Bacillusinaquosorum and application thereof

Abstract

The invention provides a strain of Bacillus inaquosorum and application of the strain of Bacillus inaquosorum, and relates to the technical field of microorganisms. The invention provides a strain of Bacillus inaquosorum cmc7, and the preservation number of the strain is CGMCC (China General Microbiological Culture Collection Center) No. 34133; the strain has good starch degradation capability and cellulose degradation capability, can be used for degrading plant straws, also has the capability of preventing and treating plant diseases, and has great potential in agricultural applications such as straw returning, plant disease prevention and treatment and the like. A metabolite prepared from the cmc7 strain also has antibacterial ability, the antibacterial ability is positively correlated with the concentration of the metabolite, and the cmc7 strain can be used for preventing and treating pathogenic bacteria or preparing products for preventing and treating pathogenic bacteria.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms, and particularly relates to a Bacillus inaquosorum strain and application thereof. Background Art

[0002] The resource utilization of plant straw and the prevention and control of pathogenic fungal diseases are crucial for ensuring sustainable agricultural development. As agricultural waste, plant straw primarily consists of carbohydrates such as cellulose, hemicellulose, and starch. Traditional incineration not only wastes resources but also causes serious environmental pollution. While returning straw to the fields is an ideal resource, its natural degradation process is limited by the efficiency of microbial decomposition, often resulting in high levels of straw residue and long decomposition cycles. Furthermore, undegraded straw can easily become a host for pathogenic fungi, increasing the risk of soil-borne diseases.

[0003] The development and application of microbial agents offer new avenues for addressing these challenges. However, existing technologies have yet to effectively integrate straw degradation and disease control capabilities within a single bacterial strain. Therefore, achieving the organic integration of straw resource utilization and green control of pathogenic fungi within a single bacterial strain system remains a pressing technical challenge in this field. Summary of the Invention

[0004] In view of this, the object of the present invention is to provide a Bacillus inaquosorum and its application, wherein the Bacillus inaquosorum has good cellulose degradation and starch decomposition properties, and also has the functions of straw degradation and plant disease prevention and control.

[0005] In order to achieve the above object, the present invention provides the following technical solutions:

[0006] The invention provides a Bacillus inaquosorum strain. The Bacillus inaquosorum strain is named cmc7 and has a preservation number of CGMCC No.34133.

[0007] The present invention also provides a fermentation broth comprising the cmc7.

[0008] The present invention also provides a method for preparing the fermentation broth, comprising inoculating the cmc7 into a culture medium and culturing the culture medium to obtain the fermentation broth.

[0009] Preferably, the culture medium comprises NB liquid medium.

[0010] Preferably, the culture temperature is 25-30° C., and the culture time is 2-4 days.

[0011] The present invention also provides a metabolite, and the preparation method of the metabolite comprises: centrifuging the fermentation broth to obtain the supernatant, freeze-drying to obtain a fermentation extract, purifying it using XAD-16 macroporous adsorption resin, and eluting with eluent to obtain the metabolite.

[0012] Preferably, the elution is performed successively with 600 mL of water, 600 mL of 30% methanol aqueous solution and 600 mL of 100% methanol solution.

[0013] The present invention also provides the use of the Bacillus inaquosorum, the fermentation broth, the preparation method or the metabolite in any one or more of the following:

[0014] S1, degrade cellulose,

[0015] S2, degraded starch,

[0016] S3. Degrade straw.

[0017] The present invention also provides the use of the Bacillus inaquosorum, the fermentation liquid, the preparation method or the metabolite in preparing a product for preventing and controlling pathogens or preventing and controlling plant diseases.

[0018] Preferably, the pathogen includes Alternaria alternate, and the plant disease includes brown spot disease.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] The present invention provides a strain of Bacillus inaquosorum, designated cmc7, which has been deposited with the China General Microbiological Culture Collection Center under the accession number CGMCC No. 34133. This strain has excellent starch-degrading and cellulose-degrading capabilities, making it suitable for degrading plant straw. Furthermore, this strain has the ability to prevent and control plant diseases, showing great potential in agricultural applications such as returning straw to fields and preventing and controlling plant diseases.

[0021] The present invention also provides a metabolite of Bacillus inaquosorum, which is prepared by the cmc7 strain. The antibacterial ability of the metabolite is positively correlated with its concentration and has significant antibacterial ability. The metabolite can be used to prevent and control pathogens or prepare products for preventing and controlling pathogens.

