阅读说明：本技术 一种丝状真菌分离纯化专用培养皿装置 (Special culture dish device of filamentous fungi separation and purification ) 是由 陈健 周翔 于 2019-09-29 设计创作，主要内容包括：一种丝状真菌分离纯化专用培养皿装置,包括样品培养皿和分离纯化盖,样品培养皿设有底托,在底托周边设有隔离外壁,在底托的正中设有中心小环,中心小环内为样品室,在中心小环内倒入培养基,所述分离纯化盖下表面设有分离室,分离室由中间分隔环分隔而成,分离室设有分离层选择性培养基层,所述中间分隔环设置在中心小环与隔离外环之间；中间分隔环的侧部上开有分离孔。可直接将高污染本底土壤样品直接定植在样品培养环中,利用反向孢子弹射方法实现细菌和真菌的分离,通过同心环装置隔离可以实现对真菌和放线菌的分离,其中利用气生菌丝爬避性和菌丝穿透性实现真菌和放线菌在分离环中分离。可用于高污染细菌本底的真菌分离和纯化分析。(A culture dish device special for separation and purification of filamentous fungi comprises a sample culture dish and a separation and purification cover, wherein the sample culture dish is provided with a bottom support, an isolation outer wall is arranged at the periphery of the bottom support, a central small ring is arranged in the center of the bottom support, a sample chamber is arranged in the central small ring, a culture medium is poured into the central small ring, a separation chamber is arranged on the lower surface of the separation and purification cover and is formed by separating a middle separating ring, the separation chamber is provided with a separation layer selective culture medium layer, and the middle separating ring is arranged between the central small ring and the isolation outer ring; the side part of the middle separating ring is provided with a separating hole. The high-pollution background soil sample can be directly planted in the sample culture ring, bacteria and fungi are separated by using a reverse spore ejection method, the fungi and actinomycetes can be separated by using a concentric ring device, and the fungi and the actinomycetes are separated in the separation ring by using aerial hypha creeping property and hypha penetrability. Can be used for fungus separation and purification analysis of high-pollution bacterial background.)

1. the utility model provides a special culture dish device of filamentous fungi separation and purification which characterized in that: comprises a sample culture dish and a separation and purification cover,

the sample culture dish is provided with a bottom support, an isolation outer wall is arranged on the periphery of the bottom support, a central small ring is arranged in the center of the bottom support, a sample chamber is arranged in the central small ring, a culture medium layer is arranged in the central small ring, the height of the culture medium layer is 2/3 of the height of the central small ring,

The lower surface of the separation and purification cover is provided with a separation chamber which is formed by separating a middle separating ring,

The separation chamber is provided with a separation layer selective culture medium layer,

The middle separating ring is arranged between the central small ring and the isolating outer ring, the height of the middle separating ring is 1.5 ~ 2 times of the height of the central small ring, and the distance between the bottom edge of the middle separating ring and the upper edge of the central small ring is 1 ~ 1.5.5 times of the height of the central small ring;

The side part of the middle separating ring is provided with a separating hole.

2. the culture dish device special for filamentous fungi separation and purification according to claim 1, wherein: the culture medium is water agar or semisolid agar.

3. the culture dish device special for filamentous fungi separation and purification according to claim 1, wherein: the separating layer culture medium is a separating selective culture medium which takes a PDA culture medium as a substrate and is added with VB1, a target test substrate and allelochemicals.

4. The culture dish device special for filamentous fungi separation and purification according to claim 3, wherein: the allelochemicals are one or more of plant extract, 3-hexen-1-ol, phellandrene, 2-carene, alpha-pinene or camphene, and the adding amount of the allelochemicals is 1 mu g of the allelochemicals added per 1000ml of the culture medium substrate.

5. The culture dish device special for filamentous fungi separation and purification according to claim 1, wherein the middle separating ring is provided with an inner ring and an outer ring, and the interval between the inner ring and the outer ring is 10 ~ 20 mm.

