阅读说明：本技术 一种高效筛选病原菌拮抗菌的培养皿及其使用方法 (Culture dish for efficiently screening pathogenic bacteria antagonistic bacteria and use method thereof ) 是由 张萍华 张艳军 陈宝婷 苏盼 于 2019-12-06 设计创作，主要内容包括：本发明公开了一种高效筛选病原菌拮抗菌的培养皿和方法,其中培养皿包括皿盖l、皿盖2和皿体,材料是玻璃或聚碳酸酯,同时培养皿体中间含有一个多微孔结构的隔断层。本发明还提供了一种高效筛选病原菌拮抗菌的方法,包括：在培养皿A面和B面分部倒入分离菌种和培养病原菌所需的固体培养基,然后取适当稀释梯度的稀释液涂布至A面培养基,并在适宜条件下培养一定天数；将病原菌涂布至B面培养基,继续在适宜条件下培养,并观察B面抑菌圈位置,然后挑选A面上抑菌圈对应位置的菌种,即可用于复筛实验。因此该高效筛选病原菌拮抗菌的培养皿结构简单,提供的方法可以将菌种分离和初筛实验同时进行,从而大大提高筛选效率,降低筛选成本。(The invention discloses a culture dish and a method for efficiently screening antagonistic bacteria of pathogenic bacteria, wherein the culture dish comprises a dish cover l, a dish cover 2 and a dish body, the culture dish body is made of glass or polycarbonate, and the middle of the culture dish body contains a partition layer with a microporous structure. The invention also provides a method for efficiently screening pathogenic bacteria antagonistic bacteria, which comprises the following steps: pouring solid culture mediums required for separating strains and culturing pathogenic bacteria into the surface A and the surface B of the culture dish in parts, then coating diluent with appropriate dilution gradient to the surface A culture medium, and culturing for a certain number of days under appropriate conditions; and coating the pathogenic bacteria on a B surface culture medium, continuously culturing under a proper condition, observing the position of a B surface inhibition zone, and then selecting strains at the corresponding position of the inhibition zone on the A surface, so that the strains can be used for a re-screening experiment. Therefore, the culture dish for efficiently screening the antagonistic bacteria of the pathogenic bacteria has a simple structure, and the provided method can simultaneously carry out strain separation and preliminary screening experiments, thereby greatly improving the screening efficiency and reducing the screening cost.)

1. The utility model provides a culture dish of high-efficient screening pathogenic bacteria antagonistic bacteria which characterized in that: the culture dish includes ware lid l, ware lid 2 and the ware body, wherein contains a microporous structure's partition layer in the middle of the culture dish body, and the culture dish tow sides all can use.

2. The culture dish for efficiently screening antagonistic bacteria against pathogenic bacteria according to claim 1, wherein: the culture dish is made of glass or polycarbonate.

3. The culture dish for efficiently screening antagonistic bacteria against pathogenic bacteria according to claim 1, wherein: the culture dish is circular.

4. The culture dish for efficiently screening antagonistic bacteria against pathogenic bacteria according to claim 1, wherein: the partition layer of the microporous structure in the middle of the dish body is a glass sand core or other materials with the same function.

5. The culture dish for efficiently screening antagonistic bacteria against pathogenic bacteria according to claim 1, wherein: the culture dish can be modified by using the existing plastic culture dish.

6. The use method of the culture dish for efficiently screening pathogenic bacteria antagonistic bacteria according to claim 1, comprising the steps of: pouring a solid culture medium required by separating strains into the surface A of the culture dish, and pouring a solid culture medium required by culturing pathogenic bacteria into the surface B of the culture dish; diluting the collected sample by using a gradient dilution method, then coating the diluent with a proper dilution gradient on a culture medium on the surface A of a culture dish, and culturing under proper conditions; taking out the culture dish for culturing for a certain number of days, coating the pathogenic bacteria on the surface B of the culture dish, and continuously culturing under proper conditions; observing the bacteriostasis condition of the surface B, selecting the strain at the corresponding position of the surface A, storing the strain in a slant culture medium, and using the strain for subsequent re-screening.

Technical Field

The invention belongs to the technical field of biology, and particularly relates to a culture dish for efficiently screening antagonistic bacteria of pathogenic bacteria and a using method thereof.

Background

There are a variety of pathogenic bacteria in the natural environment that can cause plant, animal and human diseases. Although scientists have invented a plurality of medicaments which can effectively control pathogenic bacteria, long-term use of a single medicament can easily cause drug resistance of the pathogenic bacteria, thereby reducing the control effect. Screening for pathogenic bacteria antagonistic bacteria and discovery of new agents is therefore a long and important task.

The conventional screening process of the antagonistic bacteria mainly comprises the following steps: strain separation, primary screening and secondary screening. Firstly, separating strains by adopting a dilution coating plate method, and inoculating all obtained single colonies to a slant for preservation; secondly, inoculating the obtained single colony to a corresponding culture medium plate, and performing primary screening by using an agar block method; and finally, re-screening the preliminarily screened antagonistic bacteria by using an oxford cup method to finally obtain the optimal antagonistic bacteria. The existing bacteria separation and primary screening methods are time-consuming and labor-consuming and have low efficiency.

Disclosure of Invention

In order to overcome the defects, the invention aims to provide a culture dish for efficiently screening pathogenic bacteria antagonistic bacteria and a using method thereof, so that the screening work efficiency is improved, and the experiment cost is reduced.

In order to achieve the above object, the present invention provides a culture dish for efficiently screening antagonistic bacteria against pathogenic bacteria, which is characterized in that: the culture dish comprises a dish cover l, a dish cover 2 and a dish body, wherein the middle of the culture dish body comprises a partition layer with a microporous structure; the front and back sides of the culture dish can be used, wherein one side is used for strain separation, and the other side is used for culturing corresponding pathogenic bacteria.

The invention also provides a use method of the culture dish for efficiently screening the pathogenic bacteria antagonistic bacteria, which comprises the following steps: pouring a solid culture medium required by separating strains into the surface A of the culture dish, and pouring a solid culture medium required by culturing pathogenic bacteria into the surface B of the culture dish; diluting the collected sample by using a gradient dilution method, then coating the diluent with a proper dilution gradient on a culture medium on the surface A of a culture dish, and culturing under proper conditions; taking out the culture dish for culturing for a certain number of days, coating the pathogenic bacteria on the surface B of the culture dish, and continuously culturing under proper conditions; observing the position of the inhibition zone on the surface B, selecting the strain at the corresponding position on the surface A, storing the strain in a slant culture medium, and performing subsequent re-screening.

In conclusion, the culture dish for efficiently screening the pathogenic bacteria antagonistic bacteria has a simple structure, and the provided method can simultaneously carry out strain separation and preliminary screening experiments, so that the screening efficiency is greatly improved, and the screening cost is reduced.

Drawings

FIG. 1 is a culture dish for efficiently screening antagonistic bacteria of pathogenic bacteria.

FIG. 2 plastic petri dish modified double-sided petri dish.

FIG. 3B shows the inhibition zone of bacterial blight of rice.

FIG. 4 shows the antibacterial activity of antagonistic bacteria obtained by preliminary screening.

Detailed Description