[0022] Biodeposit Information:

[0023] The Bacillus inaquosorum of the present invention has a strain name of cmc7, is classified as Bacillus inaquosorum, and is deposited in the General Microbiology Center of the China Culture Collection Administration of Microorganisms, abbreviated as CGMCC. The strain deposit number is CGMCC No. 34133, the deposit date is April 8, 2025, and the deposit address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a diagram of the colony growth morphology of the cmc7 strain in Example 1 on the culture medium.

[0025] Figure 2 This is the phylogenetic tree of the cmc7 strain in Example 1 constructed using MEGA 11 software.

[0026] Figure 3 This is a graph showing the test results of the cellulose degradation function of the cmc7 strain in Example 2.

[0027] Figure 4 This is a graph showing the starch degradation function test results of the cmc7 strain in Example 2.

[0028] Figure 5 This is a graph showing the filter paper disintegration test results of the cmc7 strain in Example 2.

[0029] Figure 6 This is a diagram showing the inhibitory effect of the cmc7 strain on the red star pathogen Alternaria alternate in Example 2.

[0030] Figure 7 This is a diagram showing the inhibitory effect of different concentrations of the metabolites of the cmc7 strain on the red star pathogen Alternaria alternate in Example 3. DETAILED DESCRIPTION

[0031] The invention provides a Bacillus inaquosorum strain. The Bacillus inaquosorum strain is named cmc7 and has a preservation number of CGMCC No.34133.

[0032] In the present invention, the cmc7 was isolated from the humus layer soil of dead branches and fallen leaves in the tobacco research institute forest and was deposited in the China General Microbiological Culture Collection Center on April 8, 2025.

[0033] The colony morphology of the cmc7 cultured on the NA solid culture medium is a light yellow circular colony with a matte and raised surface.

[0034] The present invention also provides a fermentation broth comprising the cmc7. In the present invention, the fermentation broth has the ability to degrade cellulose and starch, and further has the ability to degrade straw while also having the function of preventing and treating plant diseases. In the present invention, the pathogenic bacteria include Alternaria alternata.

[0035] In the present invention, the method for preparing the fermentation broth comprises inoculating the cmc7 in a culture medium to obtain a fermentation broth. The culture medium is NB liquid culture medium. The culture temperature is 25 to 30°C, more preferably 26 to 28°C, and most preferably 28°C. The culture time is preferably 2 to 4 days, more preferably 3 days. As an optional embodiment, the fermentation broth is OD 600 200 μL of the bacterial suspension of the cmc7 strain with a value of about 0.3 was inoculated into 500 mL of NB liquid culture medium and fermented at 180 rpm and 28° C. for 3 days to obtain the fermentation broth of the cmc7 strain.

[0036] The present invention also provides a metabolite, comprising a method for preparing the metabolite, comprising: centrifuging the fermentation broth, extracting the supernatant, lyophilizing the supernatant, obtaining a fermentation extract, purifying the extract using an XAD-16 macroporous adsorption resin, and eluting the extract with an eluent to obtain the metabolite. Preferably, the elution is performed sequentially with 600 mL of water, 30% methanol, and 100% methanol. The 100% methanol phase eluate is collected, evaporated, and weighed to obtain the purified metabolite.

[0037] In the present invention, when using the fermentation broth to prepare the metabolites, the centrifugation speed is preferably 6,000 to 10,000 rpm, more preferably 8,000 rpm. The lyophilization of the fermentation supernatant into a lyophilized powder is performed using conventional lyophilized powder preparation methods, without further limitation. In the present invention, when purifying the fermentation extract obtained above, preferably XAD-16 macroporous adsorption resin is used; more preferably, the glass column containing the XAD-16 macroporous adsorption resin has an inner diameter of 800 mm × 40 mm. Prior to purification, the fermentation extract is dissolved in distilled water at a mass volume ratio of 1 g:10 mL and loaded onto the XAD-16 macroporous resin wet-packed in a glass column (800 mm × 40 mm inner diameter). Elution is then performed sequentially using 600 mL of water, 600 mL of 30% methanol solution (water:methanol = 7:3), and 600 mL of 100% methanol solution (anhydrous methanol, purchased from China National Pharmaceutical Group Co., Ltd.), at a flow rate of approximately 3 mL / min. The 100% methanol eluate was collected, evaporated and weighed to obtain the purified metabolites.

