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Method for removing bacterial contamination in fungi

A bacterial contamination and fungus technology, applied in the field of microorganisms, can solve the problems of large restrictions on antibiotics and insignificant bacteriostatic effect, and achieve the effect of excellent bacterial removal effect, avoiding the use of antibiotics, and short experimental period.

Pending Publication Date: 2020-01-17
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a method for removing bacterial contamination in fungi, especially bacterial contamination in fungi under solid medium plate culture conditions, for the lack of an efficient and stable bacterial removal system at present, which is greatly limited by antibiotics, and the bacteriostatic effect is not obvious. removal of

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  • Method for removing bacterial contamination in fungi
  • Method for removing bacterial contamination in fungi
  • Method for removing bacterial contamination in fungi

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Test materials:

[0038] Rice blast pathogen: Magnaporthe oryzae Guy-11 was used as a strain of Magnaporthe oryzae, which was preserved by the Fungal Disease Room of the Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences. In this example, Magnaporthe oryzae Guy-11 was contaminated by Escherichia coli (E. coli for short).

[0039] Specifically follow the steps below:

[0040] (1) Preparation of medium: According to the growth rate of Magnaporthe grisea in the medium suitable for fungal culture, the CM medium used to remove bacterial contamination in fungi was screened. Among them, the medium (CM) for plate culture: 10g glucose, 2g peptone, 1g yeast extract, 1g casamino acid, 50mL nitrogen salt (20×Nitrate salts; trace elements; vitamins), 18g agar, ddH2O constant volume to 1000mL, 1M NaOH solution to adjust the pH to 6.5, sterilized by high pressure steam at 121℃ for 15min.

[0041] (2) Pre-treatment of the culture medium: Cut ou...

Embodiment 2

[0048] Select the rice blast fungus Guy-11 polluted by agrobacterium as object, and other are with embodiment 1, carry out agrobacterium removal experiment to blast fungus Guy-11 bacterial strain, experimental result is as follows figure 2 As shown, there is no difference in effect between T1 and T2. The blast fungus Guy-11 was purified after treatment, and its growth rate on the medium, colony behavior, sporulation and other traits were all restored and improved. T1 saves the use of antibiotics relative to T2. After T3 treatment, there was some improvement, but the fungal colonies could not be completely purified. After T4 treatment, the bacterial contamination and culture traits of Magnaporthe grisea Guy-11 strain were not improved, so it could no longer be used for scientific research and teaching. Genomic DNA was extracted from strains T1, T2, T3, and T4, and PCR detection was performed using Agrobacterium-specific primers. The results showed that fragments of Agrobacte...

Embodiment 3

[0050] The Magnaporthe grisea 2539 bacterial strain contaminated by Agrobacterium was selected as the object, and the others were the same as in Example 1, and the Agrobacterium removal experiment was carried out on the Magnaporthe oryzae 2539 bacterial strain. Experimental results such as image 3 As shown, there is no difference in effect between T1 and T2, and the treated Magnaporthe grisea 2539 strain grows well on the medium, the colony is purified, and the traits such as sporulation are restored. After T3 treatment, the strains were partially improved, but the fungal colonies could not be completely purified. After T4 treatment, the bacterial contamination and culture traits of Magnaporthe grisea 2539 strain were not improved, so it could no longer be used for scientific research and teaching. Genomic DNA was extracted from strains T1, T2, T3, and T4, and PCR detection was performed using Agrobacterium-specific primers. The results showed that fragments of Agrobacteriu...

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Abstract

The invention discloses a method for removing bacterial contamination in fungi. The method comprises the following steps: (1) carrying out high temperature sterilization on a prepared fungus culture medium, pouring the sterilized fungus culture medium into a sterile culture dish, and cutting one or more than one small hole on a culture medium flat plate after the culture medium is solidified; (2)selecting samples in proper size and placing the samples in the small hole; (3) covering a sterile cover glass, and slightly pressing the periphery of the cover glass to make the cover glass closely contact with the culture medium so as to ensure that no air bubbles are left between the culture medium and the cover glass; (4) putting the culture medium obtained in the step (3) into a constant temperature and humidity incubator for culture; and (5) picking new fungus hyphae growing around the cover glass, and re-inoculating the fungus hyphae to a new fungus culture medium for culture, thereby obtaining fungus colonies. The invention has the advantages of excellent bacteria removing effect, no rebound, wide applicability, simple operation, small workload and short experimental period.

Description

technical field [0001] The invention belongs to the technical field of microbes, in particular to a method for removing bacterial contamination in fungi. Background technique [0002] In the process of fungal cultivation, including the cultivation of edible fungi, the preservation, cultivation and utilization of beneficial fungal strains, and the isolation, cultivation and research of pathogenic fungi, the contamination of miscellaneous bacteria is ubiquitous. Common microorganisms that cause pollution include bacteria, fungi , mycoplasma and mites. Fungi such as molds and yeasts grow rapidly under culture conditions, and the contamination they cause is easily observed, and the strains can also be identified based on their colony characteristics on the medium and microscopic observations, while bacterial contamination exists A certain incubation period, even after several generations of culture, it will appear, so the removal of bacteria is more difficult and important. ...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N1/02C12R1/77C12R1/645
CPCC12N1/02C12N1/14
Inventor 王教瑜邱海萍施笑笑柴荣耀张震毛雪琴王艳丽孙国昌
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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