A petri dish for displaying microorganisms and a method of making the same

Abstract

The application provides a culture dish for displaying microorganisms and a manufacturing method thereof, and relates to the field of microorganisms.The culture dish for displaying microorganisms comprises a culture dish, a culture medium located in the culture dish and having microorganisms cultured on the surface thereof, a jelly wax layer covering the surface of the culture medium, and an epoxy resin layer covering the surface of the side of the jelly wax layer away from the culture medium.The culture medium with the microorganisms cultured is encapsulated by the jelly wax layer, the jelly wax layer has good sealing performance and stability for the culture medium, and has high transparency, is not easy to be contaminated, and does not require strict storage conditions.The jelly wax layer is further encapsulated by the epoxy resin layer, no reaction occurs between the two layers, the contact surface is tightly attached, the transparency is high, the surface layer is hard and not easy to be damaged, the problem of large elasticity, easy contamination and inconvenience in carrying of the jelly wax layer is overcome, and long-term storage and repeated use of the culture dish can be ensured.

Description

Technical Field

[0001] This invention relates to the field of microbiology, and more specifically to a petri dish for displaying microorganisms and a method for preparing the same. Background Technology

[0002] With the widespread use and promotion of various microbial fertilizers such as agricultural microbial agents, bio-organic fertilizers, compound microbial fertilizers, and soil remediation agents, soil problems such as soil compaction, low soil organic matter, soil salinization, and soil microbial imbalance have been largely solved. At the same time, the biological control effect of microbial fertilizers in the field of crop pest and disease control has the advantages of high efficiency, long-lasting effect, and green environmental protection, which has made it increasingly valued by the industry and recognized by farmers.

[0003] The functions of microbial fertilizer strains are mainly divided into growth-promoting, biocontrol, and decomposition / composting functions. Different strains have different functions. For example, strains with growth-promoting functions include Bacillus subtilis, Bacillus megaterium, and Aquatic Rahn's bacterium; strains with pest and disease control functions include Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus thuringiensis, and other Bacillus species, as well as Trichoderma, Streptomyces, and Paecilomyces lilacinus; and strains with decomposition / composting functions include Bacillus, Aspergillus, and yeast.

[0004] Besides those in the agricultural field, such as microbial research institutions and fertilizer manufacturers, who are familiar with microorganisms, microbial fertilizer distributors, retailers, and especially farmers who are the direct users of microbial fertilizers are gradually paying more attention to the microbial strains used in microbial fertilizers and their functions, and have put forward an urgent need for more convenient and intuitive ways to see and understand the microbial strains.

[0005] In the field, the effects of microbial fertilizers can be indirectly observed through soil improvement indicators, crop growth, disease control coefficients, and fruit quality and yield. However, the core of microbial fertilizers lies in the functional microbial strains. To allow promoters and users to more intuitively see the form and function of these microorganisms, it is necessary to create culture dishes with microbial colonies and qualitative testing capabilities for direct demonstration. However, ordinary microbial culture dishes are prone to drying out and contamination at room temperature and require storage in a 4°C freezer. Although contamination is less likely in a freezer, the culture medium is prone to leaching water, so even under such stringent conditions, preservation is not possible for extended periods. More importantly, they cannot be directly used for demonstration and explanation in fields, agricultural fairs, or exhibitions. The demonstration function of microbial culture dishes has certain limitations.

[0006] This problem exists not only in agriculture but also in education. When students learn about microorganisms, they are usually limited to abstract descriptions in textbooks and cannot directly observe the morphology and function of microorganisms. Even if microorganisms are cultured in petri dishes in advance, they cannot be reused, which is a great limitation. Summary of the Invention

[0007] Therefore, the technical problem to be solved by the present invention is to overcome the defects of existing microbial culture dishes that are prone to drying out and contamination at room temperature, and prone to water separation when stored in a low-temperature refrigerator, making them inconvenient to carry and unable to be reused. Thus, the present invention provides a culture dish for displaying microorganisms and a method for making the same.

[0008] To achieve the above objectives, the present invention provides the following technical solution:

[0009] In a first aspect, the present invention provides a culture dish for displaying microorganisms, comprising:

[0010] Petri dish;

[0011] A culture medium is located inside the petri dish, and microorganisms are cultured on its surface.

[0012] A jelly-like wax layer covers the surface of the culture medium;

[0013] An epoxy resin layer covers the surface of the jelly wax layer facing away from the culture medium.

[0014] Furthermore, the thickness of the culture medium is 3.5~6.5mm; the thickness of the jelly wax layer is 4.0~6.0mm; and the thickness of the epoxy resin layer is 3.0~5.5mm.

[0015] Furthermore, the microorganisms include fungi and / or bacteria.

[0016] Furthermore, the microorganisms include at least one of Bacillus amyloliquefaciens, Bacillus megaterium, Bacillus belesiensis, Bacillus subtilis, Aquatic Larynella, Pseudomonas proteus, Streptomyces violaceum, Streptomyces leucosus, Streptomyces rubrum, Streptomyces olvarioliticum, Streptomyces loucheri, Saccharomyces mullerii, and Trichoderma harzianum.

[0017] Furthermore, the culture medium is selected from any one of NA medium, Gao's No. 1 medium, YPD medium, PDA medium, and rhizobium medium.

[0018] Furthermore, the composition of the NA medium is as follows: 10.0g peptone, 3.0g beef meal, 5.0g NaCl, 20.0g agar, 1L distilled water, pH 7.0~7.2.

[0019] Furthermore, the composition of the Gao's No. I culture medium is as follows: 20.0g soluble starch, 0.5g NaCl, 1.0g KNO3, 0.5g K2HPO4, 0.5g MgSO4·7H2O, 0.01g FeSO4·7H2O, 20.0g agar, 1L distilled water, pH 7.2~7.4.

[0020] Furthermore, the YPD culture medium consists of: 10.0g yeast extract, 20.0g peptone, 20.0g glucose, 20.0g agar, 1L distilled water, and natural pH.

[0021] Furthermore, the PDA culture medium consists of: 1L potato filtrate, 20.0g glucose, 20.0g agar, and natural pH. The potato filtrate is prepared from 200g of potatoes.

[0022] Furthermore, the composition of the rhizobium culture medium is as follows: 1.0g yeast extract, 0.5g K2HPO4, 0.2g MgSO4·7H2O, 0.1g NaCl, 10.0g mannitol, 5mL 0.5% Congo red, 20.0g agar, 1L distilled water, pH 6.8~7.0.

[0023] Furthermore, the culture medium is selected from any one of the following: starch-solubilizing function qualitative detection medium, fat-solubilizing function qualitative detection medium, protein-solubilizing function qualitative detection medium, cellulose-solubilizing function qualitative detection medium, aniline blue medium, phosphorus-solubilizing function qualitative detection medium, and potassium-solubilizing function qualitative detection medium.

[0024] Furthermore, the composition of the qualitative detection medium for starch-solubilizing function is as follows: 10.0g peptone, 3.0g beef powder, 5.0g NaCl, 10.0g soluble starch, 20.0g agar, 1L distilled water, pH 7.0.

[0025] Furthermore, the preparation method of the qualitative detection medium for lipolysis function is as follows: Weigh 10.0g of peptone, 5.0g of NaCl, and 0.1g of CaCl2·2H2O into 800mL of distilled water, dissolve them completely, adjust the pH to 7.2, add 20.0g of agar, make up to 1L, sterilize at 121℃ for 30min, cool to 65℃, add 10mL of Tween80 sterilized separately at 121℃ for 30min, and mix well.

