CN112063725B - Chicken shin length and shin diameter related AGO3 gene molecular marker and application - Google Patents
Chicken shin length and shin diameter related AGO3 gene molecular marker and application Download PDFInfo
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Abstract
The invention discloses AGO3 gene molecular markers related to chicken shin length and shin diameter and application thereof. The molecular marker is located in the 5' -flanking region of AGO3 gene on chromosome 23 of chicken genome and comprises the following sites: the G & gtA mutation at 4329959bp, the C & gtT mutation at 4330089bp, the A & gtT mutation at 4330286bp, the C & gtT mutation at 4330299bp, the T & gtC mutation at 4330510bp and the T & gtC mutation at 4330562bp on a chicken genome, i.e., galusgallus 6 version 23 chromosome; when in application, one or more sites are selected to breed the length or diameter of the chicken shin. Therefore, the molecular markers can be used for breeding strains of breeding hens with the length of the shank or the diameter of the shank, which meet different market demands, so that the breeding is more efficient, and the market competitiveness can be increased.
Description
Technical Field
The invention relates to the technical field of molecular marker assisted selection and animal genetic breeding, in particular to an AGO3 gene molecular marker related to the length and diameter of a chicken shin and application thereof.
Background
The high-quality broiler chicken is a unique germplasm resource in China, has good meat quality, and has unique requirements on body and appearance in different regions. In some areas, the traditional Chinese medicine composition is suitable for the patients with thick and short shin, short trunk and tail, wide chest, firm and compact body and full carcasses; in some areas, the body is thin and long, the foot is high, the shank is thin, the tail feather is developed, and the body is upwarped. Therefore, the length and the thickness of the shin are important standards for measuring high-quality broilers. The shank length and shank circumference (or shank diameter) can be modified to meet different market requirements by Marker Assisted Selection (MAS). g.a.ankra-Badu et al mapped QTLs affecting chicken shin length and shin circumference on chromosomes 1, 4 and 26.
Disclosure of Invention
In order to meet different market demands and breed chicken lines with different shank lengths or shank diameters, the invention mainly aims to provide a molecular marker of AGO3 gene related to the shank length.
Another purpose of the invention is to provide a molecular marker of AGO3 gene related to the diameter of a chicken shank.
The third purpose of the invention is to provide the application of AGO3 gene molecular marker related to the length and diameter of the chicken shin in identifying the length or diameter of the chicken shin or in genetic breeding of chicken.
The fourth purpose of the invention is to provide a method for breeding a long-shin type chicken strain by utilizing AGO3 gene molecular marker related to the length of the chicken shin.
The fifth purpose of the invention is to provide a method for breeding a short-shin type chicken line by utilizing AGO3 gene molecular marker related to the length of the shin of the chicken.
The sixth purpose of the invention is to provide a method for breeding a thin-shin type chicken line by utilizing AGO3 gene molecular marker related to the diameter of the chicken shin.
The seventh purpose of the invention is to provide a method for breeding a coarse-shin chicken strain by utilizing AGO3 gene molecular marker related to the diameter of the chicken shin.
An eighth object of the present invention is to provide a method for genetic improvement of breeding hens.
The ninth purpose of the invention is to provide a breeding method of the long and thick chicken shank strain.
The tenth purpose of the invention is to provide a breeding method of the thin and short chicken shank line.
According to a first aspect of the present invention, there is provided a molecular marker of a chicken shank length-related AGO3 gene, wherein the molecular marker site is located in the 5' -flanking region of AGO3 gene on chromosome 23 of chicken genome.
In certain embodiments, the molecular markers of the chicken shin length-related AGO3 gene include the following sites: the G & gtA mutation at 4329959bp, the C & gtT mutation at 4330089bp, the A & gtT mutation at 4330286bp, the C & gtT mutation at 4330299bp, the T & gtC mutation at 4330510bp and the T & gtC mutation at 4330562bp on 23 chromosome on the chicken genome gallus gallus 6 version 23 chromosome; when in application, one or more sites are selected for breeding the chicken shin length.
According to another aspect of the invention, a molecular marker of the chicken shank diameter related AGO3 gene is provided, wherein, the molecular marker locus is located in the 5' -flanking region of AGO3 gene on chromosome 23 of chicken genome.
