CN112746109A

CN112746109A - SNP molecular marker combination related to weight and body size of Nandan Yao chicken and application thereof based on whole genome sequencing screening

Info

Publication number: CN112746109A
Application number: CN202110053154.7A
Authority: CN
Inventors: 杨秀荣; 杨祝良; 邓继贤
Original assignee: Guangxi University
Current assignee: Guangxi University
Priority date: 2021-01-15
Filing date: 2021-01-15
Publication date: 2021-05-04
Anticipated expiration: 2041-01-15
Also published as: CN112746109B

Abstract

The invention provides SNP molecular marker combinations which are screened based on whole genome sequencing and are related to the weight and the body size of Nandan Yao chicken, the SNP molecular marker combinations consist of 67 SNP markers, the information of the SNP molecular marker combinations is shown in Table 1, the SNP molecular marker combinations can be used for molecular marker assisted breeding of the Nandan Yao chicken, individuals can be directly selected and bred on the genome level at the early stage, phenotypic information is not depended on, the selection efficiency can be obviously improved, the breeding process is accelerated, the intermediate breeding cost is saved, the breeding efficiency is improved, the breeding cost is reduced, and the application prospect is wide.

Description

SNP molecular marker combination related to weight and body size of Nandan Yao chicken and application thereof based on whole genome sequencing screening

Technical Field

The invention belongs to the technical field of molecular biology, and relates to a whole genome sequencing-based SNP molecular marker combination related to the body weight and the body size of Nandan Yao chicken and application thereof.

Background

Guangxi has rich local chicken germplasm resources, the Nandan Yao chicken belongs to a meat and egg dual-purpose type variety and is originally produced in the lake Li of the Nandan county in Guangxi and the Yao village of two national villages with elevation of 800-. The skin color is mostly white, a few are black skin, and the muscle color is mostly white. The cock feathers are mainly golden yellow and brownish red, the yellow and black are secondary, the hen feathers are upright with two single crowns of Ma black and Ma Huang, 6-8 crown teeth, black beak or slate cyan, red face, crown and flesh droop, and red ear leaves or blue green. Compact body type, firm and tender meat, good reproductive performance, resistance to coarse feeding, easy feeding, wide adaptability, strong disease resistance and the like, and the Yao chicken is fine and delicious and has little subcutaneous fat. In recent years, the market demand of the Nandan Yao chicken is rapidly promoted, but the breeding progress and the auxiliary means are still laggard.

The weight and size characters are important phenotypic characters in animal genetic breeding, have close relation with important economic characters, and are also marks for measuring the health condition of chickens. The living weight is an important index for the appearance of the broiler chickens on the market, and the body size characters (shin circumference, shin length, oblique body length and the like) are taken as quantitative indexes capable of accurately reflecting the body type and appearance of the broiler chickens, so that the living weight also has an important function in the genetic breeding process.

A large number of livestock and poultry varieties have been successfully cultivated at home and abroad by utilizing the traditional breeding method. However, in the traditional breeding, offspring is mainly selected according to phenotype by virtue of breeding experience, and the genotype of a target gene cannot be directly selected, so that the breeding is long in time consumption, the phenotype is complex to measure, and the selection efficiency and accuracy are low. At present, molecular biology technology is applied to breeding, breeding can be carried out on a molecular level, and the selection efficiency of breeding can be greatly improved and the breeding time can be shortened by molecular marker-assisted breeding or genetic modification breeding (transgenic breeding). By effectively utilizing molecular markers to assist breeding, the breeding process of the Nandan Yao chicken on the characters related to the weight and the body size can be accelerated, and the production and application values are improved.

In early studies on complex phenotypes, QTL mapping was mainly used to find associations between phenotypic and genomic variations. Since the 90 s of the 20 th century, the QTL linkage analysis method is widely applied to the research of traits such as chicken growth, carcass, meat quality and the like. Although the method is a classical gene positioning method, the method is more suitable for genetic research of single-gene diseases or single-gene control traits, so that the method has limited detection effect on traits with complex diseases and low heritability, the obtained QTL has a large confidence interval, possibly comprises hundreds of genes, and is not beneficial to accurate positioning of subsequent functional genes, and meanwhile, the repeatability in different groups is poor.

