A method for rapid detection and identification of bacterial wilt caused by R. solanacearum infection

A technology for the identification of R. solanacearum and solanaceae, which is applied in the field of rapid detection and identification of bacterial wilt, can solve the problems of rare plant pathogenic microorganisms and no identification of R. solanacearum, and achieve accurate and reliable identification results and adaptability. Strong, low price effect

Active Publication Date: 2012-02-15
PLANT PROTECTION RES INST OF GUANGDONG ACADEMY OF AGRI SCI +1
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Problems solved by technology

The identification process of the above two identification systems takes more than 18 hours, and it is necessary to purchase expensive imported special identification materials and equipment, and the system is mainly suitable for the identification of human, animal and environmental microorganisms, and is rarely used for the identification of plant pathogenic microorganisms , there is no report on the identification of Solanaceae bacterial wilt in the current literature

Method used

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  • A method for rapid detection and identification of bacterial wilt caused by R. solanacearum infection
  • A method for rapid detection and identification of bacterial wilt caused by R. solanacearum infection
  • A method for rapid detection and identification of bacterial wilt caused by R. solanacearum infection

Examples

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Embodiment 1

[0032] Example 1: Detection and identification of known disease samples

[0033] 1. Experimental materials: Eggplant and tomato disease-like tissues caused by the infection of R. solanacearum and known healthy eggplant tissues were used as materials.

[0034] 2. Experimental method:

[0035] (1) Rinse the stem base and root of the diseased sample to be tested with water, and take a vascular bundle tissue with a size of about 1.5-2 mm × 2.5-3 mm from the stem base of the diseased sample to be tested, and 8 samples to be tested include 4 eggplants diseased sample and 4 tomato diseased samples, and another healthy eggplant sample was taken as a control;

[0036] (2) Use a sterile pipette to draw 100 μL of sterilized water, drop it on a clean glass slide, and use sterilized tweezers to put the diseased tissue into the 100 μL sterilized water drop on the clean glass slide. Forcefully press the diseased tissue into the water droplets, and let it stand for about 5 minutes; observe ...

Embodiment 2

[0042] Embodiment 2: detection and identification test to unknown disease sample

[0043] 1. Experimental materials: 10 different plant bacterial wilt samples or asymptomatic healthy samples collected from fields in multiple regions were tested.

[0044] 2. Experimental method:

[0045] 10 different plant samples collected from multiple regions in Guangdong were sampled, and a total of 17 samples were obtained and numbered. The plant species, collection locations, and symptoms corresponding to each number of samples are shown in Table 1. The method used for detection and identification is the same as that used in Example 1, and is compared and verified with the traditional method for isolation and identification of plant pathogenic bacteria.

[0046] The specific steps of the traditional method for isolation and identification of plant pathogenic bacteria include: 1. Disinfect the surface of the diseased tissue to be examined with 75% alcohol; Streak and culture at 28-30°C f...

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Abstract

The invention discloses a method for rapidly detecting and identifying the bacterial wilt disease caused by R. solanacearum infection. The method is composed of three stages of plant disease sample preparation, naked eye observation and PCR verification, the detection and identification can be completed within 4 hours, and the cost of detection and identification is low. This method breaks through the limitations of the traditional plant bacterial disease identification period, such as long period, strong specialization and high cost, and also overcomes the shortcomings of imported microbial identification systems that require special equipment and consumables, high cost and long identification period. By organically combining the special symptoms of disease samples with molecular biotechnology, the results are accurate and reliable, so as to meet the requirements of rapid and accurate identification of plant diseases in actual production.

Description

technical field [0001] The invention belongs to the technical field of plant disease identification, in particular to a rapid detection and identification method for bacterial wilt caused by Ralstonia solanacearum infection. Background technique [0002] For the detection and identification of bacterial wilt caused by R. solanacearum infection of Solanaceae, there are three existing technologies: the traditional method for isolation and identification of plant pathogenic bacteria, the BIOLOG microbial identification system and the Mérieux microbial identification method. [0003] The traditional identification method of plant bacterial wilt mainly includes the steps of isolating pathogenic bacteria from disease samples, obtaining pure culture of isolated strains, observing the morphological characteristics of colonies and bacteria, measuring physiological and biochemical characteristics, and determining pathogenicity. It takes at least one month or more, the workload is heav...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12Q1/04G01N27/447C12R1/01
Inventor 何自福佘小漫虞皓罗方芳
Owner PLANT PROTECTION RES INST OF GUANGDONG ACADEMY OF AGRI SCI
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