CN115109723B

CN115109723B - Bacillus californicus ST-1 and application thereof in pear disease control or storage and preservation

Info

Publication number: CN115109723B
Application number: CN202210719933.0A
Authority: CN
Inventors: 赵锦芳; 李敏; 龙同; 黄满; 高娃; 罗昕; 段先莉
Original assignee: Hubei University of Technology
Current assignee: Hubei University of Technology
Priority date: 2022-06-23
Filing date: 2022-06-23
Publication date: 2023-06-23
Anticipated expiration: 2042-06-23
Also published as: CN115109723A

Abstract

The invention provides bacillus caldarius ST-1 and application thereof in pear disease control or preservation and fresh-keeping, which is named bacillus caldarius ST-1 (Bacillus cabrialesii ST-1), and the preservation number is CCTCC NO: m2022671. The bacillus Bacillus cabrialesii ST-1 has the advantages that the generated volatile gas and thallus fermentation liquid have broad-spectrum resistance to plant pathogenic fungi, have remarkable antibacterial effect on pear rot and pear ring rot, have the antibacterial rates of 92.43 percent and 72.23 percent respectively, and have wide application prospects in the aspects of pear disease control, storage and preservation. Meanwhile, the strain has strong antibacterial activity on Fusarium, and can also be used for preventing and controlling cotton wilt and sweet potato diseases.

Description

Bacillus californicus ST-1 and application thereof in pear disease control or storage and preservation

Technical Field

The invention relates to the technical fields of microbiology, biochemistry and fermentation engineering, in particular to bacillus caldarius ST-1 and application thereof in pear disease control or storage and preservation.

Background

Pears are the third largest fruit industry in China, which is inferior to citrus and apples, and the cultivation area, yield, export quantity, variety number and the like of pears are in the forefront of the world. Korla pear (pear for short) (Pyrus sinkiangensi) is mainly planted in Korla and Ackesu region of Xinjiang. The disease problem of the bergamot pears in the cultivation process is particularly prominent and the development of the bergamot pear planting industry is severely restricted under the influence of factors such as periodic freeze injury, improper management and the like. The pear rot belongs to typical branch diseases, is caused by apple black rot peel pear variety (Valsa mali var. Pyri), and is one of the most important fungal diseases in pear cultivation and production. The pear garden has wide occurrence area, is easy to spread infection, and is aggravated year by year, the average incidence rate of the pear garden is 50% -66%, and the serious pear garden can reach more than 94%. The method can be interacted with rot germs of other hosts, so that the actual prevention and control difficulty is high. Bergamot pears are also vulnerable to pathogenic microorganisms in the postharvest storage period, ring rot is an important disease of pears in the storage period, and can cause rot of fruits in the storage period. The ring spot is one of main diseases in the field, can infect branches, leaves and fruits, can cause destructive damage to an orchard when serious, affects the yield and fruits of the orchard, and causes loss of economic benefits.

At present, the pear diseases are mainly controlled chemically, and the storage and fresh-keeping methods are also mainly treated by chemical reagents or smeared by edible film coating agents. With the increasing attention of people on chemical pesticide residues and fruit safety, the development of an environment-friendly biological control method is a key for pear disease control.

Therefore, there is a need to develop a biological control method.

Disclosure of Invention

The invention aims to provide bacillus caldarius ST-1 and application thereof in pear disease control or storage and preservation, and volatile gas and thallus fermentation liquor generated by the bacillus caldarius ST-1 have broad-spectrum resistance to plant pathogenic fungi, have remarkable antibacterial effects on pear rot and pear ring rot, and have wide application prospects in pear disease control, storage and preservation.

In a first aspect of the present invention, there is provided a strain of Bacillus Carlsbergensis ST-1, wherein the strain of Bacillus Carlsbergensis ST-1 is classified under the name Bacillus cabrialesii ST-1, and the accession number is: cctccc NO: m2022671.

In a second aspect of the present invention, there is provided a fermentation broth comprising:

fermenting the bacillus californicus ST-1 to obtain fermentation liquor or bacterial suspension;

or spray drying the fermentation liquor to obtain the dry powder microbial inoculum.

Further, the preparation method of the bacterial suspension comprises the following steps:

inoculating ST-1 to LB solid plate for overnight culture, picking single colony to LB liquid culture medium for culture, centrifuging, collecting thallus, washing, and suspending the thallus with PBS buffer solution to concentration of (0.5-5) x 10 ⁸ CFU/mL。

Further, the culture conditions of the strain ST-1 include: the temperature is 28-32 ℃ and the pH is 5-9.