In certain embodiments, the chicken shank diameter-related AGO3 gene molecular markers include the following sites: the C > T mutation at 4330299bp on the chromosome 23 of the chicken genome gallus gallus 6 version, the T > C mutation at 4330510bp on the chromosome 23 and the T > C mutation at 4330562bp on the chromosome 23; when in application, one or more sites are selected for breeding the chicken shank diameter.
According to a third aspect of the present invention, there is provided a use of molecular markers of AGO3 gene related to chicken shank length and shank diameter for identifying chicken shank length or shank diameter or for genetic breeding of chicken.
According to a fourth aspect of the present invention, there is provided a method for breeding a long-shin type chicken line by using molecular marker of AGO3 gene related to chicken shin length, wherein the method comprises the following steps:
1) detecting molecular markers influencing the length of the chicken shin at 4329959bp, 4330089bp, 4330286bp, 4330299bp, 4330510bp and 4330562bp on the chromosome 23 of the chicken;
2) GG type with molecular marker at 4329959 bp; CT pattern at 4330089 bp; AA at 4330286 bp; CT type at 4330299; TT type at 4330510; TT type at 4330562; selecting individuals of one genotype or a combination of several genotypes.
3) The selected individuals are used as breeding hens to breed the long-shin type strain.
In some embodiments, breeding a short-shin chicken line comprises selecting an F2 generation holomorphic family of apricot broiler x recessive white rock chicken crosses.
According to a fifth aspect of the present invention, there is provided a method for breeding a short-shin chicken line by using chicken shin length-related AGO3 gene molecular marker, wherein the method comprises the following steps:
1) detecting molecular markers influencing the length of the chicken shin at 4329959bp, 4330089bp, 4330286bp, 4330299bp, 4330510bp and 4330562bp on the chromosome 23 of the chicken;
2) the AA type of the molecular marker at 4329959 bp; TT type at 4330089 bp; TT type at 4330286 bp; TT type at 4330299; type CC at 4330510; type CC at 4330562; selecting individuals of one genotype or a combination of several genotypes;
3) breeding the selected individuals as breeding hens to breed the short-shin type strains.
In some embodiments, breeding a short-shin chicken line comprises selecting an F2 generation holomorphic family of apricot broiler x recessive white rock chicken crosses.
According to the sixth aspect of the invention, a method for breeding a thin-shin chicken line by utilizing the molecular marker of AGO3 gene related to the diameter of a chicken shin is provided, wherein the method comprises the following steps:
1) detecting molecular markers influencing the diameter of the chicken shin at 4330299bp, 4330510bp and 4330562bp on the chromosome 23 of the chicken;
2) type CC molecularly labeled at 4330299; type CC at 4330510; type CC at 4330562; selecting individuals of one genotype or a combination of several genotypes;
3) the selected individuals are used as breeding hens to breed and breed the thin-shin type strains.
In some embodiments, breeding a thin-shin chicken line comprises selecting an F2 generation holomorphic family of apricot broiler x recessive white rock chicken crosses.
According to the seventh aspect of the invention, the method for breeding the coarse-shin type chicken line by utilizing the molecular marker of AGO3 gene related to the diameter of the chicken shin is provided, wherein the method comprises the following steps:
1) detecting molecular markers influencing the diameter of the chicken shin at 4330299bp, 4330510bp and 4330562bp on the chromosome 23 of the chicken;
2) type CT with molecular marker at 4330299; TT type at 4330510; TT type at 4330562; selecting individuals of one genotype or a combination of several genotypes;
3) the selected individuals are used as breeding hens to breed and breed the strains with thick shins.
In some embodiments, breeding a rough-shank chicken line comprises selecting an F2 generation holomorphic family of apricot-spotted chickens x recessive-white rock chickens cross.
According to an eighth aspect of the present invention, there is provided a method for genetic improvement of breeding hens, wherein the method comprises the steps of:
determining molecular markers influencing the shank length or shank diameter of the breeding hens on the positions of 4329959bp, 4330089bp, 4330286bp, 4330299bp, 4330510bp and 4330562bp on the chromosome 23 of the breeding hens in the core group of the breeding hens, and making corresponding selections in the breeding hen seed selection process according to the genotypes of the sites of the breeding hens; so as to increase the frequency of the dominant allele of the locus generation by generation, thereby improving the character of the chicken shin of the offspring.
