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WO2022134554A1 - Procédé de marquage moléculaire snp et amorce pour le criblage de la qualité d'œufs d'oies - Google Patents

Procédé de marquage moléculaire snp et amorce pour le criblage de la qualité d'œufs d'oies Download PDF

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WO2022134554A1
WO2022134554A1 PCT/CN2021/106546 CN2021106546W WO2022134554A1 WO 2022134554 A1 WO2022134554 A1 WO 2022134554A1 CN 2021106546 W CN2021106546 W CN 2021106546W WO 2022134554 A1 WO2022134554 A1 WO 2022134554A1
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genotype
egg
snp molecular
goose
individuals
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高广亮
王启贵
吴睿
赵献芝
许国洋
张克山
王海威
谢友慧
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Chongqing Academy of Animal Sciences
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/124—Animal traits, i.e. production traits, including athletic performance or the like
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • the invention relates to a SNP molecular marker method and primer for screening goose egg quality, and mainly relates to the field of molecular biology.
  • Sichuan White Goose is an excellent local breed in China. It has the characteristics of strong stress resistance, good adaptability, roughage resistance, strong disease resistance and roughage resistance. It provides meat, eggs, down, and fatty livers for consumers. Widely bred throughout China and further used for the improvement and creation of other breeds. Egg quality traits are important economic traits and complex quantitative traits, and they are all affected by genetics, nutrition, feeding methods, and external environment. Using biological and other methods for research, molecular markers such as duck body size and egg shell color have been discovered and widely used in molecular marker-assisted selection, which has greatly promoted research progress in poultry egg quality. The reproduction of Sichuan white geese is seasonal.
  • the present invention proposes a SNP molecular marker method and primers for screening goose egg quality, and the use of the molecular marker method to select and breed the egg quality can quickly and accurately identify the target traits at the gosling stage. Breeding of individual individuals greatly improves the efficiency of selection and reduces the cost of feeding.
  • the technical scheme of the present invention is: design for the following 7 kinds of egg quality 32 molecular marker SNPs: egg weight, egg specific gravity, egg relative density, egg shell strength, egg shell thickness, egg shell weight of Sichuan White Goose and yolk weight; primers include PCR upstream and downstream amplification primers and single-base extension primer probe sequences.
  • step S2 PCR reaction of the DNA in step S1, 5 ⁇ l of system: 10*buffer 0.5 ⁇ l, Mg 2+ 0.4 ⁇ l, dNTP 0.1 ⁇ l, Hotstar 0.2 ⁇ l, upstream and downstream primer mix 1 ⁇ l, triple distilled water 1.8 ⁇ l, DNA to be detected Sample 1 ⁇ l (20ng-50ng).
  • PCR amplification procedure pre-denaturation at 95°C for 2 min, 45 cycles (denaturation at 95°C for 30s, annealing at 56°C for 30s, extension at 72°C for 60s), extension at 72°C for 5 min, and storage at 25°C.
  • S3 SAP enzyme digestion reaction: prepare the total volume of SAP enzyme Mix reaction 2*460 ⁇ l, triple distilled water 1.53*460 ⁇ l, SAPBuffer 0.17*460 ⁇ l, SAPEnzyme 0.3*460 ⁇ l according to the following sequence, according to the following procedure, Digest with SAP enzyme in a PCR machine: 40 min at 37 °C, 5 min at 85 °C, and store at 25 °C.
  • S4 single-base extension reaction: prepare a single-base extension reaction Mix in the following order: reaction system 2*460 ⁇ l, triple distilled water 0.619*460 ⁇ l, 10*iplexbuffer0.2*460 ⁇ l, Terminatormix 0.2*460 ⁇ l , single base extension probe 0.94*460 ⁇ l, single base extendase 0.041*460 ⁇ l.
  • the single-base extension reaction was performed in a PCR machine according to the following procedure: pre-denaturation at 94°C for 30s, 94°C for 5s, 52°C for 5s, 80°C for 5s, 72°C for 3 min, and 25°C ⁇ .