Compared with a QTL linkage analysis method, Genome-wide association studies (GWAS) have the characteristics of detecting SNP from the whole Genome level and analyzing the association integrally, the association of individual SNP is not considered singly any more, and the method has the advantages of more reliable results and the like. GWAS was originally proposed by Risch et al, and the concept thereof is to apply millions of Single Nucleotide Polymorphisms (SNPs) in a genome as molecular genetic markers, perform control analysis or correlation analysis on the whole genome level, and discover a new strategy of genetic variation affecting complex traits through comparison, and the new strategy becomes one of the methods with strong fine localization of the target traits of livestock and poultry.

Disclosure of Invention

The invention aims to provide a SNP molecular marker combination which is screened based on whole genome sequencing and is related to the body weight and the body size of Nandan Yao chicken in order to overcome the defects of the prior art.

Another object of the present invention is to provide the use of the above SNP marker combination.

In order to realize the purpose of the invention, the invention provides an SNP molecular marker combination related to the body weight and the body size of the Nandan Yao chicken, which consists of 67 SNP molecular markers, wherein the number of the SNP molecular markers is respectively as follows: chr, chr, The nucleotide sequences of chr2_112111693 and chr3_110246950 are respectively shown as SEQ ID NO. 1-67, the information of the nucleotide sequences is shown as table 1, the base sequences of 200bp before and after each SNP marker are provided in table 1, and base variation exists at the 100bp position.

The invention also provides application of the SNP molecular marker combination in molecular marker assisted breeding of Nandan Yao chicken.

The invention also provides application of the SNP molecular marker combination in preparation of a detection kit.

The invention also provides a method for detecting the genotype of the Nandan Yao chicken by using a molecular biology technology, which comprises the following steps: and respectively designing primers according to nucleotide sequences of two flanks of 67 SNP molecular marker loci, and carrying out genotyping on the Nandan Yao chicken material to be detected by using the primers, so as to identify the genotype of the Nandan Yao chicken to be detected.

The invention also provides application of the method in molecular marker assisted breeding of the Nandan Yao chicken.

Through the technical scheme, the invention has the following beneficial effects:

the SNP molecular marker combination related to the weight and the body size of the Nandan Yao chicken provided by the invention can be used for molecular marker assisted breeding of the Nandan Yao chicken, individuals can be directly selected and bred at the genome level in the early stage, phenotype information is not relied on, the selection efficiency can be obviously improved, the breeding process is accelerated, the intermediate breeding cost is saved, the breeding efficiency is improved, the breeding cost is reduced, and the application prospect is wide.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. It is intended that all modifications or alterations to the methods, procedures or conditions of the present invention be made without departing from the spirit or essential characteristics thereof.

Animals, instruments, reagents, and kits used in the following examples are commercially available;

example 1 screening of SNP molecular markers related to body weight and body size of Nandan Yao chicken based on whole genome sequencing

(1) Population selection and phenotype collection: feeding Nandan Yao chickens to 120 days old at an animal experiment base of Guangxi university, stopping feeding for 24 hours, and determining relevant properties such as body weight, body slant length, keel length, chest depth, chest width, hip width, shin length, shin circumference and the like by referring to agricultural industry standard NY/T823-2004 of the people's republic of China; finally, phenotypic data of 300 Nandan Yao chickens (149 cocks and 151 hens) were collected.

(2) Phenotype data sorting and analysis: performing statistics and analysis on the phenotype data collected in the step (1) by using R software, wherein the statistics and analysis comprise minimum values, maximum values, average values, standard deviations and Coefficient of Variation (CV), and the results are shown in Table 2;

TABLE 2 statistical analysis of Nandan Yao chicken phenotype assay data

(3) DNA extraction and sequencing: collecting blood of a test chicken, extracting genome DNA by referring to a phenol simulation method in molecular cloning experimental guideline IV, detecting the purity of the DNA by an ultraviolet spectrophotometer, sending the qualified DNA to Beijing Nuo He sourcing science and technology limited company for whole genome re-sequencing, wherein a sequencing platform is Illumina PE150, the depth of the re-sequencing is more than 5 x, filtering off-machine data of the sequencing and obtaining effective information, and the filtering standard mainly comprises the following steps: 1) filtering out reads containing the linker sequence; 2) removing the pair of paired reads when the content of N in the single-ended sequencing read exceeds 10% of the length proportion of the read; 3) this pair of paired reads is removed when the number of low mass (< ═ 5) bases contained in the single ended sequencing read exceeds 50% of the length proportion of the read. After obtaining the valid data, further sequencing quality evaluation is carried out on the valid data, and the error rate distribution condition is checked. The sequencing error rate and accuracy are shown in table 3. And finally obtaining high-quality data 2999.5 Gb.