In certain embodiments, the breeding hens comprise a hybrid line of apricot blossom chicks x recessive white rooks.
According to the ninth aspect of the invention, a breeding method of a long and thick chicken shank line is provided, wherein the method comprises the following steps:
1) detecting molecular markers influencing the shank character of the chicken on 4330299bp, 4330510bp and 4330562bp on the chromosome 23 of the chicken;
2) type CT with molecular marker at 4330299; TT type at 4330510; TT type at 4330562; selecting individuals of one genotype or a combination of several genotypes;
3) the selected individuals are used as breeding hens to breed long and thick-shank strains.
According to the tenth aspect of the invention, a breeding method of a short and thin chicken shank line is provided, wherein the method comprises the following steps:
1) detecting 4330510bp and 4330562bp molecular markers affecting the shank character on chicken chromosome 23;
2) TT type molecularly labeled at 4330510; TT type at 4330562; selecting individuals of one genotype or a combination of two genotypes;
3) the selected individuals are used as breeding hens to breed short and thin-shank strains.
The invention has the beneficial effects that:
1. the prejudice of the prior art is overcome, the QTL which influences the length or the diameter of the shin of the chicken is positioned on the No. 1, 4 and 26 chromosomes, but the SNP locus of the invention is in the 5' -flanking region of the No. 23 chromosome AGO3 gene and has obvious correlation with the length or the diameter of the shin, and the strains which meet different consumption requirements of the chicken, namely the length or the diameter of the shin, can be bred more efficiently.
2. According to different regions and different consumption requirements, the SNP loci disclosed by the application can be utilized to breed chicken shank strains meeting various requirements: for example, not only can the strain of long shank type or short shank type or the strain of thick shank type or the strain of thin shank type be bred, but also the strain of thin shank type or short thick shank type or the strain of long thick shank type or the strain of short thin shank type can be bred in a targeted manner, thereby more comprehensively serving the market and improving the market competitiveness of the chicken breeding.
3. The genotype at the 4329959bp, 4330089bp, 4330286bp, 4330299bp, 4330510bp and 4330562bp on the chromosome 23 disclosed by the invention can be used for efficiently breeding the long-shin type strain.
4. The short-shin type strain can be efficiently bred by utilizing the genotypes of the 4329959bp, 4330089bp, 4330286bp, 4330299bp, 4330510bp and 4330562bp on the No. 23 chromosome.
5. The invention discloses genotypes of 4330299bp, 4330510bp and 4330562bp on chromosome 23, which can efficiently breed thin-shin strains.
6. The invention discloses genotypes of 4330299bp, 4330510bp and 4330562bp on chromosome 23, which can efficiently breed crude tibia strains.
7. The invention discloses genotypes of 4330299bp, 4330510bp and 4330562bp on chromosome 23, which can efficiently breed long and thick shank strains.
8. By utilizing the genotypes of 4330510bp and 4330562bp on the chromosome 23 disclosed by the invention, the thin and short shank type strain can be efficiently bred.
9. The molecular markers can be applied to genetic improvement of breeding hens, breeding breeders with different shin types can be efficiently and accurately bred, and market demands can be met better.
Detailed Description
The present invention will be described in further detail below.
1 materials of the experiment
The F2 full sibling family (apricot flower chicken x recessive white rock chicken) with deep phenotype record is selected, the apricot flower chicken is a slow-growing native variety, the recessive white rock chicken is an exotic fast-growing broiler variety, and the process of forming the F2 family is described in detail in Lei et al, 2005.
Cloning and identification of SNP site of 5' -flanking region of chicken AG03 Gene
2.1 cloning of SNP site of 5' -flanking region of Chicken AG03 Gene
DNA samples from 10F 2 resource populations (apricot-spotted chickens X recessive white rock broilers) were randomly selected from the DNA samples to be used as pool-mixed templates, and SNPs of the AG 035' flanking region were identified by PCR and sequencing. For SNP genotyping, all DNA samples were PCR and sequenced, and the resulting sequences were then analyzed using the SeqMan program of DNAStar software to identify the genotype of each individual.