Further, the formula of the culture medium is as follows: 13.1g/L sucrose, 17.0g/L bran + peptone (1:2) and 2.0g/L potassium dihydrogen phosphate.

In a third aspect of the invention, there is provided a volatile gas prepared from the bacterial suspension of claim 2 or 3, comprising: coating the bacterial suspension on an LB solid plate, placing the LB solid plate in a polyethylene film bag for sealing, standing at 25 ℃ under the condition of HR 60% -80%, and detecting by adopting headspace solid-phase microextraction combined with GC-MS.

In a fourth aspect of the invention, the application of the bacillus californicus ST-1 or the zymophyte agent or the volatile gas in pear disease control is provided.

Further, pathogenic fungi of the bergamot pear disease include bergamot pear rot pathogen Valsa mali var. Pyri and bergamot pear ring rot pathogen Botryosphaeria dothidea.

In a fifth aspect of the invention, the application of the bacillus californicus ST-1 or the zymophyte agent or the volatile gas in the control of apple rot, cotton wilt or sweet potato diseases is provided.

The bacillus calmette-guerin ST-1 has wide bacteriostasis spectrum, can obviously inhibit important diseases in the growth period and the postharvest storage period of bergamot pears, and has stable effect. The bacterial gas volatile matter or bacterial suspension or fermentation liquor can be applied to biological control of pear rot and ring rot, and is also suitable for control of apple rot, cotton wilt and various diseases of sweet potato.

One or more technical solutions in the embodiments of the present invention at least have the following technical effects or advantages:

the bacillus californicus ST-1 and the application thereof in pear disease prevention or storage and preservation provided by the invention have broad-spectrum resistance to plant pathogenic fungi by volatile gas and thallus fermentation liquor generated by the bacillus californicus ST-1, have obvious antibacterial effects on pear rot and pear ring rot, have antibacterial rates of 92.43% and 72.23%, and have wide application prospects in pear disease prevention and preservation. Meanwhile, the strain has strong antibacterial activity on Fusarium, and can also be used for preventing and controlling cotton wilt and sweet potato diseases. The strain provided by the invention has the advantages of stable performance, simple preparation method, low cost, high viable count and large comprehensive development potential, can be used for preparing functional biocontrol agents and microbial agents for fruit preservation, and has a wide application prospect.

The preservation date of the bacillus caldarius ST-1 is 2022, 5 and 19 days, and the preservation number is CCTCC NO: m2022671. The classification is Bacillus cabrialesii ST-1, the preservation unit name is China center for type culture Collection, and the address is the university of Wuhan in Wuhan, hubei province, china, post code: 430072.

drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for the description of the embodiments will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a photograph showing colonies of Bacillus californicus Bacillus cabrialesii ST-1 isolated in the present invention on LB solid medium;

FIG. 2 is a total ion flow chromatogram of Bacillus californicus ST-1 volatile gas.

FIG. 3 shows antagonism of phytopathogenic fungi by Bacillus Carlsbergensis ST-1 volatile gases; and (3) injection: a: watermelon fusarium wilt Fusarium oxysporum; b: wheat scab Fusarium graminearum; c: sweet potato stem rot germ Fusarium proliferatu; d: rhizoctonia cerealis Botryosphaeria dothidea; e: botrytis cinerea; f: sweet potato root rot fungi Fusarium Solani; g: sweet potato vine cutting bacteria Fusarium bulbigenum; h: pyri rot pathogen Valsa mali var.

FIG. 4 is an antagonism of a Bacillus californicus ST-1 bacterial suspension or broth against a phytopathogenic fungus; and (3) injection: a: botrytis cinerea; b: gibberella wheat germ; c: watermelon fusarium wilt bacteria; d: sweet potato stem rot germ; e: black spot germ of sweet potato.

FIG. 5 shows the control of post harvest leaf scald disease in bergamot pear by bacillus californicus ST-1 volatile gas or bacterial suspension or fermentation broth; a: treating gas volatile matters; b: and (5) treating fermentation liquor.

Detailed Description

The advantages and various effects of the present invention will be more clearly apparent from the following detailed description and examples. It will be understood by those skilled in the art that these specific embodiments and examples are intended to illustrate the invention, not to limit the invention.