TABLE 3 genome DNA resequencing results statistics of Nandan Yao chicken

(4) And (3) genome data alignment: comparing and analyzing genome data obtained by sequencing through biological information analysis software BWA and SAMtools; the comparative reference genome is the fifth version of the reference gene (GCA _000002315.3) of chicken (ftp:// ftp. ensembl. org/pub/release-92/fasta/gallius _ gallius/dnas /), parameter "mem-t 4-k 32-M"; removing duplication by utilizing an rmdup parameter of SAMtools software according to a comparison result; the population sample average alignment was 98.75%, the average sequencing depth to the genome (excluding the gap region) was 8.38 ×, for detailed information see table 4;

TABLE 4 Nandan Yao chicken genome data comparison statistics

(5) SNP quality control and filtration: carrying out group SNP detection by using a mpieup module of SAMtools software, and obtaining SNPs with high quality through the following quality control during detection: 1) filtering SNPs with the quality value of Q20, namely the sequencing error rate of more than 1%; 2) if the distance between the two SNPs is detected to be within 5bp, removing the SNPs; 3) the support number (coverage depth) of SNPs is between 1/3 and 5 times the average depth. 10949206 SNPs are preliminarily obtained through the steps; then, carrying out primary filtration on the SNP information of the population by using VCFtools, wherein the filtration parameter is '-means DP 5-thin 10'; after being converted into the PLINK file format, the file is further filtered through the PLINK, and the filtering parameter is "-geno 0.1-mind 0.1-maf 0.05-hwe 1 e-6". Finally, 9080580 SNPs on the autosome were obtained for subsequent association analysis.

(6) Genome wide association analysis (GWAS): performing principal component analysis by using GCTA software, then performing association analysis by using GEMMA software in combination with phenotype information and genome SNP information, rejecting individuals with phenotype values scattered outside a standard deviation of +/-3 times of the mean value within a certain trait, and adding the first three values and the sex of the principal components serving as covariates into a mixed linear model, wherein the model is as follows:

y＝Wa+xβ+u+∈；u～MVN_n(0,λτ^-1K),∈～MVN_n(0,τ^-1I_n)

wherein y is a phenotype vector, W is a fixed effect matrix, alpha is a coefficient vector corresponding to a fixed effect, x is an SNP marker vector, beta is an effect value of the SNP marker, u is a random effect vector, e is a residual error, and tau^-1Is the residual variance, λ is the ratio between two variance components, K is the matrix of relationships, I_nIs an identity matrix;

(7) screening and extracting SNP molecular markers related to the weight and body size traits of the Nandan Yao chicken: the number of independent SNP sites on the genome was obtained using the "- -indep-pair 2550.2" parameter of PLINK (947270), with 1/947270 as the multiple test threshold. The sites reaching the significant association level are extracted through R software, 67 SNP molecular marker sites are preferably selected, the information of the sites is shown in table 1, the sites are respectively associated with the oblique body length, the living body weight, the chest depth, the chest width, the hip width, the keel length, the shin circumference and the shin length of the Nandan Yao chicken, and the sites can be used for breeding the weight and the body size characters of the Nandan Yao chicken.

When the Nandan Yao chicken is bred, the SNP molecular markers can be utilized, primers are designed on nucleotide sequences of two side wings of the Nandan Yao chicken, blood is collected and genome DNA is extracted when the chicken is 3-4 weeks old, the primers are utilized to carry out genotyping on Nandan Yao chicken materials to be tested, individuals with target character genotypes are reserved, and the breeding speed can be accelerated.

