CN116200506B - SNP molecular marker combination related to egg weight and application thereof - Google Patents
SNP molecular marker combination related to egg weight and application thereof Download PDFInfo
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Abstract
The invention relates to the technical field of molecular markers, in particular to an SNP molecular marker combination related to egg weight and application thereof. The SNP molecular marker combination provided by the invention avoids the problem of poor breeding accuracy of single molecular marker, can more accurately breed the laying hen with higher egg weight, and has obvious egg weight regulating and controlling effect on the whole period of raising the laying hen; can help to save production and breeding cost and accelerate genetic progress, and has great economic application value and scientific research value.
Description
Technical Field
The invention relates to the technical field of molecular markers, in particular to an SNP molecular marker combination related to egg weight and application thereof.
Background
Egg laying characteristics are always important characteristics of poultry breeding, including egg laying number, egg laying rate, egg weight and the like. Egg weight is an important index for evaluating egg laying performance, egg quality and cultivation profit, and is also an important factor influencing consumer selection in the market sales link. In addition, the egg processing needs to obtain the maximum output with the least investment, and the egg weight needs to be improved for the maximum effective utilization rate, so that great room is still provided for improving the egg weight of broiler chickens or local varieties in China. The genetic power of the egg weight is higher, and the breeding of the laying hen according to the egg weight index ensures that the laying hen reaches the maximum egg weight as early as possible, maintains better egg laying strength and does not increase the weight is the key point of research.
With the development of a high-throughput genotyping platform, the egg weight can be genetically improved by a molecular marking means. Genetic variations associated with egg weight may be correlated with genetic markers, thereby increasing egg weight through selection of the markers.
However, the current molecular marking means do not have wide practicability and have poor breeding accuracy.
Disclosure of Invention
In order to solve the problems, the invention provides an SNP molecular marker combination related to egg weight and application thereof. The SNP molecular marker combination provided by the invention can fully play the role of molecular markers, has the advantages of high accuracy and wide practicability compared with single-site molecular markers, can be applied to early selection of egg weight and auxiliary breeding of chickens, can help to save production cost and accelerate genetic development, and has great economic application value and scientific research value.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an SNP molecular marker combination related to egg weight, which comprises a first SNP molecular marker and a second SNP molecular marker;
the first SNP molecular marker is as follows: the nucleotide of the rs13553045 locus is A or C;
the second SNP molecular marker is as follows: the nucleotide at the rs16210999 locus is C or A.
The invention also provides application of the SNP molecular marker combination in genetic breeding of laying hens.
Preferably, the method comprises the steps of: the rs13553045 locus and the rs16210999 locus are the laying hen with AA genotype, and the egg weight is higher than that of the laying hen with other genotypes.
The invention also provides a method for breeding the laying hen with the highest egg weight, which comprises the following steps:
determining the genotype of the laying hen to be bred; the genotype is the genotype of the SNP molecular marker combination according to the technical scheme;
and selecting laying hens with the positions rs13553045 and rs16210999 being AA genotypes as the laying hens with the highest egg weight.
Preferably, the genotyping method comprises assaying using a gene analysis chip.
Preferably, the gene analysis chip comprises 600KAffymetrixAxiom Chicken Genotyping Array.
The beneficial effects are that:
the invention provides an SNP molecular marker combination related to egg weight, which comprises a first SNP molecular marker and a second SNP molecular marker; the first SNP molecular marker is as follows: the nucleotide of the rs13553045 locus is A or C; the second SNP molecular marker is as follows: the nucleotide at the rs16210999 locus is C or A. The SNP molecular marker combination provided by the invention avoids the problem of poor breeding accuracy by using a single molecular marker, can more accurately breed the laying hen with higher egg weight, and has obvious egg weight regulating and controlling effect on the whole period of raising the laying hen; can help to save production cost and accelerate genetic progress, and has great economic application value and scientific research value.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments will be briefly described below.
