Application method of bacillus subtilis metabolite as growth regulator of Aciporiopsis reeds

Abstract

The invention discloses an application of a bacillus subtilis liquid fermentation metabolite as a growth regulator of an edible fungus Ariobotrys reeds. The bacillus subtilis metabolite serving as the growth regulator of the edible fungus Ariobotrys reeds has the following advantages that the technological operation is simple, the bacillus subtilis metabolite can be used for preparation and cultivation of Ariobotrys reeds strains only through solid-liquid separation or directly after liquid fermentation, the preparation period is short (the bacillus subtilis metabolite can be used after being fermented for 5 days), and industrialized production is easy. After the bacillus subtilis metabolite is added into the culture medium of the edible fungus Acilia reeds, the average growth rate of Acilia reeds mycelia is higher than that of a control group, and the Acilia reeds are easier to culture.

Description

Technical Field

[0001] The invention belongs to the technical field of edible fungus cultivation and relates to an application and method of a liquid fermentation metabolite of Bacillus subtilis of the genus Bacillus as a growth regulator for the edible fungus Phragmites australis. Background Art

[0002] The cultivation process of edible fungi includes the preparation of fungus strains, preparation of culture medium, inoculation and cultivation, and fruiting of mushrooms. Hygrophoropsis phragmiticola ), a small genus in the order Boletales, now classified as its own independent family, Ceropegiaceae, and a new species. Since ancient times, farmers around Qilihai, Tianjin, have harvested and eaten Ceropegia repens during the mushroom-bearing season, either fresh or dried. Its flavor is extremely delicious, far surpassing other mushrooms. In recent years, excessive human collection has severely impacted the natural reproduction and growth of wild Ceropegia repens, resulting in a very small number of wild Ceropegia repens. Artificial domestication and cultivation have become an important measure to save the wild resources of Ceropegia repens. A search revealed no patent publications related to the present invention's patent application. Summary of the Invention

[0003] The present invention aims to provide an application and method of a liquid fermentation metabolite of Bacillus subtilis of the genus Bacillus as a growth regulator for the edible fungus Phragmites australis.

[0004] The technical solution adopted by the present invention to solve the technical problem is: Application of liquid fermentation metabolites of Bacillus subtilis as growth regulators for the edible fungus Phragmites australis.

[0005] Furthermore, the liquid fermentation metabolite of Bacillus subtilis is prepared by culturing in the following liquid culture medium, and the culture medium includes the following components in parts by weight: see Table 1.

[0006] Table 1 Medium for preparing liquid metabolites of Bacillus subtilis Reagent name Dosage / per 100mL Peptone 1g yeast powder 0.5g Sodium chloride 1g Alternatively, the liquid fermentation metabolite of Bacillus subtilis can be used as a growth-promoting regulator for the mycelium of Pseudomonas aeruginosa and / or as a cultivation growth regulator for Pseudomonas aeruginosa; Alternatively, the liquid fermentation culture of Bacillus subtilis requires solid-liquid separation (such as solid-liquid separation by centrifugation or plate-and-frame filtration) to obtain liquid Bacillus subtilis metabolites; Alternatively, when in use, the liquid fermentation metabolites of Bacillus subtilis are directly added to the culture medium of Pseudomonas australis and sterilized together for use.

[0007] Furthermore, 0.1-1 mL of liquid fermentation metabolites of Bacillus subtilis was added to the culture medium of every 100 g of Pseudomonas reedii.

[0008] A method for applying a liquid fermentation metabolite of Bacillus subtilis as a growth regulator for the edible fungus Phragmites australis, characterized by comprising the following steps: (1) Cultivating and preparing liquid fermentation metabolites of Bacillus subtilis; (2) adding the liquid fermentation metabolites of Bacillus subtilis obtained in step (1) to the culture medium of Pseudomonas aeruginosa, mixing and sterilizing the culture medium, and then the culture medium can be used for culturing the edible fungus Pseudomonas aeruginosa; The liquid fermentation metabolite of Bacillus subtilis can be used as a growth regulator for the edible fungus Phragmites australis.