Throughout the specification, unless specifically indicated otherwise, the terms used herein should be understood as meaning as commonly used in the art. Accordingly, unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification will control.

Unless specifically indicated otherwise, the various raw materials, reagents, instruments, equipment, etc., used in the present invention are commercially available or may be obtained by existing methods.

The bacillus caldarius ST-1 and the application thereof in pear disease control or preservation and fresh-keeping will be described in detail below with reference to examples and experimental data.

Example 1 isolation, purification and characterization of Strain ST-1

1. Isolation of Strain ST-1

Isolation and purification of strains: collecting the rhizosphere soil of forest plants in Hefeng county of Hubei province, enriching and culturing, and separating bacteria by a dilution plate method. Single colony with different forms, colors and sizes is selected, purified by streaking a 3-5 generation plate, transferred to an LB culture medium inclined plane and preserved at 4 ℃.

Inoculating the single bacterial colony into LB liquid culture medium, shake culturing at 30deg.C and 200rpm for 36-72 hr to obtain fermentation liquid. Two holes are uniformly and symmetrically drilled on a PDA solid flat plate at a position which is 2.0cm away from a central point, a strain suspension or fermentation liquor is inoculated, a 5mm fungus cake of bergamot pear rot germ and bergamot pear ring germ is inoculated at the central point, then the temperature is cultured for 5-7d at 28 ℃, a strain with antagonistic capability is screened by the existence and non-existence of a bacteriostasis ring and the size of the bacteriostasis ring, a strain with better antagonistic effect is obtained, named ST-1, and the strain is placed in a glycerol pipe and stored at the temperature of minus 20 ℃.

2. Identification of Strain ST-1

(1) Morphological characterization of strain ST-1: bacterial colony of strain ST-1 on LB solid plate is irregular round, bacterial body is grey white, opaque, rough in surface and zigzag in edge, as shown in figure 1; after gram staining, the cells were observed under a microscope to be bluish-purple like a long rod, and were gram-positive.

(2) Physiological and biochemical characteristics of strain ST-1: the growth temperature of the strain ST-1 is 25-37 ℃, the pH is 5-9, the optimal growth temperature is 30 ℃, and the optimal pH is 7.

TABLE 1 physiological and biochemical characteristics of ST-1

Physiological and biochemical index	ST-1	Physiological and biochemical index	ST-1
				VP	+	Starch hydrolysis	+
Gram staining	+	Gelatin liquefaction	+
				Nitrate reduction	+	Glucose	+
Acidified glucose	-	Arabinose (Arabic sugar)	+
				Indole compounds	-	Maltose	+
Contact enzyme	+	Sucrose	+
				Beta-galactosidase	+	Mannose	+
Urease enzyme	-	Lactose and lactose	-

(3) Determination and analysis of the 16S rDNA sequence of the strain: the genome of ST-1 was extracted with a bacterial genome extraction kit, and the 16S rDNA sequence was amplified with bacterial universal primers 27F and 1492R using this as a template. The amplified product was subjected to 1% agarose gel electrophoresis, and the result showed that the fragment size of the PCR product was about 1.5kb, and the PCR product was sent to Shanghai Biotechnology Co.Ltd for sequencing. BLAST comparison analysis of the sequencing results at NCBI shows that the sequence homology of the strain and Bacillus cabrialesii is 99.6%, and the strain is determined to be bacillus caldarius Bacillus cabrialesii by combining the morphology and the physiological and biochemical characteristics of the strain. The strain is preserved in China Center for Type Culture Collection (CCTCC) M2022671.

Example 2 optimization of Bacillus Carlsberg Bacillus cabrialesii ST-1 Medium

The single factor experiment is adopted to research the conditions of carbon source, nitrogen source, inorganic salt ion and carbon nitrogen on ST-1 growth, the carbon source with the greatest influence on ST-1 benefit is sucrose, the optimal nitrogen source is yeast extract powder, and the optimal economic benefit is wheat bran and peptone, wherein the raw materials with the optimal economic benefit are wheat bran and peptone, and the proportion is 1:2. determining that saccharose, bran, peptone and monopotassium phosphate influence ST-1 growth by adopting a Plackett-Burrnan experiment, obtaining approximate center values of influence factors by a climbing experiment, obtaining 13.1g/L saccharose, 17g/L bran+peptone (1:2) and 2.0g/L monopotassium phosphate by a response surface experiment, and obtaining the theoretical value of spore numberUp to a maximum of 59.74 x 10 ⁸ CFU/mL, through fermentation verification, the actual value of the spore number reaches the maximum under the condition, and the maximum is 59.58 x 10 ⁸ CFU/mL, 1.85×10 spores before medium optimization ⁸ The CFU/mL ratio was 32.21%.

