CN103609527A - Green-shin recessive white-feather chicken strain breeding method - Google Patents
Green-shin recessive white-feather chicken strain breeding method Download PDFInfo
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Abstract
The invention discloses a green-shin recessive white-feather chicken strain breeding method and belongs to the technical field of molecular breeding. A method of molecular assistant selection is adopted to identify genotype of a chicken dominant white-feather locus, the genotype of the locus can be quickly judged, a green-shin recessive white-feather chicken strain with the genotype of ii can be produced, troublesomeness in testcross breeding can be effectively avoided, generation interval can be shortened, breeding process can be accelerated, and breeding cost can be lowered. the green-shin recessive white-feather chicken strain bred by adopting the method is tender in meat quality, delicious in meat flavor and high in production performance, can be taken as commercial chickens to be directly put into production, and can be matched with green-shin jute feather for application.
Description
Technical field
The present invention is specifically related to a kind of selection of blue or green shank and recessive white feather chicken strain, belongs to molecular breeding technical field.
Background technology
Along with growth in the living standard, people's consumption idea changes mass type into from the scalar type in past.People are also more and more higher to the requirement of chicken meat.The local chicken of China has the advantages that meat is good, but its growth rate is slower, and feed conversion rate is low.Though and external fast large-scale broiler chicken fast growth, feed conversion rate are high, its meat is poor.For addressing this problem the large-scale broiler chicken of ，Jiang state extra income and domestic local high quality meat chicken is hybridized, the meat that can either improve the large-scale broiler chicken of state's extra income also can be accelerated growth rate and the productivity of local high quality meat chicken.But due to the impact of the long-term eating habit of China people, the white feather chicken of Fast Growth is not accepted in chicken, the especially Shelter in South China Cities of coloured plumages such as Chinese Consumer's preference jute plumage, black plumage.The demand that meets Chinese Consumer's in order to make to hybridize the phenotype of rear chicken, the Recessive Chicken that just need to isozygoty, the plumage look that can keep coloured plumage after the coloured plumage hybridization of Recessive Chicken productivity Gao，Qie Yu China, for being significant in China's high-quality a breed of chicken.
Summary of the invention
The selection that the object of this invention is to provide a kind of blue or green shank and recessive white feather chicken strain.
In order to realize above object, the technical solution adopted in the present invention is:
A selection for blue or green shank and recessive white feather chicken strain, comprises the following steps:
(1) utilize blue or green shank and recessive white feather chicken as male parent, white Leghorn is hybridized as female parent, produces F1 generation;
(2) F1 generation selfing obtains F2 generation, and the F2 that selects and remain is for the white plumage male and female of blue or green shin chicken;
(3) utilize AS-PCR to measure F2 for the genotype of the white plumage male and female of blue or green shin chicken, the dominant Bai Yu site of selecting and remain is the genotypic individuality of ii, eliminating dominant Bai Yu site is II and the genotypic individuality of Ii, and obtaining dominant Bai Yu site is the white plumage male and female of the genotypic blue or green shin of ii chicken;
(4) getting dominant Bai Yu site is the selfing of the white plumage male and female of the genotypic blue or green shin of ii chicken, utilize the method mensuration self progeny's of step (3) genotype, repeat above-mentioned steps to genetic stability, obtaining dominant Bai Yu site is the genotypic blue or green shank and recessive white feather chicken strain of ii.
In described step (1), blue or green shank and recessive white feather chicken is the blue or green shin white feather chicken after blue or green shin jute plumage local chicken sudden change, and through the artificial RFLP-PCR method of introducing PvuII, measuring dominant Bai Yu site is ii genotype.
