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Method for rapidly distinguishing columnar apple seedlings by using DNA molecular marker

A technology of DNA molecules and apples, applied in the field of molecular genetics and breeding, can solve problems such as difficult identification and differentiation

Active Publication Date: 2016-12-07
QINGDAO AGRI UNIV
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Problems solved by technology

[0005] The object of the present invention is to overcome the defect that 1-2 year old columnar apple hybrid seedlings and columnar apple varieties are difficult to distinguish and distinguish in the seedling stage, and to provide a method for quickly distinguishing columnar apple seedlings by using DNA molecular markers, That is, a DNA molecular marker tightly linked with the control apple columnar gene Co, and a method for detecting the molecular marker by using PCR technology

Method used

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  • Method for rapidly distinguishing columnar apple seedlings by using DNA molecular marker
  • Method for rapidly distinguishing columnar apple seedlings by using DNA molecular marker
  • Method for rapidly distinguishing columnar apple seedlings by using DNA molecular marker

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Embodiment 1

[0055] In this example, 25 columnar apple types and 31 common apple types stored in the Jiaozhou Agricultural Science and Technology Demonstration Garden of Qingdao Agricultural University are used as test materials. The 25 columnar types include the hybrid F1 population of 'Fuji' apple and columnar apple 23 columnar hybrid individuals, 2 columnar apple varieties 'Lujia 4' and 'Weisekexu'; 31 common apple types including 29 hybrid F1 populations of 'Fuji' and columnar apples Common hybrid individuals, two common varieties 'Fuji' and 'Xu'.

[0056] The specific operation steps are as follows:

[0057] 1. Extraction of Apple Genomic DNA by CTAB Method

[0058] 1. Take a 2ml centrifuge tube, add 1.2ml CTAB and 50μL β-mercaptoethanol, and preheat it in a 65°C water bath, take about 0.5g of young apple leaves, add liquid nitrogen to quickly grind them into fine powder, and transfer to the above In a centrifuge tube containing CTAB, oscillate to mix, place in a water bath at 65°C ...

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Abstract

The invention relates to a method for rapidly distinguishing columnar apple seedlings by using a DNA molecular marker. According to the method, DNA of an apple leaf to be detected is used as a template, two pairs of oligodeoxyribonucleotide sequences Co1 and Co2) are used as primer pairs, PCR reaction amplification is performed on the DNA molecular marker linked with a columnar gene, and electrophoretic detection is performed on amplified products; if electrophoretic results of the amplified products of the two pairs of primers show that at least one pair of primers has a DNA specific band corresponding to the DNA molecular marker, the corresponding apple seedlings are columnar apple seedlings; if the electrophoretic results of the amplified products of the two pairs of primers show that no primer has corresponding DNA specific band, the corresponding apple seedlings are general apple seedlings. By the method, columnar apple seedlings in 1-2-year-old apple seedlings can be conveniently, rapidly and accurately distinguished, so that the breeding efficiency is greatly improved and the breeding process is accelerated; meanwhile, true and false columnar apple seedlings can be simply and quickly identified, so that disputes caused by the false seedlings are avoided.

Description

Technical field: [0001] The invention relates to a method for quickly distinguishing cylindrical apple seedlings by using DNA molecular markers, and belongs to the technical field of molecular genetic breeding. Background technique: [0002] Columnar apple (Columnar apple) has the characteristics of short internodes, axillary buds germinate into a large number of short branches, little or no lateral extension of new shoots, and a natural single-stem shape, which is different from ordinary types and ordinary short branches type or dwarf type. This tree is compact and requires very little pruning, making it ideal for high density planting and mechanized management. Promoting this tree type will greatly save labor and production costs, and meet the development requirements of labor-saving cultivation of apples. Studies have shown that apple columnar traits are quality traits controlled by a dominant single gene (Co). Therefore, columnar apple is an ideal genetic resource for...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/686C12Q1/6895C12Q2600/13C12Q2600/158C12Q2565/125
Inventor 张玉刚孙欣柏素花孙晓红侯鸿敏刘源霞祝军戴洪义
Owner QINGDAO AGRI UNIV
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