[0026] Furthermore, the preparation method of the protein functional qualitative detection medium is as follows: Weigh 2.0g of casein, moisten with 5mL of 0.5mol / L NaOH, add 150mL of distilled water, stir in a boiling water bath until completely dissolved, add distilled water to 800mL, then add 3.0g of beef extract, 5.0g of NaCl, and 0.1g of CaCl2·2H2O, dissolve thoroughly, add 20.0g of agar, mix evenly, bring the volume to 1L with distilled water, set the pH to natural, and sterilize at 121℃ for 30min.

[0027] Furthermore, the composition of the qualitative detection medium for cellulose dehydrogenation function is as follows: CMC-Na 10.0g, (NH4)2SO4 4.0g, K2HPO4 6.5g, MgSO4·7H2O 0.5g, peptone 1.0g, agar 20.0g, water 300mL, pH natural.

[0028] Further, the preparation method of the aniline blue medium is as follows: weigh 10.0g of yeast powder and 20.0g of glucose into 800mL of distilled water, dissolve them completely, add 20.0g of agar, make up to 1L, set the pH to natural, sterilize at 121℃ for 30min, cool to 65℃, add 10mL of aniline blue stock solution sterilized separately at 121℃ for 30min, and mix well.

[0029] Furthermore, the composition of the qualitative detection medium for phosphorus solubility is: 10.0g glucose,

[0030] 0.5g (NH4)2SO4, 0.3g NaCl, 0.3g KCl, 0.3g MgSO4·7H2O, 0.03g FeSO4·7H2O, 0.03g MnSO4·H2O, 0.2g egg lecithin, 1g CaCO3, 0.5g yeast powder, 20.0g agar, 1L distilled water, pH 7.0~7.2.

[0031] Furthermore, the composition of the potassium solubilization function qualitative detection medium is as follows: glucose 10.0g, Na2HPO4 0.2g, MgSO4·7H2O 0.2g, NaCl 0.2g, CaSO4 0.2g, CaCO3 5.0g, potassium feldspar powder 2.5g, agar 20.0g, distilled water 1L, pH natural.

[0032] Furthermore, the culture dish is made of glass or plastic.

[0033] Secondly, the present invention provides a method for preparing the culture dish displaying microorganisms, comprising the following steps:

[0034] (1) Pour the sterilized culture medium into the sterilized petri dish and let it solidify;

[0035] (2) Inoculate and activate microorganisms on the surface of the culture medium, and then culture them;

[0036] (3) Clean air blown until there are no water stains on the edge and surface of the culture medium;

[0037] (4) Pour melted jelly wax onto the surface of the culture medium and let it solidify to form a jelly wax layer;

[0038] (5) Pour the prepared epoxy resin adhesive onto the surface of the jelly wax layer and let it solidify to form an epoxy resin layer.

[0039] Furthermore, in step (4), the temperature of the melted jelly wax is 70~75°C.

[0040] Further, in step (5), the preparation method of the epoxy resin adhesive includes: taking 3 parts of the A glue of the crystal epoxy resin dripping, bathing it in a water bath at 65°C for 15 minutes, taking 1 part of the B glue of the crystal epoxy resin dripping and pouring it into the A glue, and stirring evenly.

[0041] Furthermore, step (2) includes a step of performing qualitative detection of microbial function after culturing the microorganisms.

[0042] Furthermore, the qualitative detection of microbial functions includes: qualitative detection of starch-dissolving function, qualitative detection of fat-dissolving function, qualitative detection of protein-dissolving function, qualitative detection of cellulose-dissolving function, qualitative detection of lignin-dissolving function, qualitative detection of phosphorus-dissolving function, qualitative detection of potassium-dissolving function, and qualitative detection of antibacterial function.

[0043] Furthermore, the method for qualitative detection of starch-dissolving function includes: preparation of staining solution: dilute Lugol's iodine solution stock solution 25 times with sterile distilled water to prepare 25× Lugol's iodine solution, add 7% concentrated hydrochloric acid to 25× Lugol's iodine solution and mix well; pour the prepared staining solution into the culture dish for culturing microorganisms, cover the culture medium, soak and stain for 3 minutes, and then pour off the iodine solution.

[0044] Further, the method for qualitative detection of cellulose desiccation function includes: preparation of rinsing solution: sterilize distilled water, cool to room temperature, and add streptomycin sulfate to a final concentration of 50 μg / mL; preparation of Congo red staining solution: sterilize 0.2% Congo red solution, cool to room temperature, and add streptomycin sulfate to a final concentration of 50 μg / mL; preparation of rinsing solution: sterilize 1% NaCl solution, cool to room temperature, and add streptomycin sulfate to a final concentration of 50 μg / mL; rinse the culture dish containing microorganisms with the prepared rinsing solution until the spores around the colonies are rinsed clean; cover the culture medium with the prepared Congo red staining solution, stain for 4 hours, and discard the staining solution; soak and rinse the stained culture dish twice with the prepared rinsing solution, 7-10 min each time, and discard the rinsing solution.

[0045] Furthermore, methods for activating microorganisms include the three-zone streak method and the point analysis method.

[0046] Unless otherwise specified, the sterilization methods for different petri dishes and culture media shall be those conventionally chosen in the art, such as:

[0047] The sterilization method for glass petri dishes is: moist heat sterilization at 121℃ for 30 minutes;

[0048] The sterilization method for plastic petri dishes is: ethylene oxide sterilization;

[0049] The sterilization methods for the culture medium are: 121℃ moist heat sterilization for 30 min, or 121℃ moist heat sterilization for 15 min, or 115℃ moist heat sterilization for 30 min.

[0050] Different microorganisms require different culture media and inoculation, activation, and culturing methods. Unless otherwise specified, conventional methods used in this field are employed, such as:

[0051] Bacteria such as Bacillus amyloliquefaciens, Bacillus megaterium, Bacillus belye, and Bacillus subtilis were cultured using NA medium and activated by the three-zone streak method, and cultured at 30°C for 48 hours.

[0052] Fungi were cultured using PDA and YPD media, and activated by electrolysis at 28°C for 4–10 days.

[0053] Streptomyces was cultured using Gao's No. 1 medium, and the activation method was the three-zone streak method. The culture was carried out at 28°C for 7 days.

[0054] Aquatic Rahn's bacteria were cultured using rhizobium culture medium and activated using the three-zone streak method, and cultured at 35°C for 24 hours.

[0055] The technical solution of this invention has the following advantages:

[0056] 1. The microbial culture dish provided by this invention encapsulates the culture medium containing microorganisms through a jelly wax layer. The jelly wax layer provides excellent sealing and stability for the culture medium, has high transparency, is not easily contaminated, and does not require stringent storage conditions. Furthermore, the jelly wax layer is further encapsulated by an epoxy resin layer. There is no reaction between the two layers, the contact surfaces adhere tightly, the transparency is high, and the surface is hard and not easily damaged. This overcomes the problems of the jelly wax layer being highly elastic, easily soiled, and inconvenient to carry, ensuring long-term storage and repeated use of the culture dish. Therefore, this culture dish has the advantages of being resistant to direct touch, easy to carry, can be displayed in any setting, is not easily contaminated, and can be reused. It solves the defects of ordinary culture dishes, which are easily contaminated and dry out in a short period, requiring constant remaking, and require sealing film to prevent opening and touching for observation.