In described step (3), utilize AS-PCR to measure F2 for the genotypic method of the white plumage male and female of blue or green shin chicken, comprise the following steps: the chicken complete genome DNA to be measured that comprises PMEL17 gene of take is template, take primer pair P1 as primer PCR amplification chicken PMEL17 gene I allelomorph, take primer pair P2 as primer PCR amplification chicken PMEL17 gene i allelomorph; The fragment that amplification is obtained is carried out agarose gel electrophoresis, identifies the genotype in the dominant Bai Yu of chicken site according to agarose gel electrophoresis result;
Described primer pair P1 is:
Upstream primer, P1-F:5 '-CCGGCCTCTACTGCCTCAACGTCTCGT-3 ' (as shown in SEQ ID No.1),
Downstream primer, P1-R:5 '-GTGCCCAGAGCTTCGGCGATGAGCGGTG-3 ' (as shown in SEQ ID No.2);
Described primer pair P2 is:
Upstream primer, P2-F:5 '-CCACGCTCACCGTGGGGCTGCTGCTCAT-3 ' (as shown in SEQ ID No.3),
Downstream primer, P2-R:5 '-GAGGGGGTGAGTGCTGTGTTCTC-3 ' (as shown in SEQ ID No.4).
Described pcr amplification reaction system is: 2 * Taq PCR MasterMix, 12.5 μ L, ddH
2o 9.5 μ L, P1-F 0.4 μ L, P1-R 0.6 μ L, P2-F 0.6 μ L, P2-R 0.4 μ L, DNA profiling 1.0 μ L.
Described 2 * Taq PCR MasterMix contains Taq archaeal dna polymerase, dNTPs, magnesium chloride and PCR reaction buffer, is commercial goods, can be purchased from companies such as the precious biology in Sheng Gong, Dalian, Shanghai, hundred Tykes, Beijing, Beijing Kang Wei.
Described pcr amplification reaction program is: 94 ℃ of denaturation 4min; 94 ℃ of sex change 30s, 64 ℃ of annealing 30s, 72 ℃ are extended 15s, 35 circulations; 72 ℃ are extended 10min.
The mass concentration of described Ago-Gel is 1.5%.
The voltage of described agarose gel electrophoresis is 120V, and electrophoresis time is 40min.
The described method of identifying the dominant white plumage loci gene type of chicken is: II genotype shows as 322bp and 224bp band; Ii genotype shows as 322bp, 224bp and 144bp band; Ii genotype shows as 322bp and 144bp band.
Preferably, the dominant Bai Yu site that step (4) is obtained is that the genotypic blue or green shank and recessive white feather chicken strain of ii and the inter breed crossing of blue or green shin yellow pockmarked feather chicken are supporting, produces coloured plumage broiler chicken.
Described blue or green shin yellow pockmarked feather chicken kind is gu-shi chicken, the Ji, Huaibei, limit fiber crops chicken, Huainan Spotted-brown Chickens, Wuhua chicken, Chongren Chicken, Lang Yaji, Lushi chicken, Land of Peach Blossoms chicken, precious jade chicken, camellia chicken etc.
Beneficial effect of the present invention:
The present invention adopts the method for molecule assisted Selection to identify the genotype in the dominant Bai Yu of chicken site, can determine fast the genotype in this site, producer gene type is the blue or green shank and recessive white feather chicken strain of ii, can effectively avoid the loaded down with trivial details of test cross seed selection, shorten the generation interval, accelerate breeding process, reduce and cultivate cost.Adopt blue or green its fine and tender taste of shank and recessive white feather chicken strain of the inventive method seed selection, meat flavour is delicious, and productivity is high, both can be used as commercial chicken and has directly put into production, and can also carry out supporting application with blue or green shin jute plumage.
In the present invention, for detection of the primer of the white plumage loci gene type of chicken PMEL17 gene dominant, be to insert (CTGGGCACC-) design allelomorph I Auele Specific Primer P1-R for PMEL17 gene exon10 9bp; For PMEL17 gene exon10, without 9bp, insert design allelomorph i Auele Specific Primer P2-F.Primer P1-R and the P1-F I allelomorph that can only increase like this, the clip size of amplification is 224bp; Primer P2-R and the P2-F i allelomorph that can only increase, the clip size of amplification is 144bp.In addition, because the combination of upstream primer P1-F and downstream primer P2-R also can be increased, the clip size of amplification is 322bp, the impact that this amplification is not inserted by 9bp.
