Method for measuring content of live spores in fungal microbial pesticide quickly
A microbial pesticide, rapid determination technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as cumbersome operations, inability to distinguish target colonies and colonies of miscellaneous bacteria, affecting measurement results, etc., to achieve accurate detection. High accuracy, simple and rapid measurement, and easy operation
Inactive Publication Date: 2013-01-30
农业部农药检定所
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Problems solved by technology
The above method has the following main disadvantages: the operation is cumbersome, it is difficult to distinguish fungal spores and auxiliary particles under a microscope, and due to the short incubation time, some spores have not yet germinated, which affects the measurement results
[0005] One consideration is to directly obtain the content of live spores by plate counting method. However, if the fungi are operated according to the plate colony counting method commonly used for bacterial counting, the colonies of different spores will grow together and form sheets. Large colonies will affect the measurement results; if the colonies are too small, the counts cannot be seen clearly, and the target colonies and colonies of miscellaneous bacteria cannot be distinguished
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[0031] Example: using figure 1 According to the method shown, the viable spore content of Trichoderma wettable powder, Paecilomyces lilacinus wettable powder, Paecilomyces lilacinus liquid, and Beauveria bassiana parent drug were respectively determined. As embodiment 1 to embodiment 4.
[0032] The determination steps are as follows:
[0033] (1) Add 38g of potato dextrose agar powder (PDA) into 1000mL of distilled water, heat to boil, divide into 500mL Erlenmeyer flasks, after autoclaving, cool to about 60°C and prepare 0.25‰, 0.5‰, Sodium deoxycholate PDA plates with 1%, 2%, 5‰ content;
[0034] (2) Under aseptic conditions, stir the sample evenly, accurately weigh 3.0 g of the sample, dissolve it in 27.0 mL of sterile water containing 0.05% Tween20, soak it for 30 min, shake it for 30 min, and obtain a 10-fold diluted sample solution, Mark it as No. 0, and then perform serial dilutions, and set 3 replicate samples for each dilution;
[0035] (3) Take 100 μL of the abov...
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The invention discloses a method for measuring the content of live spores in a fungal microbial pesticide quickly. The method comprises the following steps of: (1) adding 0.25 to 5 weight percent of sodium deoxycholate into a fungal culture medium, and preparing flat plates; (2) soaking fungal microbial pesticide samples in sterilization water containing Tween 20 or polyethylene glycol octylphenol ether, and shaking; (3) diluting in a multiple gradient mode, coating the samples of different dilutability on the flat plate respectively and uniformly, and culturing at room temperature for 48 to 120 hours; (4) counting colony counts, wherein the flat plates on which the colony counts are 30 to 300 are effectively-counted flat plates; and (5) counting the content of the live spores, namely C is equal to T*N*10, wherein in a formula, C is the content of the fungal pesticide live spores, the unit is colony forming unit (CFU) / gram, T is dilution times corresponding to the counted flat plates, and N is average colony counts on a plurality of effectively-counted flat plates of the same dilution times. By the method, the content of the live spores of the fungal microbial pesticide can be measured simply, conveniently and quickly, and the detection accuracy is high.
Description
technical field [0001] The invention relates to a detection method of biological pesticides, in particular to a method for determining the content of live spores in fungal microbial pesticides. Background technique [0002] Microbial pesticides generally include bacterial, fungal and viral microbial pesticides. It is a pesticide developed by directly using the natural active substances produced by bacteria, fungi and viruses or the living organism itself to prevent and control plant diseases and insect pests. This type of pesticide has the advantages of strong selectivity, safety to humans, livestock, crops and the natural environment, no natural enemies, and no resistance. The amount of bacteria (especially the amount of viable bacteria) in microbial pesticides is an important factor that determines product quality and field control effects, so the amount of viable bacteria is an important technical indicator for the quality detection of microbial pesticide products. [0...
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IPC IPC(8): C12Q1/06
Inventor 袁善奎姜辉崔晓岚王一喆林荣华瞿唯钢胡承勇曲甍甍周艳明
Owner 农业部农药检定所
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