[0057] 2. Experimental verification shows that the culture dishes for displaying microorganisms provided by this invention can be stored for a long time at low temperature (4~10℃), room temperature (15~30℃), or even high temperature (35~45℃), and also have excellent antifreeze properties.

[0058] 3. The microbial culture dish provided by this invention can not only be placed and displayed in the laboratory for a long time, but also helps sales personnel, distributors, farmers, and other consumers to see and understand the morphology, types, and different functions of functional strains in microbial fertilizers more directly and intuitively. On the one hand, it is beneficial for manufacturers to introduce their products in depth and promote sales, such as as a tool for online live broadcasts or on-site demonstrations at trade fairs, greatly promoting the rapid promotion of microbial fertilizers and significantly improving their sales efficiency. On the other hand, it can also be used in fields in different seasons and regions to explain to farmers, making it easier for farmers and other consumers to understand and accept microbial fertilizers, thus promoting their application. Therefore, the application of this culture dish increases the promotion and application area of ​​fertilizers, thereby improving soil problems, preventing the occurrence of pests and diseases, improving the quality of agricultural products, and increasing farmers' income.

[0059] 4. The culture dish for displaying microorganisms provided by this invention can be used as a teaching aid in the classroom, becoming a means for teachers to explain microorganisms. This allows students to understand microbial strains not only by the abstract descriptions of microorganisms in books, but also to directly observe the morphology and function of microorganisms, thereby increasing students' interest in exploring the field of microorganisms.

[0060] 5. The microbial culture dish provided by the present invention is simple to make, easy to operate, and low in cost, making it easy to promote and apply on a large scale in industry.

[0061] 6. The microbial culture dishes provided by this invention, through optimization of the qualitative detection method for starch solubility, solve the defect that the blue color of the culture medium obtained by conventional qualitative detection methods fades in a short time, resulting in unstable qualitative results; through optimization of the qualitative detection method for cellulose solubility, solve the defect that the culture medium obtained by conventional qualitative detection methods is contaminated by spores scattered by the functional bacteria, making it difficult to observe qualitative results. The above improvements ensure the stability of the culture dishes and are more conducive to the long-term display of microbial functions and the repeated use of the culture dishes. Attached Figure Description

[0062] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0063] Figure 1 This is an external view of the petri dish prepared according to Embodiment 1 of the present invention;

[0064] Figure 2This is a diagram of the internal structure of the petri dish prepared according to Example 1 of the present invention;

[0065] Figure 3 This is an external view of the petri dish prepared according to Embodiment 6 of the present invention;

[0066] Figure 4 This is a diagram of the internal structure of the petri dish prepared in Example 6 of the present invention.

[0067] Figure 5 This is an external view of the petri dish prepared in Embodiment 13 of the present invention;

[0068] Figure 6 This is a diagram of the internal structure of the petri dish prepared in Example 13 of the present invention. Detailed Implementation

[0069] The following embodiments are provided to better understand the present invention and are not limited to the preferred embodiments described. They do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0070] The crystal epoxy resin epoxy resin was purchased from Dongguan Tegu New Materials Co., Ltd., model TK-6830.

[0071] The transparent jelly wax was purchased from Foshan Xiaohuoba Biotechnology Co., Ltd., and is transparent, smokeless, and odorless.

[0072] The disposable petri dishes were purchased from Ruisu Experimental Equipment Store in Haimen City; they were 9cm in diameter and 2cm deep.

[0073] The culture medium raw materials were purchased from Beijing TransGen Biotech Co., Ltd.

[0074] The incubator was purchased from Shanghai Yuejin Medical Instrument Co., Ltd., model LRHS-400-II;

[0075] The clean bench was purchased from the medical equipment factory of Shanghai Boxun Industrial Co., Ltd., model SW-CJ-2FD.

[0076] The strains used are: Bacillus amyloliquefaciens, Bacillus megaterium, Streptomyces violaceum, Saccharomyces pulveratum, Trichoderma harzianum, Streptomyces leucosus and Streptomyces rubrum and griseus, Bacillus belye, Streptomyces olvarii, Bacillus subtilis, Aquatic Rahn's bacterium, Pseudomonas proteus, and Streptomyces loucheri. All of these strains are commercially available.

[0077] Where specific experimental steps or conditions are not specified in the embodiments, they can be performed according to the conventional experimental steps or conditions described in the literature in this field. All raw materials or instruments used are commercially available conventional products, including but not limited to those used in the embodiments of this application.

[0078] Example 1

[0079] This embodiment provides a culture dish for displaying the colony morphology of Bacillus amyloliquefaciens, and its preparation method is as follows:

[0080] (1) Prepare NA medium according to the following formula: 10.0g peptone, 3.0g beef powder, 5.0g NaCl, 20.0g agar, 1L distilled water, pH 7.0; Pour the NA medium sterilized at 121℃ for 30min and cooled to about 60℃ into a disposable plastic petri dish sterilized with ethylene oxide, to a thickness of 5mm, and allow it to solidify naturally;

[0081] (2) Bacillus amyloliquefaciens was activated into NA medium using the three-zone streak method with a sterile inoculation loop and incubated upside down in a 30°C incubator for 48 hours;

[0082] (3) Place the culture dish with a single colony on the laminar flow hood and blow it with clean air for 30 minutes until there are no water stains on the edge and surface of the culture medium;

[0083] (4) Melt the transparent jelly wax in a stainless steel beaker using an electromagnetic frequency of 300W, let it cool naturally to 70~75℃, slowly pour the melted jelly wax from one side of the petri dish until it covers the surface of the culture medium, with a thickness of 5mm, let it cool naturally for 5 minutes until it solidifies, forming a jelly wax layer.

[0084] (5) Prepare epoxy resin adhesive as follows: Take 3 parts of A glue of crystal epoxy resin and pour it into a clean glass beaker. Heat it in a water bath at 65°C for 15 minutes. After A glue is transparent and free of bubbles, take 1 part of B glue of crystal epoxy resin and pour it into A glue. Stir slowly along the beaker wall with a glass rod for 5 minutes to obtain a transparent, uniform, bubble-free epoxy resin adhesive. Slowly pour the prepared epoxy resin adhesive from one side of the petri dish until it covers the surface of the jelly wax layer with a thickness of 4.5 mm. Let it air dry for 48 hours to solidify and form an epoxy resin layer.

[0085] The petri dish prepared in Example 1 is as follows: Figure 1 and 2 As shown, the petri dish consists of the following parts: petri dish, NA medium, jelly wax layer, and epoxy resin layer. Among them, Bacillus amyloliquefaciens is cultured in NA medium, jelly wax layer completely covers the surface of NA medium, and epoxy resin layer completely covers the surface of jelly wax layer.

[0086] Example 2

[0087] This embodiment provides a culture dish for displaying the colony morphology of Bacillus megaterium, and its preparation method is as follows:

[0088] (1) Prepare NA medium according to the following formula: 10.0g peptone, 3.0g beef powder, 5.0g NaCl, 20.0g agar, 1L distilled water, pH 7.0; Pour the NA medium sterilized at 121℃ for 30min and cooled to about 60℃ into a disposable plastic petri dish sterilized with ethylene oxide, to a thickness of 4mm, and allow it to solidify naturally;

[0089] (2) Bacillus megaterium was activated into NA medium using the three-zone streak method with a sterile inoculation loop and incubated at 30°C for 48 hours.