Sequence listing

<110> Guangxi university

<120> SNP molecular marker combination related to weight and body size of Nandan Yao chicken and application thereof based on whole genome sequencing screening

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atctttgctt tgaagtgagg tttttgtttg tgatgtaacy aggaagtgtt attccagcag 120

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atttaagggt tggcagtgaa atcaaagttg ttctggagat cctggcatac ctgaggtctt 180

ccaaaggtaa gcagcttctt 200

<210> 8

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<212> DNA

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ttctgcaaag gtcagcttag acgtgcagct tctttcaaag gtgctgcatt gggggtttca 60

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cacagatacc ggaagcagct catacccacg tgtgctgttt gtagcttaca tgtctgctga 180

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<213> Gallus gallus domesticus

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tattcaggta actagtgaaa taatatataa tgatttctga taacttcaga tgcctggaca 180

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<210> 11

<211> 200

<212> DNA

<213> Gallus gallus domesticus

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caggaggtgg aaggggaatg ggatcccagt gctcctccag gacactgtcg ctgcaggagc 60

ccagcaggac caggacactg cgtgtagcca gctctgactw tctctgttgc ctcctctagc 120

tcaaggcaca gcagcaccac tgtacttact cctgttccac acccagcttt cactctgtgc 180

tcacaggtgt ggctggaaga 200

<210> 12

<211> 200

<212> DNA

<213> Gallus gallus domesticus

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tatagctatt gctgcacctt catgttcttt actttggggt gctaagcaag ccaaacagta 60

tgttacttta ctgcaatgta gctatgacct ttctttgcgy gagatgtgat tgatactttc 120

acagttcctg atgttaaaaa acaggaagac acgaggatcc tatgcaaatt taaggaatta 180

aggcttagtc ctagttaaca 200

<210> 13

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 13

atgtcaaaag acacttgatt ttcttacaca atattcactt acaatctttt tgtaattcac 60

aaataggaca agaaaggtat acgaaaagga tctcactggm ctgtttcacg tgctgttatt 120

ctttatgaat tccatcctgc tacgtgagat tctcagcctt gcaacaccca ttagtcaaac 180

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<210> 14

<211> 200

<212> DNA

<213> Gallus gallus domesticus

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ccaatgtggt tttcttttat ttggattcac caggcttaaa acttaactaa ctgtactatc 180

tataaaaaga aatgttctgg 200

<210> 15

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 15

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aagagaaacg attcaaaatg caataccgag aaaggaaagw ataaacgaac aaaatccgta 120

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<211> 200

<212> DNA

<213> Gallus gallus domesticus

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<211> 200

<212> DNA

<213> Gallus gallus domesticus

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ctgctgccat ctgtctcaca gcaacaacat gtaaaggaat attggtggga aggtttaacc 180

tctactgcca tgccaccaac 200

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<211> 200

<212> DNA

<213> Gallus gallus domesticus

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gccttttcat ggcctgccag cagaactgca ggtgctattr aaagatgtaa ctacacagaa 120

cagataatta gctcggcttg tcacacttac ctggaacggc tgtcttagaa gccaaggctt 180

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<211> 200

<212> DNA

<213> Gallus gallus domesticus

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<210> 20

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<212> DNA

<213> Gallus gallus domesticus

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tgtgattaca aataagtaag aaataagtaa aaagccctct ggcttttcag atcctaagtt 180

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<211> 200

<212> DNA

<213> Gallus gallus domesticus

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atccctccat gagggccagg gcctgaaaat gtaaggcttc tttcaattgt ctgaagtcct 180

cacctacttc ataagtcaga 200

<210> 22

<211> 200

<212> DNA

<213> Gallus gallus domesticus

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cttttagtgc aaaatatttt aactgaatgg gtagtatttt agattaatat catgttttgt 180

ggtgtttttt tttttaaagc 200

<210> 23

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 23

tatctgtgct agcaatttgt caccagtgta cttcagaaac ctccttcaga aactctgctg 60

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aaatgtaagg cttctttcaa ttgtctgaag tcctcaccta cttcataagt cagatgatct 180

gtagcaaaca ccacaaagtt 200

<210> 24

<211> 200

<212> DNA

<213> Gallus gallus domesticus

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gaccacagag gaattgcttt ggcagggccg tgaggatgaa agtgatattc tggtccctgc 60

