion of the molecular marker on the chromosome 1 of the genome of the goat, selecting the insertion type individual as the parent to increase the hair length of the offspring cashmere goat.

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