Method for counting plant pathogenic oomycetes zoospores

A technology of zoospores and plant pathogens, applied in the field of microbiology and plant pathology, can solve problems such as difficult to count accurately, affect the accuracy of results, and cannot be accurately quantified in chemical substance inhibition experiments, so as to improve accuracy and precision, The method is simple and reliable

Pending Publication Date: 2019-07-05
YUNNAN AGRICULTURAL UNIVERSITY
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

Yet this invention has the following disadvantages: (1) is only suitable for catching spores or pollen in the air, and cannot be applied to spores in water or soil, and the scope of use is relatively limited
(2) Unable to accurately count spores
Yet, there are following two problems in this invention: (1) higher requirement is arranged for operating time
If the operation time is long, zoospore flagella will fall off and form immobile resting spores, which will cause greater interference to the process of leaf discs attracting zoospores, thus affecting the accuracy of results
(2) This method needs to prepare host plants and prepare leaf discs in advance, the operation process is relatively cumbersome, and the versatility is not high...

Method used

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  • Method for counting plant pathogenic oomycetes zoospores
  • Method for counting plant pathogenic oomycetes zoospores
  • Method for counting plant pathogenic oomycetes zoospores

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Experimental program
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Embodiment 1

[0038] In order to verify the accuracy of the counting method described in the present invention, this embodiment uses a known concentration of Phytophthora capsici zoospore suspension to carry out a verification test.

[0039] The specific method is as follows:

[0040] ①Prepare the zoospore suspension of Phytophthora capsici with a concentration of 2×10 5 pieces / ml.

[0041] ②Use a pipette to draw 40 μL of zoospore suspension into the groove of the concave glass slide, and add 10 μL of diallyl trisulfide aqueous solution of 6 different concentrations respectively, so that the final concentrations of diallyl trisulfide are respectively 0ppm, 10ppm, 50ppm, 100ppm, 500ppm, 1000ppm. Place the concave glass slide on the stage of the microscope, find the zoospores with a 4x objective lens, and observe with a 10x objective lens.

[0042] ③ Open the camera software of the microscope and set the camera parameters as follows:

[0043] Microscope Camera Model: Mingmei MC50; Microsc...

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Abstract

The invention relates to the fields of microorganisms and phytopathology, and particularly discloses a method for counting plant pathogenic oomycetes zoospores. According to the invention, the cameraphase parameter is set, so that the two morphological characteristics of the zoospores, namely blue spores and orange spores, can be clearly observed, thereby realizing the counting of the zoospores.According to the method, the zoospores and the resting spores can be counted simultaneously, so that the related experiments of the oomycetes zoospores are facilitated, and the accuracy and the precision of experimental results are improved. The method disclosed by the invention is simple and reliable, and all microscopes with imaging equipment can be suitable for counting oomycetes zoospores, sothat an evaluation index is provided for evaluating the severity of plant diseases and the pesticide effect of the bactericide.

Description

technical field [0001] The invention relates to the field of microorganisms and plant pathology, in particular to a counting method / statistical method of plant pathogenic oomycete zoospores. Background technique [0002] Phytopathogenic oomycetes infect plants by releasing zoospores with flagella into the soil water, which are attracted by the host root exudates to swim to new host plants, then stop forming restospores, and germinate to produce germ tubes to infect plants. Due to the hydrophilic nature of phytopathogenic oomycetes, zoospores have become an important medium for oomycetes to infect plants. In view of this, the number of oomycete zoospores has become an important indicator for the severity of plant diseases and the evaluation of fungicides. However, zoospores have a strong ability to move. Therefore, how to count the zoospores of oomycetes in water and obtain the ratio of zoospores to restospores is a technical problem to be solved urgently in the research of o...

Claims

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Application Information

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IPC IPC(8): G06K9/00G06K9/46C12Q1/06
CPCC12Q1/06G06V20/693G06V20/698G06V10/56
Inventor 刘屹湘吴家庆朱书生张贺杨敏黄惠川梅馨月杜飞何霞红李成云朱有勇
Owner YUNNAN AGRICULTURAL UNIVERSITY
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