Example 3 volatile gases produced by Bacillus ST-1 have remarkable bacteriostatic Activity against various pathogenic fungi

1. Culture of strains

Taking out ST-1 stored in a glycerol tube, activating, inoculating to LB solid plate, culturing overnight, picking single colony to LB liquid medium, culturing at 30deg.C and 200rpm for 10-12 hr, centrifuging, collecting thallus, washing with sterile water for 3 times, and suspending the thallus weight to 1.0X10 by 0.2mol/L PBS buffer with pH of 7.0 ⁸ CFU/mL, 100. Mu.L was pipetted onto LB plates.

And (3) carrying out passage activation on the botrytis cinerea, the gibberella wheat, the fusarium wilt of watermelons, the stem rot pathogen of sweet potatoes, the root rot pathogen of sweet potatoes, the vine rot pathogen of sweet potatoes, the black spot disease of sweet potatoes, the rot pathogen of pear trees and the ring spot pathogen of pear trees on a PDA culture medium, and carrying out dark culture at 28 ℃.

The LB medium (w/v) consists of: yeast extract powder 0.5%, peptone 1%, sodium chloride 0.5%, agar 2%, pH natural, and sterilizing at 121deg.C for 20min.

The PDA culture medium is as follows: 200g of potato, 15g of glucose, 18g of agar and natural pH, and sterilizing for 20min at 121 ℃.

2. Double-layer flat plate method for measuring antibacterial effect of ST-1 volatile gas

Activating ST-1, inoculating into LB liquid medium, culturing, preparing bacterial suspension to 1.0x10 ⁸ CFU/mL, 100. Mu.L was pipetted onto LB plates for further use. A fungus cake is prepared by using a 5mm puncher, is connected to the center of a PDA flat plate, is then placed on the ST-1 bacteria flat plate in an inverted mode, and the contact area of the two flat plates is sealed by using a preservative film of 3 cm. The blank control group was obtained by placing the plate inoculated with the pathogenic fungi upside down on a blank LB plate. Each treatment was repeated 4 times and incubated at 28℃and colony diameter of each treated pathogenic fungus was tested when the blank control was grown on a full plate. The rate of inhibition of the pathogenic fungi by each treatment was calculated. The specific results are shown in Table 2.

TABLE 2ST-1 antibacterial ratio of volatile gases against different fungi

Note that: the different lowercase letters in the table indicate significant differences between the same column data (p < 0.05); different capital letters indicate that there is a very significant difference (p < 0.01) between the same column of data.

The data were statistically analyzed for the bacteriostatic rate using SPSS 26.0 software.

The results show that: the volatile gas of the bacillus Bacillus cabrialesii ST-1 strain has inhibition effect on 8 plant pathogenic fungi, the average inhibition rate reaches more than 70%, wherein the inhibition rate on botrytis cinerea is the highest and is 97.78%; the inhibition rate of the sweet potato stem rot in the 3 sweet potato diseases is 90.63%; the bacteriostasis rates of the pyritic rot germ and the pyritic ring germ are 92.43 percent and 72.23 percent respectively.

The bacterial strain B.canlialei LM-73 (with the preservation number of CCTCC NO: M2021686) screened from the blueberry rhizosphere in the earlier stage of the patent applicant has the bacteriostasis rate of 76.47 percent on the pear rot disease, has lower inhibition rate of 32.58 percent on the pear ring rot disease, obviously improves the bacteriostasis rate of both the pear rot disease and the pear ring rot disease, has wider bacteriostasis spectrum, has better inhibition effect on Fusarium, and has the average bacteriostasis rate of more than 60 percent on 3 diseases of sweet potatoes.

