A method for complementary breeding of gray chickens using Anyi gray chickens

Abstract

The present invention discloses a method for complementary breeding of Wahu grey chickens using Anyi Wahu grey chickens, belonging to the technical field of poultry genetic breeding and reproduction. The complementary breeding method includes: selecting black-feathered slow-feathering Anyi Wahu grey chickens as the first paternal line and other mature strains as the maternal line, crossing the first paternal line with the maternal line to obtain the parental generation female parent; selecting grey-white feathered fast-feathering Anyi Wahu grey chickens as the terminal paternal line and crossing them with the parental generation female parent to obtain the commercial generation of Wahu grey chickens. The complementary breeding method provided by the present invention solves problems such as the separation of feather colors in the supply of pure lines and the loss of Wahu grey chicken germplasm resources caused by the supply of pure lines; moreover, according to actual needs, the present invention can replace the mature strains in the complementary breeding mode, thereby producing Wahu grey chickens with different growth rates to adapt to different consumer markets, and realizing a virtuous cycle of the protection and development and utilization of Anyi Wahu grey chickens.

Description

Technical Field

[0001] The present invention relates to the technical field of poultry genetic breeding and reproduction, and particularly relates to a supporting breeding method for producing Wahu grey chickens by using Anyi Wahu grey chickens. Background Art

[0002] Anyi Wahu grey chicken is a local breed characterized by a grey feather phenotype. After several generations of phenotypic selection, the feather color still has serious segregation. The offspring will show grey feathers (dark grey and greyish white), black feathers and a small number of yellowish-brown feather individuals, with about 70% of the grey-feathered offspring. The original breeding farm has tried to cross grey-feathered chickens with other breeds of different feather colors, but there is still serious segregation in the offspring, with about 45% grey feathers, about 45% black feathers and about 5% yellowish-brown feathers. Since consumers prefer grey-feathered (dark grey) individuals, the prices of black-feathered and greyish-white feather individuals are much lower. In order to ensure the relative consistency of the feather color of the commercial generation, the original breeding farm can only provide mixed seedlings by pure breeding of the original species at present, but about 30% of them still do not meet the requirements. The method of providing pure lines is extremely unfavorable for the protection of the Anyi Wahu grey chicken breed. And the method of providing pure lines also has the following defects:

[0003] (1) The method of providing pure lines leads to too high a cost of chick seedlings. Since the original species has not been systematically selected and bred, the growth performance and reproductive performance (especially the egg-laying performance) of the original Anyi Wahu grey chickens are low, resulting in a high breeding cost. Moreover, the slaughter weight of the commercial generation of broilers does not match the mainstream consumer market, which is not conducive to the development and growth of the Anyi Wahu grey chicken industry.

[0004] (2) The misidentification rate is high and it causes great harm to the chick seedlings. Due to the five-grey characteristics of the Wahu grey chickens, the current misidentification rate of sex identification is as high as 15%-20%, and the harm to the chick seedlings during the identification process is obvious. The mortality rate of the identified chicken flock within 7 days is 5 percentage points higher than that of the non-identified chicken flock.

[0005] (3) There is currently no effective method to reduce the proportion of non-grey feather trait individuals. When the original breeding farm of Anyi Wahu grey chickens conducts breeding and selection, grey-feathered individuals are retained in each generation and other feather color individuals are eliminated. After 8 generations of breeding and selection, this large-scale segregation phenomenon still exists and there is no effective solution. Summary of the Invention

[0006] The purpose of the present invention is to provide a supporting breeding method for producing Wahu grey chickens by using Anyi Wahu grey chickens, so as to solve the problems existing in the above-mentioned prior art. The breeding method produces Wahu grey chickens that not only maintain the grey feather trait of the commercial generation of broilers but also ensure good meat quality, and solves the problems such as feather color segregation in the existing pure line supply and the loss of the Wahu grey chicken germplasm resources.