  • S5 resin purification: add 16 ⁇ l of triple-distilled water to the 384-well plate of the reaction product, centrifuge at 2000 rpm in a centrifuge for 3 min; add resin, perform resin purification reaction on an inversion shaker for 35 min, and desalt; After the reaction is completed, centrifuge at 2000 rpm for 3 min in a centrifuge; place the desalted sample on the sample target and crystallize naturally.
  • S6 mass spectrometry detection and data analysis: the reaction results obtained from D1-D6 are detected on a nucleic acid time-of-flight mass spectrometer, the mass spectrometry peaks are detected by the Typer4.0 software, and the genotypes of the target sites of each sample are interpreted according to the mass spectrometry peaks .
  • PCR forward and reverse primers and the single-base extension probe sequences are as follows:
  • the scheme uses molecular markers to select and breed 7 kinds of egg qualities, which can quickly and accurately select and breed individuals with target traits at the gosling stage, which greatly improves the breeding efficiency. efficiency and reduce the cost of feeding.
  • FIG. 1 is an analysis diagram of the population structure of Sichuan white geese according to an embodiment of the present invention.
  • Fig. 2 is the GWAS correlation analysis diagram of egg quality traits in the embodiment of the present invention, note: A average egg weight; B egg shape index; C egg relative density; D egg shell strength; E egg shell weight; F egg shell thickness; G egg yolk weight.
  • Fig. 3 is the QQ diagram of the quality traits of goose eggs according to the embodiment of the present invention, note: A average egg weight; B egg shape index; C egg relative density; D egg shell strength; E egg shell weight; F egg shell thickness; G egg yolk weight .
  • FIG. 4 is a sequence list of primers for 32 molecular markers of 7 egg qualities according to the embodiment of the present invention.
  • Fig. 5 is a graph showing the influence of genotypes of 32 SNP molecular markers on 7 egg quality traits of goose according to the embodiment of the present invention.
  • the letters in brackets are genotypes.
  • the shoulders of the data of the same line are marked with the same letter, which means the difference is not significant (P>0.05), and different letters means the difference is significant (p ⁇ 0.05); * means the difference is significant (p ⁇ 0.05), ** means the difference is extremely significant (p ⁇ 0.01). ), "/" means no genotype; "-" is not the genotype of gene deletion.
  • Fig. 6 is the genotype diagram of SNP19 in Sichuan white goose population according to the embodiment of the present invention, Note: Nocall: detection failure individual; G: GG genotype; A: AA genotype;
  • Fig. 7 is a genotype diagram of SNP20 in Sichuan white goose population according to the embodiment of the present invention, Note: Nocall: individuals who failed to detect; G: GG genotype; T: TT genotype.
  • FIG. 8 is a genotype diagram of SNP22 in Sichuan white goose population according to the embodiment of the present invention, Note: Nocall: individuals who failed to detect; C: CC genotype; A: AA genotype.
  • Figure 9 is a genotype diagram of SNP29 in the Sichuan white goose population according to the embodiment of the present invention, Note: Nocall: individuals who failed to detect; G: GG genotype; T: TT genotype.
  • Figure 10 is a genotype diagram of SNP46 in the Sichuan white goose population according to the embodiment of the present invention, Note: Nocall: individuals failed in detection; A: AA genotype; G: GG genotype.
  • Figure 11 is a genotype diagram of SNP52 in the Sichuan white goose population according to the embodiment of the present invention, Note: Nocall: individuals who failed to detect; T: TT genotype; C: CC genotype.
  • Figure 12 is a genotype diagram of SNP61 in the Sichuan white goose population according to the embodiment of the present invention, Note: Nocall: individuals failed in detection; G: GG genotype; T: TT genotype.
  • Figure 13 is a genotype diagram of SNP62 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; G: GG genotype; C: CC genotype.
  • Figure 14 is a genotype diagram of SNP66 in Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; T: TT genotype; C: CC genotype.