The PCR amplification system is shown in Table 1 below, and the reaction procedure was pre-denaturation at 94 ℃ for 3min, 32 cycles (denaturation at 94 ℃ for 30s, annealing at 58 ℃ for 30s, and extension at 72 ℃ for 30s), final extension at 72 ℃ for 5min, and storage at 16 ℃. Wherein the primer sequence is Forward primer: AG03-F, ATTGAGGCTGTGGATGAA; reverse primer: AG03-R, GCTATTGGAGGTGATATTGT.
TABLE 1 PCR reaction System
2.2 identification of SNP site of 5' -flanking region of chicken AGO3 Gene
6 SNPs were identified in the 5' -flanking region of the AGO3 gene (as shown in Table 2), and F2 families (apricot blossom chickens X recessive white rock broilers) were genotyped and correlated. These 6 SNPs were designated g.4329959G > A, g.4330089C > T, g.4330286A > T, g.4330299C > T, g.4330510T > C and g.4330562T > C, respectively, based on the position of the transcription initiation site, and the corresponding sites were located at 4329959, 4330089, 4330286, 4330299, 4330510 and 4330562, respectively, on chromosome 23 of the chicken genome.
Table 2 identification of 6 SNPs in the 5' flanking region of the Chicken AG03 Gene
Note: 1. the Site in DNA is marked with the first base position (4, 329, 236) of the AG03DNA sequence as position 0 and upstream as "-".
2. The Site in Chr23 is marked with the chicken Gallus Gallus 6 reference genome.
2.3 SNP sites of the 5' flanking region of the Chicken AG03 Gene and Association analysis with growth traits
2.3.1 Association analysis method
Association analysis between SNPs and growth traits was performed using the GLM program (General Linear Models Procedure) of SAS software. The model is as follows:
Yijkl＝μ+Si+Gj+Hk+Fl+eijkl
wherein Y represents a trait phenotype value, mu is a population mean, S is a gender effect, G is a genotype effect, H is an incubation batch effect, F is a family effect, and e represents a random residual error.
2.3.2 SNP sites of the 5' flanking region of the Chicken AG03 Gene and correlation analysis results with Chicken shank Length or shank diameter
The results show that 6 SNPs in table 3 were significantly (P < 0.05) or very significantly (P < 0.01) associated with different day ages of shank length, shank diameter as follows: the g.4329959G & gtA locus is obviously related to the 42-day-old shin length (P is less than 0.05), wherein the GG genotype shin length is the longest, and the AA genotype shin length is the shortest; the g.4330089C & gtT locus is obviously related to the shank length (P & lt 0.05), wherein the CT genotype shank length is the longest, and the TT genotype shank length is the shortest; the site g.4330286A > T is obviously related to the tibia length of 49 days old (P is less than 0.01), wherein the AA genotype is the longest and the TT genotype is the shortest; g.4330299C > T site is significantly related to 49 days old shin length (P < 0.05) and 49 days old shin diameter (P < 0.05), wherein CT genotype shin length is longest, shin diameter is largest, TT genotype shin length is shortest, shin diameter is smallest; the g.4330510T > C locus is significantly related to the shank length at 49 days (P < 0.05) and the shank diameter at 49 days (P < 0.01), wherein the TT genotype is the longest shank length and the largest shank diameter, and the CC genotype is the shortest shank length and the smallest shank diameter; g.4330562T > C is significantly correlated with a shank length of 49 days old (P < 0.01) and with a shank diameter of 49 days old (P < 0.01), wherein TT genotype is longest in shank length and largest in shank diameter, and CC genotype is shortest in shank length and smallest in shank diameter; specific results are shown in table 3 below (where SL represents shank length, SD represents shank diameter, and the numbers represent age in days):
TABLE 3 correlation results of SNPs of the 5' -flanking region of the Chicken AG03 Gene with the Chicken shank Length or shank diameter
Note:a，bP＜0.05；A，Bp < 0.01.P < 0.05 shows significant relationship, and P < 0.01 shows significant relationship.
