Method for rapidly detecting bacterial canker of tomatoes and special primer for method

A technology for tomato canker and pathogens, applied in the field of rapid detection of tomato canker pathogens, can solve the problems that threaten tomato yield, quality fruit resistance, economic loss, etc.

Inactive Publication Date: 2015-05-27
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Once the disease occurs without effective measures to control it, it will seriously threaten the yield, quality and resistance of post-harvest fruits of tomatoes, causing serious economic losses.

Method used

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  • Method for rapidly detecting bacterial canker of tomatoes and special primer for method
  • Method for rapidly detecting bacterial canker of tomatoes and special primer for method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1, the acquisition of tomato canker bacterium template DNA

[0020] The tomato canker bacterium was cultivated in the modified 523 medium, shaken at 25°C for 3 days, collected the bacteria, added 100 μL of sterile water, heated at 95°C for 10 min, and centrifuged at 1000 r / min for 5 min to make a template.

Embodiment 2

[0021] Embodiment 2, tomato canker bacterium LAMP amplification and detection

[0022] The reaction system is: 2 μL template DNA in 25 μL total system, 0.4 mol / L Betaine betaine, MgSO 4 2.0 mmol / L, 4 U of Bst DNA polymerase, 1.0 mmol / L of dNTPs, 1.5 μL each of primer 1 and primer 2, 0.5 μL each of primer 3 and primer 4, 1×Isothermal Amplification Buffer.

[0023] The reaction program was as follows: incubation at 65°C for 40 min, inactivation at 80°C for 20 min.

[0024] The detection method is: take 5 μL LAMP amplification products and use 1.5% agarose gel electrophoresis to detect the amplification results (see figure 1 ).

[0025] Dilute SYBR Green I dye with 10×TBE, add it to the remaining amplification product at a ratio of 1 / 100, and observe the color change (see figure 2 ).

Description of drawings

[0026] figure 1 It is the result of 1.5% agarose gel electrophoresis detection of the LAMP amplification product of P. tomato canker.

[0027] figure 2 It is...

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PUM

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Abstract

The invention discloses a method for rapidly detecting bacterial canker of tomatoes and a special primer for the method. A special primer is a primer group which is composed of nucleotide sequences in a sequence 1, a sequence 2, a sequence 3 and a sequence 4 in a sequence table. The method for detecting the bacterial canker of tomatoes comprises the following steps: with a standard strain DNA (deoxyribonucleic acid) of the bacterial canker of tomatoes as a template, carrying out constant-temperature PCR (polymerase chain reaction) amplification under guide of the primer group composed of the nucleotide sequences in the sequence 1, the sequence 2, the sequence 3 and the sequence 4 in the sequence table of the claim 1; and detecting that a gradient strip is formed on the amplification product through agarose gel electrophoresis detection, and directly dyeing the amplification product by virtue of SYBR Green I, so that the amplification product is saffron. The invention provides a group of primers for rapidly amplifying a specific region of bacterial canker genome of tomatoes. The primers play an important role in rapid detection and authentication in the bacterial canker of tomatoes in fields and quarantine departments.

Description

technical field [0001] The invention relates to a method for rapidly detecting tomato canker bacterium and special primers thereof. Background technique [0002] Bacterial canker of tomato is a disease of tomato ( Lycopersicon esculentum Mill.) is one of the most serious diseases in production. Since the disease was first discovered on greenhouse tomatoes in Michigan, USA in 1909, it has been widely distributed in various tomato producing areas in the United States, and has gradually become a worldwide disease. At present, more than 60 countries in the Americas, Europe, Asia, Africa and Oceania have reported the occurrence of tomato canker. Once the disease occurs without effective measures to control it, it will seriously threaten the yield, quality and resistance of post-harvest fruits of tomato, causing serious economic losses. [0003] The records of tomato canker disease in my country began in 1954. At present, the disease is found in Beijing, Heilongjiang, Jilin, Li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/01
CPCC12Q1/6844C12Q1/689C12Q2531/119
Inventor 张娜杨文香赵赛刘大群
Owner HEBEI AGRICULTURAL UNIV.
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