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gggtttaagt gaccagcttt ccgctgggtc aaactatcac tgatgaagct ttgaggttcc 180

tcctgttcta ggccagagca 200

<210> 25

<211> 200

<212> DNA

<213> Gallus gallus domesticus

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ttccccatca cccccccccc caacccccct ccatggggct ccagatcccc aaagaaatca 60

ttgcagctgc acaaatagtg attaataaat cctgcttccy ccatcaaggg aggaaatctt 120

tattcatatg tatcctataa actaattttt gcatctttgc aataaatcca tctcccagcc 180

accagctctg ctctgccttc 200

<210> 26

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 26

cataaagtgg cttgggttgg aagagacctc aaggatcatc aagttccaac tcccttctgc 60

aggcagggtt gccaaccact agatcaagta ctagatcagk gccagggccc tatccaactt 120

ggcctttaac gcctccagag tattttccat gagacccatc atttaggtag ggcaagcaag 180

agactaccag gccagcccca 200

<210> 27

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 27

agcagtatta taggttggaa tgctgtttac aggtatcagg actttgggga tcaggtttga 60

gttctgtgga gctgtaacag ctgttataac ctgtgccacy ccaggctgaa tcactggggt 120

cacaaccgta ctacctacat cggctgcact gcaaacaacc gaatggctcg ctttggactc 180

agaaactgcc ggtggtgggt 200

<210> 28

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 28

aagagtggct gtagtgttta cgggattcat ggcagaaaac agccacatga tggagaacag 60

ccctaggggc agtttcctaa ctatggagat gttacagagy agctttgcat acagtttaag 120

tttaaggctt catccaaact aatcaaaacc agtgagtatc ttttcacaga ctgaacaggc 180

tttggatttt ggcttattca 200

<210> 29

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 29

ttgggtctgg tacatttaca actgaagggc agaaaatgtc ttagcactac ttatttgttg 60

ttaaaatgtg aagagtgaca caccgcaaaa aaataataay aactaatgtc tgaggaaaac 120

atgctgtcaa gattcagaaa gttcacaaga aagctgttct actcggtgtg gcattttaaa 180

caaaagatct tcttctcttc 200

<210> 30

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 30

gagagctcag cactgcagga gccacccact gggactcaca gccctacaac caggcagcag 60

ttctgcctca ggaagggtct ccatgtgtaa aggcttaack tcccccgcaa cccacttatc 120

ctgagactga agtcttactg aactctaagc acttcgacca actttaacca cagtattgta 180

gaagaaaatg aacaaaatac 200

<210> 31

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 31

aataaatagt ggaatcccaa agttcttttg caagactcca acttgtattt tgaacttcac 60

actataaatt catctcaatt tgctgctttt aagtagctgy ttctcaaggc ccttgtagta 120

tggctttgtc atttaaattg tattgatttt ggttttttac acctctcagt gttgtcattt 180

tctttcatgt tttctttata 200

<210> 32

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 32

gattttaagc ctggattacc ctaatgcttt cttatcaata gatcattaag cctgtttgaa 60

cgcactctcc ataggtagaa atttactttc agtgctaacr tctacaagga ttgaataaag 120

cttttagtgg agtagaagtt atactttaac gatggaaatc ttaatcaaat aataaaattg 180

gctgatgatt ggctctgtca 200

<210> 33

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 33

gaaaagtgtc ttgcatggag aatctacata tgggtaagct gcaataacaa tgaatcaggt 60

agtaccactt ctccagaaga gagcagttca acagtacaay aaaaatgtta acggagctgg 120

actgaccaac ttgggtaaat gtgttctacc tgtctcaaaa gttcattctc catgttgaac 180

tgaagacgtt tattatttta 200

<210> 34

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 34

gctacaaata atggaaatct acttgactag acatgagatt aaaaaaactg aaactaccct 60

ataaggaaat ctgaatggaa tgctttattt cgggctgcty tcatcagcag ctgggcttca 120

ggacacagtc tttattcatt catatatgcc cagtcttcac gcccacaaac cctgatgggc 180

tgagatgcta cctggtacac 200

<210> 35

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 35

tttttttttt acctaaatat attacttggt tatgctttgt agaaacaaga ttaatgcaaa 60

tagctcaata ttatctgggc tgagacaaga caagcagaty gccagtctat ctccccacta 120

tgccaaagga tgcccactat gccattaggc tcatcaaact ttccagcttt atctgcgaaa 180

gattttgtga tagtgaagca 200

<210> 36

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 36