FIG. 1 is an egg weight curve of egg weight SNP molecular markers of different genotypes combined at different ages of a layer.
Detailed Description
The invention provides an SNP molecular marker combination related to egg weight, which comprises a first SNP molecular marker and a second SNP molecular marker;
the first SNP molecular marker is as follows: the nucleotide of the rs13553045 locus is A or C;
the second SNP molecular marker is as follows: the nucleotide at the rs16210999 locus is C or A.
The rs13553045 locus is the corresponding rs number on the NCBI website, and the corresponding position is the 170621072 th bit of the 1 st chromosome sense strand in the sequence information of the version Galus_gap-6.0 of the chicken reference genome; the rs16210999 site is the corresponding rs number on NCBI website, and the corresponding position is 1825734 th bit of the 28 th chromosome sense strand in the chicken reference genome gallus_gap-6.0 version sequence information.
According to the SNP molecular marker combination provided by the invention, two dominant sites on the whole genome are combined in a modularized manner (namely, the layer chicken with the heaviest egg weight is screened according to genotypes of the two SNP sites), so that the problem that the production potential of the egg cannot be fully developed by single molecular marker breeding is avoided, and the egg weight of the egg can be more accurately bred.
The invention also provides application of the SNP molecular marker combination in genetic breeding of laying hens. In the present invention, the application preferably includes: the rs13553045 locus and the rs16210999 locus are the laying hen with AA genotype, and the egg weight is higher than that of the laying hen with other genotypes.
The genotype of the rs13553045 locus is the genotype formed by the base of the rs13553045 locus and the locus of the corresponding allele; the genotype of the rs16210999 locus is the genotype of the base composition of the rs16210999 locus and the locus of the corresponding allele.
The SNP molecular marker combination provided by the invention can be applied to early selection of egg weight and genetic breeding of laying hens, can help to save production cost and accelerate genetic progress, and has great economic application value and scientific research value.
The invention also provides a method for breeding the laying hen with the highest egg weight, which comprises the following steps:
determining the genotype of the laying hen to be bred; the genotype is the genotype of the SNP molecular marker combination according to the technical scheme;
and selecting laying hens with the positions rs13553045 and rs16210999 being AA genotypes as the laying hens with the highest egg weight.
In the present invention, the method for determining the genotype preferably comprises determining by using a gene analysis chip. The source of the gene analysis chip is not particularly required, and the gene analysis chip is commercially available and well known to those skilled in the art. The gene analysis chip in the embodiment of the invention preferably adopts 600KAffymetrixAxiom Chicken GenotypingArray of Freon company in the United states, and the commodity catalog number is: 902148.
the invention discovers a brand new molecular marker which is obviously related to the weight of eggs, fully considers the interaction relation between two molecular markers, and when the rs13553045 locus is AA genotype and the rs16210999 locus is AA genotype, the egg weight of the egg chicken is higher and is 3.21 g higher than the average value (average value of all genotype combinations); if one of the molecular markers is selected for breeding (the rs13553045 locus is AA genotype or the rs16210999 locus is AA genotype), the egg weight under the molecular marker cannot be achieved.
For further explanation of the present invention, the following describes in detail a SNP molecular marker set related to egg weight and its application provided in the present invention with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Confirmation of rs13553045 site SNP and rs16210999 site SNP and egg weight relativity
1. Egg weight measurement
The egg weight of 40 week old layer of F2 group of 1446 Heilong and Dongxiang green shell layer was measured.
2. SNP detection
1. Genomic DNA extraction
Taking blood from chicken to be tested by chicken wing vein, anticoagulating with ACD anticoagulant, performing digestion treatment with protease (purchased from biological engineering (Shanghai) stock Co., ltd., product number B600169-0002), extracting DNA by phenol imitation method, sterilizing with double distilled water, dissolving; the ACD anticoagulant consists of the following components in concentration: 13.2g/L of sodium citrate, 4.8g/L of citric acid and 14.7g/L of glucose.