[0009] Furthermore, the metabolites of the liquid fermentation of Bacillus subtilis in step (2) can be used as a growth-promoting regulator for the mycelium of Pseudomonas aeruginosa and / or as a cultivation growth regulator for Pseudomonas aeruginosa.

[0010] Furthermore, when the liquid fermentation metabolite of Bacillus subtilis is used as a growth regulator for the mycelium of Pseudomonas aeruginosa, 0.1-1 mL of the liquid fermentation metabolite of Bacillus subtilis is added to the culture medium of 100 g of the mycelium of Pseudomonas aeruginosa; When the Bacillus subtilis liquid fermentation metabolite agent is used as a growth regulator for cultivation of reed wax mushroom, 0.1-1 mL of the Bacillus subtilis liquid fermentation metabolite is added to every 100 g of reed wax mushroom culture material; Alternatively, the culture temperature of the mycelium of Pseudomonas aeruginosa is 25° C.-28° C., and the culture time is 7-14 days; Alternatively, the cultivation time of the fruiting body of Phragmites australis is 50-60 days from the start of spawning.

[0011] Furthermore, the culture medium of the mycelium of Pseudomonas aeruginosa comprises the following components in percentage by weight: Wheat grain 2-10%, glucose 1-3%, peptone 0.1-0.3%, agar 1-2%, KH2PO4 0.1-0.3%, MgSO4·7H2O 0.1-0.2%, Bacillus subtilis metabolite solution 0.1-1% (v / w 总 ); Alternatively, in step (1), solid-liquid separation is required after liquid fermentation culture of Bacillus subtilis to obtain liquid Bacillus subtilis metabolites.

[0012] A method for preparing a reed wax agaricus strain using liquid fermentation metabolites of Bacillus subtilis comprises the following steps: adding the liquid fermentation metabolites of Bacillus subtilis to a culture medium of reed wax agaricus, pouring a plate, inoculating a reed wax agaricus block, and culturing at 25°C for 7 to 12 days. The resulting strain is a reed wax agaricus strain, which can be used as a primary strain of the edible fungus reed wax agaricus; The liquid fermentation metabolite of Bacillus subtilis can be used as a growth regulator for the edible fungus Phragmites australis.

[0013] The beneficial effects achieved by the present invention are: 1. The Bacillus subtilis metabolites of the present invention have the following advantages as growth regulators for the edible fungus Phragmites australis: the process is simple to operate. After liquid fermentation, only solid-liquid separation is required, or the product can be directly used for cultivation of Phragmites australis. This results in a short preparation cycle (it can be used after 5 days of fermentation, compared to 7 days currently), making it easy to manufacture on a factory scale. Adding the Bacillus subtilis metabolites to the culture medium for the edible fungus Phragmites australis causes the mycelium of Phragmites australis to grow strong and dense, making it easier to cultivate. Adding the Bacillus subtilis metabolites to the culture medium for the edible fungus Phragmites australis accelerates the growth of the fungus and facilitates fruiting. Thus, the Bacillus subtilis metabolites can be used to prepare solid cultures of the edible fungus Phragmites australis and as growth regulators for Phragmites australis cultivation. 2. The Bacillus subtilis metabolites of the present invention, which are derived from liquid fermentation, solve technical problems such as weak vigor in the preparation of solid cultures of Phragmites australis and the difficulty in domesticating and cultivating the fungus. The reed wax cap mushroom is a new species. Its wild growth cycle in the Qilihai wetland is short and its nutritional requirements are quite demanding. Currently, only one research unit has reported on its cultivation method, and widespread adoption has been delayed. Adding Bacillus subtilis metabolites enhances its fruiting, promoting its absorption of nutrients from the substrate and, in other words, its vegetative and reproductive growth. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 This is a photo of the plate culture results of Bacillus subtilis isolated in the present invention; Figure 2 This is a microscopic examination diagram of Bacillus subtilis in the present invention; Figure 3 This is a photo of the liquid fermentation results of the Bacillus subtilis metabolites of the present invention; Figure 4 The photos of hyphae growth results of the control group and the addition of Bacillus subtilis metabolites in the examples disclosed in the present invention (measured after 10 days of culture); Figure 5 This is a graph showing the effect of adding different concentrations of Bacillus subtilis metabolites on mycelial growth in the examples disclosed in the present invention (measured after 10 days of culture); DETAILED DESCRIPTION

[0015] For a better understanding of the present invention, the present invention is further described in detail below with reference to the embodiments. However, the scope of protection claimed by the present invention is not limited to the scope represented by the embodiments.