[0090] Steps (3) to (5) refer to steps (3) to (5) of Example 1.

[0091] Example 3

[0092] This embodiment provides a petri dish for displaying the colony morphology of purple Streptomyces, and its preparation method is as follows:

[0093] (1) Prepare Gao's No. I culture medium according to the following formula: soluble starch 20.0g, NaCl 0.5g, KNO3 1.0g, K2HPO4 0.5g, MgSO4·7H2O 0.5g, FeSO4·7H2O 0.01g, agar 20.0g, distilled water 1L, pH 7.2; pour Gao's No. I culture medium sterilized at 121℃ for 30min and cooled to about 60℃ into disposable plastic petri dishes sterilized with ethylene oxide, with a thickness of 6mm, and allow it to solidify naturally;

[0094] (2) The purple Streptomyces was activated into Gao's No. 1 medium using the three-zone streak method with a sterile inoculation loop and cultured at 28°C for 7 days.

[0095] Steps (3) to (5) refer to steps (3) to (5) of Example 1.

[0096] Example 4

[0097] This embodiment provides a culture dish for displaying the colony morphology of the fungal strain *Miller's yeast*, and its preparation method is as follows:

[0098] (1) Prepare YPD medium according to the following formula: 10.0g yeast powder, 20.0g peptone, 20.0g glucose, 20.0g agar, 1L distilled water, pH natural; sterilize at 115℃ for 20min, pour the YPD medium cooled to about 60℃ into sterilized disposable plastic petri dishes with a thickness of 4.0mm, and let it solidify naturally;

[0099] (2) Powdered Miller's yeast was activated into YPD medium using a sterile inoculation loop with three-zone streak, and cultured at 28℃ for 5 days;

[0100] Steps (3) to (5) refer to steps (3) to (5) of Example 1.

[0101] Example 5

[0102] This embodiment provides a petri dish for displaying the colony morphology of the fungal strain Trichoderma harzianum, and its preparation method is as follows:

[0103] (1) Prepare PDA culture medium according to the following formula: 1L potato filtrate, 20.0g glucose, 20.0g agar, pH natural, potato filtrate is made from 200g potatoes; sterilize at 121℃ for 15min, and pour the PDA culture medium, which is cooled to about 60℃, into sterilized disposable plastic petri dishes with a thickness of 6mm, and let it solidify naturally;

[0104] (2) Use a sterile inoculation loop to pick up spores of Trichoderma harzianum and inoculate them into the center of PDA medium. Incubate at 28°C for 10 days until the mycelium is yellow-green and covers the entire culture dish.

[0105] Steps (3) to (5) refer to steps (3) to (5) of Example 1.

[0106] Example 6

[0107] This embodiment provides a petri dish for demonstrating the starch-hydrolyzing function of Streptomyces leucosus, and its preparation method is as follows:

[0108] (1) Prepare the starch-releasing function qualitative test medium according to the following formula: 10.0g peptone, 3.0g beef powder, 5.0g NaCl, 10.0g soluble starch, 20.0g agar, pH 7.0; pour the starch-releasing function qualitative test medium sterilized at 121℃ for 30min and cooled to about 60℃ into a disposable plastic petri dish sterilized with ethylene oxide, to a thickness of 4mm, and allow it to solidify naturally;

[0109] (2) Prepare the staining solution according to the following formula: dilute the original Lugol's iodine solution 25 times with distilled water sterilized at 121℃ for 30 min to prepare 25× Lugol's iodine solution; add 7% concentrated hydrochloric acid to the 25× Lugol's iodine solution and mix well;

[0110] (3) Inoculate a 6 mm diameter Streptomyces leucovora mycelium cake at the center of the starch-dissolving function qualitative test medium and incubate at 28℃ for 7 days until a transparent ring with a diameter of about 2 cm appears around the colony.

[0111] (4) Pour the staining solution prepared in step (2) into the culture dish after step (3) to cover the entire culture medium, soak and stain for 3 minutes, pour off the iodine solution, and obtain a starch-degrading qualitative culture dish with a transparent hydrolysis ring of about 2 cm in diameter around the colony that does not turn blue.

[0112] (5) Place the petri dish obtained in step (4) on a clean bench and blow it with clean air for 1.5 hours until there are no water stains on the edge and surface of the culture medium;

[0113] Steps (6) to (7) refer to steps (4) to (5) of Example 1.

[0114] The petri dish prepared in Example 6 is as follows Figure 3 and 4 As shown, the petri dish consists of the following parts: a petri dish, a starch-solubilizing function qualitative detection medium, a jelly wax layer, and an epoxy resin layer. Streptomyces is cultured in the starch-solubilizing function qualitative detection medium, and a clear starch-solubilizing transparent zone appears around the bacteria. The jelly wax layer completely covers the starch-solubilizing function qualitative detection medium, and the epoxy resin layer completely covers the surface of the jelly wax layer. This culture medium can be used to demonstrate the starch-solubilizing function of Streptomyces.

[0115] Example 7

[0116] This embodiment provides a culture dish for demonstrating the lipolysis function of Streptomyces griseus, and its preparation method is as follows:

[0117] (1) Prepare the medium for qualitative detection of fat-dissolving function as follows: Weigh 10.0g of peptone, 5.0g of NaCl, and 0.1g of CaCl2·2H2O into 800mL of distilled water, dissolve them completely, add 20.0g of agar, adjust the pH to 7.2, bring the volume to 1L, sterilize at 121℃ for 30min, cool to 65℃, add 10mL of Tween80 sterilized at 121℃ for 30min separately, mix well; pour the medium for qualitative detection of fat-dissolving function, cooled to about 60℃, into a sterilized disposable plastic petri dish, 4.5mm thick, and allow it to solidify naturally;

[0118] (2) A 6 mm diameter Streptomyces red and gray mold cake was inoculated at the center of the medium for qualitative detection of fat-dissolving function and cultured at 28℃ for 7 days. An opaque halo of saponified crystals appeared around the colony.

[0119] (3) Place the petri dish obtained in step (2) on a clean bench and blow it with clean air for 30 minutes until there are no water stains on the edge and surface of the culture medium;

[0120] Steps (4) to (5) refer to steps (4) to (5) of Example 1.

[0121] Example 8

[0122] This embodiment provides a culture dish for demonstrating the protein-degrading function of Bacillus belesii, and its preparation method is as follows:

[0123] (1) Prepare the protein decomposition function qualitative test medium as follows: Weigh 2.0g of casein, moisten with 5mL of 0.5mol / L NaOH, add 150mL of distilled water, stir in a boiling water bath until completely dissolved, add distilled water to 800mL, then add 3.0g of beef extract, 5.0g of NaCl, and 0.1g of CaCl2·2H2O, dissolve thoroughly, add 20.0g of agar, mix well, and bring the volume to 1L with distilled water. Set the pH to natural and sterilize at 121℃ for 30min. Pour the protein decomposition function qualitative test medium, cooled to about 60℃, into sterilized disposable plastic petri dishes to a thickness of 5.0mm, and allow to solidify naturally.

[0124] (2) Activated Bacillus belye was inoculated at the center of the protein functional qualitative test medium and cultured at 30℃ for 4 days. A protein-clear zone with a diameter of about 1.5 cm appeared around the colony.

[0125] (3) Place the petri dish obtained in step (2) on a clean bench and blow it with clean air for 30 minutes until there are no water stains on the edge and surface of the culture medium;

[0126] Steps (4) to (5) refer to steps (4) to (5) of Example 1.