[0007] To achieve the above purpose, the present invention provides the following scheme:

[0008] The present invention provides a method for producing a complete set of seeds of tile-gray chickens by using Anyi tile-gray chickens, comprising the following steps:

[0009] Select the black-feathered slow-feathered Anyiwa gray chicken as the first paternal line and other mature lines as maternal lines, cross the first paternal line with the maternal line to obtain the parental female;

[0010] The Anyi Wahui chicken with gray and white feathers and fast feathering was selected as the terminal paternal line and hybridized with the parental female to obtain the commercial generation of Wahui chicken.

[0011] Furthermore, the black-feathered slow-feathered Anyiwa gray chicken is a homozygous slow-feathered Anyiwa gray chicken; the gray-white-feathered fast-feathered Anyiwa gray chicken is a homozygous fast-feathered Anyiwa gray chicken; and the other mature strains include strains that do not carry the dominant white feather mutant allele.

[0012] Furthermore, the other mature strains include recessive white-feathered Rock chicken or jute chicken.

[0013] Furthermore, molecular markers are used to assist in the selection of homozygous slow-feathering Anyi Wahui chickens or homozygous fast-feathering Anyi Wahui chickens.

[0014] Furthermore, the steps of using molecular marker-assisted selection are as follows:

[0015] Detecting the SNP site of Anyi Wahui chicken at the 742 bp of the sequencing sequence shown in SEQ ID NO: 1, and analyzing the sequencing data of the SNP site;

[0016] If the analysis result is R, it is determined to be a homozygous slow-feathering Anyi Wahui chicken; if it is G, it is determined to be a heterozygous slow-feathering Anyi Wahui chicken; eliminating R and G will result in a homozygous fast-feathering Anyi Wahui chicken.

[0017] Furthermore, the sequencing data were analyzed using SequencingAnalysis Software v7.0.

[0018] The present invention discloses the following technical effects:

[0019] The supporting seed production method provided by the present invention has the following advantages:

[0020] (1) It can solve the problem of color separation in the existing pure-line breeding; (2) It can solve the risk problem of the loss of the source of Wahui chickens caused by pure-line breeding; (3) This matching mode can replace line X in the matching mode according to actual needs, so as to produce Wahui chickens with different growth rates to adapt to different consumer markets, and realize a virtuous cycle of the protection, development and utilization of Anyi Wahui chickens; (4) This matching mode introduces mature lines that have been highly systematically selected and bred, with high production and reproductive performance, and can reduce the cost of chick seedlings; (5) Only two pure lines need to be retained, reducing the breeding cost of the conservation farm. (6) The Wahui chickens produced by this matching mode have 75% of the blood relationship of the original Anyi Wahui chickens, ensuring the flavor of the commercial broilers; (7) The Wahui chickens produced by the matching breeding method of the present invention can identify the sexes of commercial broilers according to the characteristics of slow and fast feathers, improving the breeding efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the following will briefly introduce the drawings required in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0022] Figure 1 It is a schematic diagram of the genotype at the target locus; (a) is a slow-feathering homozygote, and (b) is a slow-feathering heterozygote. DETAILED DESCRIPTION OF THE INVENTION

[0023] The various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and implementation schemes of the present invention.

[0024] It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention. In addition, for the numerical ranges in the present invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any intermediate value within any stated value or stated range, as well as each smaller range between any other stated value or intermediate value within the stated range, is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0025] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0026] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0027] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0028] The present invention is based on the conclusions drawn by the applicant's statistical analysis of the seedling emergence data of the previous original breeding farm and the subsequent hybridization experiments. The original breeding farm has always adopted the method of self-pollination of gray feather (dark gray) male and female chickens. The seedlings produced by this method are divided into black feathers, dark gray feathers (standard feather color of tile-gray chickens) and gray-white feathers, and the ratio is close to 1:2:1. The above results show that the dark gray feather trait may be controlled by heterozygotes. On this basis, the applicant used gray-white feathers and black positive and negative cross experiments, and the results showed that all the male and female offspring were dark gray feathers. This provides a way to solve the problem of feather color separation of tile-gray chickens in the present invention. In order to improve the production performance of gray-feathered Anyi tile-gray chickens, the present invention introduces the X strain with excellent production performance (whose feather color is a non-gray feather strain and does not carry a dominant white feather mutation allele) in the matching system scheme, and replaces the original pure line mating method with the matching system to produce tile-gray chicken chicks. The matching system method not only maintains the gray feather traits of commercial broilers, but also ensures good meat quality and growth performance, and realizes a virtuous cycle of protection and development and utilization of Anyi tile-gray chickens. The invention aims to provide a matching scheme, which is a three-line matching scheme, which can ensure that all commercial seedlings are gray feather (dark gray) offspring, and the commercial generation contains 75% Anyi Wahui chicken blood source, which can ensure the meat quality of the commercial generation. The method is as follows:

[0029] Step 1: Establishment of the black-feathered slow-feathering line (A line) of Anyi Wahui chicken

[0030] (1) From the original population, select black-feathered Anyiwa gray chickens and hens as the basic population;

[0031] (2) Using molecular marker-assisted selection, fast-feathering roosters and fast-feathering hens were eliminated to establish a pure black-feathered slow-feathering line;

[0032] Step 2: Establishment of the Grayish-white and Fast-feathering Line (Line B) of Anyi Variegated Gray Chickens

[0033] (1) In the original breed population, select grayish-white Anyi variegated gray chicken cocks and hens as the foundation flock;

[0034] (2) Using the method of marker-assisted selection, eliminate slow-feathering cocks and hens to establish a grayish-white and fast-feathering pure line;

[0035] Step 3: Crossbreeding and Matching

[0036] The matching pattern is: Grayish-white Anyi variegated gray chicken ♂×(Black Anyi variegated gray chicken ♂×X line ♀)♀. For the X line in this matching pattern, its feather color is a non-gray feather line and does not carry the dominant white feather mutant allele, and there are no requirements for shank color and skin color traits. Under this matching pattern, all commercial broilers are gray-feathered, and the sex of chicks can be self-distinguished according to feather speed, with fast-feathering being hens and slow-feathering being cocks.

[0037] Specifically as follows:

[0038] Example 1: Using Anyi variegated gray chickens and recessive white Rock chickens to produce "Three Grays" Anyi variegated gray chickens

[0039] The purpose of this example is to verify the production of chick seedlings with the characteristics of "Three Grays" using recessive white Rock chickens.

[0040] Step 1: Select black-feathered and slow-feathering homozygous Anyi variegated gray chicken cocks.

[0041] The specific steps are as follows:

[0042] (1) From the 180-day-old Anyi variegated gray chicken population, select cocks with black feathers, blue shanks, and white skin.

[0043] (2) Genotype the above black-feathered individuals by sequencing, and select individuals with the target locus (the sequencing sequence is shown in SEQ ID NO: 1) as the R genotype, and select 3 individuals (named A). The specific method: Use Sequencing Analysis Software v7.0 to analyze the sequencing data, set "Mixed Bases Settings." to 90%, and if the analysis result is "R", keep it (if the analysis result is "R", it is determined as a slow-feathering homozygote, and if it is "G", it is determined as a slow-feathering heterozygote).

[0044]

[0045] The partial sequences marked with background shading in the above sequences are the upstream and downstream primers for sequencing. R is the SNP locus, representing G / A.

[0046] The PCR reaction system is as follows: 10 μL of 2×Eco Taq PCR SuperMix, 0.2 μL each of the forward and reverse primers, 1 μL of template DNA, and supplemented with ddH2O to 20 μL.

[0047] The PCR reaction program is as follows: pre-denaturation at 95°C for 5 min; denaturation at 95°C for 10 sec, annealing at 50 - 60°C for 30 sec, extension at 72°C for 90 sec, for a total of 35 cycles; finally, extension at 72°C for 7 min and storage at 16°C.

[0048] Figure 1 It is a schematic diagram of the genotype at the target site; (a) is the slow-feathering homozygote, and (b) is the slow-feathering heterozygote.

[0049] Step 2: Cross the 3 roosters selected in Step 1 with 30 hens of Recessive White Rock chickens (named X1), and select 30 hens from the hybrid offspring for the next cross.