[0016] The raw materials used in the present invention, unless otherwise specified, are conventional commercial products. The methods used in the present invention, unless otherwise specified, are conventional methods in the art. The quality of each substance used in the present invention is the quality of conventional use.

[0017] Application of liquid fermentation metabolites of Bacillus subtilis as growth regulators for the edible fungus Phragmites australis.

[0018] Preferably, the liquid fermentation metabolite of Bacillus subtilis is prepared by culturing in the following liquid culture medium, wherein the culture medium comprises the following components in parts by weight: see Table 2.

[0019] Table 2 Medium for preparing liquid metabolites of Bacillus subtilis Reagent name Dosage / per 100mL Peptone 1g yeast powder 0.5g Sodium chloride 1g Alternatively, the liquid fermentation metabolite of Bacillus subtilis can be used as a growth-promoting regulator for the mycelium of Phragmites australis and / or as a cultivation growth regulator for Phragmites australis; Alternatively, the liquid fermentation culture of Bacillus subtilis requires solid-liquid separation (such as solid-liquid separation by centrifugation or plate-and-frame filtration) to obtain liquid Bacillus subtilis metabolites; Alternatively, when in use, the liquid fermentation metabolites of Bacillus subtilis are directly added to the culture medium of Pseudomonas australis and sterilized together for use.

[0020] Preferably, 0.1-1 mL of Bacillus subtilis liquid fermentation metabolites is added to the culture medium of every 100 g of Phragmites australis.

[0021] A method for applying a liquid fermentation metabolite of Bacillus subtilis as a growth regulator for the edible fungus Phragmites australis, characterized by comprising the following steps: (1) Cultivating and preparing liquid fermentation metabolites of Bacillus subtilis; (2) adding the liquid fermentation metabolites of Bacillus subtilis obtained in step (1) to the culture medium of Pseudomonas aeruginosa, mixing and sterilizing the culture medium, and then the culture medium can be used for culturing the edible fungus Pseudomonas aeruginosa; The liquid fermentation metabolite of Bacillus subtilis can be used as a growth regulator for the edible fungus Phragmites australis.

[0022] Preferably, the metabolites of the liquid fermentation of Bacillus subtilis in step (2) can be used as a growth-promoting regulator for the mycelium of Pseudomonas aeruginosa and / or as a cultivation growth regulator for Pseudomonas aeruginosa.

[0023] Preferably, when the Bacillus subtilis liquid fermentation metabolite agent is used as a growth-promoting regulator for the mycelium of Pseudomonas aeruginosa, 0.1-1 mL of the Bacillus subtilis liquid fermentation metabolite is added to the culture medium of 100 g of Pseudomonas aeruginosa mycelium; When the liquid fermentation metabolite of Bacillus subtilis is used as a growth regulator for the cultivation of Phragmites australis, 0.1-1 mL of the liquid fermentation metabolite of Bacillus subtilis is added to every 100 g of the culture medium of Phragmites australis; Alternatively, in step (1), solid-liquid separation is required after liquid fermentation culture of Bacillus subtilis to obtain liquid Bacillus subtilis metabolites.

[0024] A method for preparing a reed wax agaricus strain using liquid fermentation metabolites of Bacillus subtilis comprises the following steps: adding the liquid fermentation metabolites of Bacillus subtilis to a culture medium of reed wax agaricus, pouring a plate, inoculating reed wax agaricus blocks, and culturing at 25°C for 7 to 12 days. The obtained strain is the reed wax agaricus strain, which can be used as a primary strain of the edible fungus reed wax agaricus; The liquid fermentation metabolite of Bacillus subtilis can be used as a growth regulator for the edible fungus Phragmites australis.

[0025] Alternatively, the liquid fermentation culture of Bacillus subtilis requires solid-liquid separation to obtain liquid Bacillus subtilis metabolites.

