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In-situ culture method for actinomyces in soil

A technology of in-situ culture and actinomycetes, which is applied in the field of microbial culture, can solve the problems of laborious and cumbersome work, high requirements for observation and counting, and small colonies, so as to reduce workload, avoid easily destroying the ecological environment of microorganisms, and have wide applicability Effect

Pending Publication Date: 2020-06-09
XINJIANG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] There are many shortcomings in the existing in situ culture technology of actinomycetes at home and abroad, such as: heat treatment, chemical treatment, addition of antibiotics and other methods are used in the separation process to remove the dominant but non-target strains in the environment, and the cultured colonies The present invention aims to provide a method for in-situ cultivation of actinomycetes in soil, which is prepared by mixing and sterilizing soil samples and culture medium. Substrate, after diluting the soil sample, inverting the plate culture, you can get a colony formed by the growth of a single actinomycete individual, and establish an effective isolation method for actinomycetes

Method used

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  • In-situ culture method for actinomyces in soil
  • In-situ culture method for actinomyces in soil
  • In-situ culture method for actinomyces in soil

Examples

Experimental program
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Effect test

Embodiment 1

[0032] A method for cultivating actinomycetes in situ in soil, comprising the following steps:

[0033] (1) Soil sample treatment: in a sterile environment, take a dry soil sample and sieve it to remove impurities and increase the density of the soil sample, collect the sieved soil sample, and seal it for later use;

[0034](2) Weigh 2000-2050 parts of unsterilized soil samples and 200-250 parts of culture medium prepared in step (1), place them in a centrifuge tube, and carry out high-pressure high-temperature sterilization at 121°C for 30 minutes. Steam enters, so as not to wet the soil sample, and prepare the culture substrate;

[0035] (3) Dilution: In the culture medium prepared in step (2), add 200-300 parts of unsterilized soil samples prepared in step (1), and mix well;

[0036] (4) Cultivation: Pour all the diluted soil samples in step (3) into a sterile petri dish, use a sterile applicator to smooth the soil samples, and add 30%-50% to the petri dish by weight perce...

Embodiment 2

[0046] In a sterile environment, take dry soil samples and sieve them for later use; weigh 2000 parts of soil samples and 250 parts of culture medium, place them in a centrifuge tube, and mix well; if the soil sample has high fertility, no culture medium can be added , directly weighed 2250 parts of soil samples, and then carried out autoclaving and high temperature sterilization at 121°C for 30 minutes to prepare the culture medium; add 200 parts of unsterilized soil samples to the culture medium and mix them thoroughly; the mixed soil samples Pour it all into a sterile petri dish, use a sterile applicator to level the soil sample, add 30%-50% sterile water or culture medium to the petri dish, use the applicator to make a second leveling, and seal it with a Membrane seal to prevent water evaporation, 25-35 ℃, culture 3-30 days.

Embodiment 3

[0048] In a sterile environment, take a dry soil sample and sieve it for later use; weigh 2000 parts of the soil sample and 200 parts of the culture medium, put them in a centrifuge tube, and mix them well; if the soil sample has high fertility, no culture medium can be added , directly weighed 2200 parts of soil samples, and then carried out autoclaving and high temperature sterilization at 121°C for 30 minutes to prepare the culture medium; add 200 parts of non-sterilized soil samples to the culture medium and mix them thoroughly; the mixed soil samples Pour it all into a sterile petri dish, use a sterile applicator to level the soil sample, add 30%-50% sterile water or culture medium to the petri dish, use the applicator to make a second leveling, and seal it with a Membrane seal to prevent water evaporation, 25-35 ℃, culture 3-30 days.

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Abstract

The invention discloses an in-situ culture method for actinomyces in soil. The method comprises the following steps: performing mixing and sterilization on 2025 parts of a soil sample and 225 parts ofa culture medium, preparing a culture matrix, diluting 250 parts of the soil sample, and performing plate pouring culture, so as to obtain a bacterial colony formed by single actinomyce individuals through growth. An effective isolation method of actinomyces is established, the method ensures that the workload is effectively reduced in the situation that mose actinomyce resource information is obtained; in addition, methods such as thermal treatment, chemical treatment and antibiotic addition are not implemented to the soil sample in the method, dominant but not target strains in the environment can be eliminated, therefore, the problem that microbial ecological environments can be easily damaged in a common method can be avoided, in-situ culture of the actinomyces is achieved, the numberof actinomyces which can be cultured is increased, great progress of actinomyces isolation technologies is made, and wide applicability to actinomyce resource protection and development is achieved.

Description

technical field [0001] The invention relates to the technical field of microorganism culture, and specifically, the invention relates to the technical field of a method for in-situ culture of actinomycetes in soil. Background technique [0002] Actinomyces are Gram-positive bacteria with branched mycelium. Actinomyces can be used as producers of antibiotics, vitamins, enzymes and enzyme inhibitors. They are a class of bacteria with important economic value and multiple uses. microorganism. But at present, the actinomycetes isolated by people only account for about 10% of all actinomycetes in the soil. In nature, most of them are uncultured microorganisms that cannot be isolated by traditional culture methods. Finding an isolation method suitable for actinomycetes is one of the important ways to mine uncultivated microbial resources and discover new species of actinomycetes and bacteria producing new active substances. Microorganisms are of great importance in terms of the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/04
CPCC12N1/20
Inventor 郭飞木沙江·托乎提马正海张民伟牛丽娟苟渔林富顺
Owner XINJIANG UNIVERSITY
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