  • Figure 15 is a genotype diagram of SNP67 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: failed individual; A: AA genotype; G: GG genotype;
  • Figure 16 is a genotype diagram of SNP69 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; C: CC genotype; T: TT genotype.
  • Figure 17 is a genotype diagram of SNP88 in Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; G: GG genotype; A: AA genotype.
  • Figure 18 is a genotype diagram of SNP93 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; G: GG genotype; T: TT genotype.
  • Figure 19 is a genotype diagram of SNP98 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; G: GG genotype; T: TT genotype.
  • Figure 20 is a genotype diagram of SNP105 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; G: GG genotype; T: TT genotype.
  • Figure 21 is a genotype diagram of SNP116 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; C: CC genotype; T: TT genotype.
  • Figure 22 is a genotype diagram of SNP121 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; A: AA genotype; G: GG genotype.
  • Figure 23 is a genotype diagram of SNP132 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; C: CC genotype; T: TT genotype.
  • Figure 24 is a genotype diagram of SNP134 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; A: AA genotype; G: GG genotype.
  • Figure 25 is a genotype diagram of SNP137 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; G: GG genotype; T: TT genotype.
  • Figure 26 is a genotype diagram of SNP152 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; C: CC genotype; A: AA genotype.
  • Figure 27 is a genotype diagram of SNP153 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; A: AA genotype; G: GG genotype.
  • Figure 28 is a genotype diagram of SNP165 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; A: AA genotype; G: GG genotype.
  • Figure 29 is a genotype diagram of SNP171 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; G: GG genotype; A: AA genotype.
  • Figure 30 is a genotype diagram of SNP173 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; A: AA genotype; G: GG genotype.
  • Figure 31 is a genotype diagram of SNP175 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; T: TT genotype; A: AA genotype.
  • Figure 32 is a genotype diagram of SNP177 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; C: CC genotype; T: TT genotype.
  • Figure 33 is a genotype diagram of SNP179 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; C: CC genotype; T: TT genotype.
  • Figure 34 is a genotype diagram of SNP203 in the Sichuan white goose population according to the embodiment of the present invention. Note: Nocall: individuals who failed the test; A: AA genotype; G: GG genotype.
  • Figure 35 is the genotype of SNP206 in the Sichuan white goose population according to the embodiment of the present invention, Note: Nocall: individual failed to detect; T: TT genotype; C: CC genotype.
  • Figure 36 is the genotype of SNP212 in the Sichuan white goose population according to the embodiment of the present invention, Note: Nocall: individual failed to detect; G: GG genotype; A: AA genotype.
  • Figure 37 is the genotype of SNP219 in the Sichuan white goose population according to the embodiment of the present invention, Note: Nocall: individual failed to be detected; A: AA genotype; G: GG genotype.
  • whole genome DNA is extracted by collecting venous blood of 209 goose wings, followed by whole genome resequencing (11G data volume, 10X) of DNA, and BWA software is used to compare the whole genome data to the new genome constructed by this research group,
  • the GATK method was used to extract SNPs.
  • 217 gvcf files were merged with gatk software, and a total of 16,687,310 initial SNPs were obtained, and then the vcf files were converted into plink soft PED and MAP format files for subsequent GWAS analysis.
  • 9,279,339 SNPs and 209 individuals passed the quality control. Genome-wide association analysis was done using GEMMA software based on the mixed linear model.
  • the mixed linear model In the mixed linear model, both fixed effects (such as SNP, population structure, etc.) and random effects (such as kinship) can be considered, which can reduce the false positives of GWAS analysis results.
  • GWAS Genome-wide association analysis
  • y W ⁇ +x ⁇ + ⁇
  • ⁇ the coefficient vector corresponding to the covariate matrix
  • ⁇ the effect size of the SNP
  • ⁇ represents the random residual.
  • 40 SNP molecular markers were screened for 7 egg qualities (egg weight, egg specific gravity, egg relative density, egg shell strength, egg shell thickness, egg shell weight and yolk weight).
  • the present invention uses 209 Sichuan white geese (female geese) groups in Anfu waterfowl experimental base in Rongchang District, Chongqing as experimental materials.
