Method for trapping soil microorganisms

A technology of soil microorganisms and microorganisms, which is applied in the field of microbial cultivation, can solve the problems of special microorganisms that are difficult to cultivate, cannot obtain all information of microorganisms, and is not suitable for growth, etc., and achieve the effect of expanding the separation depth, slowing down the reproduction speed, and eliminating the influence of shading

Pending Publication Date: 2021-02-19
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, when nutrient-rich beef extract peptone was used as a medium to isolate soil microorganisms in the laboratory, a large number of nutrient-poor microorganisms were not suitable for growth.
[0006] Secondly, due to the dilution of sterile water in the conventional plate culture method,

Method used

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  • Method for trapping soil microorganisms
  • Method for trapping soil microorganisms
  • Method for trapping soil microorganisms

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] In order to clarify the efficiency of microbial trap enrichment method D7 (referred to as enrichment method D7, the same below) for cultivating microorganisms, we compared it with the conventional culture method CK in terms of the number of cultivable colonies and the species of cultivable bacteria.

[0060] 1. Enrichment method D7:

[0061] S1: microbial trapping,

[0062] (1) After the soil sample is removed of impurities and larger lumps, put it into a glass petri dish with a diameter of 120 mm, the thickness of the soil layer is about 1.5 cm, and use distilled water to moisten the soil with a titration bottle;

[0063] (2) if figure 2 , 3 As shown, to prepare the microorganism trapping device, first apply glue on the edge of the microporous membrane a with a diameter of 50 mm and a pore size of 0.45 μm, and place a stainless steel flat-bottomed gasket b on the microporous membrane a; then add 3mL solid medium c, solid medium c contains 1.2% gellan gum and 1.0% v...

Embodiment 2

[0075] In order to clarify the culture temperature and humidity suitable for the bacteria isolated by the enrichment method D7, soil bacteria were cultivated according to the enrichment method D7 at different culture temperatures, and the number of cultivable bacteria isolated under different conditions was compared; by adding different amounts of Sterile water simulates low, medium and high humidity conditions, and analyzes the influence of humidity on the number of bacteria isolated.

[0076] Embodiment 2 is except that following test and result are different, all the other operation steps are identical with embodiment 1.

[0077] 1. The influence of culture temperature on the number and types of culturable bacteria

[0078] The culture devices were respectively placed at 25°C, 30°C, 37°C and room temperature (the temperature is not stable, culture at 20-35°C.

[0079] 2. Effect of soil moisture on the number and types of cultivable bacteria

[0080] The production of the ...

Embodiment 3

[0083] Further, in order to clarify the influence of trapping medium and separation medium on the number of cultivable bacteria, agar and gellan gum were used as trapping media to trap soil bacteria, and the influence of the two media on the number of cultivable bacteria was analyzed; The microbiological isolation medium was used to cultivate bacteria, and the differences in cultivable bacteria isolated from different media were compared, which provided a reference for the efficient use of the enrichment method D7 to isolate soil bacteria.

[0084] Embodiment 3 is except that following test and result are different, all the other operation steps are identical with embodiment 1.

[0085] 1. Effects of different trapping media on the number of cultivable bacteria

[0086] (1) When making the microorganism trapping device, a solid medium c is set to treat with 1.2% gellan gum and 1.2% agar as the supporting medium respectively;

[0087] (2) The culture device is placed in a 28°C i...

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Abstract

The invention discloses a method for trapping soil microorganisms. The method comprises the following steps: firstly, carrying out microorganism culture, putting soil into a glass culture dish, wetting the soil with distilled water, manufacturing a microorganism trapping device containing a solid culture medium, putting the device into the wet soil in the culture dish, manufacturing a microorganism culture device, and putting the microorganism culture device into an incubator for culturing; and then carrying out microorganism separation, mashing the solid culture medium, adding sterile water for standing, carrying out gradient dilution, coating an oligotrophic culture medium with a bacterium solution, carrying out blow-drying and culturing in the incubator, observing bacterial colony growth, and carrying out storage after purification and identification. According to the method, the soil microorganisms are trapped and domesticated in the soil through the solid culture medium, so that the soil microorganisms including bacteria which are difficult to culture and low in propagation speed can be obtained, and the separation depth of the soil microorganisms is expanded. The microorganisms obtained with the method can be used for morphological observation, classification and identification of the microorganisms, plant disease control, pesticide degradation, environmental protection and the like.

Description

technical field [0001] The invention relates to the technical field of microorganism cultivation, in particular to a method for trapping soil microorganisms. Background technique [0002] The soil has the nutrients, water, air, pH, osmotic pressure, temperature and other conditions required for the growth and development of various microorganisms, so the soil is a good environment for microorganisms to live, and contains almost all microbial populations, so the soil is the most abundant. "Strain Resource Library". The amount of microorganisms contained in the soil is huge, especially bacteria. It is estimated that each gram of soil contains 4000-7000 different bacterial species, with a total of about 1 billion (Yanyan, Chen Jian, Yanchangjiang, etc. using molecular biology Methods for technical identification of soil microorganisms [J]. Journal of Northeast Agricultural University, 2008,39(003):129-133.). [0003] Soil microorganisms are rich in species, but due to the lim...

Claims

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Application Information

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IPC IPC(8): C12N1/02C12M1/16C12M1/12C12M1/00C12R1/07C12R1/01C12R1/38C12R1/06
CPCC12M21/16C12M33/14C12N1/02
Inventor 卢灿华杨应美姜宁马俊红盖晓彤雷丽萍晋艳殷红慧高朝阳赵兴能夏振远
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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