CN107447009A - A kind of molecular labeling related to chicken feed utilization ratio character and its application - Google Patents
A kind of molecular labeling related to chicken feed utilization ratio character and its application Download PDFInfo
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- CN107447009A CN107447009A CN201710736580.4A CN201710736580A CN107447009A CN 107447009 A CN107447009 A CN 107447009A CN 201710736580 A CN201710736580 A CN 201710736580A CN 107447009 A CN107447009 A CN 107447009A
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Abstract
The invention discloses a kind of molecular labeling related to chicken feed utilization ratio character for belonging to technical field of gene detection and its application.The SNP marker corresponds in NCBI the chicken reference gene group Gallus_gallus chromosome positive-sense strands the 9th, 226,493 of 5.0 version sequence information 5 announced, and base is C or T herein.The SNP marker of the present invention is related to chicken feed utilization ratio character, it is a new molecular labeling, by determining that the genotype of chicken SNP site to be measured carries out Seedling selection to chicken feed utilization ratio character, production cost can be saved and accelerate genetic progress, a breed of chicken is preferably served, there is very big Economic Application value and scientific research value.
Description
Technical field
The invention belongs to technical field of gene detection, and in particular to a kind of molecule related to chicken feed utilization ratio character
Mark and its application.
Background technology
The production efficiency for improving poultry is always one of significant challenge of domestic fowl farming.Wherein feed conversion rate is also referred to as feed
Remuneration is to influence the important one side of production efficiency.For laying hen, feed conversion rate (FCR) and remaining feeding graded effect
(RFI) be evaluate laying hen production efficiency leading indicator.And RFI is to be more applicable for evaluating egg feedstuff utilization compared with FCR
Batch mark of efficiency, it is defined as actual feed intake and predicts the difference between feed intake.
Certain progress is had been achieved with using selection of the traditional breeding way to RFI, but for laying hen, feed efficiency
Room for promotion it is also very big.By molecular labeling means from genetically can further lifting egg feedstuff efficiency.It is related to RFI
Hereditary variation can be associated with genetic marker, so as to improve RFI by the selection to mark.Whole-genome association
It is one of main means of hereditary connection between phenotype and genotype of identifying.Heredity connection between identification phenotype and genotype
System, it is easy to carry out Seedling selection to food utilization efficiency character, saves production cost and accelerate genetic progress, preferably serve
A breed of chicken.Therefore, the research of the molecular labeling related to chicken feed utilization ratio character is significant.
The content of the invention
The present invention carries out early stage choosing to facilitate in order to accurately determine the genotype of chicken to be measured to food utilization efficiency character
Select, save production cost and simultaneously accelerate genetic progress, propose a kind of molecular labeling related to chicken feed utilization ratio character and its
Using.Concrete technical scheme is as follows：
A kind of SNP marker related to chicken feed utilization ratio character, SNP (SNP) molecule
Mark corresponds in NCBI the chicken reference gene group Gallus_gallus-5.0 version sequences No. 5 chromosome justice of information announced
The 9th, 226,493, chain, base is C or T herein (corresponding No. rs is rs315135692 on NCBI websites).
Application of the described SNP marker in chicken genetic breeding.
The method of one breeding hen feed utilization ratio character Seedling selection, it is that chicken is raised according to the genotype of above-mentioned SNP site
Expect that utilization ratio character carries out Seedling selection, comprise the following steps：
(1) genomic DNA of chicken to be measured is extracted；
(2) genotype of 9,226,493rd SNP site of No. 5 chromosome positive-sense strands of chicken to be measured is detected；
(3) Seedling selection is carried out to chicken feed utilization ratio character based on the genotype of SNP site, wherein, TT genotype
Chicken feed utilization ratio is better than CT types and CC genotype, and CT genotype chicken feeds utilization ratio is better than CC genotype.
The method of the genomic DNA of extraction chicken to be measured described in step (1) is：Chicken wings venous blood collection is carried out to chicken to be measured,
Through cracking, protease digestion processing after carrying out anti-freezing processing with anti-coagulants, method extraction genomic DNA is then imitated using phenol, to go out
Bacterium distilled water dissolves.
Genotype uses Axiom in step (2)TMGenome-Wide Chicken Genotyping Array are detected.
The genotype of 9,226,493rd SNP marker of No. 5 chromosome positive-sense strands of chicken to be measured described in step (2) is
CC, CT or TT.