  • the Sichuan White Goose female geese in the laying period (30-60 weeks) were used as the research animals (209), and the female geese were raised in individual egg-laying cages, and the male and female geese were bred at a ratio of 1:4.
  • From 35 weeks to 60 weeks, in order to accurately record the quality traits of goose eggs each goose is systematically marked with foot number and wing number after hatching.
  • the above Sichuan white goose is weighed and recorded after hatching.
  • each individual was transferred to an individual cage (600mm ⁇ 800mm ⁇ 900mm) for feeding finger 65 weeks (off period), and with free drinking water and feeding standard feed, record and count the egg laying cycle (28-65 weeks)
  • the average egg weight of each Sichuan white goose was collected.
  • 3 consecutive goose eggs were collected from the above 209 geese, and the eggshell weight, yolk weight, and eggshell weight were weighed using an electronic balance.
  • the egg specific gravity was measured by the NaCl solution floating method, and 9 gradient concentrations were set (1.068, 1.072, 1.076, 1.086, 1.092, 1.096, 1.100g/cm ); Utilize the vernier caliper to measure the blunt part, the tip and the middle part of the eggshell, and get the average value of the above three to calculate the eggshell thickness; Utilize the vernier caliper to detect the egg length diameter and the short diameter, and calculate the egg shape index; Eggshell strength was measured using an eggshell dynamometer (EFG-0502, Robotmation Corporation).
  • the present invention performs genome-wide association analysis (GWAS) on the egg quality traits of the above-mentioned 209 geese and all SNP sites, and uses GEMMA software to complete the analysis based on a mixed linear model.
  • SNP molecular markers related to shell weight traits 5 molecular markers related to eggshell thickness traits, and 11 SNP molecular markers related to egg yolk weight traits.
  • the Sichuan white goose samples in the following examples are all from the Anfu Waterfowl Experimental Base, Rongchang District, Chongqing City, and all individuals are hatched in the same batch and raised in the same environment. After entering the breeding period, single-cage breeding was adopted, and the individual egg quality (egg weight, egg shape index, egg specific gravity, egg shell lightness, egg shell weight, egg shell thickness and egg yolk weight) of Sichuan White Goose was measured based on the recording of 7 indicators. data.
  • egg quality egg weight, egg shape index, egg specific gravity, egg shell lightness, egg shell weight, egg shell thickness and egg yolk weight
  • the present invention performs genome-wide association analysis (GWAS) on the egg quality traits and all-in SNP sites of the above-mentioned 209 geese, and uses GEMMA software to complete the analysis based on a mixed linear model.
  • GWAS results screened 7 SNP molecular markers related to egg quality traits (Figure 2-3).
  • the whole genome DNA of the above 209 geese was selected as the experimental material, and time-of-flight mass spectrometry was performed for the above SNP loci.
  • Primer design According to the SNP site, primer design is carried out by Agena's AssayDesign3.1 software, as shown in Figure 4.
  • Synthesis of primers and quality inspection Synthesize the primers synthesized by the company, and conduct quality inspection by matrix-assisted laser desorption ionization time-of-flight mass spectrometer (MALDI-TOF) to check whether the actual molecular weight is consistent with the theoretical molecular weight, and whether the purity of the primers meets the experimental requirements. .
  • the specific operation is as follows: draw 2ul of each extension primer synthesized into Mix, draw 2ul from the extension primer Mix and add it to 40ulddH2O, and perform mass spectrometry detection.
  • PCR amplification procedure pre-denaturation at 95°C for 2 min, 45 cycles (denaturation at 95°C for 30s, annealing at 56°C for 30s, extension at 72°C for 60s), extension at 72°C for 5 min, and storage at 25°C.
  • the PCR product was digested with SAP enzyme: the total volume of the SAP enzyme Mix reaction was prepared in the following order: 2*460 ⁇ l, three-distilled water 1.53*460 ⁇ l, SAPBuffer 0.17*460 ⁇ l, SAPEnzyme 0.3*460 ⁇ l, according to the following procedures, in PCR Perform SAP enzyme digestion in the instrument: 37°C for 40 min, 85°C for 5 min, and store at 25°C.