[0026] Specifically, the relevant preparation and application results are as follows: A method and application of a liquid fermentation metabolite of Bacillus subtilis as a growth regulator for the edible fungus Phragmites australis. The preparation method comprises the following steps: 1) Cultivation and preparation of metabolites from liquid fermentation of Bacillus subtilis; the liquid culture medium used for liquid fermentation of Bacillus subtilis is shown in Table 1, and the results of liquid fermentation are shown in Figure 2 .from Figure 2 As can be seen, the liquid culture of Bacillus subtilis is effective. 2) The liquid fermentation metabolite solution obtained in step 1) is subjected to solid-liquid separation to obtain a metabolite solution. 3) The Bacillus subtilis metabolite solution obtained in step 2) is used as a growth regulator for mycelial culture and fruiting body cultivation of Pseudomonas aeruginosa.

[0027] The culture medium for the mycelium of Pseudomonas reedii comprises the following components in parts by weight: 2-10% wheat grain, 1-3% glucose, 0.1-0.3% peptone, 1-2% agar, 0.1-0.3% KH2PO4, 0.1-0.2% MgSO4·7H2O, 0.1-1% (v / w) of a Bacillus subtilis metabolite solution. 总 ).

[0028] The culture temperature of reed wax mushroom mycelium is 25℃ and the culture time is 7-14 days. Figure 3 、 Figure 4 The results of an experiment on the growth of Pseudomonas aeruginosa mycelium using different additions of Bacillus subtilis metabolites showed that after 10 days of culture in a constant temperature incubator, the mycelial radius of the 0.1% and 1% additions was significantly greater than that of the control group (the control group did not contain Bacillus subtilis liquid fermentation metabolites and only used the common culture medium for Pseudomonas aeruginosa mycelium). This indicates that Bacillus subtilis metabolites significantly affect the growth of Pseudomonas aeruginosa mycelium.

[0029] The process of the present invention is simple to operate, has a short preparation cycle, and is easy to manufacture in a factory. Adding a Bacillus subtilis metabolite to the culture medium of the edible fungus Psoralea corylifolia causes the mycelium of Psoralea corylifolia to grow thicker and denser, making it easier to cultivate. Adding a Bacillus subtilis metabolite to the culture medium of the edible fungus Psoralea corylifolia causes the mycelium to grow faster and produce more easily. The Bacillus subtilis metabolite acts as a growth regulator, promoting nutrient absorption and cell division during the growth of Psoralea corylifolia.

[0030] The method of the present invention can enhance the vitality of the reed wax mushroom and facilitate fruiting. The Bacillus subtilis metabolites significantly promote the growth of the reed wax mushroom, and the method is easy to implement in factories and small-scale farmers' workshops. The Bacillus subtilis metabolites act as growth regulators, promoting nutrient absorption and cell division during the growth process of the reed wax mushroom.

[0031] It should be noted that in the embodiments of the present invention, v / w (volume to mass ratio) refers to the ratio of mL to g or L to kg, that is, the dimensions are mL to g or L to kg.

[0032] Although the embodiments of the present invention are disclosed for illustrative purposes, those skilled in the art will understand that various substitutions, changes and modifications are possible without departing from the spirit and scope of the present invention and the appended claims. Therefore, the scope of the present invention is not limited to the contents disclosed in the embodiments.

Claims

1. Application of liquid fermentation metabolites of Bacillus subtilis as growth regulators for the edible fungus Phragmites australis.

2. The use according to claim 1, characterized in that: The liquid fermentation metabolite of Bacillus subtilis is prepared by culturing in the following liquid culture medium, wherein the culture medium comprises the following components in parts by weight: Alternatively, the liquid fermentation metabolite of Bacillus subtilis can be used as a growth-promoting regulator for the mycelium of Phragmites australis and / or as a cultivation growth regulator for Phragmites australis; Alternatively, the liquid fermentation culture of the Bacillus subtilis requires solid-liquid separation to obtain liquid Bacillus subtilis metabolites; Alternatively, when using, the liquid fermentation metabolites of Bacillus subtilis are directly added to the culture medium of Pseudomonas australis and sterilized together before use.