[0038] The present invention also provides the use of the Bacillus inaquosorum, the fermentation broth, the preparation method or the metabolite in any one or more of the following:

[0039] S1, degrade cellulose,

[0040] S2, degraded starch,

[0041] S3. Degrade straw.

[0042] In the present invention, the straw includes one or more of tobacco straw, wheat straw and rice straw.

[0043] The present invention also provides the use of the Bacillus inaquosorum, the fermentation broth, the preparation method, or the metabolite in preparing a product for preventing and treating pathogens or preventing and treating plant diseases. In the present invention, the pathogen includes Alternaria alternate, and the plant disease includes brown spot disease.

[0044] The brown spot pathogen Alternaria alternate in the present invention can infect a variety of plants, such as tobacco, tomato, pepper, eggplant, etc.

[0045] In the present invention, the cmc7 metabolite extract has an inhibitory effect on the red star pathogen Alternaria alternate CX06. As the concentration increases, the inhibitory effect also increases. 50 It is 0.498mg / mL.

[0046] The present invention does not specifically limit the product type. In the product, the Bacillus inaquosorum c-terminal, the fermentation broth, or the metabolite can be the sole active ingredient, or other active ingredients may be included. The product of the present invention can be used alone or in combination with other excipients and / or products. The present invention does not specifically limit the concentration of the bacterial solution for specific applications and can be conventionally selected based on actual needs.

[0047] In the present invention, the soil of the humus layer of dead branches and leaves in the forest was taken from the Tobacco Research Institute of the Chinese Academy of Agricultural Sciences in November 2023.

[0048] In the present invention, in the following embodiments:

[0049] The Cas02 strain: Bacillus amyloliquefaciens, the strain name is Cas02, the classification name is Bacillus amyloliquefaciens, deposited in the General Microbiology Center of the China Microorganism Culture Collection Administration, referred to as CGMCC, the strain deposit number is CGMCC No.15514, and the deposit time is: March 26, 2018. The deposit address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing. The Bacillus amyloliquefaciens Cas02 is Bacillus amyloliquefaciens Cas02, with a deposit number of CGMCCNO.15514. It has been disclosed in the prior art, see the literature: Chu DP, Ilyas N, Peng LJ, Wang XQ, Wang DK, Xu ZC, Gao Q, Tan XL, Zhang CS, LiY Q, Yuan Y, 2022. Genomic insights on fighting bacterial wilt by a novel Bacillus amyloliquefaciens strain Cas02. Microbial Biotechnology, 15(4): 1152-1167.

[0050] The YG3 strain: Stenotrophomonas maltophilia; the strain name is YG3, the classification name is Stenotrophomonas maltophilia, and it is deposited in the General Microbiology Center of the China Culture Collection Administration of Microorganisms, with the strain deposit number CGMCC No. 34131, and the deposit date is April 8, 2025. The deposit address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0051] The htf4 strain: Delftia acidovorans; the strain name is htf4, classified as Delftia acidovorans, preserved in the General Microbiology Center of the China Culture Collection Administration of Microorganisms, abbreviated as CGMCC, the strain collection number is CGMCC No.34132, the preservation time is: April 8, 2025, and the preservation address is: No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing.

[0052] The NH1 / 13 strain: Brachybacterium horti, the strain name is NH1 / 13, the classification name is Brachybacterium horti, it is preserved in the General Microbiology Center of the China Culture Collection Administration of Microorganisms, abbreviated as CGMCC, the strain collection number is CGMCC No. 29851, the preservation time is: February 2, 2024, and the preservation address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0053] In the present invention, the Alternaria alternate CX06 described in the following examples is a known strain, see literature: Sui X, Han X, Cao J, Li Y, Yuan Y, Gou J, Zheng Y, Meng C, Zhang C. Biocontrol potential of Bacillus velezensis EM-1 associated with suppressive rhizospheresoil microbes against tobacco bacterial wilt. Front Microbiol. 2022Aug 23; 13: 940156.

[0054] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; the materials and reagents used are commercially available reagents and materials unless otherwise specified.