  • the products were prepared in the following order for the single-base extension reaction Mix: Reaction name total volume 2*460 ⁇ l: triple distilled water 0.619*460 ⁇ l, 10*iplexbuffer 0.2*460 ⁇ l, Terminatormix enzyme 0.2*460 ⁇ l, single-base extension probe 0.94 *460 ⁇ l, single base elongase 0.041*460 ⁇ l.
  • the single-base extension reaction was performed in a PCR machine according to the following procedure: pre-denaturation at 94°C for 30s, 45 external cycles (denaturation at 94°C for 5s, 5 internal cycles: 5s at 52°C and 5s at 80°C, extension at 72°C for 3 min) , stored at 25°C
  • the reaction product was subjected to resin purification, put into a 384-well plate, added 16 ⁇ l of triple-distilled water, and centrifuged at 2000 rpm in a centrifuge for 3 min; added resin, performed a resin purification reaction on an inversion shaker for 35 min, and desalted; after the reaction was completed Centrifuge at 2000 rpm for 3 min in a centrifuge; place the desalted sample on the sample target and crystallize naturally;
  • Mass spectrometry detection and data analysis The reaction results obtained above were detected on a nucleic acid time-of-flight mass spectrometer, the mass spectrometry peaks were detected by the Typer 4.0 software, and the genotype of each sample target site was interpreted according to the mass spectrometry peak map.

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Abstract

La présente invention concerne le domaine technique des marqueurs moléculaires SNP, et fournit un procédé de marqueur moléculaire pour mesurer le taux de qualification d'éclosion des oeufs d'oies, comprenant les étapes suivantes : obtention d'un locus SNP du génome entier en comparant les données de reséquençage du génome entier à une séquence du génome de référence d'une oie, et obtention de sept marqueurs moléculaires SNP candidats de la caractéristique de taux de qualification d'éclosion des oeufs d'oies au moyen d'un procédé d'analyse d'association du génome entier ; et détection des polymorphismes des sept marqueurs moléculaires SNP en utilisant un procédé de spectrométrie de masse par temps de vol, et calcul des fréquences de génotype des marqueurs moléculaires SNP. Un bloc d'haplotype est également associé de manière significative à un taux de qualification de production d'œufs. Un marqueur moléculaire SNP de polymorphisme et une paire d'amorces comprennent au moins l'un des sept marqueurs moléculaires SNP. Selon les marqueurs moléculaires SNP généraux de l'oie blanche du Sichuan obtenus par le procédé de criblage, sept marqueurs moléculaires SNP efficaces sont fournis, une banque de marqueurs moléculaires SNP de l'oie blanche du Sichuan est complétée, et les individus ayant un taux d'éclosion des oeufs élevé ou faible peuvent être rapidement détectés en utilisant le procédé, réduisant ainsi le temps et les coûts de l'élevage conventionnel des oies.
PCT/CN2021/106546 2020-12-23 2021-07-15 Procédé de marquage moléculaire snp et amorce pour le criblage de la qualité d'œufs d'oies Ceased WO2022134554A1 (fr)

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CN116287309A (zh) * 2023-03-10 2023-06-23 四川农业大学 鉴定鹅繁殖障碍的分子标记、引物、pcr方法和应用
CN116590432A (zh) * 2023-06-15 2023-08-15 重庆市畜牧科学院 一种与鹅肉品质性状相关的snp分子标记
CN117165659A (zh) * 2023-09-13 2023-12-05 上海市农业科学院 鹅双酶切简化基因组测序建库试剂盒及建库方法和应用

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CN112592983B (zh) * 2020-12-23 2023-08-11 重庆市畜牧科学院 一种筛选鹅蛋品质的snp分子标记引物
CN116515956A (zh) * 2023-04-25 2023-08-01 重庆市畜牧科学院 基于gwas筛选鹅产蛋分子标记及选择方法

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