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Molecular markers and applications related to excellent lactation traits in Mediterranean buffalo

A technology for lactation traits and water buffalo, which is applied in the field of water buffalo molecular marker screening and auxiliary selection, can solve the problems of low economic benefits of buffalo breeding, and achieve the effect of not being disturbed by environmental conditions

Active Publication Date: 2022-02-22
HUAZHONG AGRI UNIV
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] There are rich breeds of buffalo at home and abroad, but there is no specialized lactating breed / strain at present. Even the breeds such as Moura, Nile-Lafite, and Mediterranean buffalo with high average yield per unit area have an average milk yield of only Holstein cows. 1 / 5-1 / 8 of that, resulting in low economic benefits of buffalo farming (Jiang et al 2010)

Method used

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  • Molecular markers and applications related to excellent lactation traits in Mediterranean buffalo
  • Molecular markers and applications related to excellent lactation traits in Mediterranean buffalo
  • Molecular markers and applications related to excellent lactation traits in Mediterranean buffalo

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1: Identification of ApoB_SNP1 to ApoB_SNP6 markers associated with lactation traits of Mediterranean buffaloes and primer design for detecting genotypes at ApoB_SNP1 to ApoB_SNP6 loci

[0066] 1. Test object

[0067] The trial selected 350 Mediterranean buffalo whose DNA samples came from Italian pastures; all lactation records were provided by the Italian Breeding Association (AIA) and the Italian Agricultural Research Council (CAR).

[0068] 2. Primer Design

[0069] According to the buffalo ApoB gene sequence in GenBank (accession number: NC_037556.1) as a template, use Primer5.0 primer design software to design primer pairs as shown in the fragments of SEQ ID NO: 7-14, and entrust Beijing Qingke Biotechnology Co., Ltd. Co., Ltd. (Wuhan) synthesis.

[0070] 3. PCR amplification reaction

[0071] Using the Mediterranean buffalo genomic DNA as a template, the PCR amplification reaction was carried out according to the following procedures: the PCR reaction ...

Embodiment 2

[0080] Example 2: Application of markers ApoB_SNP1 to ApoB_SNP6 in association analysis of lactation traits in Mediterranean buffaloes

[0081] In order to determine whether the detected ApoB_SNP1 to ApoB_SNP6 markers are related to the differences in lactation phenotypes of Mediterranean buffaloes, Mediterranean buffaloes (350 heads) were selected as test materials in this example. The polymorphism was detected by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS), and the correlation between different genotypes and lactation traits of buffalo was analyzed. The general linear model in SAS9.4 software was used for the association analysis between genotype and phenotype values. The SNPs were the same as the haplotype association analysis model, and the analysis model was: Y ijkl =u+G i +P j +S k +F l +PS jk +PF jl +SF kl +e. Among them, Y ijkl is the observed value of the trait; u is the total mean value of the trait; G i is th...

Embodiment 3

[0091] Example 3: Identification of haplotype combinations of ApoB_SNP1 to ApoB_SNP6 loci with excellent lactation traits

[0092] (1) Construction of haplotype and haplotype combination

[0093] Haploview software was used to analyze the haplotypes of ApoB_SNP1 to ApoB_SNP6, and at the same time, the genotype data of all individual ApoB_SNP1 to ApoB_SNP6 sites were input into the PHASE program to calculate the genotype of each individual, and at the same time calculate the composition between the sites. The degree of linkage disequilibrium.

[0094] Haplotype block analysis is attached Figure 7 shown. It was found that SNP sites g.-1718A>G, g.-1823G>C, g.-2063A>C participated in forming a haplotype block 1 (haplotype block), SNP sites g.37375G>A and g.40787A >G participates in a haplotype block 2, both of which are in linkage disequilibrium.

[0095] (2) Association analysis of different haplotype combinations and milk production traits

[0096] Calculate the haplotype ...

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Abstract

Molecular markers and applications associated with superior lactation traits in Mediterranean buffalo. The present invention provides SNP markers related to lactating traits of buffaloes cloned from ApoB gene and used alone or in combination. The SNP markers are selected from SNP1-6, and their nucleotide sequences are shown in SEQ ID NO: 1-6. Show. The AA genotype of SNP1, the GG genotype of SNP2, the AA genotype of SNP3, the GG genotype of SNP4, the GG genotype of SNP5 and the GG genotype of SNP6 were dominant genotypes. The present invention also provides two dominant haplotype combinations block1 and block2. The SNP markers provided by the invention are closely related to the milk production traits of Mediterranean buffaloes, and the milk production traits of the subject can be quickly determined by detecting the molecular markers, so as to achieve the purpose of selecting target traits, and have the advantages of being fast, accurate, and free from environmental interference.

Description

technical field [0001] The invention belongs to the technical field of buffalo molecular marker screening and auxiliary selection. It specifically involves the SNP markers and their haplotype combinations and applications related to the excellent lactation traits of Mediterranean buffaloes. Background technique [0002] Buffaloes are the second largest dairy producing livestock in the world, accounting for 12% of total global milk production (Zicarelli 2020). Buffalo milk is rich in nutritional value, and its total protein, medium-chain fatty acid, conjugated linoleic acid, retinol and vitamin E content are all higher than those of Holstein milk. It is a high-quality raw material for processing and producing a variety of high-end dairy products, such as cheese, Yogurt and cream etc. (Mahmood and Sumaira 2010). Therefore, in the past two decades, the buffalo industry has received more and more national attention, and the buffalo herd has grown at a rate of 2% per year (Ahma...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888C12Q2600/156C12Q2600/124C12Q2600/172
Inventor 滑国华曹家瑄林雨昕孙晖杨利国李华钊吴祺鹏
Owner HUAZHONG AGRI UNIV
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