[0055] Example 1

[0056] 1. Isolation of Bacillus inaquosorum

[0057] 10 g of soil from the humus layer of dead branches and leaves in the tobacco forest was added to 90 mL of sterilized ultrapure water, shaken at 28°C and 180 rpm for 30 min, mixed thoroughly and then allowed to stand. 1 mL of the suspension was taken for gradient dilution. 100 μL of the diluted suspension was spread on NA solid culture medium and cultured in an incubator at 28°C for 5 days. The colonies on the NA solid culture medium were streaked and purified repeatedly for 5 times. The purified single strain was cultured on the NA solid culture medium. The colony morphology on the culture medium was a matte and raised light yellow round colony (such as Figure 1 The strain number is cmc7.

[0058] 2. Identification of Bacillus inaquosorum

[0059] Genomic DNA from the cmc7 strain was extracted and PCR amplified using 2× Taq Plus Master Mix II DNA polymerase from Novozymes, using the extracted cmc7 genomic DNA as the template and 16S rRNA universal primers. The resulting PCR product was sent to Beijing Qingke Biotechnology Co., Ltd. for sequencing.

[0060] The sequences of the 16S rRNA universal primers are: upstream primer (27F): 5'-AGAGTTTGATCCT GGCTCAG-3' (SEQ ID NO. 1), downstream primer (1492R): 5'-TACGGTTACCTTG TTACGACTT-3' (SEQ ID NO. 2). The PCR amplification system is shown in Table 1, and the PCR amplification program used is shown in Table 2.

[0061] Table 1 PCR amplification system

[0062] Reagent name Dosage / μL DNA template (genomic DNA of NH1 / 13) 1 Upstream primer 1 Downstream primer 1 2×TaqPlusMasterMixⅡ 12.5 <![CDATA[ddH2O]]> 9.5

[0063] Table 2 PCR amplification program

[0064]

[0065] The 16S rRNA amplified sequence (1424 bp) of the cmc7 strain is as follows:

[0066]

[0067] Gene sequence comparison and analysis were performed using EzBioCloud (https: / / www.ezbiocloud.net / ) and NCBI Blast, and similar bacterial sequences with high similarity to the amplified sequence of cmc7 strain were downloaded and phylogenetic trees were constructed using MEGA 11 software. Figure 2 As shown, the cmc7 strain was most closely related to Bacillus_inaquosorum|GCA_000332645.1 at 99.93%. The results confirmed that the strain cmc7 belonged to Bacillus inaquosorum.

[0068] The strain cmc7 was deposited in the General Microbiology Center of the China Culture Collection Administration, abbreviated as CGMCC, on April 8, 2025. The culture deposit number is CGMCC No. 34133, and the deposit address is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0069] Example 2

[0070] 1.Bacillus inaquosorum degradation function test

[0071] (1) Bacillus inaquosorum has the ability to degrade cellulose

[0072] The cellulose degradation test of cmc7 strain was carried out using 2g / L CMC-Na as indicator and NA solid medium as basal medium. 600 2.5 μL of a bacterial suspension with a pH value of approximately 0.3 was inoculated in the center of a plate containing an indicator and incubated in the dark at 28°C for 48 hours. After removing the bacterial colonies of the cmc7 strain, the plate was stained with 1g / L Congo red for 30 minutes, and then decolorized with 1mol / L NaCl solution. The color change of the plate was observed, and the cellulose degradation ability of the cmc7 strain was evaluated based on the diameter of the color change circle. Other strains screened at the same time were tested. Figure 3 The results showed that the cmc7 strain had a stronger ability to degrade sodium carboxymethyl cellulose than the Cas02 strain (Bacillus amyloliquefaciens), the yg3 strain (Stenotrophomonas maltophilia), the htf4 strain (Delftia acidovorans), and the NH1 / 13 strain (Brachybacterium horti).

[0073] (2) Bacillus inaquosorum has the function of degrading starch

[0074] The specific experimental method is as follows: 2g / L soluble starch was used as substrate and NA solid medium was used as basal medium to test the starch degradation of the strain. 600 2.5 μL of the bacterial suspension of the cmc7 strain with a value of about 0.3 was inoculated in the center of the plate added with the indicator and incubated in the dark at 28°C for 48 hours. After removing the bacterial colonies of the cmc7 strain, the plate was stained with iodine-potassium iodide solution (1 g / L) as a stain. The color change of the plate was observed, and the starch degradation ability of the cmc7 strain was evaluated based on the diameter of the color change circle. At the same time, other strains screened by this laboratory were tested. Figure 4 The results showed that the cmc7 strain had a stronger ability to degrade starch than the Cas02 strain (Bacillus amyloliquefaciens), the yg3 strain (Stenotrophomonas maltophilia), the htf4 strain (Delftia acidovorans), and the NH1 / 13 strain (Brachybacterium horti).