[0127] Example 9

[0128] This embodiment provides a culture dish demonstrating the cellulose-degrading function of *Streptomyces olfactoryum*, and its preparation method is as follows:

[0129] (1) Prepare the cellulose dehydrogenation function qualitative test medium according to the following formula: CMC-Na 10.0g, (NH4)2SO4 4.0g, K2HPO4 6.5g, MgSO4·7H2O 0.5g, peptone 1.0g, agar 20.0g, distilled water 300mL, pH natural; pour the cellulose dehydrogenation function qualitative test medium sterilized at 121℃ for 30min and cooled to about 60℃ into a sterilized disposable plastic petri dish, with a thickness of 5.5mm, and allow it to solidify naturally;

[0130] (2) Prepare the rinsing solution according to the following formula: 1L of distilled water, sterilize at 121℃ for 30min, cool to room temperature and add streptomycin sulfate to a final concentration of 50μg / mL;

[0131] (3) Prepare Congo red staining solution according to the following formula: 0.2% Congo red solution, sterilize at 121℃ for 30 min, cool to room temperature and add streptomycin sulfate to a final concentration of 50 μg / mL;

[0132] (4) Prepare the rinsing solution according to the following formula: 1% NaCl solution, sterilize at 121℃ for 30 min, cool to room temperature and add streptomycin sulfate to a final concentration of 50 μg / mL;

[0133] (5) Inoculate the center of the cellulose decomposition function qualitative test medium with a 6 mm diameter olive color-producing Streptomyces mycelium cake and incubate at 28℃ for 9 days.

[0134] (6) Rinse the spore powder around the colony with the rinsing solution prepared in step (2) for 2 minutes until the spore powder is rinsed clean;

[0135] (7) Pour the Congo red staining solution prepared in step (3) into the culture medium after rinsing in step (6), cover the culture medium with the staining solution, stain for 4 hours, and then discard the staining solution.

[0136] (8) Pour the rinsing solution prepared in step (4) into the culture medium stained in step (7), soak twice, 10 min each time, and discard the rinsing solution; obtain a culture dish for qualitative detection of fibrosis function with a transparent hydrolysis zone of about 2 cm around the colony that does not turn red.

[0137] (9) Place the petri dish obtained in step (8) in a clean bench and blow it with clean air for 1.5 hours until there are no water stains on the edge and surface of the culture medium;

[0138] Steps (10) to (11) refer to steps (4) to (5) of Example 1.

[0139] Example 10

[0140] This embodiment provides a culture dish for demonstrating the lignin-degrading function of Bacillus subtilis, and its preparation method is as follows:

[0141] (1) Prepare aniline blue medium as follows: Weigh 10.0g of yeast powder and 20.0g of glucose into 800mL of distilled water, dissolve them completely, add 20.0g of agar, make up to 1L, set the pH to natural, sterilize at 121℃ for 30min, cool to 65℃, add 10mL of aniline blue stock solution sterilized at 121℃ for 30min separately, mix well; pour the aniline blue medium cooled to about 55℃ into sterilized disposable plastic petri dishes, with a thickness of 4.5mm, and let it solidify naturally;

[0142] (2) Activated Bacillus subtilis was inoculated at the center of the aniline blue medium and cultured at 30℃ for 7 days. A 1.5cm aniline blue decolorization zone appeared around the colony.

[0143] (3) Place the petri dish obtained in step (2) on a clean bench and blow it with clean air for 30 minutes until there are no water stains on the edge and surface of the culture medium;

[0144] Steps (4) to (5) refer to steps (4) to (5) of Example 1.

[0145] Example 11

[0146] This embodiment provides a culture dish for demonstrating the phosphate-solubilizing function of aquatic Ranunculella lataniae, and its preparation method is as follows:

[0147] (1) Prepare the qualitative test medium for phosphorus solubilization function according to the following formula: 10.0g glucose, 0.5g (NH4)2SO4, 0.3g NaCl, 0.3g KCl, 0.3g MgSO4·7H2O, 0.03g FeSO4·7H2O, 0.03g MnSO4·H2O, 0.2g egg lecithin, 1.0g CaCO3, 0.5g yeast powder, 20.0g agar, 1L distilled water, pH 7.0; pour the phosphorus solubilization function qualitative test medium sterilized at 115℃ for 30min and cooled to about 60℃ into a sterilized disposable plastic petri dish, with a thickness of 6mm, and allow it to solidify naturally;

[0148] (2) Draw a cross on the back of a 90mm diameter petri dish. Using the intersection of the cross as the center, inoculate aquatic Laenella bacteria at two points 25mm away from the center of the cross with a sterile inoculation needle. Incubate at 30℃ for 7 days. A transparent zone with a diameter of about 1.5cm will appear around the colony.

[0149] (3) Seal the petri dishes with transparent rings (to prevent them from drying out at high temperatures) and place them in an 80°C oven for 4 hours to weaken the phosphate-solubilizing function of the bacteria and prevent the phosphate-solubilizing rings from gradually increasing in size;

[0150] (4) Place the high-temperature treated phosphorus-solubilizing culture dish on a clean bench and blow it with clean air for 30 minutes until there are no water stains on the edge and surface of the culture medium;

[0151] Steps (5) to (6) refer to steps (4) to (5) of Example 1.

[0152] Example 12

[0153] This embodiment provides a culture dish for demonstrating the potassium-solubilizing function of a strain of *Pseudomonas proteus*, and its preparation method is as follows:

[0154] (1) Prepare the potassium solubilization function qualitative test medium according to the following formula: glucose 10.0g, Na2HPO4 0.2g, MgSO4·7H2O 0.2g, NaCl 0.2g, CaSO4 0.2g, CaCO3 5.0g, potassium feldspar powder 2.5g, agar 20g, distilled water 1L, pH natural; pour the potassium solubilization function qualitative test medium sterilized at 115℃ for 30min and cooled to about 60℃ into a sterilized disposable plastic petri dish, with a thickness of 4.5mm, and allow it to solidify naturally;

[0155] (2) Draw a cross on the back of a 90mm diameter petri dish. Using the intersection of the cross as the center, inoculate Pseudomonas proteoglycans at two points 25mm away from the center of the cross with a sterile inoculation needle. Incubate at 30℃ for 7 days. A potassium-free transparent zone with a diameter of about 2cm will appear around the colony.

[0156] (3) Place the culture dish with the potassium-free transparent ring on the ultra-clean bench and blow it with clean air for 30 minutes until there are no water stains on the edge and surface of the culture medium;

[0157] Steps (4) to (5) refer to steps (4) to (5) of Example 1.

[0158] Example 13

[0159] This embodiment provides a culture dish demonstrating the antibacterial function of Streptomyces louchei, and its preparation method is as follows:

[0160] (1) Prepare PDA culture medium according to the following formula: 1L potato filtrate, 20.0g glucose, 20.0g agar, pH natural, potato filtrate is made from 200g potatoes; Pour the PDA culture medium sterilized at 121℃ for 15min and cooled to about 60℃ into sterilized disposable plastic petri dishes with a thickness of 6mm, and let it solidify naturally;

[0161] (2) Using the confrontation method, a cross was drawn on the back of a 90 mm diameter petri dish. With the intersection of the cross as the center, two 6 mm diameter activated Streptomyces loucheri mycelium were inoculated at two points 25 mm away from the center of the cross. One 6 mm diameter Alternaria solanacearum (the pathogen of early blight of tomato) mycelium was inoculated at the center of the circle as the treatment group. The petri dishes inoculated only with Alternaria solanacearum mycelium without Streptomyces loucheri served as the control group. The petri dishes were placed at 28 °C for 7 days until Alternaria solanacearum covered the entire culture medium in the control group, while the treatment group showed a clear inhibition zone.

