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Molecular marker identification method for bacterial angular leaf spot resistant gene of cucumber

A technology of molecular markers and identification methods, which is applied in the field of plant biology, can solve the problems of accelerating the process of cucumber breeding and unclear molecular mechanisms, and achieve the effects of improving breeding efficiency, being easy to master, and reducing production costs

Inactive Publication Date: 2011-06-22
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the molecular mechanism of the interaction between cucumber and bacterial angular spot is still unclear, and the structure and function of many pathogenic genes and cucumber disease resistance genes are still unclear. Molecular markers have important practical application value for accelerating the process of cucumber breeding

Method used

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  • Molecular marker identification method for bacterial angular leaf spot resistant gene of cucumber
  • Molecular marker identification method for bacterial angular leaf spot resistant gene of cucumber

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Effect test

Embodiment 1

[0019] The molecular marker identification method of cucumber anti-bacterial angular leaf spot gene, the method comprises the following steps:

[0020] Use the DNA of a cucumber variety or strain with unknown resistance to bacterial angular spot disease as a template, and use CSWCT24A as a primer to perform PCR amplification, and perform 3% agarose gel electrophoresis on the amplified product or 7.5% agarose gel electrophoresis on the amplified product. Denatured polyacrylamide gel electrophoresis test, if the amplified band type is consistent with the known resistance to bacterial angular spot of cucumber Dongnong 803, then the cucumber variety or line with unknown resistance to bacterial angular spot For a variety or strain resistant to bacterial angular leaf spot, the PCR amplification program is: 94°C pre-denaturation for 5 minutes, 94°C denaturation for 30 seconds, 57°C annealing for 1 minute, 35 cycles, 72°C extension for 1 minute, 72°C extension for 10 minutes, Finally ...

Embodiment 2

[0024] In the molecular marker identification method of the cucumber anti-bacterial angular leaf spot gene, in the PCR amplification, the PCR reaction system of every 20 μl is as follows: 10×buffer: 2 μl, 2mM dNTP: 2 μl, Primer 1 of 10 pmol / μl: 0.8 μl, 10 pmol / μl Primer 2: 0.8 μl, 25 mM MgCl 2 : 2.5 μl, 5 U / μl TaqDNA polymerase: 0.2 μl, 60 ng / μl template DNA: 1 μl, ddH2O: 10.7 μl.

Embodiment 3

[0026] Application of molecular marker identification method of cucumber bacterial angular leaf spot resistance gene in assisted selection of disease-resistant cucumber materials.

[0027] Using the new cucumber hybrid varieties Dongnong 804 and Dongnong 805 bred by the cucumber research group of the College of Horticulture of Northeast Agricultural University as materials; extracting the DNA of a single plant as a template, and using a specific molecule of cucumber anti-bacterial angular leaf spot provided by the present invention The marker CSWCT24A was used as a primer for PCR amplification. The results showed that the PCR amplification band patterns of Dongnong 804 and Dongnong 805 were completely consistent with the amplified band pattern of the disease-resistant control Dongnong 803, and the results of PCR detection were consistent with those of the field bacteria. The concordance rate of leaf spot resistance identification was 100%, which proved that CSWCT24A is a practi...

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Abstract

The invention relates to a molecular marker identification method for the bacterial angular leaf spot resistant gene of cucumbers, which comprises: performing polymerase chain reaction (PCR) amplification by using the DNA of a cucumber variety or strain of which the bacterial angular leaf spot resistance is unknown as a template and chimpanzee sanctuary and wildlife conservation trust (CSWCT)24A as a primer; detecting the product of the amplification by electrophoresis in 3-percent gelose gel or 7.5-percent denaturant polyacrylamide gel; and if the band type obtained by amplification is consistent with the band of a known bacterial angular leaf spot resistant cucumber Dongnong 803, determining the cucumber variety or strain of which the bacterial angular leaf spot resistance is unknown as a bacterial angular leaf spot resistant variety or strain. The PCR amplification process comprises: pre-denaturing at 94 DEG C for 5 minutes, denaturing at 94 DEG C for 30 seconds, annealing at 57 DEG C for 1 minute, circulating for 35 times, extending at 72 DEG C for 1 minute, extending at 72 DEG C for 10 minutes, and storing at 4 DEG C. The method is used for identifying the bacterial angular leaf spot resistant gene of cucumbers.

Description

Technical field: [0001] The invention relates to a molecular marker identification method of a cucumber anti-bacterial angular leaf spot gene, belonging to the field of plant biotechnology. Background technique: [0002] Cucumber bacterial angular spot (Pseudomopnas syringae pv. lachrymans) is an important bacterial disease. The pathogenic bacteria is Pseudomonas syringae pathogenic variety of cucumber, which belongs to the genus Pseudomonas of the thin-walled phylum. At the beginning of the disease, there are small water-soaked spots, which are limited by the veins after expansion. The lesions are polygonal, light yellow to brown, with yellow halos on the edges. When the disease is severe, the lesions cover the leaves, causing the leaves to dry and curl. The disease occurs in both protected land and open field production, and the open field is the most harmful, resulting in severe production reduction. With the continuous expansion of cucumber planting area in recent year...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 秦智伟武涛王丽莉周秀艳辛明
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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