[0050] Step 3: Select "gray and white feather" fast-feathering homozygous Anyi Wuhui roosters.

[0051] The specific steps are as follows:

[0052] (1) From the Anyi Wuhui chicken population at 180 days old, select roosters with gray and white feathers, blue feet, and white skin.

[0053] (2) Refer to Step 1, genotype the above individuals by sequencing, eliminate individuals with the R genotype and G genotype at the target site, and select 3 roosters (named B1) from the remaining population. The specific method: genotype the target site by sequencing. Use Sequencing Analysis Software v7.0 to analyze the sequencing data, set "Mixed Bases Settings." to 90%, and eliminate individuals with the parsing results of "R" and "G".

[0054] Step 4: Cross the 3 roosters selected in Step 3 with the 30 hens selected in Step 2, observe the feather color of the offspring, and separate them by male and female.

[0055] (1) Collect breeding eggs, and incubate 100 breeding eggs after 7 days.

[0056] (2) Count the feather color of the hatched chicks. A total of 85 chicks hatched, all with gray feathers (as shown in Table 1), and they meet the three gray characteristics - gray feathers, gray beaks, and gray feet.

[0057] (3) Sex identification: Select 10 slow-feathering and 10 fast-feathering chicks according to the phenotype, dissect and observe the sex. The results show that all fast-feathering chicks are hens, and all slow-feathering chicks are roosters.

[0058] Table 1 Statistical table of feather color phenotypes

[0059] Trait Grey feather Grey-white feather Black feather Quantity (85) 85 0 0 Proportion 100％ 0 0

[0060] Example 2 Production of "Five-Grey" Wuhui Chickens Using the Original Breed of Anyi Wuhui Chickens and the High-Yield Yellow-Feathered Strain

[0061] This example makes full use of the advantages of the mature strain in terms of high egg production and growth rate to breed chicks with the appearance characteristics of Five-Grey. The steps are as follows:

[0062] Step 1: Select black-feathered slow-feathering homozygous Anyi Wuhui roosters.

[0063] The specific steps are as follows:

[0064] (1) From the Anyi Wuhui chicken population at 180 days old, select roosters with black feathers, blue shanks, and white skin.

[0065] (2) Referring to the method of molecular marker-assisted selection in Example 1, genotype the above black-feathered individuals by sequencing, select and retain individuals with the target locus of R genotype, and select 3 (named A).

[0066] Step 2: Cross the roosters selected in Step 1 with the Huangma chicken strain N2 (named X2) introduced from the original Anyi Wuhui chicken farm, and retain the female chicks for later use.

[0067] Cross the 3 roosters selected in Step 1 with 30 Huangma chicken hens, and select 30 hens from the hybrid offspring for the next cross.

[0068] Step 3: Select grey-white feathered, grey-skinned, grey-crowned, grey-beaked, and grey-shanked fast-feathering homozygous Anyi Wuhui roosters.

[0069] The specific steps are as follows:

[0070] (1) From the "Five-Grey" strain of the Anyi Wuhui chicken population at 180 days old, select roosters with grey-white feathers, grey skin, grey crowns, grey beaks, and grey shanks.

[0071] (2) Refer to Step 1, genotype the above individuals by sequencing, eliminate individuals with the target locus of R genotype and G genotype, and select 3 roosters (named B2) from the remaining population. Specific method: Genotype the target locus by sequencing. Use Sequencing Analysis Software v7.0 to analyze the sequencing data, set "Mixed Bases Settings." to 90%, and eliminate individuals with the analysis results of "R" and "G".

[0072] Step 4: Cross the roosters selected in Step 3 with the hens selected in Step 2, observe the feather color of the offspring, and separate them by gender.

[0073] (1) Collect breeding eggs, and incubate 100 breeding eggs after 7 days.

[0074] (2) Count the feather colors of the hatched chicks. A total of 84 chicks hatched, all with grey feathers (as shown in Table 2), and all had the five grey characteristics, namely grey feathers, grey skin, grey crowns, grey beaks, and grey feet.