3. Detection of ST-1 volatile bacteriostatic components using headspace solid-phase microextraction in combination with GC-MS

Step 1: activating bacillus Bacillus cabrialesii ST-1, picking single colony, inoculating the single colony into LB culture medium with the content of 50mL (250 mL) of the culture medium, culturing at 30 ℃ for 24h at 200r/min to obtain seed liquid, sucking the seed liquid into LB liquid fermentation culture medium according to the inoculum size with the final concentration of 5%, placing the seed liquid into a headspace sample injection bottle, and culturing the headspace bottle in a constant-temperature shaking table with the temperature of 30 ℃ and 200r/min for 60h.

Step 2: the headspace sample injection bottle is placed at 40 ℃ for 1h of balance, and the microextraction head is placed in a gas chromatography sample injection port for activation for 10min at 230 ℃. And after the headspace bottle is balanced, inserting an activated microextraction head into the headspace bottle, and performing solid-phase extraction on volatile components collected in the headspace bottle. Microextraction head solid phase extraction using divinylbenzene/carbon molecular sieves/polydimethylsiloxane.

Step 3: and (3) inserting the extraction head into a headspace sample injection bottle for extraction for 20min, rapidly taking out the extraction head after adsorption is finished, inserting the extraction head into a GC-MS sample injection port for analysis for 2min, and then carrying out GC-MS analysis.

Air quality conditions: the column was an HP-5MS (30 mm. Times.0.25 μm) elastic quartz capillary column. The chromatographic column is heated by a system, the initial temperature is 50 ℃, the temperature is kept for 5min, and then the temperature is increased to 250 ℃ by 10 ℃ and kept for 5min. The temperature of the column box is 250 ℃, the carrier gas is high-purity He (more than or equal to 99.99%), the pre-column pressure is 7.93psi, the carrier gas flow is 1mL/min, and the sample injection amount is 1 mu L. Mass spectrometry conditions: the ion source is EI source, the ion source temperature is 230 ℃, the sample inlet temperature is 250 ℃, the quadrupole rod temperature is 150 ℃, the electron energy is 70eV, the emission current is 34.6Ma, and the solvent delay is 6min. And comparing and identifying the components with the database (National Institute of Standards and technology) NIST after detection, and selecting the components with the similarity of more than 80 percent for component analysis.

TABLE 3 ST-1 volatile gas Components

-: no antibacterial effect exists; +: 10-20% of antibacterial effect; a ++21-50% bacteriostatic effect; ++ 51-90% antibacterial effect; ND: no data

4. Method for measuring bacteriostasis of fermentation liquor by plate counter method

And (5) selecting various plant pathogenic fungi hyphae, and growing on a PDA culture medium for 5-10 days for later use. Punching the cultured fungi by using a puncher to prepare a 5mm fungus cake, and connecting the fungus cake with the center of a PDA flat plate. Two holes are uniformly and symmetrically drilled on the PDA solid plate at the position 2.0cm away from the central point, the strain suspension or fermentation liquor is inoculated, and the PDA solid plate is placed in the dark at 28 ℃ for 5-7d. Each treatment was repeated 4 times and the colony diameter of each treatment with suppressed pathogenic fungi was tested.

Example 4 application of Bacillus Carlsberg Bacillus cabrialesii ST-1 in control of post harvest leaf scald disease

1. Prevention and treatment of leaf scald disease of pear after picking by ST-1 fermentation liquor

Step 1: selecting healthy Korla bergamot pears with the weight close to that of the bergamot pears, sterilizing, cleaning, drying for standby, and using a sterile puncher to equidistantly punch 3 inoculation holes with the depth of 3mm at the neck of the bergamot pears.

Step 2: the bacterial cake of the Rhizoctonia cerealis is prepared by using a sterile puncher, and the size is 5mm.

Step 3: 50 μl of ST-1 bacterial suspension was placed in three inoculation wells at the "neck" of bergamot pear, and 6 replicates of each group were run with equal volumes of sterile deionized water as a blank.

Step 4: group A bergamot pears: punching is not performed, pathogenic bacteria are not inoculated, and the group B bergamot pears: punching, inoculating no pathogenic bacteria, and performing group C bergamot pears: punching and inoculating pathogenic bacteria cakes, and not inoculating fermentation liquid, wherein the group D bergamot pears are: punching and inoculating pathogenic bacteria cakes, and simultaneously inoculating fermentation liquor, wherein each group is parallel to 6. And (3) placing the treated bergamot pears into a polyethylene film self-sealing bag for sealing, and storing at 25 ℃ under the condition of 60% -80% of HR. And observing the disease conditions of the bergamot pears in the control group and the experimental group, photographing and recording the data of each treatment group.

