CN115044666A - SNP molecular marker for predicting sow second-birth syndrome and application thereof - Google Patents

Abstract

The invention relates to an SNP molecular marker for predicting sow 'second-birth syndrome' and application thereof, belonging to the technical field of molecular marker detection. The invention discovers that the SNP molecular marker of the 43594186bp site on the No. 12 chromosome of the pig is related to the second-birth syndrome of the sow for the first time, and when the genotype is AA, the incidence rate of the second-birth syndrome of the sow is higher; when the genotype is AC or CC, the sow 'second-birth syndrome' has low incidence rate. By utilizing the SNP molecular marker, the occurrence condition of the 'two-fetus syndrome' can be rapidly predicted in the early stage of the breeding pig, and the SNP molecular marker can be used for breeding the breeding pig with low occurrence rate of the 'two-fetus syndrome'. The invention has important significance for the breeding of sows without the 'two-birth syndrome'.

Description

A SNP molecular marker for predicting second birth syndrome in sows and its application

technical field

The invention relates to a SNP molecular marker for predicting "second birth syndrome" of sows and its application, and belongs to the technical field of molecular marker detection.

Background technique

Primitive sows are more prone to dystocia, long labor, postpartum infection, poor breast development, small litter size in second litter, repeated estrus after weaning, insufficient milk, etc., which is called "second birth syndrome". Large-scale pig farms with high management level have a low incidence of "second-birth syndrome" in sows, about 10%, while small-scale farms with relatively poor management levels have a second-birth syndrome incidence of up to 30%. % or even higher. The emergence of second-parity syndrome seriously affects the reproductive performance of sows, and causes a higher culling rate of second-parity sows, which brings greater economic losses to pig farms.

Domestic and foreign researchers have done a lot of research on the causes of second-birth syndrome in sows. It is generally believed that the incidence of second-birth syndrome is related to factors such as breeding age, nutritional level, and lactation time. However, even in large-scale farms with a high level of management, about 10% of sows still suffer from "second birth syndrome". It has been reported that some sows have slow growth and development, especially the development of the skeleton, pelvis and birth canal, which has a greater impact on the second birth syndrome. Therefore, this research team speculates that although the incidence of "second child syndrome" in sows is related to the above-mentioned level of feeding management, "second child syndrome" may also be related to the expression of certain genes, and for the first time proposes a scientific feeding method. Although management can reduce the incidence of second birth syndrome in sows, it cannot completely solve the problem of second birth syndrome in sows. If molecular markers, especially genomic selection (GS) technology, are applied to sows with second birth syndrome This research will screen out the SNP molecular markers related to the second-birth syndrome in sows, which will greatly improve the accuracy of the breeding of sows without the second-birth syndrome. Second birth syndrome problem.

At present, GWAS has been widely used in candidate gene screening related research, such as the related research on the age of Duroc sows at puberty, the screening of pig inguinal/scrotal hernia susceptibility genes, the research on growth traits of Duroc pigs, and the whole genome of lean meat mass in different breeds of pigs. Association analysis, etc.

But so far, there is no research on molecular markers related to pig "second birth syndrome" using GWAS technology at home and abroad.

SUMMARY OF THE INVENTION

The main purpose of the present invention is to provide a SNP molecular marker for predicting the "second birth syndrome" of sows and its application. The present invention finds for the first time that the SNP molecular marker located at the 43594186 bp site on the pig chromosome 12 is related to the occurrence of "second child syndrome" in sows. Molecular marker-assisted breeding work for breeding pigs without "two-child syndrome".

To achieve the above object, the present invention adopts the following technical solutions:

The first aspect of the present invention provides a SNP molecular marker for predicting "two-birth syndrome" in sows, the SNP molecular marker is located at the 43594186 bp site on chromosome 12 of the pig reference genome version 10.2, and this site exists A/C base mutation.

The second aspect of the present invention provides the application of the SNP molecular marker described in the first aspect above in predicting the "second-birth syndrome" of sows and/or selecting breeding pigs with a low incidence of "second-birth syndrome".

A third aspect of the present invention provides a primer combination for detecting the SNP molecular marker described in the first aspect, the upstream primer sequence is: ACCCAAACAGAGGCAACACAG;

The downstream primer sequence is: TTTAGCACCACCAGAGTTTGAGG.