Accompanying drawing explanation
Fig. 1 is the PCR electrophoretogram in each sample PMEL17 gene dominant Bai Yu site in the embodiment of the present invention 1;
Note: swimming lane is distinguished the electrophoresis banding pattern of representation DNA Marker I and genotype ii, ii, Ii, II, II, ii, Ii, ii and Ii from left to right.
Embodiment
Following embodiment is only described in further detail the present invention, but does not form any limitation of the invention.
Embodiment 1
The selection of blue or green shank and recessive white feather chicken strain in the present embodiment, comprises the following steps:
(1) utilize blue or green shank and recessive white feather chicken in gu-shi chicken colony to make male parent, white Leghorn is hybridized as female parent, produces F1 generation;
(2) F1 generation selfing obtains F2 generation, and the F2 that selects and remain is for the white plumage male and female of blue or green shin chicken;
(3) utilize the method for AS-PCR to measure F2 for the genotype of the white plumage male and female of blue or green shin chicken, specifically comprise the following steps:
1. the collection of sample
F2, for the blue or green shin white feather chicken wing venous blood collection of selecting and remain, is added to 1/3 anticoagulant, by Proteinase K method, extract blood DNA, DNA Sample storage is standby in 4 ℃ of Refrigerator stores;
2. design of primers
Take PMEL17 gene order (GenBank Accession No.AY636129) as template design primer is to P1 and P2, as follows;
Described primer pair P1 is:
Upstream primer, P1-F:5 '-CCGGCCTCTACTGCCTCAACGTCTCGT-3 ',
Downstream primer, P1-R:5 '-GTGCCCAGAGCTTCGGCGATGAGCGGTG-3 ';
Described primer pair P2 is:
Upstream primer, P2-F:5 '-CCACGCTCACCGTGGGGCTGCTGCTCAT-3 ',
Downstream primer, P2-R:5 '-GAGGGGGTGAGTGCTGTGTTCTC-3 ';
3. pcr amplification
Take primer pair P1 and P2 has set up 25.0 μ L amplification systems as primer: 2 * Taq PCR MasterMix(is purchased from Beijing hundred Imtech) 12.5 μ L, ddH
2o 9.5 μ L, P1-F 0.4 μ L, P1-R 0.6 μ L, P2-F 0.6 μ L, P2-R 0.4 μ L, DNA profiling 1.0 μ L;
Response procedures is: 94 ℃ of denaturation 4min; 94 ℃ of sex change 30s, 64 ℃ of annealing 30s, 72 ℃ are extended 15s, 35 circulations; 72 ℃ are extended 10min;
4. the detection of pcr amplification product and result are judged
Get 10.0 μ L pcr amplification afterproducts, 1.5% agarose gel electrophoresis (electrophoretic voltage: 120V; Electrophoresis time: 40min) point sample, after electrophoresis finishes, with gel imaging system, Ago-Gel to be taken a picture and detected, electrophoresis pattern is referring to Fig. 1; According to the stripe size occurring in electrophoresis pattern, judge: when there is 322bp and 224bp fragment, be dominant white plumage homozygote (II genotype); When there is the fragment of 322bp and 144bp, be wild type homozygote (ii genotype); When there is the fragment of 322bp, 224bp and 144bp, it is dominant white plumage heterozygote (Ii genotype);
(4) according to genotype result of determination, the dominant Bai Yu site of selecting and remain is the genotypic individuality of ii, and eliminating dominant Bai Yu site is II and the genotypic individuality of Ii, and obtaining dominant Bai Yu site is the genotypic male and female chicken of ii;
(5) getting dominant Bai Yu site is the selfing of the white plumage male and female of the genotypic blue or green shin of ii chicken, utilize the method mensuration self progeny's of step (3) genotype, repeat above-mentioned steps to genetic stability, obtaining dominant Bai Yu site is the genotypic blue or green shank and recessive white feather chicken strain of ii;
(6) dominant Bai Yu site step (5) being obtained is that the genotypic blue or green shank and recessive white feather chicken strain of ii is supporting as male parent and the inter breed crossing of blue or green shin yellow pockmarked feather chicken, produces coloured plumage broiler chicken.