[0162] (3) Place the culture dishes of the treatment group and the control group in a clean bench and blow them with clean air for 30 minutes until there are no water stains on the edge and surface of the culture medium;

[0163] Steps (4) to (5) refer to steps (4) to (5) of Example 1.

[0164] The petri dish prepared in Example 13 is as follows: Figure 5 and 6As shown, both the treatment and control group culture dishes consisted of the following components: a culture dish, PDA medium, a jelly wax layer, and an epoxy resin layer. In the treatment group, *Streptomyces loucherei* and *Alternaria solanacearum* were cultured in the PDA medium, with a clear inhibition zone around *Streptomyces loucherei*. The jelly wax layer completely covered the PDA medium, and the epoxy resin layer completely covered the surface of the jelly wax layer. In the control group, *Alternaria solanacearum* was cultured in the PDA medium, and the jelly wax layer completely covered the PDA medium, and the epoxy resin layer completely covered the surface of the jelly wax layer. This culture medium can be used to demonstrate the antibacterial function of *Streptomyces loucherei*.

[0165] Experiment Example 1: Storage Stability Experiment of Petri Dishes

[0166] Several petri dishes prepared according to Example 1 were used as the experimental group, while petri dishes prepared according to steps (1) to (3) of Example 1 were used as the control group;

[0167] The culture dishes of the experimental group and the control group were treated as follows:

[0168] The culture dishes were placed at temperatures of -20℃, -10℃, -4℃, 4℃, 10℃, 15℃, 20℃, 25℃, 30℃, 35℃, 38℃, and 45℃, and the conditions of each group of culture dishes were observed after 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 2 days, 5 days, 10 days, 20 days, 30 days, 1 month, 2 months, 3 months, 6 months, and 12 months.

[0169] The final results are summarized as follows:

[0170] (1) The culture dishes prepared in Example 1 can be kept at -20℃ for 12 hours, -10℃ for 24 hours, and -4℃ for 2 days without freezing. Even after being frozen solid for a long time and then thawed, they still maintain their original shape. In contrast, the culture dishes in the control group freeze solid after being placed at -4℃ for 12 hours. After thawing, there is more moisture on the surface of the culture medium, and the morphology of the bacteria is destroyed. This proves that the culture dishes provided by the present invention have certain antifreeze properties and can be stored and used in colder regions.

[0171] (2) The petri dishes prepared in Example 1 can be stored at low temperature (4~10℃) and room temperature (15~30℃) for more than one year, while the petri dishes of the control group dried out or became contaminated within 30 days. This proves that the petri dishes provided by the present invention are more suitable for storage at low temperature and room temperature, which is beneficial for users in different regions to store and use for a long time in different seasons.

[0172] (3) The petri dishes prepared in Example 1 can be stored at high temperature (35~45℃) for up to 1 year, while the petri dishes in the control group dried out or became contaminated within 15 days. This proves that the petri dishes provided by the present invention have good heat resistance.

[0173] Experiment Example 2: Screening Experiment of Petri Dish Sealing Materials

[0174] (1) Experiment on the sealing effect of crystal epoxy resin dripping on culture medium

[0175] The experimental method is as follows:

[0176] Commonly used culture media for preparing bacterial strains include: NA medium, Gao's No. 1 medium, YPD medium and PDA medium. Pour them into sterilized glass petri dishes and let them solidify before use.

[0177] Take 3 parts of the A component of the crystal epoxy resin epoxy resin and pour it into a clean glass beaker. Incubate it in a water bath at 65°C for 15 minutes. Once the A component is transparent and free of bubbles, take 1 part of the B component of the crystal epoxy resin epoxy resin and pour it into the A component. Slowly stir the glass rod along the beaker wall for 5 minutes to obtain a transparent, uniform, and bubble-free epoxy resin.

[0178] Pour the prepared epoxy resin into petri dishes containing NA medium, Gao's No. 1 medium, YPD medium and PDA medium respectively, to a thickness of 4.5 mm, and let it stand at room temperature for 48 hours to solidify.

[0179] Experimental results: Although epoxy resin glue solidifies after being poured onto the surface of the culture medium, the curing effect of the transparent epoxy resin glue is affected by the moisture on the surface of the culture medium, resulting in a white cured colloid that completely covers the culture medium below and cannot be observed. Therefore, epoxy resin glue cannot be used as a sealing material for direct contact with the culture medium.

[0180] (2) Experiment on the sealing effect of gelatin wax on culture medium

[0181] The experimental method is as follows:

[0182] Commonly used culture media for preparing bacterial strains include: NA medium, Gao's No. 1 medium, YPD medium and PDA medium. Pour them into sterilized glass petri dishes and let them solidify before use.

[0183] Melt the transparent jelly wax in a stainless steel beaker using an electromagnetic flux of 300W, let it cool naturally to 70-75℃, and then pour it into petri dishes containing NA medium, Gao's No. 1 medium, YPD medium and PDA medium respectively, to a thickness of 6.0 mm. Let it stand at room temperature for 10 minutes to solidify.

[0184] Experimental results:

[0185] a. Transparency

[0186] The jelly-like wax layer is highly transparent, making the underlying culture medium clearly visible.

[0187] b. Sealing

[0188] Pour 3 mm of purified water into the petri dish that has been sealed with gel wax, cover it, and let it stand at room temperature for 2 hours. Observe whether the water has seeped into the culture medium.

[0189] Results: No water seeped into the culture medium, proving that the jelly wax could adhere tightly to the petri dish and had good sealing properties.

[0190] c. Stability

[0191] The raw material of jelly wax was placed in an open container and placed at different temperatures of -20℃, 4℃, 25℃, 35℃ and 45℃ for one month to study whether the raw material would lose weight when exposed to air.

[0192] Result: No weight loss occurred, proving that the raw material can be stored for a long time and has high stability.

[0193] (3) Experiment on the sealing effect of crystal epoxy resin dripping on the jelly wax layer

[0194] Jelly wax material is elastic and easily gets dirty, making it difficult to handle and carry. To facilitate use and long-term preservation, the jelly wax layer is further melted and sealed.

[0195] Further experiments were conducted based on the culture dishes obtained from the above-mentioned experiment on the sealing effect of jelly wax on the culture medium:

[0196] Pour the prepared epoxy resin into a petri dish, cover the surface of the jelly wax layer with a thickness of 4.5 mm, and let it stand at room temperature for 48 hours to solidify.

[0197] Results: The jelly wax layer and the crystal epoxy resin epoxy layer made good contact with each other, with no reaction. The contact surfaces adhered tightly, with high transparency. The surface was hard and not easily damaged, making it easy to carry and preserve for a long time.

[0198] Experiment Example 3: Screening Experiment for Melting and Sealing Temperature of Jelly Wax

[0199] The melting temperature of the jelly wax can reach up to 135℃. Since high temperatures may have a significant impact on the morphology of the microorganisms, in order to ensure sealing efficiency without affecting the observation of microbial morphology and qualitative functional test results, this experiment explored a gradient sealing temperature, as follows:

[0200] Bacillus amyloliquefaciens was activated by streaking three zones onto NA medium with a sterile inoculation loop and incubated at 30°C for 48 hours.

