Preparation and cultivation method for ustilaginoidea virens thin-wall conidia capable of preventing pollution

A rice smut fungus and a culture method technology, which is applied in the field of plant pathology, can solve the problems of high pollution probability, material pollution in petri dishes, and miscellaneous bacteria pollution on culture plates, and achieves the effects of convenient operation and pollution prevention.

Pending Publication Date: 2017-06-20
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The air around the culture dish is in a state of constant flow for sporulation culture in this type of incubator, and the probability of contamination of the materials in the culture dish is quite high. Usually, more than 50% of the culture plates will be contaminated by bacteria after 5 days of cultivation, even if some No obvious colonies of penicillium and other contaminating bacteria can be seen on the culture plate, but the probability of contamination that is invisible to the naked eye is still relatively high
Therefore, it is technically difficult to rule out the possibility of contamination by using a petri dish to prepare and cultivate conidia of rice smut

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0028] The preparation and cultivation of conidia of rice false smut bacteria on agar medium all adopt conventional technical specifications, that is, the culture technology of using culture dish in combination with agar medium is used for preparation and cultivation; there is no use of triangular flasks as culture utensils so far. , The spore preparation technology of the spore-forming culture of Aspergillus oryzae on the agar medium. The possible reason is the long-term dependence and habitual use of the petri dish, and it is generally believed that the spore production of pathogenic bacteria requires a good aeration environment, and the petri dish can just meet the good aeration.

[0029] After the common triangular flask is put on the bottle stopper, although form the poor closed environment of aeration, the contriver repeatedly tests and finds, the rice false smut bacterium can form conidia on the potato sucrose agar medium in this relatively closed environment; And The a...

Embodiment 1

[0038] Apply a kind of method for preparing and cultivating the conidia of the rice smut bacteria strain Uv-105 for preventing pollution of the present invention, and prepare and cultivate the conidia of the rice smut bacteria strain Uv-105, and operate as follows:

[0039] 1. Use a 250mL common triangular flask as a device for preparing spores, and clean the triangular flask for later use.

[0040] 2. Preparation of triangular flask type sporulation medium: use potato sucrose medium as sporulation medium, the proportion of this medium is: potato 100g, sucrose 10g, agar 20g, water 1000mL; 1 Prepared Erlenmeyer flasks, each filled with 30mL, after routine autoclaving and high-temperature sterilization, it will become the Erlenmeyer flask-type culture medium for future use.

[0041] 3. Transplantation of the mother thallus: use the conidia liquid of the rice smut bacteria strain Uv-105 as the mother thallus, and use a pipette to transplant 50 μL of the mother spore liquid into t...

Embodiment 2

[0045] Apply a method for preparing and cultivating conidia of the rice smut bacteria strain Uv-110 by applying a method for preparing and cultivating the conidia of the rice smut bacteria strain Uv-110 to prevent pollution, and implement according to the steps 1 to 5 of Example 1, the difference is only The bacterial strain of step 3 is Uv-110; As a result, the spores on a bottle of colonies were eluted with 15 mL of sterile water, and the concentration of spores obtained was 4.01×10 6 Individual / mL, parenchyma liquid without contamination by bacteria.

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PUM

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Abstract

The invention discloses a preparation and cultivation method for ustilaginoidea virens thin-wall conidia capable of preventing pollution. A conical flask can be used for effectively obstructing the characteristics of common microorganisms, a common conical flask is adopted to manufacture a utensil for culturing spores, and a potato cane sugar culture medium is taken as a spore production culture medium to prepare and cultivate the high-quality ustilaginoidea virens thin-wall conidia. The preparation and cultivation method comprises the following steps: 1) cleaning the common conical flask for standby use; 2) taking the potato cane sugar culture medium as the spore production culture medium to prepare a conical flask type culture medium; 3) transplanting matrix thalluses; 4) carrying out spore production culture under a temperature condition of 28DEG C; 5) carrying out elution by bacteria free water to collect new-generation thin-wall conidia. The preparation and cultivation method has the following advantages: 1) shake cultivation equipment is not required, and obtained conidia liquid finished product contains few mycelia, culture media, thallus metabolism excretion products and the like; 2) after the conical flask is provided with a conventional bottle stopper, common microorganisms can be prevented from entering, the conical flask can be favorably prevented from being polluted by infectious microbes, so that pollution can be effectively prevented, and a high-quality spore finished product can be obtained; 3) an operation for collecting the spores in the conical flask is more convenient than the operation for collecting the spores in a culture dish. According to the protection of the conical flask, spore production bacterial colonies can be put for a period of time at will without pollution, and a spore production state is kept so as to be convenient to connect with a working procedure which needs spores.

Description

technical field [0001] The present invention relates to phytopathological techniques. Specifically, it is a method for preparing and cultivating conidia of the rice false smut fungus to prevent pollution. Background technique [0002] Rice false smut is an important disease of rice, which can cause great losses to rice production. The disease is caused by the infection of rice smut fungus (Ustilaginoidea virens). Conidia are the asexual propagules of rice false smut, which play an important role in the disease cycle and epidemic of rice false smut. In the laboratory, conidia are also an indispensable material for the study of rice false smut. Studies related to the formation and development of spores, the survival adaptability or resistance to adverse environments, and the recognition and interaction between spores and hosts need to be used. Materials in the form of spores; relevant research on the development of new prevention and control drugs, such as the toxic effect ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N3/00C12R1/645
CPCC12N3/00
Inventor 张君成蒲相君张丽丽
Owner GUANGXI UNIV
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