Claims (10)
1. a selection for blue or green shank and recessive white feather chicken strain, is characterized in that: comprise the following steps:
(1) utilize blue or green shank and recessive white feather chicken as male parent, white Leghorn is hybridized as female parent, produces F1 generation;
(2) F1 generation selfing obtains F2 generation, and the F2 that selects and remain is for the white plumage male and female of blue or green shin chicken;
(3) utilize AS-PCR to measure F2 for the genotype of the white plumage male and female of blue or green shin chicken, the dominant Bai Yu site of selecting and remain is the genotypic individuality of ii, eliminating dominant Bai Yu site is II and the genotypic individuality of Ii, and obtaining dominant Bai Yu site is the white plumage male and female of the genotypic blue or green shin of ii chicken;
(4) getting dominant Bai Yu site is the selfing of the white plumage male and female of the genotypic blue or green shin of ii chicken, utilize the method mensuration self progeny's of step (3) genotype, repeat above-mentioned steps to genetic stability, obtaining dominant Bai Yu site is the genotypic blue or green shank and recessive white feather chicken strain of ii.
2. the selection of blue or green shank and recessive white feather chicken strain according to claim 1, is characterized in that: in described step (1), blue or green shank and recessive white feather chicken is the blue or green shin white feather chicken after blue or green shin jute plumage local chicken sudden change.
3. the selection of blue or green shank and recessive white feather chicken strain according to claim 1, it is characterized in that: in described step (3), utilize AS-PCR to measure F2 for the genotypic method of the white plumage male and female of blue or green shin chicken, comprise the following steps: the chicken complete genome DNA to be measured that comprises PMEL17 gene of take is template, take primer pair P1 as primer PCR amplification chicken PMEL17 gene I allelomorph, take primer pair P2 as primer PCR amplification chicken PMEL17 gene i allelomorph; The fragment that amplification is obtained is carried out agarose gel electrophoresis, identifies the genotype in the dominant Bai Yu of chicken site according to agarose gel electrophoresis result;
Described primer pair P1 is:
Upstream primer, P1-F:5 '-CCGGCCTCTACTGCCTCAACGTCTCGT-3 ',
Downstream primer, P1-R:5 '-GTGCCCAGAGCTTCGGCGATGAGCGGTG-3 ';
Described primer pair P2 is:
Upstream primer, P2-F:5 '-CCACGCTCACCGTGGGGCTGCTGCTCAT-3 ',
Downstream primer, P2-R:5 '-GAGGGGGTGAGTGCTGTGTTCTC-3 '.
4. the selection of blue or green shank and recessive white feather chicken strain according to claim 3, is characterized in that: described pcr amplification reaction system is: 2 * Taq PCR MasterMix, 12.5 μ L, ddH
2o 9.5 μ L, P1-F 0.4 μ L, P1-R 0.6 μ L, P2-F 0.6 μ L, P2-R 0.4 μ L, DNA profiling 1.0 μ L.
5. the selection of blue or green shank and recessive white feather chicken strain according to claim 3, is characterized in that: described pcr amplification reaction program is: 94 ℃ of denaturation 4min; 94 ℃ of sex change 30s, 64 ℃ of annealing 30s, 72 ℃ are extended 15s, 35 circulations; 72 ℃ are extended 10min.
6. the selection of blue or green shank and recessive white feather chicken strain according to claim 3, is characterized in that: the mass concentration of described Ago-Gel is 1.5%.