[0201] Streptomyces leucosus was activated by streaking three zones onto Gao's No. 1 medium using a sterile inoculation loop and incubated at 28°C for 7 days.

[0202] Powdered Miller's yeast was activated by streaking in three zones onto YPD medium using a sterile inoculation loop and cultured in a constant temperature incubator at 28℃ for 4 days.

[0203] Trichoderma harzianum was activated by spot inoculation at the center of YPD medium using a sterile inoculation needle and cultured in a constant temperature incubator at 28℃ for 10 days.

[0204] Once colonies have formed in each culture medium, they are ready for use.

[0205] Different temperatures of gelatinous wax were poured into the above culture medium, and the results were observed as follows:

[0206] Table 1. Screening results for different sealing temperatures

[0207]

[0208] As shown in Table 1, the jelly wax has good fluidity and high melting efficiency at 70-75℃. Furthermore, the morphology of the colonies after melting is not significantly different from or does not change from the original morphology before melting. Therefore, the optimal melting temperature for jelly wax is 70-75℃.

[0209] Example 4: Improved Qualitative Detection Method for Starch-Clearing Function

[0210] Conventional methods:

[0211] (1) Medium for qualitative detection of starch-releasing function: 10.0g peptone, 3.0g beef powder, 5.0g NaCl, 10.0g soluble starch, 20.0g agar, pH 7.0; Sterilize at 121℃ for 30min and cool to about 60℃. Pour the medium for qualitative detection of starch-releasing function into a sterile disposable plastic petri dish with a thickness of 4mm and allow it to solidify naturally.

[0212] (2) Prepare the staining solution according to the following formula: dilute the original Lugol's iodine solution 25 times with distilled water to prepare 25× Lugol's iodine solution;

[0213] (3) Inoculate a 6 mm diameter Streptomyces leucovora mycelium cake at the center of the starch-dissolving function qualitative test medium and incubate at 28℃ for 7 days until a transparent ring with a diameter of about 2 cm appears around the colony.

[0214] (4) Pour the staining solution prepared in step (2) into the culture dish after step (3) to cover the entire culture medium, soak and stain for 3 minutes, pour off the iodine solution, and obtain a culture dish for qualitative detection of starch-dissolving function with transparent hydrolysis rings of about 2 cm in diameter around the colonies that do not turn blue.

[0215] Results of conventional methods: Due to the instability of the complex formed by iodine molecules and starch, the blue color around the starch-dissolving transparent ring gradually disappears over time, and the qualitative test results show that the morphology is maintained for no more than 48 hours.

[0216] Based on the aforementioned problems with conventional qualitative detection methods for starch-releasing function, this experiment improved the staining steps of the above methods and explored the optimal concentration of concentrated hydrochloric acid added to the staining solution to determine the optimal staining solution composition, ensuring that the qualitative results remain unchanged over a long period. The specific methods and results are as follows:

[0217] The culture dishes obtained according to the above conventional method step (3) were stained according to the treatment methods in Table 2:

[0218] Table 2. Treatment effects of staining solutions with different hydrochloric acid concentrations

[0219]

[0220] Therefore, the final method for preparing the staining solution is as follows: dilute the original Lugol's iodine solution 25 times with distilled water sterilized at 121℃ for 30 minutes to prepare 25× Lugol's iodine solution; add 7% concentrated hydrochloric acid to the 25× Lugol's iodine solution and mix well.

[0221] By optimizing the qualitative detection method for starch-releasing function, the defect of unstable qualitative results caused by the blue fading of the culture medium obtained by conventional qualitative detection methods in a short period of time has been solved, so that the qualitative results can be maintained in a long-term observable state.

[0222] Example 5: Improved Qualitative Detection Method for Cellulose Function

[0223] Conventional methods:

[0224] (1) Prepare the cellulose dehydrogenation function qualitative test medium according to the following formula: CMC-Na 10.0g, (NH4)2SO4 4.0g, K2HPO4 6.5g, MgSO4·7H2O 0.5g, peptone 1.0g, agar 20.0g, distilled water 300mL, pH natural; pour the cellulose dehydrogenation function qualitative test medium sterilized at 121℃ for 30min and cooled to about 60℃ into a sterilized disposable plastic petri dish, with a thickness of 5.5mm, and allow it to solidify naturally;

[0225] (2) A 6 mm diameter Streptomyces olfaction mycelium was inoculated at the center of the cellulose decomposition function qualitative test medium and cultured at 28℃ for 9 days.

[0226] (3) Stain with 0.2% Congo red for 10 min, then rinse with 1% sodium chloride solution for 10 min.

[0227] Results of conventional methods: The culture dishes prepared by conventional methods for qualitative detection of cellulose desiccation function were lightly stained and, after being placed at 28°C for 2 days, were contaminated with the spores of Streptomyces olfragilis, resulting in colonies covering the culture dishes and affecting the observation of qualitative results.

[0228] Based on the aforementioned problems with conventional qualitative detection methods for cellulose desorption, this experiment added a rinsing step to the conventional method to completely wash away the spores and prevent contamination from the spores themselves. Simultaneously, antibiotics were added to the staining and rinsing solutions to reduce the possibility of subsequent contamination by other bacteria. The specific details are as follows:

[0229] Rinse solution preparation: 1L of distilled water, sterilized at 121℃ for 30min, cooled to room temperature, and streptomycin sulfate added to a final concentration of 50μg / mL;

[0230] Preparation of Congo red staining solution: 0.2% Congo red solution, sterilize at 121℃ for 30 min, cool to room temperature and add streptomycin sulfate to a final concentration of 50 μg / mL;

[0231] Rinse solution preparation: 1% NaCl solution, sterilized at 121℃ for 30 min, cooled to room temperature and streptomycin sulfate added to a final concentration of 50 μg / mL;

[0232] Rinse the culture dish with rinsing solution to remove spores around the colonies for 2 minutes until the spores are completely removed; cover the culture medium with staining solution and stain for 4 hours; discard the staining solution; soak and rinse the stained culture dish twice with rinsing solution for 10 minutes each time, and discard the rinsing solution; obtain culture dishes for qualitative detection of cellulose dehydrolysis function by showing a transparent hydrolysis zone of about 2 cm around the colonies that does not turn red.

[0233] The culture dishes prepared by this method for qualitative detection of cellulose desiccant function remained uncontaminated after being placed at 28℃ for 2 days, and maintained a state of being uncontaminated by its own bacteria and other miscellaneous bacteria for a long time. The qualitative functional characteristics were stable and easy to observe. This method solves the defect of conventional qualitative detection methods where the culture medium is contaminated by spores of functional bacteria, making it difficult to observe the qualitative results.

[0234] The commercial application effects of the microbial display petri dishes provided by this invention are as follows: In 2021, by using the petri dishes provided by this invention as display tools for microbial strains in sales channels, the acceptance of microbial fertilizers by distributors and farmers was enhanced, leading to increased sales volume. Specifically, the sales performance of microbial fertilizers in various regions of the Mumei Tuli Group's sales department (the percentage increase in total sales compared to 2020) is as follows: North China region increased by 8.6%; Northeast region increased by 16.5%; Northwest region increased by 4.2%; South China region increased by 20.04%; and Shandong region increased by 12.6%. It is evident that using the microbial display petri dishes provided by this invention has significantly improved the company's profitability.