[0075] (3) Sex identification: Select 10 slow-feathering and 10 fast-feathering chicks according to their phenotypes, and dissect them to observe their sexes. The results show that all fast-feathering chicks are hens, and all slow-feathering chicks are roosters.

[0076] Table 2 Statistical table of feather color phenotypes

[0077] Trait Grey feather Grey-white feather Black feather Quantity (84) 84 0 0 Proportion 100％ 0 0

[0078] Control Example 1 Direct mating of purebreds 1

[0079] This scheme is one of the schemes currently used in the purebred farm.

[0080] Step 1: Select 10 roosters with grey feathers, grey beaks, and grey crowns, and 100 hens.

[0081] Step 2: Artificial insemination, collect eggs, and count the traits of the offspring

[0082] (1) Collect breeding eggs, and incubate 300 breeding eggs after 7 days.

[0083] (2) Count the traits of the hatched chicks. A total of 265 chicks hatched, all with black skin, among which 130 had grey feathers, 68 had grey and white feathers, and 67 had black feathers.

[0084] Table 3 Statistical table of feather color phenotypes

[0085] Trait Grey feather Grey-white feather Black feather Quantity (265) 130 68 67 Proportion 49.0％ 25.7％ 25.3％

[0086] Control Example 2 Direct mating of purebreds 2

[0087] This scheme is one of the schemes currently used in the purebred farm.

[0088] Step 1: Select 10 roosters with grey feathers, grey crowns, grey beaks, grey skin, and grey feet, and 100 hens.

[0089] Step 2: Artificial insemination, collect eggs, and count the traits of the offspring

[0090] (1) Collect breeding eggs, and incubate 300 breeding eggs after 7 days.

[0091] (2) Count the traits of the hatched chicks. A total of 255 chicks hatched, and the results show that 125 had grey feathers, 63 had grey and white feathers, and 67 had black feathers.

[0092] Table 4 Statistical table of feather color phenotypes

[0093] Trait Grey feather Grey-white feather Black feather Quantity (255) 125 63 67 Proportion 49.0％ 24.7％ 26.3％

[0094] The embodiments described above are only descriptions of the preferred embodiments of the present invention, and do not limit the scope of the present invention. Without departing from the spirit of the present invention, various deformations and improvements made by those of ordinary skill in the art to the technical solutions of the present invention shall fall within the protection scope determined by the claims of the present invention.

Claims

1. A method for supporting seed production of Wahuij chickens using Anyi Wahuij chickens, characterized in that, The following steps are involved: Select the black-feathered slow-feathered Anyiwa gray chicken as the first paternal line and other mature lines as maternal lines, cross the first paternal line with the maternal line to obtain the parental female; Select the Anyi Wahui chicken with grey and white feathers as the terminal paternal line, cross it with the parental female line to obtain the commercial Wahui chicken line; The black-feathered slow-feathered Anyiwa gray chicken is a homozygous slow-feathered Anyiwa gray chicken; the gray-white-feathered fast-feathered Anyiwa gray chicken is a homozygous fast-feathered Anyiwa gray chicken; the other mature strains include strains that do not carry the dominant white feather mutant allele; The other mature strains are recessive white feather Rock chicken or jute chicken.

2. The supporting seed production method according to claim 1, characterized in that Molecular marker-assisted selection was used to select homozygous slow-feathering Anyi Wahui chicken or homozygous fast-feathering Anyi Wahui chicken.

3. The supporting seed production method according to claim 2, characterized in that, The steps of utilizing molecular marker assisted selection are as follows: Detecting the SNP site of Anyi Wahui chicken at the 742 bp of the sequencing sequence shown in SEQ ID NO: 1, and analyzing the sequencing data of the SNP site; If the analysis result is R, it is determined to be a homozygous slow-feathering Anyi Wahui chicken; if it is G, it is determined to be a heterozygous slow-feathering Anyi Wahui chicken; eliminating R and G will result in a homozygous fast-feathering Anyi Wahui chicken.

4. The supporting seed production method according to claim 3, characterized in that The sequencing data were analyzed using Sequencing Analysis Software v7.0.

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