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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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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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 acquisition of stress-resistant new varieties through molecular assisted technology, among which the development of linkage markers is the premise of this work

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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Embodiment 1

[0034] The SNP molecular marker of apple rootstock cold resistance identification of the present invention adopts the following method to obtain:

[0035] 1. Reagents and instruments used

[0036] Genomic DNA (gDNA) of individuals in the test population was extracted using the Plant Genomic DNA Kit (TIANGEN) kit, which was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.; restriction enzymes for enzyme digestion EcoRI Purchased from Sigma-Aldrich Company in the United States; reagents required for post-processing of enzyme-digested samples: adapters, ATP, ligase, blunt-end enzyme, etc. were purchased from Illumina Company in the United States; The IlluminaHiseq 2500 sequencing platform (USA) was used.

[0037] 2. SNP molecular marker acquisition method

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

[0039] The hybrid offspring of apple cold-resistant rootstock "60-160" (low temperature semi-lethal temperature -37.42C°) x non-cold-resistant root...

Embodiment 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 above-mentioned PCR primers for amplifying the SNP molecular markers are fixed on the SNP gene chip, and the gene chip is manufactured by Affymetrix Company of the United States. The genomic DNA of the test population was extracted, KASP reaction was performed on the SNP gene chip, the SNP molecular marker was amplified, and the cold resistance performance of the test population was quickly identified through the genotype of the SNP molecular marker. The specific operation is as follows:

[0059] Collect 182 "60-160" × "M9" mapping population strains, use the Plant Genomic DNA Extraction Kit (PlantGenomic DNA Kit, Beijing Tiangen Company) to extract the genomic DNA of the test population, and perform KASP on the SNP gene chip produced reaction. A total of 1 microliter of KASP amplification reaction system, including 20 nano...

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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...

Claims

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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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