Chicken feed utilization ratio described in step (3) is the remaining feeding graded effect of 37-40 week old.
TT genotype described in step (3) refers to the homozygote that No. 5 bit bases of chromosome positive-sense strand the 9,226,493rd are T；
CC genotype refers to the homozygote that the site base is C；CT genotype refers to the heterozygote that the site base is C and T.
Beneficial effects of the present invention are：The SNP marker of the present invention is related to chicken feed utilization ratio character, is one
New molecular labeling, by determining that the genotype of chicken SNP site to be measured carries out Seedling selection, energy to chicken feed utilization ratio character
Enough save production cost and accelerate genetic progress, preferably serve a breed of chicken, there is very big Economic Application value and section
Grind value.
Brief description of the drawings
Fig. 1 is different genotype and the result figure of food utilization efficiency association analysis in embodiment 2.
Embodiment
Following examples facilitate a better understanding of the present invention, but not limited to this.In addition to specified otherwise, in each embodiment
The equal conventional commercial of equipment and reagent used can obtain.
Colony to be measured comes from Jiangsu Inst. of Fowls Science, and Bai Laihang pure lines and blue-shelled egg layer pure lines are carried out
Reciprocal cross, produce F2 and be used for follow-up phenotype record and whole-genome association for colony.
Embodiment 1：The confirmation of rs14512566 sites SNP marker and food utilization efficiency correlation
Experiment material：1534 Jiangsu Inst. of Fowls Science Bai Laihang and blue-shelled egg layer hybridization F2 generation individuals.
First, food utilization efficiency determines
The remaining feed intake effect value (RFI) of each individual 37 week old to 40 week old of measure, feeds conventional egg during this period
Chicken in egg-laying period feed (table 1).Specifically assay method is：Record feed intake individual in the stage (feed intake, FI) and egg
Weight (egg mass, EM).Simultaneously at the beginning and end of this stage, the body weight of each individual is determined, it is average for calculating it
Body weight (mean BW, MBW), metabolic body size (MBW0.75) and daily gain (daily BW gain, BWG).Finally, according to as follows
Formula calculates each individual RFI phenotypes：
RFI=FI- (b0+b1MBW0.75+b2BWG+b3EMD)
Wherein RFI is the remaining feed intake of each individual, and FI is feed intake, MBW0.75For metabolic body size, BWG is to increase day by day
Weight, EMD are egg size, b0For intercept, b1, b2, b3For regression coefficient.RFI calculating is completed by R linear model fitting function.
Table 1 tests chicken feed composition and trophic level (air bells dry basis)
Premix provides for every kilogram of feed：The 715IU of vitamin A 7, vitamin D32 755IU, vitamin E
8.8IU, vitamin K32.2mg, vitamin B10.65mg, vitamin B22.21mg, pantothenic acid 3.51mg, nicotinic acid 19.8mg, pyrrole
Tremble alcohol 3.25mg, biotin 0.20mg, folic acid 0.28mg, vitamin B120.02mg, manganese 65mg, iron 60mg, copper 8mg, zinc
80mg, iodine 1.0mg, selenium 0.3mg.
2nd, the detection of SNP marker
1st, extracting genome DNA
1) 20 μ l ACD anti-freezing chicken blood samples are taken, add 500 μ l cell pyrolysis liquids effect 30min, the μ l of 10% Proteinase K 10,
55 DEG C of water-baths are stayed overnight.
ACD anti-coagulants：1.32% (m/v) sodium citrate, 0.48% (m/v) citric acid, 1.47% (m/v) glucose.
Proteinase K：Purchased from Sangon Biotech (Shanghai) Co., Ltd..
2) postdigestive blood is added to the Tris saturated phenols (PH8.0) of equivalent volumes, slowly reverse centrifuge tube 10min,
12000rpm is centrifuged 10 minutes, with the careful Aspirate supernatant of bigbore pipette tips into clean centrifuge tube.
3) it is repeated once previous step work.If supernatant is limpid transparent, this step can be omitted.
4) phenol of equivalent is added:Chloroform (1:1), slowly reverse centrifuge tube 10 minutes, 12000rpm are centrifuged 10 minutes, with big
The careful Aspirate supernatant of pipette tips of bore is into clean centrifuge tube.