6. the culture dish device special for filamentous fungi separation and purification according to claim 5, wherein: the separation holes on the inner ring are diagonally opened by 2, and the separation holes on the outer ring are diagonally opened by 4.

7. The culture dish device special for filamentous fungi separation and purification as claimed in claim 1, 5 or 6, wherein the separation hole is opened from the bottom edge of the middle separation ring to 2/3 of the middle separation height, and the length of the separation hole is 8 ~ 12 mm.

Technical Field

The invention relates to biotechnology detection, in particular to a culture dish device special for separation and purification of filamentous fungi.

Background

Fungi separation and purification research generally comes from samples with high environmental pollution background, wherein a large amount of bacillus is always accompanied with growth of filamentous fungi when the soil samples are separated by a conventional streaking and diluting method, so that strain separation and purification work are difficult.

Disclosure of Invention

The invention aims to solve the technical problem of the prior art and provides a culture dish device special for separation and purification of filamentous fungi without repeated streaking separation or dilution separation.

The technical problem to be solved by the invention is realized by the following technical scheme, and the culture dish device special for separation and purification of the filamentous fungi is characterized in that: comprises a sample culture dish and a separation and purification cover,

The sample culture dish is provided with a bottom support, an isolation outer wall is arranged on the periphery of the bottom support, a central small ring is arranged in the center of the bottom support, a sample chamber is arranged in the central small ring, a culture medium layer is arranged in the central small ring, the height of the culture medium layer is 2/3 of the height of the central small ring,

The lower surface of the separation and purification cover is provided with a separation chamber which is formed by separating a middle separating ring,

The separation chamber is provided with a separation layer selective culture medium layer,

the middle separating ring is arranged between the central small ring and the isolating outer ring, the height of the middle separating ring is 1.5 ~ 2 times of the height of the central small ring, and the distance between the bottom edge of the middle separating ring and the upper edge of the central small ring is 1 ~ 1.5.5 times of the height of the central small ring;

The side part of the middle separating ring is provided with a separating hole.

The technical problem to be solved by the invention can be further realized by the following technical scheme, wherein the culture medium is water agar or semisolid agar. After the culture medium is solidified, directly pressing the soil to be detected into a sample chamber ring paved with the culture medium, and dripping a drop of sterile water or dilute water agar to prevent the soil from drifting;

the technical problem to be solved by the invention can be further realized by the following technical scheme, wherein the separation layer culture medium is a separation selective culture medium which takes a PDA culture medium as a substrate and is added with VB1, a target test substrate and allelochemicals.

the technical problem to be solved by the invention can be further realized by adopting the following technical scheme that the allelochemicals are one or more of plant extract, 3-hexen-1-ol, phellandrene, 2-carene, alpha-pinene or camphene, and the adding amount of the allelochemicals is 1 mu g of the culture medium substrate added per 1000 ml.

the technical problem to be solved by the invention can be further solved by the following technical scheme that the middle separation ring is provided with an inner ring and an outer ring, and the distance between the inner ring and the outer ring is 10 ~ 20 mm.

The technical problem to be solved by the invention can be further realized by the following technical scheme that the separation holes on the inner ring are diagonally opened by 2, and the separation holes on the outer ring are diagonally opened by 4.

The technical problem to be solved by the invention can be further realized by the following technical scheme that the separation hole is opened from the bottom edge of the middle separation ring to the position 2/3 of the middle separation height, and the length of the separation hole is 8 ~ 12 mm.

compared with the prior art, the method can directly fix the high-pollution background soil sample in the sample culture ring, realize the separation of bacteria and fungi by using a reverse spore ejection method, realize the separation of the fungi and the actinomycetes by the isolation of a concentric ring device, and realize the separation of the fungi and the actinomycetes in the separation ring by using the creeping property and the penetrability of aerial hyphae. Can be used for fungus separation and purification analysis of high-pollution bacterial background.

Drawings

FIG. 1 is a schematic structural view of the present invention;

Fig. 2 is a view showing the structure of the spacer ring.

Detailed Description

the following further describes particular embodiments of the present invention to facilitate further understanding of the present invention by those skilled in the art, and does not constitute a limitation to the right thereof.

