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Primer sequence for identifying cucumber ampho disease and its identification method

A cucumber downy mildew, primer sequence technology, applied in the field of DNA sequence, to achieve the effect of great application value

Inactive Publication Date: 2007-03-28
TIANJIN RES INST OF VEGETABLE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Molecular marker technology has been reported on the search for molecular markers linked to target traits, but molecular markers closely linked to genes related to cucumber downy mildew resistance have not been reported yet.

Method used

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  • Primer sequence for identifying cucumber ampho disease and its identification method
  • Primer sequence for identifying cucumber ampho disease and its identification method
  • Primer sequence for identifying cucumber ampho disease and its identification method

Examples

Experimental program
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Effect test

Embodiment 1

[0029] A method for identifying resistance to cucumber downy mildew

[0030] (1) Extract cucumber genomic DNA; (2) Carry out PCR amplification, put into in the special-purpose thin-walled tube of PCR amplification: described cucumber genomic DNA 20ng, the nucleotide sequence 30ng described in SEQ ID No1 in the sequence table, sequence Nucleotide sequence 30ng, dNTP0.2mM, Mg described in SEQ IDNo2 in the list 2+1.5mM, 1 times PCR buffer, 1.0 units of Taq DNA polymerase, add sterile double distilled water to 20μl, put the special thin-walled tube for PCR amplification into the PCR instrument for amplification, and the amplification conditions are: 94°C Pre-denaturation for 200 seconds, denaturation at 94°C for 60 seconds, annealing at 55°C for 50 seconds, extension at 72°C for 90 seconds, 30 cycles, and extension at 72°C for 350 seconds, the amplification is complete; (3) Gel electrophoresis analysis of PCR amplification products : The amplified product is separated by electrop...

Embodiment 2

[0032] A method for identifying resistance to cucumber downy mildew

[0033] (1) Extract cucumber genomic DNA; (2) Carry out PCR amplification, put into in the special-purpose thin-walled tube of PCR amplification: described cucumber genomic DNA 15ng, the nucleotide sequence 20ng described in SEQ ID No1 in the sequence list, sequence Nucleotide sequence 20ng, dNTP0.15mM, Mg described in SEQ IDNo2 in the list 2+ 1.2mM, 1 times PCR buffer, 0.8 units 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 180 seconds, denaturation at 94°C for 60 seconds, annealing at 50°C for 40 seconds, extension at 72°C for 60 seconds, 25 cycles, and extension at 72°C for 300 seconds. Amplification is complete: (3) Gel electrophoresis analysis of PCR amplification products : The amplified product is separated by electrophores...

Embodiment 3

[0035] A method for identifying resistance to cucumber downy mildew

[0036] (1) extract cucumber genomic DNA; (2) carry out PCR amplification, put into in the special-purpose thin-walled tube of PCR amplification: described cucumber genomic DNA 30ng, the nucleotide sequence 40ng described in SEQ ID No1 in the sequence list, sequence Nucleotide sequence 40ng, dNTP0.25mM, Mg described in SEQ IDNo2 in the list 2+ 2.0mM, 1 times PCR buffer, 1.2 units 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 59°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 electrophoresis analysis of PCR amplification products : The amplified product is separated by electrop...

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Abstract

The invention is a primer sequence for identifying resistance to cucumber frost mildew and identifying method, where the primer sequence is composed of an upper primer with a nucleotide sequence described in sequence list SEQ ID No.1 and an lower primer with that in sequence list SEQ ID No.2; includes the steps: (1) extracting a cucumber gene group DNA; (2) making PCR amplification by using the above two nucleotide sequences; (3) making gel electrophoretic analysis on the product of PCR amplification; (4) according to the relative position of all the identified samples, identifying the resistance to cucumber frost mildew, and the fitting rate of the identified result to the field resistance to diseases reaches 94.7% and it has the advantages of rapidness, accuracy, no influence by the environmental conditions, etc, and has very great application value in screening the resistance to cucumber frost mildew.

Description

technical field [0001] The invention relates to a DNA sequence in biotechnology and a method for identifying cucumber downy mildew resistance using the DNA sequence. Background technique [0002] Cucumber downy mildew [Pseudoperonospora cubensis] is a widespread worldwide plant disease. The disease is transmitted by airflow, and its occurrence and prevalence are related to the plant growth environment, especially the temperature and humidity of the air, especially the humidity. The disease mainly affects leaves, but occasionally stems, tendrils, and pedicels can also be affected. In an environment suitable for the onset of the disease, cucumber seedlings can be susceptible to the disease, and the onset of the adult plant is mostly after the plants enter flowering and fruiting. In the early stage of the disease, water-soaked light green spots appeared on the leaves, which expanded and became polygonal due to the veins, and the color changed to yellow-green, yellow, and fina...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 杜胜利鞠秀芝张桂华李淑菊魏爱民韩毅科张历
Owner TIANJIN RES INST OF VEGETABLE
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