5) isometric chloroform is added, slowly reverse centrifuge tube 10 minutes, 12000rpm is centrifuged 10 minutes, and use is bigbore
The careful Aspirate supernatant of pipette tips is into clean centrifuge tube.
6) absolute ethyl alcohol of two volumes is added, jog centrifuge tube, has treated that white hazy precip occurs stopping shaking.
7) carefully DNA precipitations are chosen with pipette tips, be placed in the centrifuge tube for filling 75% ethanol.
8) ethanol is exhausted, allows ethanol volatilization clean at room temperature, add appropriate TE buffer solutions DNA.
9) DNA dissolved is placed in -20 DEG C of preservations.
2nd, Genotyping
The genomic DNA of each chicken to be measured is taken, using the 600K Affymetrix Axiom Chicken of Affymetrix company of the U.S.
Genotyping Array (cat. no：902148), according to the signal intensity in each site, to each individual genotype
Carry out parting.
3rd, chicken feed utilization ratio character whole-genome association
Using genome-wide efficient mixed-model association (GEMMA) statistical analysis software
Mixed linear model carry out statistical analysis, specifically refer to:“Genome-wide efficient mixed-model
Analysis for association studies ", Xiang Zhou et al., Nature Genetics, VOLUME
44, NUMBER 7, JULY 2012, and " Efficient multivariate linear mixed model
algorithms for genome-wide association studies”,“Krina T Zondervan and Lon R
Cardon,PROTOCOL,VOLUME 2,NUMBER 10,OCTOBER,2012”。
Statistic analysis models are：
Y=Wa+x β+μ+ε；
Y represents individual phenotypic number；
W represents covariant；
A represents coefficient of correspondence；
X represents SNP genotype；
SNP effects corresponding to β representatives；
μ represents remaining multigentic effect；
ε represents post fit residuals effect.
Data analysis is analyzed using GeneTitan (U.S.'s Affymetrix Products) genetic chip automated processing system,
The software analyzed data is Affymetrix Power Tools.
As a result show, No. 5 position SNP markers of chromosome 9,226,493 are notable with food utilization efficiency character (RFI)
It is related.
Embodiment 2：Association analysis of the F2 for individual rs14512566C/T genotype and food utilization efficiency
Experiment material：1534 Jiangsu Inst. of Fowls Science Bai Laihang laying hens and blue-shelled egg layer hybridization F2 generations
Body.
First, food utilization efficiency determines
The remaining feed intake effect value (RFI) of each individual 37 week old to 40 week old of measure.
2nd, SNP detection
1st, extracting genome DNA
Operating procedure is the same as embodiment 1.
2nd, genotype determines
Genomic DNA is taken, using the 600K Affymetrix Axiom Chicken of Affymetrix company of the U.S.
The genotype of each individual of Genotyping Array (cat. no 902148) detections, the results are shown in Table 2.
Distribution of the site rs315135692 C/T genotype of table 2 in F2 colonies
As can be seen from Table 2, TT genotype is experimental population preponderant genotype.
3rd, genotype and food utilization efficiency association analysis
Statistical analysis is carried out using the GLM processes of SAS statistics analysis software packages, according to generalized linear model to for trying chicken group
Genotype and food utilization efficiency character carry out variance statistic analysis, P-value ＜ 0.05 show significant difference.
Statistic analysis models：
Y=μ+G+e
Y represents individual phenotypic number；
μ represents colony's average；
G represents genotype effects；
E represents post fit residuals effect.
As a result Fig. 1 and table 3 are seen.
37~40 week old residue feed intake phenotypic numbers of the different genotype of table 3 individual
| Food utilization efficiency | CC | CT | TT | P-value |
| 37-40 week old RFI | 3.47±6.25 | 1.23±6.19 | -0.70±6.61 | <0.01 |
From table 3 it can be seen that there were significant differences by the hen 37-40 week old RFI of three kinds of genotype.The feeding of TT genotype hens
Material utilization ratio is better than CT types and CC genotype individuals, and the food utilization efficiency of CT genotype hens is better than CC genotype individuals.
Claims (8)
- A kind of 1. SNP marker related to chicken feed utilization ratio character, it is characterised in that the SNP marker pair The chicken reference gene group Gallus_gallus-5.0 version sequences No. 5 chromosome positive-sense strands the 9th of information that should be announced in NCBI, 226,493, base is C or T herein.