7. the selection of blue or green shank and recessive white feather chicken strain according to claim 3, is characterized in that: the voltage of described agarose gel electrophoresis is 120V, and electrophoresis time is 40min.
8. the selection of blue or green shank and recessive white feather chicken strain according to claim 3, is characterized in that: the described method of identifying the dominant white plumage loci gene type of chicken is: II genotype shows as 322bp and 224bp band; Ii genotype shows as 322bp, 224bp and 144bp band; Ii genotype shows as 322bp and 144bp band.
9. the selection of blue or green shank and recessive white feather chicken strain according to claim 1, it is characterized in that: the dominant Bai Yu site that step (4) is obtained is that the genotypic blue or green shank and recessive white feather chicken strain of ii and the inter breed crossing of blue or green shin yellow pockmarked feather chicken are supporting, produces coloured plumage broiler chicken.
10. the selection of blue or green shank and recessive white feather chicken strain according to claim 9, is characterized in that: described blue or green shin yellow pockmarked feather chicken kind is gu-shi chicken, the Ji, Huaibei, limit fiber crops chicken, Huainan Spotted-brown Chickens, Wuhua chicken, Chongren Chicken, Lang Yaji, Lushi chicken, Land of Peach Blossoms chicken, precious jade chicken or camellia chicken.
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| CN104719253A (en) * | 2015-04-15 | 2015-06-24 | 广西南宁市富凤农牧有限公司 | Breeding method of low-speed type super-quality green-leg parental breeding chickens |
| CN104823919A (en) * | 2015-05-18 | 2015-08-12 | 贵州省畜牧兽医研究所 | Cultivation method and application of recessive white feather new strain of Xiaoxiang chicken in southeast of Guizhou province |
| CN108220408A (en) * | 2018-01-12 | 2018-06-29 | 江苏省家禽科学研究所 | A kind of selection of grain-saving type blueness shank and recessive white meat-type chickens new lines |
| CN111357714A (en) * | 2020-04-24 | 2020-07-03 | 广东天农食品有限公司 | Method for breeding green-foot partridge chickens |
| CN112063725A (en) * | 2020-09-14 | 2020-12-11 | 华南农业大学 | Chicken shin length and shin diameter related AGO3 gene molecular marker and application |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104293905A (en) * | 2014-04-11 | 2015-01-21 | 河南农业大学 | Primer, kit and detection method for detecting chicken green shin character linkage SNP locus genotype |
| CN104293905B (en) * | 2014-04-11 | 2016-06-01 | 河南农业大学 | A kind of primer for detecting the blue or green chain SNP site genotype of shin proterties of chicken, test kit and detection method |
| CN104719253A (en) * | 2015-04-15 | 2015-06-24 | 广西南宁市富凤农牧有限公司 | Breeding method of low-speed type super-quality green-leg parental breeding chickens |
| CN104823919A (en) * | 2015-05-18 | 2015-08-12 | 贵州省畜牧兽医研究所 | Cultivation method and application of recessive white feather new strain of Xiaoxiang chicken in southeast of Guizhou province |
| CN104823919B (en) * | 2015-05-18 | 2017-03-15 | 贵州省畜牧兽医研究所 | The breeding method of southeast of Guizhou Province Fructus Foeniculi chicken recessive white feather new lines and application |
| CN108220408A (en) * | 2018-01-12 | 2018-06-29 | 江苏省家禽科学研究所 | A kind of selection of grain-saving type blueness shank and recessive white meat-type chickens new lines |
| CN108220408B (en) * | 2018-01-12 | 2021-07-06 | 江苏省家禽科学研究所 | Grain-saving green-shin recessive white feather broiler new strain breeding method |
| CN111357714A (en) * | 2020-04-24 | 2020-07-03 | 广东天农食品有限公司 | Method for breeding green-foot partridge chickens |
| CN112063725A (en) * | 2020-09-14 | 2020-12-11 | 华南农业大学 | Chicken shin length and shin diameter related AGO3 gene molecular marker and application |
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