agtgggtccc cagtggtgtg gcctacttgc atcttggggg tttttattgg aggttggggg 60

atcagtagtc accctggtaa aaggtacacg gcctgcagay gttgtgatca actctggttt 120

tcatcactaa ggggacctta attgtgtccc tctggcaaat caggacacct ctttggtacc 180

tgatacagtt atacgctctc 200

<210> 37

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 37

ccctactcca tgccagagca gcagtggttg ttagagcttc tgggcttctg ttgggggtgc 60

cagtagggaa ggacccttca gccatcagtc agcaagggcy tggggactcc tggagcacgg 120

ccccatcaga caactgctgt aacccaaatc tctctctaaa atgggctaca ttcaccttat 180

gaatacacac gcacacattt 200

<210> 38

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 38

cccatccagc ctggcttgaa tgtctccagg gatggagctt tcatcacctc tctgggcaac 60

ctgtgccagt gcctcaccac ccacactgtg aagaaaacty cttccttata tctaatctaa 120

atctctctgg ttttagttgg aaaccatttc cccttgtcct atcacaacag atcccgctga 180

agagtctgtc cccttctttc 200

<210> 39

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 39

gcatggagaa tctacatatg ggtaagctgc aataacaatg aatcaggtag taccacttct 60

ccagaagaga gcagttcaac agtacaataa aaatgttaay ggagctggac tgaccaactt 120

gggtaaatgt gttctacctg tctcaaaagt tcattctcca tgttgaactg aagacgttta 180

ttattttagc atctggatac 200

<210> 40

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 40

tagctggact gaagcaaaac ttcaggagct ctataccaca aagtaacaac ctgtaaaaca 60

aaaaggaaga taagttacag caagcttagc tgtacaactr caacctcaga aatgttactc 120

tacacagatc caatttagct tcatccaaac cacaaagcct cacagattca ggcaaattta 180

tttgatcagt gcgtagaaaa 200

<210> 41

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 41

ttgctgtgtg tggaaacttg tgctggtttc tgaagtggtt aattctgcag gacagaaggc 60

cacgcttctg tgtgatgtgg tgatgaagtg agatgtagcr tcctacagct tgggatttgc 120

ttttcagtta ggtgagactt ctgcaagggg aagcattcct tcctagagcc aaggaagacc 180

cttccatggg gctttcccca 200

<210> 42

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 42

tgccagcacc tcaccaccct ctggggaaag aatttcctcc cagcatctaa actaaatctt 60

cctgctctgt tggaccccag atcggggctt gagaatcccr tgtgttggca tgagcagttg 120

ctgcgtgttc catgtgcata gtgatgcaaa ggattgtgtt gagatgcctt gtgccagtca 180

gtgttgtgtt ctggtgggtg 200

<210> 43

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 43

gtgtcattaa ttatttaact ttaccatagg cctaatttgt tgtgtagtgg aacaagtgga 60

agtctttcca ccaacttcaa actaaataag atgagattty gtaataggca gagtacacag 120

tctagacttt taaatacttc agtacttact taaaaggatt caggttgttt ggattgaatg 180

gtaataacct cagttaagtg 200

<210> 44

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 44

gccattgtat ggttctatga atatattcca aatatccgaa ggtaatgctt tgttgtattt 60

aaaacttgcg ttcttattac taaacatttc caatagatcy ggaatctcac tccatttttt 120

cttctagttt agaaaagctg gagattacac ttcagaaagt caaactttct cttagaaaca 180

tatggtgatg caagttggat 200

<210> 45

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 45

aatccttagg gtagaaatgg tctggatggc ctctgaaggc ctctgaaggc catctagtcc 60

aactcccttg caatgaacag ggacatcacg gccagatcak gttgcccagg gcctgatccg 120

ccctcacctt ttaagtctcc agggatgggg catcaaccat atcactgggt aacctgatcc 180

agtgcctcac caccctaact 200

<210> 46

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 46

cattaaaatc acattacagc tacccttgtt gtctcttaga catgtataac gttaaaaaag 60

tagttttccc ctcaagcgat gatgaaatat ccttgcttcy atgaaatatc ttgtcaaaat 120

acttaaatat acctttggct tgagcacaga agtgttgtag caatgtagct ctggcataaa 180

acattttaaa ttctttactt 200

<210> 47

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 47

cttgactgac gtcccttcca acccaagcca ttgtatggtt ctatgaatat attccaaata 60

tccgaaggta atgctttgtt gtatttaaaa cttgcgttcy tattactaaa catttccaat 120

agatctggaa tctcactcca ttttttcttc tagtttagaa aagctggaga ttacacttca 180

gaaagtcaaa ctttctctta 200

<210> 48