[0235] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A petri dish for displaying microorganisms, characterized in that, include: Petri dish; A culture medium is located inside the petri dish, and microorganisms are cultured on its surface. A jelly-like wax layer covers the surface of the culture medium; An epoxy resin layer covers the surface of the jelly wax layer facing away from the culture medium. The method for preparing the culture dish displaying microorganisms includes the following steps: (1) Pour the sterilized culture medium into the sterilized petri dish and let it solidify; (2) Inoculate and activate microorganisms on the surface of the culture medium, and then culture them; (3) Clean air blown until there are no water stains on the edge and surface of the culture medium; (4) Pour melted jelly wax onto the surface of the culture medium and let it solidify to form a jelly wax layer; (5) Pour the prepared epoxy resin adhesive onto the surface of the jelly wax layer and let it solidify to form an epoxy resin layer.

2. The culture dish for displaying microorganisms according to claim 1, characterized in that, The thickness of the culture medium is 3.5~6.5 mm; The thickness of the jelly wax layer is 4.0~6.0mm; The thickness of the epoxy resin layer is 3.0~5.5mm.

3. The culture dish for displaying microorganisms according to claim 1, characterized in that, The microorganisms include fungi and / or bacteria.

4. The culture dish for displaying microorganisms according to claim 3, characterized in that, The microorganisms include at least one of Bacillus amyloliquefaciens, Bacillus megaterium, Bacillus belesiensis, Bacillus subtilis, Aquatic Rahn's bacterium, Pseudomonas proteus, Streptomyces violaceum, Streptomyces leucosus, Streptomyces rubrum and Streptomyces oleifera, Streptomyces loucheri, Saccharomyces mullerii, and Trichoderma harzianum.

5. The culture dish for displaying microorganisms according to claim 1, characterized in that, The culture medium is selected from any one of NA medium, Gao's No. 1 medium, YPD medium, PDA medium, and rhizobium medium.

6. The culture dish for displaying microorganisms according to claim 1, characterized in that, The culture medium is selected from any one of the following: starch-solubilizing function qualitative detection medium, fat-solubilizing function qualitative detection medium, protein-solubilizing function qualitative detection medium, cellulose-solubilizing function qualitative detection medium, aniline blue medium, phosphorus-solubilizing function qualitative detection medium, and potassium-solubilizing function qualitative detection medium. The composition of the qualitative detection medium for starch-solubilizing function is as follows: 10.0g peptone, 3.0g beef powder, 5.0g NaCl, 10.0g soluble starch, 20.0g agar, 1L distilled water, pH 7.0; The preparation method of the qualitative detection medium for fat-dissolving function is as follows: Weigh 10.0g of peptone, 5.0g of NaCl, and 0.1g of CaCl2·2H2O into 800mL of distilled water, dissolve them completely, add 20.0g of agar, adjust the pH to 7.2, bring the volume to 1L, sterilize at 121℃ for 30min, cool to 65℃, add 10mL of Tween80 sterilized separately at 121℃ for 30min, and mix well; The preparation method of the protein function qualitative detection medium is as follows: Weigh 2.0g of casein, moisten with 5mL of 0.5mol / L NaOH, add 150mL of distilled water, stir in a boiling water bath until completely dissolved, add distilled water to 800mL, then add 3.0g of beef extract, 5.0g of NaCl, and 0.1g of CaCl2·2H2O, dissolve thoroughly, add 20.0g of agar, mix evenly, bring the volume to 1L with distilled water, set the pH to natural, and sterilize at 121℃ for 30min; The composition of the medium for qualitative detection of cellulose decomposition function is as follows: CMC-Na 10.0g, (NH4)2SO4 4.0g, K2HPO4 6.5g, MgSO4·7H2O 0.5g, peptone 1.0g, agar 20.0g, water 300mL, pH natural; The preparation method of the aniline blue medium is as follows: Weigh 10.0g of yeast powder and 20.0g of glucose into 800mL of distilled water, dissolve them completely, add 20.0g of agar, make up to 1L, set the pH to natural, sterilize at 121℃ for 30min, cool to 65℃, add 10mL of aniline blue stock solution sterilized separately at 121℃ for 30min, and mix well; The composition of the qualitative detection medium for phosphorus solubility is as follows: glucose 10.0g, (NH4)2SO4 0.5g, NaCl 0.3g, KCl 0.3g, MgSO4·7H2O 0.3g, FeSO4·7H2O 0.03g, MnSO4·H2O 0.03g, egg lecithin 0.2g, CaCO3 1.0g, yeast powder 0.5g, agar 20.0g, distilled water 1L, pH 7.0~7.2; The composition of the culture medium for qualitative detection of potassium solubilization function is as follows: 10.0g glucose, 0.2g Na2HPO4, 0.2g MgSO4·7H2O, 0.2g NaCl, 0.2g CaSO4, 5.0g CaCO3, 2.5g potassium feldspar powder, 20.0g agar, 1L distilled water, and natural pH.

7. The culture dish for displaying microorganisms according to claim 1, characterized in that, In step (4), the temperature of the melted jelly wax is 70~75℃.

8. The culture dish for displaying microorganisms according to claim 1, characterized in that, In step (5), the preparation method of the epoxy resin adhesive includes: taking 3 parts of the A glue of the crystal epoxy resin dripping, bathing it in a water bath at 65°C for 15 minutes, taking 1 part of the B glue of the crystal epoxy resin dripping and pouring it into the A glue, and stirring evenly.

9. The culture dish for displaying microorganisms according to claim 1, characterized in that, Step (2) further includes a step of qualitative detection of microbial function after culturing the microorganisms. The qualitative tests for microbial function include at least one of the following: qualitative tests for starch solubility, qualitative tests for fat solubility, qualitative tests for protein solubility, qualitative tests for cellulose solubility, qualitative tests for lignin solubility, qualitative tests for phosphorus solubility, qualitative tests for potassium solubility, and qualitative tests for antibacterial function.

10. The culture dish for displaying microorganisms according to claim 9, characterized in that, The method for qualitative detection of starch-releasing function includes: preparation of staining solution: dilute Lugol's iodine solution stock solution 25 times with sterile distilled water to prepare 25× Lugol's iodine solution, add 7% concentrated hydrochloric acid to 25× Lugol's iodine solution and mix well; pour the prepared staining solution into the culture dish for culturing microorganisms, cover the culture medium, soak and stain for 3 minutes, and then pour off the iodine solution.

11. The culture dish for displaying microorganisms according to claim 9, characterized in that, The method for qualitative detection of cellulose desiccation function includes: preparation of rinsing solution: sterilize distilled water, cool to room temperature, and add streptomycin sulfate to a final concentration of 50 μg / mL; preparation of Congo red staining solution: sterilize 0.2% Congo red solution, cool to room temperature, and add streptomycin sulfate to a final concentration of 50 μg / mL; preparation of rinsing solution: sterilize 1% NaCl solution, cool to room temperature, and add streptomycin sulfate to a final concentration of 50 μg / mL; rinse the culture dish containing microorganisms with the prepared rinsing solution until the spores around the colonies are rinsed clean; cover the culture medium with the prepared Congo red staining solution, stain for 4 hours, and discard the staining solution; soak and rinse the stained culture dish twice with the prepared rinsing solution, 7-10 min each time, and discard the rinsing solution.

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