2. Genotyping
Genomic DNA of each chicken to be tested was taken and the genotype of each individual was examined using 600KAffymetrixAxiom Chicken GenotypingArray (catalogue number: 902148) from Freund's company, USA.
3. Whole genome correlation analysis of laying hen egg laying characters
Statistical analysis was performed using a hybrid linear model of genome-wide efficient mixed-model association (GEMMA) statistical analysis software, and reference may be made to: [ Xiang Z, stephens M.genome-wide Efficient Mixed Model Analysis for Association Studies [ J ]. Nature Genetics,2012,44 (7): 821-824 ] and [ Zhou X, stephens M.effect multivariate linearmixed model algorithms for genome-wide association publications ].
The statistical analysis model is:
y＝Wa+xβ+μ+ε；
y represents an individual body shape value;
w represents a covariate;
a represents a corresponding coefficient;
x represents SNP genotype;
beta represents the corresponding SNP effect;
mu represents the remaining polygenic effect;
epsilon represents the residual effect.
Analysis of data was performed using the GeneTitan (product of Freon-fly Co., USA) gene chip automated processing system, and the software for data analysis was Affymetrix PowerTools.
The result shows that in the chicken reference genome Gallus-galus-6.0 version sequence information, the 170621072-position SNP locus (namely, the rs13553045 locus) of the chromosome 1 and the 1825734-position SNP locus (namely, the rs16210999 locus) of the chromosome 28 are obviously related to the egg weight of 40 weeks.
Example 2
Correlation analysis of F2 generation individual rs13553045T/G genotype and rs16210999T/C genotype and egg weight
1. Experimental materials: the F2 generation individuals are hybridized with 1446 white-legged laying hens and Dongxiang green-shell laying hens, and the F2 generation individuals are obtained in the following way:
WL/DBS group: randomly mating 6 white lead laying hens (WL) ×133 Dongxiang green shell laying hens (DBS) (-male) for producing F1 generation;
DBS/WL group: 6 Dongxiang green-shell laying hens (female parent) ×80 white-to-home laying hens (female parent) are randomly mated for producing F1 generation;
in both F1 groups, 25 males 406 females from the WL/DBS group and 24 males 233 females from the DBS/WL group were randomly selected, and the females and male chickens selected for both F1 groups were housed together to produce F2 generation via random mating.
Egg weights were determined for each individual of the F2 generation at the time of start of production (noted AFE) and 32, 36, 40, 44, 48, 52, 56, 60, 66, 72 weeks of age.
2. The SNP detection method is the same as in step two of example 1. Genotype step results are shown in tables 1 and 2.
TABLE 1 distribution of loci rs13553045A/C genotypes across the F2 population.
As can be seen from the results in Table 1, the AA genotype is the dominant genotype of the experimental population.
Table 2 distribution of loci rs16210999C/A genotype in the F2 population.
From the results in Table 2, it can be seen that the CC genotype is the dominant genotype of the experimental population.
3. Genotype combination and 40 week egg weight correlation analysis
The genotypes of the two molecular markers rs13553045 and rs16210999 are combined, and the weights of the 40-week eggs under different combinations are counted.
The results are shown in Table 3.
TABLE 3 44 week egg weight for individuals combining different genotypes
Note that: the egg weight average value (namely 48.64 corresponding to All) of the individuals with different genotypes in the table 3 is calculated as follows: the average value corresponding to each genotype combination is multiplied by the corresponding individual number, and then added, and divided by the total individual number.
As can be seen from Table 3, the layers with both chromosome 1 rs13553045 (AA) and chromosome 5 rs16210999 (AA) genotypes had higher egg weights than the average egg weight of all genotypes was 3.21 grams higher.
Average egg weights from open to 72 weeks were determined for individuals of the F2 generation different genotype combinations and the results are shown in FIG. 1 and Table 4, where AFE in FIG. 1 and Table 4 represents open week age.
