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Molecular markers for detecting cucumber corynespora leaf spot bacteria and application thereof

A technology of promoting leaf spot bacteria and molecular markers, applied in the field of molecular biology, can solve the problems of indistinguishable, high experimental cost, complicated operation, etc., and achieve the effect of practical detection, rapid detection and high sensitivity

Inactive Publication Date: 2017-09-05
SUZHOU COSMETIC MATERIALS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing immunoassay methods generally prepare monoclonal antibodies using the host-selective toxin protein of Corynespora polybasicum as the antigen, and the operation is relatively complicated and the experimental cost is relatively high.
The detection method based on computer color images may affect the detection accuracy due to the different environmental factors such as shooting status and lighting, the degree of disease and the typicality of lesions.
There are few reports on the PCR detection method of Corynespora multiprimum, only 3-4 control strains are used in the existing methods, the specificity of the primers needs to be further verified, and the detection technology is not yet perfect
At the same time, the existing PCR detection methods for Corynesporium leaf spot of cucumber failed to directly distinguish Corynesporium leaf spot from cucumber downy mildew, anthracnose, and bacterial angular spot

Method used

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  • Molecular markers for detecting cucumber corynespora leaf spot bacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] A molecular marker used for detecting Phytophthora spirochete, said molecular marker and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.5 ~6.

[0028] The 5' ends of the upstream primers of the molecular markers P1, P2 and P3 are modified by amination.

[0029] The application of the molecular marker in the preparation of a kit for detecting the leaf spot fungus on cucumber sticks.

[0030] The components of the kit include: molecular markers, dNTP, LATaq, PCR reaction buffer, MgCl 2 , double distilled water.

[0031] The steps of the kit detecting the leaf spot fungus of cucumber sticks include:

[0032] (1) Sampling: extract the genomic DNA of the cucumber leaf sample;

[0033] (2) Using the DNA extracted in step (1) as a PCR template, and using P1 as a specific primer, carry out the first round of PCR amplification;

[0034] (3) Get the amplified product of step (2), dilute it 200 times and use it as the template for the second round of PC...

Embodiment 2

[0043] A molecular marker used for detecting Phytophthora spirochete, said molecular marker and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.5 ~6.

[0044] The 5' ends of the upstream primers of the molecular markers P1, P2 and P3 are modified by amination.

[0045] The application of the molecular marker in the preparation of a kit for detecting the leaf spot fungus on cucumber sticks.

[0046] The components of the kit include: molecular markers, dNTP, LATaq, PCR reaction buffer, MgCl 2 , double distilled water.

[0047] The steps of the kit detecting the leaf spot fungus of cucumber sticks include:

[0048] (1) Sampling: extract the genomic DNA of the cucumber leaf sample;

[0049] (2) Using the DNA extracted in step (1) as a PCR template, and using P1 as a specific primer, carry out the first round of PCR amplification;

[0050] (3) Get the amplified product of step (2), dilute it 400 times and use it as the template for the second round of PC...

Embodiment 3

[0059] A molecular marker used for detecting Phytophthora spirochete, said molecular marker and its sequence are: P1, SEQ ID NO.1-2; P2, SEQ ID NO.3-4; P3, SEQ ID NO.5 ~6.

[0060] The 5' ends of the upstream primers of the molecular markers P1, P2 and P3 are modified by amination.

[0061] The application of the molecular marker in the preparation of a kit for detecting the leaf spot fungus on cucumber sticks.

[0062] The components of the kit include: molecular markers, dNTP, LATaq, PCR reaction buffer, MgCl 2 , double distilled water.

[0063] The steps of the kit detecting the leaf spot fungus of cucumber sticks include:

[0064] (1) Sampling: extract the genomic DNA of the cucumber leaf sample;

[0065] (2) Using the DNA extracted in step (1) as a PCR template, and using P1 as a specific primer, carry out the first round of PCR amplification;

[0066] (3) Get the amplified product of step (2), dilute it 500 times and use it as the template for the second round of PC...

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Abstract

The invention discloses molecular markers for detecting cucumber corynespora leaf spot bacteria and application thereof. The molecular markers with sequences comprise a molecular marker P1 with SEQ ID NO.1 to 2, a molecular marker P2 with SEQ ID NO.3 to 4, and a molecular marker P3 with SEQ ID NO.5 to 6. The molecular markers have the advantages that (1) the cucumber corynespora leaf spot bacteria can be accurately, quickly, sensitively and practically detected; (2) the sensitivity on the detection of pathogenic bacteria is high, and the molecular markers can be applied to the quick detection of the cucumber corynespora leaf spot bacteria in disease plants.

Description

technical field [0001] The invention belongs to the field of molecular biology, and relates to a method for detecting pathogenic bacteria by means of molecular biology, in particular to a molecular marker for detecting the leaf spot fungus of cucumber and its application. Background technique [0002] Corynespora multiprimum is the pathogen that causes cucumber leaf spot. The host range of Corynespora multiprimum is wide, and the modes of transmission are diverse. In recent years, investigations have found that leaf spot diseases of cucumbers, tomatoes and other vegetables caused by Corynesporum polybasicus have occurred in a large area in 11 provinces and municipalities in China, including Shandong, Hebei, Liaoning, and Inner Mongolia, causing serious economic losses. At the end of the 20th century, cucumber Corynesporium leaf spot broke out in Wafangdian City, Liaoning Province, seriously endangering the production of cucumber. At present, the disease has become one of t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/6895
Inventor 金仲恩陈晋纳欧阳智琨喻国贞张帆全春兰
Owner SUZHOU COSMETIC MATERIALS
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