- 2. application of the SNP marker in chicken genetic breeding described in claim 1.
- 3. the method for a breeding hen feed utilization ratio character Seedling selection, it is characterised in that SNP positions according to claim 1 The genotype of point carries out Seedling selection to chicken feed utilization ratio character.
- 4. according to the method for claim 3, it is characterised in that comprise the following steps：(1) genomic DNA of chicken to be measured is extracted；(2) genotype of 9,226,493rd SNP marker of No. 5 chromosome positive-sense strands of chicken to be measured is detected；(3) Seedling selection is carried out to chicken feed utilization ratio character based on the genotype of SNP site, wherein, TT genotype chicken is raised Material utilization ratio is better than CT types and CC genotype, and CT genotype chicken feeds utilization ratio is better than CC genotype.
- 5. according to the method for claim 4, it is characterised in that the genomic DNA of extraction chicken to be measured described in step (1) Method is：Chicken wings venous blood collection is carried out to chicken to be measured, handled after carrying out anti-freezing processing with anti-coagulants through cracking, protease digestion, Then method extraction genomic DNA is imitated using phenol, with the distilled water dissolving that sterilizes.
- 6. according to the method for claim 4, it is characterised in that genotype uses Axiom in step (2)TMGenome-Wide Chicken Genotyping Array are detected.
- 7. according to the method for claim 4, it is characterised in that No. 5 chromosome positive-sense strands of chicken to be measured described in step (2) the The genotype of 9,226,493 SNP sites is CC, CT or TT.
- 8. according to the method for claim 4, it is characterised in that chicken feed utilization ratio described in step (3) is 37-40 The remaining feeding graded effect of week old.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110468221A (en) * | 2019-09-19 | 2019-11-19 | 江苏省家禽科学研究所 | One kind SNP marker relevant to chicken glandular stomach number of nipples and its application |
| CN112002371A (en) * | 2020-07-31 | 2020-11-27 | 中国农业科学院北京畜牧兽医研究所 | Genome selection method for residual feed intake of white feather broilers |
| CN114742290A (en) * | 2022-03-30 | 2022-07-12 | 东北农业大学 | Method for predicting conversion efficiency of white feather broiler feed through plasma metabolite abundance modeling |
| CN115261485A (en) * | 2022-06-15 | 2022-11-01 | 安徽农业大学 | Local chicken residual feed intake related molecular marker and application thereof |
| CN117187403A (en) * | 2023-08-11 | 2023-12-08 | 江苏省家禽科学研究所 | Application of SNP genetic marker related to residual feed consumption in chicken egg producing period in chicken genetic breeding |
| CN115261485B (en) * | 2022-06-15 | 2024-05-31 | 安徽农业大学 | Local chicken residual feed intake related molecular marker and application thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110468221A (en) * | 2019-09-19 | 2019-11-19 | 江苏省家禽科学研究所 | One kind SNP marker relevant to chicken glandular stomach number of nipples and its application |
| CN112002371A (en) * | 2020-07-31 | 2020-11-27 | 中国农业科学院北京畜牧兽医研究所 | Genome selection method for residual feed intake of white feather broilers |
| CN112002371B (en) * | 2020-07-31 | 2023-09-26 | 中国农业科学院北京畜牧兽医研究所 | Genome selection method for residual feed intake of white-feather broilers |
| CN114742290A (en) * | 2022-03-30 | 2022-07-12 | 东北农业大学 | Method for predicting conversion efficiency of white feather broiler feed through plasma metabolite abundance modeling |
| CN115261485A (en) * | 2022-06-15 | 2022-11-01 | 安徽农业大学 | Local chicken residual feed intake related molecular marker and application thereof |
| CN115261485B (en) * | 2022-06-15 | 2024-05-31 | 安徽农业大学 | Local chicken residual feed intake related molecular marker and application thereof |
| CN117187403A (en) * | 2023-08-11 | 2023-12-08 | 江苏省家禽科学研究所 | Application of SNP genetic marker related to residual feed consumption in chicken egg producing period in chicken genetic breeding |
| CN117187403B (en) * | 2023-08-11 | 2024-05-17 | 江苏省家禽科学研究所 | Application of SNP genetic marker related to residual feed consumption in chicken egg producing period in chicken genetic breeding |
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