[0075] (3) Bacillus inaquosorum has the ability to degrade filter paper

[0076] In order to test the ability of the cmc7 strain to degrade cellulose, a filter paper disintegration experiment was set up, with no bacteria added as a blank control (CK). The results showed that the strain had a certain ability to disintegrate filter paper. The specific test method is as follows: a filter paper disintegration test was performed on the cmc7 strain using 0.05g filter paper strips as substrate and 50mL NB liquid culture medium as basal culture medium. Two filter paper strips were placed in each treatment. The OD 600 The bacterial suspension with a value of 0.3 was inoculated into the culture medium at a concentration of 1% and cultured at 180 rpm and 28°C for 10 days. The disintegration of the filter paper was observed and the degree of disintegration was evaluated to evaluate the cellulose degradation ability of cmc7. At the same time, other strains screened in this laboratory were tested. Figure 5The results showed that the cmc7 strain had the strongest ability to degrade filter paper compared to the Cas02 strain (Bacillus myloliquefaciens), the yg3 strain (Stenotrophomonas maltophilia), the htf4 strain (Delftia acidovorans), and the NH1 / 13 strain (Brachybacterium horti).

[0077] (4) Bacillus inaquosorum has the function of degrading straw

[0078] In order to verify the degradation effect of the cmc7 strain screened in Example 1 on real straw materials, the following experiment was set up: tobacco straw / wheat straw / rice straw powder was added to distilled water to a final concentration of 20 g / L, and then 15.15 g / L Na2HPO4·12H2O, 3 g / L KH2PO4, 0.5 g / L NaCl, and 1 g / L NH4Cl were added and sterilized for use.

[0079] OD 600 The cmc7 strain suspension with a value of about 0.3 was inoculated into the above-mentioned straw culture medium, and the fermentation medium without bacterial liquid was used as a control. After culturing at 30℃ and 180rpm for 10 days, the supernatant was discarded by centrifugation, and the straw was repeatedly rinsed with distilled water. After centrifugation, the relative degradation rate of the straw was determined by the weight loss method according to the following formula.

[0080] Y=(m1-m2) / m1*100%

[0081] Where: Y is the degradation rate; m1 is the dry mass of straw before fermentation; m2 is the dry mass of straw after fermentation.

[0082] The results are shown in Table 3.

[0083] Table 3 Relative degradation rates of different straws by bacterial suspension of cmc7 strain

[0084] Group tobacco straw wheat straw rice straw CK 16.54±0.13％b 19.00±0.00％b 17.33±0.67％b cmc7 22.06±0.29％a 24.33±0.18％a 21.67±0.51％a

[0085] (5) Bacillus inaquosorum has the function of antagonizing tobacco brown spot pathogenic fungi

[0086] The specific experimental methods are as follows:

[0087] The pathogen of tobacco brown spot disease (Alternaria alternate, CX06) was activated on potato dextrose agar (PDA) solid medium. A 5mm punch was used to take a bacterial cake of the pathogen Alternaria alternate CX06 and inoculated it into the center of the PDA medium. The OD 600 50 μL of bacterial suspensions of cmc7, Cas02, yg3, htf4, and NH1 / 13 strains (with a RI of approximately 0.3) were added to the perforations of OA oatmeal culture medium. The suspensions were incubated in the dark at 25°C for 10 days, and the antibacterial effects were observed.

[0088] Depend on Figure 6 As can be seen, the cmc7 strain has a significant inhibitory effect on A. amyloliquefaciens compared to the Cas02 strain (Bacillus amyloliquefaciens), the yg3 strain (Stenotrophomonas maltophilia), the htf4 strain (Delftia acidovorans), and the NH1 / 13 strain (Brachybacterium horti).