3. The use according to claim 2, characterized in that: Add 0.1-1 mL of Bacillus sp. to the culture medium of 100 g of Phragmites australis. Bacillus subtilis Liquid fermentation metabolites.

4. A method for using a liquid fermentation metabolite of Bacillus subtilis as a growth regulator for the edible fungus Phragmites australis, characterized in that: The steps include: (1) Cultivating and preparing liquid fermentation metabolites of Bacillus subtilis; (2) adding the liquid fermentation metabolites of Bacillus subtilis obtained in step (1) to the culture medium of Pseudomonas aeruginosa, mixing and sterilizing the culture medium, and then the culture medium can be used for culturing the edible fungus Pseudomonas aeruginosa; The liquid fermentation metabolite of Bacillus subtilis can be used as a growth regulator for the edible fungus Phragmites australis.

5. The application method according to claim 4, characterized in that: The metabolites of the liquid fermentation of Bacillus subtilis in step (2) can be used as a growth-promoting regulator for the mycelium of Pseudomonas aeruginosa and / or as a cultivation growth regulator for Pseudomonas aeruginosa.

6. The application method according to claim 5, characterized in that: When the Bacillus subtilis liquid fermentation metabolite is used as a growth-promoting regulator for the mycelium of Pseudomonas aeruginosa, 0.1-1 mL of the Bacillus subtilis liquid fermentation metabolite is added to the culture medium of 100 g of Pseudomonas aeruginosa mycelium; When the Bacillus subtilis liquid fermentation metabolite agent is used as a growth regulator for cultivation of reed wax mushroom, 0.1-1 mL of the Bacillus subtilis liquid fermentation metabolite is added to every 100 g of reed wax mushroom culture material; Alternatively, the culture temperature of the mycelium of Pseudomonas aeruginosa is 25° C., and the culture time is 7-14 days; Alternatively, the cultivation time of the fruiting body of Phragmites australis is 50-60 days from the start of spawning.

7. The use according to claim 6, characterized in that: The culture medium of the mycelium of the reed parasol mushroom comprises the following components in percentage by weight: Wheat grain 2-10%, glucose 0.1-0.3%, peptone 1-1.5%, KH2PO4 0.3-0.5%, MgSO4·7H2O 0.7-1%, Bacillus subtilis liquid fermentation metabolite solution 0.1-1% (v / w 总 ); Alternatively, the cultivation medium for Phragmites australis comprises the following components in percentage by weight: Here, v / w refers to the ratio of mL to g.

8. The application method according to any one of claims 4 to 7, characterized in that: The liquid fermentation metabolites of Bacillus subtilis in step (1) are prepared by culturing in the following liquid culture medium, wherein the culture medium comprises the following components in parts by weight: Alternatively, in step (1), solid-liquid separation is required after liquid fermentation culture of Bacillus subtilis to obtain liquid Bacillus subtilis metabolites.

9. A method for preparing reed parasitic mushrooms by utilizing liquid fermentation metabolites of Bacillus subtilis, characterized in that: The method comprises the following steps: adding liquid fermentation metabolites of Bacillus subtilis to a culture medium of Pseudomonas aeruginosa, pouring a plate, inoculating Pseudomonas aeruginosa blocks, and culturing at 25° C. for 7 to 12 days to obtain a Pseudomonas aeruginosa strain, which can be used as a primary strain of the edible fungus Pseudomonas aeruginosa. The liquid fermentation metabolite of Bacillus subtilis can be used as a growth regulator for the edible fungus Phragmites australis.

10. The method according to claim 9, characterized in that: The liquid fermentation metabolite of Bacillus subtilis is prepared by culturing in the following liquid culture medium, wherein the culture medium comprises the following components in parts by weight: Alternatively, the liquid fermentation metabolite of Bacillus subtilis can be used as a growth-promoting regulator for the mycelium of Pseudomonas aeruginosa and / or as a cultivation growth regulator for Pseudomonas aeruginosa; Alternatively, the liquid fermentation culture of the Bacillus subtilis requires solid-liquid separation to obtain liquid Bacillus subtilis metabolites.

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