Claims (6)
1. A method for breeding a long-shin type chicken line by utilizing chicken shin length-related AGO3 gene SNP molecular markers, wherein the method comprises the following steps:
1) the following AGO3 gene SNP molecular markers related to the chicken shin length are detected: g > A mutation at 4329959bp, C > T mutation at 4330089bp, A > T mutation at 4330286bp, C > T mutation at 4330299bp, T > C mutation at 4330510bp and T > C mutation at 4330562bp on a chicken genome, i.e., the gene sequence of the chicken genome, i.e., the gene sequence of the gene No. 23, the chicken genome, the gene of the gene, the gene;
2) selecting individuals having a combination of one or more of the following genotypes: the SNP molecular marker is of GG type at 4329959 bp; CT type at 4330089 bp; AA at 4330286 bp; CT at 4330299; TT type at 4330510; TT type at 4330562;
3) the selected individuals are used as breeding hens to breed the long-shin type strain.
2. A method for breeding a short-shin type chicken line by utilizing chicken shin length-related AGO3 gene SNP molecular markers, wherein the method comprises the following steps:
1) the following AGO3 gene SNP molecular markers related to the chicken shin length are detected: g > A mutation at 4329959bp, C > T mutation at 4330089bp, A > T mutation at 4330286bp, C > T mutation at 4330299bp, T > C mutation at 4330510bp and T > C mutation at 4330562bp on a chicken genome, i.e., the gene sequence of the chicken genome, i.e., the gene sequence of the gene No. 23, the chicken genome, the gene of the gene, the gene;
2) selecting individuals having a combination of one or more of the following genotypes: the SNP molecular marker is of an AA type at the 4329959bp position; TT type at 4330089 bp; TT type at 4330286 bp; TT type at 4330299; type CC at 4330510; type CC at 4330562;
3) breeding the selected individuals as breeding hens to breed the short-shin type strains.
3. A method for breeding a thin-shin type chicken line by utilizing chicken shin diameter-related AGO3 gene SNP molecular markers, wherein the method comprises the following steps:
1) the following AGO3 gene SNP molecular markers related to the chicken shin diameter are detected: c > T mutation at 4330299bp, T > C mutation at 4330510bp and T > C mutation at 4330562bp on the chromosome 23 of the chicken genome, i.e. the version 6 of the chicken genome;
2) selecting individuals having a combination of one or more of the following genotypes: the SNP molecular marker is CC type at 4330299; type CC at 4330510; type CC at 4330562;
3) the selected individuals are used as breeding hens to breed and breed the thin-shin type strains.
4. A method for breeding a rough shank type chicken line by utilizing chicken shank diameter related AGO3 gene SNP molecular markers, wherein the method comprises the following steps:
1) the following AGO3 gene SNP molecular markers related to the chicken shin diameter are detected: c > T mutation at 4330299bp, T > C mutation at 4330510bp and T > C mutation at 4330562bp on the chromosome 23 of the chicken genome, i.e. the version 6 of the chicken genome;
2) selecting individuals having a combination of one or more of the following genotypes: the SNP molecular marker is CT type at 4330299; TT type at 4330510; TT type at 4330562;
3) the selected individuals are used as breeding hens to breed and breed the strains with thick shins.
5. A method for genetic improvement of breeding hens, wherein the method comprises the steps of:
determining in a chicken core population the SNP molecular markers of AGO3 gene related to chicken shank length of breeders according to claim 1 or 2, selecting a long shank dominant genotype according to one or several of the genotypes of step 2) of claim 1 or selecting a short shank dominant genotype according to one or several of the genotypes of step 2) of claim 2;
or determining in a chicken core population the SNP molecular markers of AGO3 gene related to chicken shank diameter of breeding hens as described in claim 3 or 4, selecting a fine shank dominant genotype according to one or several of the genotypes described in step 2) of claim 3 or selecting a coarse shank dominant genotype according to one or several of the genotypes described in step 2) of claim 4;
so as to improve the frequency of the dominant allelic gene type generation by generation, thereby improving the character of the chicken shin of the offspring.
6. The method of genetic improvement of breeding hens of claim 5, wherein said breeding hens comprise a hybrid line of Almond chicken x recessive white rock chicken.
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| CN112063725A (en) | 2020-12-11 |
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