TABLE 4 effects of ST-1 fermentation broths on controlling bergamot pear disease

The experimental results show that: the incidence of bergamot pears in the blank control group inoculated with the bergamot pear Botryosphaeria dothidea is 100% in 120h, 144h and 168h, and the incidence of bergamot pears in the experimental group inoculated with the ST-1 fermentation broth is 20%, 20% and 26.7% respectively, and the control effect is above 70%, as shown in Table 4. The ST-1 bacterial suspension or fermentation liquor can effectively inhibit the generation of the bergamot pear ring rot.

2. Prevention and treatment of leaf spot after pear picking by bacillus californicus ST-1 volatile gas

Step 1: example 4 step 1.

Step 2: example 4 step 2.

Step 3: group A bergamot pears: the holes are not punched, and pathogenic bacteria are not inoculated; group B bergamot pears: punching, and not inoculating pathogenic bacteria; group C bergamot pears: punching and inoculating pathogenic bacteria cakes, and simultaneously placing 3 blank LB plates into a polyethylene film self-sealing bag for sealing; group D bergamot pears: perforating and inoculating pathogenic bacteria cake, and coating 3 with 10 ⁸ The LB plates of CFU/mL Bacillus cabrialesii ST-1 were placed in polyethylene film bags and sealed, with 6 parallel pears per group. Storing at 25 ℃ and under the environment of 60% -80% of HR.

Step 4: and (3) placing the treated bergamot pears in an incubator at 25 ℃, observing the disease conditions of the bergamot pears in a control group and an experimental group, photographing and recording the data of each treatment group.

TABLE 5-ST-1 volatile gas effects of controlling bergamot pear morbidity

The experimental results show that: the incidence of the bergamot pears of the positive control group inoculated with the bergamot pear is 93.3 and 100% at 120h and 144h respectively, the incidence of the bergamot pear of the experimental group inoculated with the ST-1 is 26.7% and 46.6%, and the control effect is more than 50%, as shown in Table 5. It is shown that the gaseous volatile matters generated by ST-1 can inhibit the occurrence of pear ring rot.

In conclusion, the volatile gas and the thallus fermentation liquor generated by the strain Bacillus cabrialesii ST-1 have broad-spectrum resistance to plant pathogenic fungi, have remarkable antibacterial effects on pear rot and pear ring rot, have antibacterial rates of 92.43% and 72.23% respectively, and have wide application prospects in the aspects of pear disease control, storage and preservation.

Meanwhile, the strain has strong antibacterial activity on Fusarium, and can also be used for preventing and controlling cotton wilt and sweet potato diseases. The strain provided by the invention has the advantages of stable performance, simple preparation method, low cost, high viable count and large comprehensive development potential, can be used for preparing functional biocontrol agents and microbial agents for fruit preservation, and has a wide application prospect.

The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principle of the present invention should be made in the equivalent manner, and the embodiments are included in the protection scope of the present invention.

Finally, it is also noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.

While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. It is therefore intended that the following claims be interpreted as including the preferred embodiments and all such alterations and modifications as fall within the scope of the invention.

It will be apparent to those skilled in the art that various modifications and variations can be made to the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present invention also include such modifications and alterations insofar as they come within the scope of the appended claims or the equivalents thereof.

Sequence listing

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Claims

1. Bacillus californicus @Bacillus cabrialesii) ST-1, wherein the bacillus caldarius ST-1 has a deposit number of: cctccc NO: m2022671.

2. A fermentation broth, the fermentation broth comprising:

a fermentation broth or a bacterial suspension obtained by fermenting bacillus caldarius ST-1 according to claim 1;

3. The fermenting bacteria agent according to claim 2, wherein the preparation method of the bacterial suspension comprises the following steps:

4. A fermentation broth according to claim 3, wherein the culture conditions of strain ST-1 comprise: the temperature is 28-32 ℃ and the pH is 5-9.

5. Use of bacillus californicus ST-1 according to claim 1 or a zymophyte according to claim 2 in preventing and controlling pear diseases or preserving pear diseases, wherein pathogenic fungi of the pear diseases are pear rot pathogensValsa mali var. pyriRhizoctonia cerealis of Hexiang pearBotryosphaeria dothidea。

6. Use of the bacillus californicus ST-1 of claim 1 or the zymophyte of claim 2 in the control of sweet potato stalk rot.