Table 4 egg weight molecular modules combination egg weights (g) at different ages of week of laying hen
| Genotype of the type | AFE | 32 weeks of | 36 weeks | For 40 weeks | 44 weeks | 48 weeks | For 52 weeks | 56 weeks | For 60 weeks | 66 weeks | 72 weeks |
| AA+AA | 37.23 | 48.15 | 49.82 | 51.85 | 52.47 | 54.22 | 55.27 | 55.85 | 56.58 | 55.35 | 56.78 |
| AA+CA | 36.77 | 47.05 | 48.88 | 50.17 | 51.00 | 52.96 | 54.08 | 54.77 | 54.87 | 54.23 | 54.34 |
| AA+CC | 35.92 | 46.31 | 47.91 | 49.04 | 49.79 | 51.75 | 52.15 | 53.51 | 53.70 | 52.94 | 53.07 |
| AC+AA | 35.80 | 46.37 | 47.82 | 50.52 | 50.66 | 52.65 | 52.09 | 54.74 | 54.24 | 54.49 | 54.00 |
| AC+CA | 35.62 | 45.82 | 47.50 | 48.58 | 49.42 | 51.28 | 51.75 | 53.20 | 52.97 | 52.56 | 52.88 |
| AC+CC | 35.34 | 44.93 | 46.43 | 47.66 | 48.54 | 50.43 | 50.75 | 51.84 | 52.00 | 51.45 | 51.59 |
| CC+AA | 33.71 | 44.05 | 45.49 | 47.64 | 46.52 | 48.91 | 48.93 | 51.90 | 50.15 | 50.00 | 50.79 |
| CC+CA | 33.91 | 44.12 | 45.58 | 46.84 | 47.73 | 49.50 | 49.72 | 50.88 | 51.44 | 50.60 | 51.24 |
| CC+CC | 34.73 | 44.27 | 45.63 | 46.54 | 47.31 | 49.54 | 49.20 | 50.91 | 50.66 | 50.21 | 49.98 |
From fig. 1 and table 4, it can be seen that the egg weight molecular module combination aa+aa has higher egg weight from the beginning of production to 72 weeks of age, and can be used for breeding laying hens with relatively higher egg weight.
In conclusion, the invention greatly avoids the defect of breeding by utilizing a single molecular marker, and the molecular module provided by the patent can fully play the role of the molecular marker to accurately improve the egg weight of the egg, thereby having great economic application value and scientific research value and being the optimization and upgrading of the molecular marker technology.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (3)
1, application of SNP molecular marker combination in egg weight auxiliary breeding of laying hens; the SNP molecular marker combination comprises a first SNP molecular marker and a second SNP molecular marker;
the first SNP molecular marker is as follows: the nucleotide of the rs13553045 locus is A or C;
the second SNP molecular marker is as follows: the nucleotide of the rs16210999 locus is C or A;
the egg weight of the laying hens with the AA genotype at the site rs13553045 and the site rs16210999 is higher than that of the laying hens with other genotypes;
the laying hen is an individual F2 generation hybridized by the white-to-home laying hen and the Dongxiang green-shell laying hen.
2. The method for breeding the laying hen with the highest egg weight is characterized by comprising the following steps of:
determining the genotype of the laying hen to be bred; the genotype is the genotype of SNP molecular marker combination; the SNP molecular marker combination comprises a first SNP molecular marker and a second SNP molecular marker; the laying hens to be bred are F2 generation individuals hybridized with the white-to-home laying hens and the Dongxiang green-shell laying hens;
the first SNP molecular marker is as follows: the nucleotide of the rs13553045 locus is A or C;
the second SNP molecular marker is as follows: the nucleotide of the rs16210999 locus is C or A;
and selecting laying hens with the positions rs13553045 and rs16210999 being AA genotypes as the laying hens with the highest egg weight.
3. The method according to claim 2, wherein the genotype determination method comprises determining using a gene analysis chip.
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