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 48

ctacagataa atcccagaag tatccaaact tctgccttca aacctcagac atctcttata 60

cttagatcaa aatctaagag tggaagtaga ccgtttacak ctttaagcag ttaaaagaag 120

aggacagcta tgtatcctta ttttatcaaa cagtgcacac ctttaaacca tcttcccctg 180

acattttttg ataacttcgc 200

<210> 49

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 49

gtaagttata tctccctctc ccccagctca gctctagttc ttccaccttc atgaaaacta 60

gagaattcag tagctccata atacatgatt gaggtactty gcagagaagg cctgagttca 120

gtacatgctg ctgaagcacc atccctcagg catgcagtgt atcagcagaa caactgtgaa 180

aagaactcag gatccttaat 200

<210> 50

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 50

caatccaagc gccttttaga ctctttcagt gttggtacct tcttccagat gccagaaaaa 60

aaaagtgtga acctgttctc agcagcagta acagcagctk ccctgctgac aagcaagcaa 120

acactgtgag aagatcatct ccaagtcact gttgtgaact ttggcaagat acagcatgtg 180

ttctgttctc cacctgctgt 200

<210> 51

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 51

gtgacccatc atctccaaat aaatgttcag agctctgtct gaaggaccgc tttcaagagg 60

ggatggagag agcgctgtga tgcacgctgg caccagcagr ttgcctggct tgcccgctca 120

ggaacaaatg gcattgaaag ctgctgtgct ggttaaataa atacctttcc tgccttcaga 180

acctactcta cttgaaggac 200

<210> 52

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 52

tatctgtgcc gcgcgtgtct gagaaaacgg cgcggagccc gtggggaaat atcccggtgg 60

gttacgggct gacagaacct tagggttgaa gaacaccggy gacaagtggc tttctgcgtg 120

gctcgtggct cgccggtgtg ctaaagcatc agagccgcag cgcagggggc tgctggcggt 180

gtctgtgctc gggcccgcgt 200

<210> 53

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 53

gatggtttta aacaaaaaga ggggagattt aggttagatt ttaggggaaa attcttagaa 60

ggtggtgagg gactggcaca ggctgctcag agaagctgtk ggggccccat cgctggagat 120

gttcaaggcc gtgttggatg gggccctggg tcacccagtc tggcaggtga caatcctgcc 180

catggcagcg gtttggaact 200

<210> 54

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 54

gtatcaaaaa caacttgttg ttgttgttgt tgttgttaca atttctttgc tgtgtctaaa 60

atagaagtag gaagtcttga aagcagatgc agctacagcr tttaacttta ccaagttaac 120

caagaagggt gaaggtaaca tactttacgt gcacgaaagg agattattat ctccagagca 180

gatatgaatg ttacagagaa 200

<210> 55

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 55

tcaagatgaa gcattttttt cccctcttct ttgagacctg gttaaactat ttggatgttg 60

ttaagcttag tatcatttat aaggataatt caagccaacr ggagactgaa tggaaaaata 120

gaattgcctg agagtatttg cagactttaa gacagtatat cttgttttaa taattaataa 180

caattttgtt ttttatctta 200

<210> 56

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 56

cacctagaag ggaagatgtt tattttaata cacatttgtt tagattcaca tttcacattt 60

caagaaggta tcaatttttg cagataatta aaagaaaaaw ttgcagataa ttaaatcata 120

tgaaccagca aacctttgtt atgttagcct aaagagagaa aagggttgtt acttttcctt 180

tgtgtgatcc ctcacaagag 200

<210> 57

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 57

agctgctggt taatcacctg ctgaagtgct aaaagtgata gcttggctct gtttcctcaa 60

gcaacggtga gctgtcaagg ctgaccagca agaaaatgty ctgcagctag gagaattccc 120

ctgccatcct atttagagcc attgcaatta ggacttgctg ctttcatctg cactgatttt 180

gctgtgactc caacagtctg 200

<210> 58

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 58

gtcaaaaagg acaaagcctc ctcagcatac ctgaggatag tcctaagagt aaaggcaaat 60

aaaacaagtc tctgtagatg gaagacagcc tgctccccar ttcatgagca aaacattgtg 120

ctgttactca agctatagtt ctagtgatcc tatttcacta ttcacacagc atcattcctt 180

tctgaaacag tcccagaaga 200

<210> 59

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 59

aaacaggaaa gtcagaaatg agctacctct tgtatatgct cctttgtgct tttcatgatg 60

atacagtctt taacaatgtg attgcatgca attttgatgy cgaaaccctc ccccatccag 120

ctctctgctg gatgctttag cagatccact gttcagcttt atacagtgtt tattttgttc 180