A fourth aspect of the present invention provides a kit for predicting "second birth syndrome" in sows, the kit comprising the primer combination described in the third aspect above.

The fifth aspect of the present invention provides the combination of the primers of the third aspect or the kit of the fourth aspect in predicting the "second child syndrome" of sows and/or selecting breeding pigs with a low incidence of "second child syndrome". application.

Compared with the prior art, the present invention has the following advantages:

The present invention finds for the first time that the SNP molecular marker at the 43594186bp site on the pig chromosome 12 is related to the occurrence of "second child syndrome" in sows. When the genotype is AA, the occurrence rate of "second child syndrome" in sows is higher; When the genotype is AC or CC, the incidence of "second birth syndrome" in sows is lower. Using the SNP molecular marker of the present invention can not only rapidly predict the occurrence of "second child syndrome" in breeding pigs at an early stage, but also be used for breeding pigs with low incidence of "second child syndrome". The present invention is of great significance for the selection and breeding of sows without "two-birth syndrome".

Description of drawings

Fig. 1 is the gel electrophoresis picture of PCR product;

Fig. 2 is the SNP molecular marker typing diagram of position 43594186 of pig chromosome 12;

Figure 3 is a bar chart for the analysis of the incidence of "two-fetal syndrome" in three genotypes of sows.

Detailed ways

It should be noted that the following detailed description is exemplary and intended to provide further explanation of the invention. 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 invention belongs.

It should be noted that the terminology used herein is for the purpose of describing specific embodiments only, and is not intended to limit the exemplary embodiments according to the present invention. As used herein, unless the context clearly dictates otherwise, the singular is intended to include the plural as well, furthermore, it is to be understood that when the terms "comprising", "comprising" are used in this specification, they indicate the presence of a characteristic , steps, operations, and their combinations.

The present invention provides a SNP molecular marker for predicting "two-birth syndrome" of sows, the SNP molecular marker is located at the 43594186 bp site on chromosome 12 of the porcine reference genome version 10.2, and this site has A/C base mutation. The present invention finds for the first time that the SNP molecular marker located at the 43594186 bp site on the pig chromosome 12 is associated with the occurrence of "second child syndrome" in sows.

The present invention provides the application of the above-mentioned SNP molecular markers in predicting the "second child syndrome" of sows and/or selecting breeding pigs with a low incidence of "second child syndrome".

Through testing, when the genotype of the 43594186bp locus on chromosome 12 is AA, the incidence of "second child syndrome" in sows is higher; when the genotype is AC or CC, the incidence of "second child syndrome" in sows is higher than Low.

When selecting breeding pigs with a low incidence of "two-child syndrome", the offspring with the SNP molecular marker C at the 43594186 bp position of chromosome 12 in the breeding sow population were continuously selected, and the 12th genotype in the breeding sow population was continuously eliminated. The SNP molecule at the 43594186 bp position of chromosome number is marked as the gene progeny of A.

The present invention also provides a primer combination for detecting the SNP molecular marker described in the first aspect, wherein the upstream primer sequence is: ACCCAAACAGAGGCAACACAG;

The downstream primer sequence is: TTTAGCACCACCAGAGTTTGAGG.

The present invention provides a kit for predicting "second birth syndrome" in sows, the kit comprising the primer combination described in the third aspect above. Preferably, the kit further includes dNTPs, Taq DNA polymerase, Mg ²⁺ , and PCR reaction buffer.

The present invention also provides the application of the above-mentioned primer combination or kit in predicting the "two-fetal syndrome" of sows and/or selecting breeding pigs with a low incidence of "two-fetal syndrome".

In order to enable those skilled in the art to understand the technical solutions of the present invention more clearly, the technical solutions of the present invention will be described in detail below with reference to specific embodiments.

Example

1000 newborn sows were used as test objects, and ear tissue samples larger than 1 mm ³ were collected for DNA extraction; the extracted DNA was used as a template, and PCR amplification was performed using specific primers labeled with SNP molecules.

The specific primers for SNP molecular markers, the upstream and downstream primer sequences are as follows:

Upstream: 5'-ACCCAAACAGAGGCAACACAG-3'

Downstream: 5'-TTTAGCACCACCAGAGTTTTGAGG-3'

The amplification procedure was: 95°C, 5min, 33×[95°C, 30s, 61°C, 30s, 72°C, 30s], 72°C, 8min, and stored at 4°C; the final PCR product was carried out in 1% agarose Electrophoresis, the results of electrophoresis are shown in Figure 1.

