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Primer sequence for judging cucumber powdery mildew resistance and judging method therefor

A technology of cucumber powdery mildew and primer sequence, which is applied in the field of identification of primer sequence and identification of cucumber powdery mildew resistance, and can solve the problems of molecular markers that have not been reported.

Inactive Publication Date: 2004-03-17
TIANJIN RES INST OF VEGETABLE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] There have been many reports on molecular marker technology in finding molecular markers linked to target traits, but molecular markers closely linked to genes related to cucumber powdery mildew resistance have not been reported yet.

Method used

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  • Primer sequence for judging cucumber powdery mildew resistance and judging method therefor
  • Primer sequence for judging cucumber powdery mildew resistance and judging method therefor
  • Primer sequence for judging cucumber powdery mildew resistance and judging method therefor

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Embodiment 1

[0018] A method for identifying cucumber powdery mildew resistance using a primer sequence for identifying cucumber powdery mildew resistance, comprising the following steps: (1) extracting the DNA of the cucumber genome, the extraction method is a conventional method; (2) carrying out PCR amplification, in Put in a thin-walled tube dedicated to PCR amplification: cucumber genomic DNA 30ng, upstream primer 5′-CAG TAA ATG AAA GAA AAG AAG-3′30ng, downstream primer 5′-ATA CAT AGC CAT ACA AAA AT-3′30ng, dNTP 0.2mM, Mg 2+ 1.5mM, 1 times PCR buffer, 1 unit of Taq DNA polymerase, add sterile double distilled water to 20μl, put the special thin-walled tube for PCR amplification into a PCR instrument for amplification, and the amplification conditions are: 94°C Pre-denaturation for 300 seconds, denaturation at 94°C for 60 seconds, annealing at 48°C for 60 seconds, extension at 72°C for 120 seconds, 35 cycles, and extension at 72°C for 420 seconds, the amplification is complete; (3) Gel...

Embodiment 2

[0020] A method for identifying cucumber powdery mildew resistance using a primer sequence for identifying cucumber powdery mildew resistance, comprising the steps of: (1) extracting the DNA of the cucumber genome, the extraction method is a conventional method; (2) performing PCR amplification, in Put into a thin-walled tube dedicated to PCR amplification: cucumber genomic DNA 15ng, upstream primer 5′-CAG TAA ATG AAA GAA AAG AAG-3′20ng, downstream primer 5′-ATA CAT AGC CAT ACA AAA AT-3′20ng, dNTP0.15mM, Mg 2+1.2 mM, 1 times PCR buffer, 1.2 units of Taq DNA polymerase, add sterile double distilled water to 20 μl, put the special thin-walled tube for PCR amplification into a PCR instrument for amplification, and the amplification conditions are: 94°C Pre-denaturation for 180 seconds, denaturation at 94°C for 60 seconds, annealing at 47°C for 40 seconds, extension at 72°C for 60 seconds, 25 cycles, and extension at 72°C for 300 seconds, the amplification is complete; (3) Gel ele...

Embodiment 3

[0022] A method for identifying cucumber powdery mildew resistance using a primer sequence for identifying cucumber powdery mildew resistance, comprising the following steps: (1) extracting the DNA of the cucumber genome, the extraction method is a conventional method; (2) carrying out PCR amplification, in Put in a thin-walled tube dedicated to PCR amplification: cucumber genomic DNA 20ng, upstream primer 5′-CAG TAA ATG AAA GAA AAG AAG-3′40ng, downstream primer 5′-ATA CAT AGC CAT ACA AAA AT-3′40ng, dNTP 0.25mM, Mg 2+ 2.0 mM, 1 times PCR buffer, 0.8 units of Taq DNA polymerase, add sterile double distilled water to 20 μl, put the special thin-walled tube for PCR amplification into a PCR instrument for amplification, and the amplification conditions are: 94°C Pre-denaturation for 240 seconds, denaturation at 94°C for 60 seconds, annealing at 53°C for 60 seconds, extension at 72°C for 100 seconds, 30 cycles, and extension at 72°C for 600 seconds, the amplification is complete; (...

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Abstract

The present invention discloses one kind of primer series for identifying cucumber powdery mildew and its identification method. The primer series consists of upstream primer 5'-CAG TAA ATG AAA GAA AAG AAG-3' and downstream primer 5'-ATA CAT AGC CAT ACA AAA AT-3'. The identification method of cucumber powdery mildew with the primer series includes the following steps: extracting cucumber genomeDNA; PCR proliferation; gel electrophoresis analysis of the PCR proliferation product; and identifying the resistance of cucumber powdery mildew according to the relative position of the identified sample on gel strip. The said method has its result possess coincidence rate with field detection result up to 94 %, and is fast, accurate and not affected by environment condition, so that it has high application value in screening cucumber powdery mildew resistance.

Description

technical field [0001] The invention relates to a DNA primer sequence in biotechnology and a method for identifying cucumber powdery mildew resistance using the DNA primer sequence. Background technique [0002] Cucumber powdery mildew (Sphaerotheca fuliginea L.) is a widespread worldwide plant disease. The disease spreads by airflow, likes temperature and humidity, and is resistant to drying. Under the conditions of suitable temperature and humidity, the pathogen spores germinate, and the disease first appears on the front or back of the lower leaves of the plant, appearing as small white powder spots, and then expands into powdery round spots. When the conditions are suitable, the white powdery spots continue to expand and connect into sheets, becoming a large area of ​​white powder with inconspicuous edges until it covers the entire leaf and looks like a layer of white hair, so it is commonly called "white hair disease". The leaves of powdery mildew gradually turn yello...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/00C12N15/11C12P19/34C12Q1/68
Inventor 杜胜利张桂华李淑菊魏爱民张历韩毅科
Owner TIANJIN RES INST OF VEGETABLE
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