[0089] Example 3

[0090] 1. Functional studies of Bacillus inaquosorum metabolites

[0091] The specific experimental methods are as follows:

[0092] OD 600 200 μL of the bacterial suspension of the cmc7 strain obtained in Example 1, with a RI of approximately 0.3, was inoculated into 500 mL of NB liquid medium and fermented at 180 rpm and 28°C for 3 days to obtain a fermentation broth of the cmc7 strain. The fermentation broth of the cmc7 strain obtained above was centrifuged at 8000 rpm for 5 minutes, and the supernatant was lyophilized to a powder to obtain a fermentation extract. The fermentation extract was partially purified using the macroporous adsorption resin XAD-16 (Shanghai Mosu Scientific Instrument Co., Ltd.). 5 g of the fermentation extract was dissolved in 50 mL of distilled water and loaded onto the XAD-16 macroporous resin wet-packed in a glass column (800 mm × 40 mm inner diameter). Elution was then performed sequentially with 600 mL of water, 600 mL of 30% methanol in water (water:methanol = 7:3), and 600 mL of 100% methanol (anhydrous methanol, purchased from China Pharmaceutical Group Co., Ltd.) at a flow rate of 3 mL / min. The 100% methanol phase eluate was collected, rotary evaporated in a 40°C water bath, and weighed to obtain the purified 100% methanol phase metabolites.

[0093] Weigh 100 mg of the above-mentioned purified 100% methanol phase metabolite into a 2 mL EP centrifuge tube, vortex and oscillate for 5 minutes with 500 μL of anhydrous methanol, and then centrifuge (8000 rpm for 5 minutes) to collect the supernatant. The precipitate after centrifugation is further extracted with 500 μL of anhydrous methanol to ensure that the active substance is completely dissolved. The extraction operation is repeated three times. After the three extractions, about 1500 mL of methanol extract obtained is dried with nitrogen, dissolved in 10 mL of 30% methanol solution, and sterilized by 0.22 μm filter membrane to obtain a purified 100% methanol phase metabolite solution.

[0094] The red star pathogen Alternaria alternate CX06 was activated and cultured in OA medium for about 7 days. The purified 100% methanol phase metabolite solution obtained above was subjected to antibacterial tests at final concentrations of 2 mg / mL, 1 mg / mL and 0.5 mg / mL respectively. The extract solution was first dissolved in OA oatmeal medium and then poured into the plate. After the plate solidified and cooled, a 5mm punch was used to take the red star fungus cake (Alternaria alternate CX06) and placed it in the center of the culture medium. It was cultured in the dark at 25°C for 7-10 days, and the antibacterial effect was observed. Figure 7 The results showed that cmc7 extract had an inhibitory effect on Alternaria alternate CX06. With the increase of concentration, the inhibitory effect also increased. 50 About 0.498mg / mL.

[0095] The results of the above examples show that the cmc7 strain has the functions of degrading cellulose, degrading straw, and preventing and controlling the pathogen of tobacco brown spot disease (Alternaria alternate, CX06). The metabolites of the cmc7 strain have an inhibitory effect on the brown spot disease pathogen Alternaria alternateCX06, and the inhibitory effect increases with increasing concentration.

[0096] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. A strain of Bacillus inaquosorum, characterized in that The Bacillus inaquosorum is named cmc7 and its deposit number is CGMCC No.34133.

2. A fermentation broth, characterized in that: Containing cmc7 as claimed in claim 1.

3. The method for preparing the fermentation broth according to claim 2, characterized in that: The method comprises inoculating the cmc7 into a culture medium and culturing the culture medium to obtain a fermentation liquid.

4. The preparation method according to claim 3, characterized in that The culture medium includes NB liquid medium.

5. The preparation method according to claim 3, characterized in that The culture temperature is 25-30° C., and the culture time is 2-4 days.

6. A metabolite, characterized in that The preparation method of the metabolite comprises: centrifuging the fermentation broth according to claim 2, taking the supernatant, freeze-drying to obtain a fermentation extract, purifying it using XAD-16 macroporous adsorption resin, and eluting with eluent to obtain the metabolite.

7. The metabolite according to claim 6, characterized in that The elution was performed successively with 600 mL of water, 600 mL of a 30% methanol aqueous solution and 600 mL of a 100% methanol solution.

8. Use of the Bacillus inaquosorum of claim 1, the fermentation broth of claim 2, the preparation method of claim 3 or 4, or the metabolite of claim 5 or 6 in any one or more of the following: S1, degrade cellulose, S2, degraded starch, S3. Degrade straw.

9. Use of the Bacillus inaquosorum according to claim 1, the fermentation liquid according to claim 2, the preparation method according to claim 3 or 4, or the metabolite according to claim 5 or 6 in preparing a product for controlling pathogens or controlling plant diseases.

10. The use according to claim 9, characterized in that The pathogenic bacteria include Alternaria alternate, and the plant diseases include brown spot disease.

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