Sanger sequencing was performed on the PCR product, and ChromasPro software was used to analyze the sequencing results; according to the sequencing peak map, the genotypes of the SNP locus could be determined as AA, AC and CC types, as shown in Figure 2.

After the sows are sexually mature, they are bred, and the incidence of second-birth syndrome in sows with different genotypes of the SNP gene locus is counted in the second parity. The statistical results are shown in Table 1:

Table 1 Incidence of second-birth syndrome in sows with different genotypes at SNP loci

Note: Different letters in a, b, and c indicate significant differences between groups (P≤0.05).

It can be seen from Table 1: when the genotype at the 43594186th position of chromosome 12 is AA, the number of sows with second birth syndrome is 0.835±0.11; when the genotype at the 43594186th position of chromosome 12 is AC, it occurs The number of sows with second-birth syndrome was 0.558±0.18; when the genotype at position 43594186 of chromosome 12 was CC, the number of sows with second-birth syndrome was 0.115±0.12. That is: when the genotype is AA, the incidence of second-birth syndrome in sows reaches 83.5%; when the genotype is AC, the incidence of second-birth syndrome in sows drops to 55.8%; and when the genotype is CC , the incidence of second birth syndrome in sows is only 11.5%.

After chi-square fitness test analysis, it was found that the incidence of AA type sow second-birth syndrome was significantly higher than that of AC type and CC type; while CC type was significantly lower than AC type (P<0.05), as shown in Figure 3 .

According to the results of the correlation analysis of the incidence of second birth syndrome and genotype in sows, it can be determined that the sow "second birth" can be determined by the detection of the SNP molecular markers after collecting ear tissue samples and extracting DNA from newly born breeding pigs. Fetal Syndrome”.

In the breeding of breeding pigs, the gene progeny with the SNP molecular marker C at the 43594186 position of the chromosome 12 in the pig herd should be continuously selected, and the gene progeny with the SNP molecular marker A at the 43594186 position of the chromosome 12 in the pig herd should be continuously eliminated. , select breeding pigs with low incidence of "second birth syndrome".

The above-mentioned embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above-mentioned embodiments, and any other changes, modifications, substitutions, combinations, The simplification should be equivalent replacement manners, which are all included in the protection scope of the present invention.

Claims (8)

1. A SNP molecular marker for predicting sow "two-birth syndrome", wherein the SNP molecular marker is located at the 43594186 bp site on chromosome 12 of the pig reference genome version 10.2, and this site exists A/C base mutation. 2. The application of the SNP molecular marker according to claim 1 in predicting the "second child syndrome" of sows and/or selecting breeding pigs with a low incidence of "second child syndrome". 3. application according to claim 2 is characterized in that, when the genotype of the 43594186bp site of chromosome 12 is AA, the incidence rate of sow "second child syndrome" is higher; when the genotype is AC or CC , the incidence of sow "second child syndrome" is low. 4. application according to claim 2, is characterized in that, when selecting breeding pigs with low incidence rate of "two-birth syndrome", continuously selecting and retaining the SNP molecular marker at the 43594186bp position of chromosome 12 in the breeding sow population For the gene progeny of C, the gene progeny of A in the SNP molecular marker at the position of 43594186bp of chromosome 12 in the breeding sow group is continuously eliminated. 5. a primer combination for detecting SNP molecular marker as claimed in claim 1, is characterized in that, its upstream primer sequence is: ACCCAAACAGAGGCAACACAG; The downstream primer sequence is: TTTAGCACCACCAGAGTTTGAGG. 6 . A kit for predicting "two-fetal syndrome" in sows, characterized in that it comprises the primer combination according to claim 5 . 7 . 7. The test kit for predicting sow "pig two-birth syndrome" according to claim 6, characterized in that, the test kit further comprises dNTPs, Taq DNA polymerase, Mg ²⁺ , and PCR reaction buffer. 8. Application of the primer combination according to claim 5 or the kit according to any one of claims 6 to 7 in predicting sow "second child syndrome" and/or selecting breeding pigs with low incidence of "second child syndrome" .

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