A culture dish device special for separating and purifying filamentous fungi, which comprises a sample culture dish 2 and a separating and purifying cover 1,

The sample culture dish is provided with a bottom support, an isolation outer wall 6 is arranged on the periphery of the bottom support, a central small ring 5 is arranged in the center of the bottom support, a sample chamber 3 is arranged in the central small ring, a culture medium layer 4 is arranged in the central small ring, the height of the culture medium layer is 2/3 of the height of the central small ring,

The lower surface of the separation and purification cover is provided with a separation chamber which is formed by separating a middle separating ring,

a separation layer selective culture medium layer 8 is arranged in the separation chamber,

the middle separating ring is arranged between the central small ring and the isolating outer ring, the height of the middle separating ring is 1.5 ~ 2 times of the height of the central small ring, and the distance between the bottom edge of the middle separating ring and the upper edge of the central small ring is 1 ~ 1.5.5 times of the height of the central small ring;

The side part of the middle separating ring is provided with a separating hole.

The culture medium is water agar or semisolid agar. After the culture medium is solidified, the soil to be detected is directly pressed into the sample chamber ring paved with the culture medium, and a drop of sterile water or dilute water agar can be dropped to prevent the soil from drifting.

The separating layer culture medium is a separating selective culture medium which takes a PDA culture medium as a substrate and is added with VB1, a target test substrate and allelochemicals.

The allelochemicals are one or more of plant extract, 3-hexen-1-ol, phellandrene, 2-carene, alpha-pinene or camphene, and the adding amount of the allelochemicals is 1 mu g of the allelochemicals added per 1000ml of the culture medium substrate.

The middle separating ring is provided with an inner ring 7 and an outer ring 9, and the interval between the inner ring and the outer ring is 10 ~ 20 mm.

the separation holes 10 on the inner ring are diagonally opened by 2, and the separation holes on the outer ring are diagonally opened by 4.

the separation hole is formed from the bottom edge of the middle separation ring to 2/3 of the middle separation height, and the length of the separation hole is 8 ~ 12 mm.

the following specifically describes the embodiments of the present invention, and 1 example is provided, however, the practice of the present invention is not limited to the following example.

Specifically, the method comprises the following steps: the sample culture dish bottom support is formed by PE injection molding, the diameter of the bottom surface is 89mm, and the height of the isolation outer wall is 35 mm.

The diameter of the small central ring is 20mm, and the height of the small central ring is 8 mm.

the top separation chamber, outer circle diameter 92mm, the height is 35 mm. The spacing between the middle separating rings is 15mm, and the height of the separating rings is 11.2mm. A separation hole is arranged between the inner ring and the outer ring of the separation ring, the separation hole is 2/3 with the middle separation height, and the length of the hole is 10 mm. The inner ring is diagonally opened by 2, and the outer ring is diagonally opened by 4.

The target separation layer culture medium is a separation selective culture medium which takes a PDA culture medium as a substrate and is added with VB1, a target test substrate and allelochemicals.

VB1, adding 6 ~ 10mg/L, adding 4 ~ 6% of cellulose for a target test substrate by mass percent, wherein the allelochemicals are one or more of plant extract, 3-hexen-1-ol, phellandrene, 2-carene, alpha-pinene or camphene, and the adding amount of the allelochemicals is 1 mu g of the culture medium substrate added per 1000 ml.

(1) The device is simple in sample preparation and convenient to test, and is suitable for separating fungi from high-pollution samples;

(2) The device can separate fungi, actinomycetes and bacteria respectively, and is also suitable for separating bacteriophage or separating microorganisms which are not polluted by the bacteriophage;

(3) The manufacturing cost is low, and the use separation efficiency is high;

(4) The method is used for separation and purification analysis of fungi of more than thirty complex samples, and the coincidence rate is more than 95 percent;

(5) Purification and isolation of bacteria or phages in a sample of isolated contaminants can be used by inversion.

In order to illustrate the present invention more clearly, the following examples are given without any limitation to the scope of the present invention.