tcgctagttt acttagcggc 200

<210> 60

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 60

aagtggtcac tgtcaaaaag gacaaagcct cctcagcata cctgaggata gtcctaagag 60

taaaggcaaa taaaacaagt ctctgtagat ggaagacags ctgctcccca attcatgagc 120

aaaacattgt gctgttactc aagctatagt tctagtgatc ctatttcact attcacacag 180

catcattcct ttctgaaaca 200

<210> 61

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 61

ctccacactc tgagtcaacg gctcatccca attcagtatc accaaacttg ctcataacac 60

cttccagtcc tacatccaag tcatttaggg aaacatcaay ggccctaaaa cagagccctg 120

cggaaccccc actgactgta atccttcagt gcccccaagc agggtgttaa ggaggttaac 180

actgagtcag gtcaagtaac 200

<210> 62

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 62

tcatcccaat tcagtatcac caaacttgct cataacacct tccagtccta catccaagtc 60

atttagggaa acatcaacgg ccctaaaaca gagccctgcr gaacccccac tgactgtaat 120

ccttcagtgc ccccaagcag ggtgttaagg aggttaacac tgagtcaggt caagtaacaa 180

acacctagta tttattaata 200

<210> 63

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 63

aacctatcta tggttgtggc accaaagcac tctaaatatt tatttatctt caggtgtgta 60

gacagtaatc aagcttaagg caatgcccct ttttttcatr agtcagcaag acccatagcc 120

aggcccaggc aggccagtga ttccaagaaa aacaaacacc ctaaacttat tcttttttga 180

aattttaact ttttttccct 200

<210> 64

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 64

ctaaattgtt aggaaaatat ggcatgtgtt gtaggtttca gaacatagat gttcctcaca 60

catgaaattt catgctaaat atcccaactt actttcaccr tagttctagc agagactttc 120

tctgtaatgg gacaaaactc aggtaaagtc acaactgtgt taaacaagta attgcagcag 180

aaacttattt tgcagagcca 200

<210> 65

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 65

ctgtttttat tcactgtgtt aaaatgatgt gaaattttga gtttcagaat aaaaaagctg 60

tttctctagc agcttattct ctgtccatgc aataactgty gccttgttca tcttaataca 120

actacccaga caatgttgta ttgattaact catatacact gtgacttgat acctaattat 180

ttcttttgtc atcttgctga 200

<210> 66

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 66

tcccagcttt tccacaggaa ggacagaaaa tgagatcagt aagtgttcct gcaaagacac 60

ttacccatta aatcccacac cataggactg gatcatatgy ggccacactc tttaagtaaa 120

catgatgaaa aaaatccaat agatgtgtta ttattttttc ccacgggctg gagagtataa 180

cacgagaaaa gcttctaaag 200

<210> 67

<211> 200

<212> DNA

<213> Gallus gallus domesticus

<400> 67

aacaaccatt tggatgtggt gctcagggcc atgactgagc ggagggctgt tagagttagg 60

gtaggatggt tggggtgggg ttggactgga tgatctttaw ggtcttttcc aacctgagcg 120

attctatgat ttctctccac accctccctt catgtattca cagacatgga taggatccat 180

tccccccttc ttttttcccc 200

Claims

1. The SNP molecular marker combination related to the weight and the body size of the Nandan Yao chicken based on whole genome sequencing screening is characterized by consisting of 67 SNP markers, wherein the number of the SNP markers is respectively as follows: chr, chr, The nucleotide sequences of chr2_112111693 and chr3_110246950 are shown as SEQ ID NO. 1-67 respectively, and the information of the sequences is shown as table 1.

2. The application of the SNP molecular marker combination of claim 1 in the molecular marker assisted breeding of Nandan Yao chicken.

3. The method for detecting the genotype of the Nandan Yao chicken by utilizing the molecular biology technology is characterized by comprising the following steps: the primers are designed according to the nucleotide sequences on two flanks of the 67 SNP molecular marker loci of claim 1, and the primers are used for genotyping the Nandan Yao chicken material to be detected, so that the genotype of the Nandan Yao chicken to be detected is identified.

4. The method of claim 3, applied to molecular marker assisted breeding of Nandan Yao chicken.