SNP molecular marker for cold-resistance identification of apple rootstocks, primers and application of SNP molecular marker

A technology of molecular markers and cold resistance, applied in the field of crop genetics and breeding, can solve the problems of long cycle, achieve accurate identification, reduce time and economic cost

Active Publication Date: 2018-06-01
SHANDONG INST OF POMOLOGY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, at present, it takes a long time to obtain new varieties through conventional breeding, and more and more attention has been paid to the acquis

Method used

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  • SNP molecular marker for cold-resistance identification of apple rootstocks, primers and application of SNP molecular marker
  • SNP molecular marker for cold-resistance identification of apple rootstocks, primers and application of SNP molecular marker
  • SNP molecular marker for cold-resistance identification of apple rootstocks, primers and application of SNP molecular marker

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[0033] Example 1

[0034] The SNP molecular marker for identification of cold resistance of the apple rootstock of the present invention is obtained by the following method:

[0035] 1. Reagents and instruments used

[0036] The Plant Genomic DNA Kit (TIANGEN) was used to extract genomic DNA (gDNA) of individuals in the test population, which was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.; restriction enzymes were used for restriction enzymes EcoRⅠ Purchased from Sigma-Aldrich, USA; reagents required for post-processing of digested samples: adapters, ATP, ligase, blunt-end enzyme, etc., were purchased from Illumina, USA; sequencing was performed at Beijing Nuohe Zhiyuan Bioinformatics Co., Ltd., sequencing Use IlluminaHiseq 2500 sequencing platform (USA).

[0037] 2. SNP molecular marker acquisition method

[0038] (1) Construction of "60-160×M9" genetic linkage map

[0039] The hybrid progeny of apple cold-resistant rootstock "60-160" (low-temperature semi-leth...

Example Embodiment

[0057] Example 2

[0058] In order to efficiently, quickly and accurately detect the SNP molecular marker of the present invention, it can be made into a SNP gene chip. The SNP gene chip is fixed with the PCR primers for amplifying the SNP molecular markers, and the gene chip is made by Affymetrix, USA. Extract the genomic DNA of the test population, perform KASP reaction on the SNP gene chip, amplify the SNP molecular markers, and quickly identify the cold resistance of the test population through the genotype of the SNP molecular markers. The specific operations are as follows:

[0059] Collect 182 "60-160"×"M9" mapping population lines, use Plant Genomic DNA Kit (PlantGenomic DNA Kit, Beijing Tiangen Company) to extract the genomic DNA of the test population, and perform KASP on the produced SNP gene chip reaction. KASP amplification reaction system totals 1 microliter, containing 20 ng template DNA, 0.14 microliter of a mixture of 3 primers (T allele primer, G allele primer ...

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Abstract

The invention discloses an SNP molecular marker for cold-resistance identification of apple rootstocks, primers and an application of the SNP molecular marker, and the molecular marker is a T-to-G transversion type. An amplification primer and an SNP gene chip are developed in accordance with the molecular marker, so that early identification of cold-resistance characters of an apple rootstock individual is achieved. According to the SNP molecular marker and the SNP gene chip provided by the invention, rapid and accurate identification of cold-resistance identification of the apple rootstockscan be achieved, so that time and economic cost of apple rootstock hybrid seedling breeding are reduced. The invention offers a marker resource for cold-resistance molecular marker-assisted breeding of the apple rootstocks and lays a foundation for breeding a new cold-resistance germplasm via genetic engineering means; therefore, an important theoretical significance and a high practical application value are guaranteed.

Description

technical field [0001] The present invention relates to a SNP (Single Nucleotide Polymorphism) molecular marker and its primer for identifying the cold resistance of apple rootstocks, and also relates to a SNP gene chip and a method for quickly identifying the cold resistance of apple rootstocks, belonging to crop genetics and breeding technology field. Background technique [0002] Due to the frequent occurrence of extreme weather due to abnormal climate, it has a great impact on agricultural production, and low temperature has become an important environmental factor restricting the efficiency of agricultural production in my country. Abiotic stresses such as chilling injury and freezing injury caused by low temperature are the main limiting factors affecting plant growth and reproduction. In the process of plant breeding, the ability to resist low temperature stress needs to be used as an important indicator to evaluate and measure the value and potential of new germplas...

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11C40B40/06
CPCC12Q1/6895C12Q2600/13C12Q2600/156C12Q2600/172
Inventor 王海波李林光程来亮常源升孙家正何平李慧峰
Owner SHANDONG INST OF POMOLOGY
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