CN115109723A

CN115109723A - Bacillus cassiae ST-1 and application thereof in prevention and control or storage and fresh keeping of pear diseases

Info

Publication number: CN115109723A
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: 2022-09-27
Anticipated expiration: 2042-06-23
Also published as: CN115109723B

Abstract

The invention provides a Bacillus calfsii ST-1 strain and application thereof in preventing and treating diseases of bergamot pears or storing and refreshing, which is named as Bacillus calfsii ST-1(Bacillus calbrielisii ST-1), and the preservation number of the Bacillus calfsii ST-1 strain is CCTCC NO: m2022671. The Bacillus cabrialisii ST-1 generates volatile gas and thallus fermentation liquor which have broad-spectrum resistance to plant pathogenic fungi, has obvious bacteriostatic action on the pear rot and the pear ring rot, has bacteriostatic rates of 92.43 percent and 72.23 percent respectively, and has wide application prospect in the aspects of preventing, controlling, storing and refreshing the pear diseases. Meanwhile, the strain has strong bacteriostatic activity on fusarium, and can also be used for preventing and treating cotton wilt and sweet potato diseases.

Description

Bacillus cassiae ST-1 and application thereof in prevention and control or storage and fresh keeping of pear diseases

Technical Field

The invention relates to the technical field of microbiology, biochemistry and fermentation engineering, in particular to a bacillus cassiicola ST-1 and application thereof in preventing and treating diseases of bergamot pears or storing and refreshing.

Background

The pear is the third fruit industry in China, which is second only to citrus and apple, and the cultivation area, the yield, the export quantity, the variety quantity and the like of the pear are all in the forefront of the world. Korla bergamot pears (bergamot pears for short) (Pyrus sinkiangensi) are mainly planted in Korla and Acksui areas, Ba, Xinjiang. The disease problem of the bergamot pears in the cultivation process is particularly prominent under the influence of factors such as periodic freezing injury and improper management, and the development of the bergamot pear planting industry is severely restricted. The bergamot pear rot belongs to a typical branch disease, is caused by apple black-rot shell pear varieties (Valsa mali var. pyri), and is one of the most important fungal diseases in pear tree cultivation and production. The occurrence area is wide, the infection is easy to spread, the occurrence rate is increased year by year, the average incidence rate of the pear orchard is 50-66%, and the average incidence rate of severe pear orchard can reach more than 94%. Because the bacteria can be infected with rot germs of other hosts in an interactive way, the actual prevention and control difficulty is higher. The bergamot pears are also easily damaged by pathogenic microorganisms in the storage period after picking, and the ring spot is an important disease in the storage period of the pears, can cause fruit decay after picking and causes the fruit decay in the storage period. The ring spot is also one of the main diseases in the field, can infect branches, leaves and fruits, and can cause destructive damage to an orchard in severe cases, thereby affecting the yield and fruits and causing the loss of economic benefit.

At present, the disease of the bergamot pear is mainly controlled chemically, and the storage and preservation method of the bergamot pear mainly adopts chemical reagent treatment or edible film coating agent coating. With the increasing concern of people on chemical pesticide residue and fruit safety, the development of an environment-friendly biological control method is the key point of the disease control of bergamot pears.

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

Disclosure of Invention

The invention aims to provide a bacillus cassiae ST-1 and application thereof in preventing, controlling, storing and refreshing bergamot pear diseases, wherein volatile gas and thallus fermentation liquor generated by the bacillus cassiae ST-1 have broad-spectrum resistance to plant pathogenic fungi, have obvious bacteriostatic action on bergamot pear rot and bergamot ring rot, and have wide application prospect in the aspects of preventing, controlling, storing and refreshing bergamot pear diseases.

In the first aspect of the invention, a Bacillus caldarius ST-1 is provided, the Bacillus caldarius ST-1 is classified and named as Bacillus calbrialesii ST-1, and the preservation number is as follows: CCTCC NO: m2022671.

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

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

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

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

inoculating ST-1 to an LB solid plate for overnight culture, selecting a single colony to an LB liquid culture medium for culture, centrifuging and collecting thalli, washing, and then suspending the thalli to a concentration of (0.5-5) multiplied by 10 by using PBS buffer solution 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 of sucrose, 17.0g/L of bran and peptone (1:2) and 2.0g/L of potassium dihydrogen phosphate.

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

In the fourth aspect of the invention, the application of the bacillus kadsurae ST-1 or the fermentation inoculum or the volatile gas in the prevention and control of pear diseases is provided.

Further, the pathogenic fungi of the bergamot pear disease include the bergamot pear rot pathogen vala mali var. pyri and bergamosphaeria dothidea.

In the fifth aspect of the invention, the Bacillus calfsii ST-1 or the fermentation inoculant or the volatile gas is applied to the control of apple rot, cotton wilt or sweet potato diseases.

The bacillus cassiidii ST-1 has a wide antibacterial spectrum, can obviously inhibit important diseases in the growing period and the storage period after picking of bergamot pears, and has a stable effect. The bacterial gas volatile matter or bacterial suspension or fermentation liquor can be applied to biological control of the pear rot and ring rot, and is also suitable for control of apple rot, cotton wilt and various diseases of sweet potatoes.

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

the Bacillus calmette-guerin ST-1 and the application thereof in preventing and treating fragrant pear diseases or storing and refreshing, which are provided by the invention, have broad-spectrum resistance to plant pathogenic fungi, have obvious bacteriostasis on fragrant pear rot and fragrant pear ring rot, and have wide application prospects in preventing and treating fragrant pear diseases and storing and refreshing, wherein the bacteriostasis rate is 92.43% and 72.23% respectively. Meanwhile, the strain has strong bacteriostatic activity on fusarium, and can also be used for preventing and treating cotton wilt and sweet potato diseases. The strain has stable performance, simple preparation method of bacterial suspension, low cost, high viable count and great comprehensive development potential, can be used for preparing functional biocontrol agents and microbial agents for fruit preservation, and has wide application prospect.

The preservation date of the bacillus karezii ST-1 is 2022, 5 months and 19 days, and the preservation number is CCTCC NO: m2022671. The classification name is Bacillus cabrialeisi ST-1, the name of the preservation unit is China center for type culture Collection, the address is Wuhan university in Wuhan city, Hubei province, China, and the postcode is as follows: 430072.

drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the description of the embodiments are briefly introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on the drawings without creative efforts.

FIG. 1 is a photograph showing the colony of Bacillus caldarius Bacillus cabrialeii ST-1 isolated in the present invention on LB solid medium;

FIG. 2 is a chromatogram of total ion flux of volatile gases of Bacillus caldarius ST-1.

FIG. 3 is the antagonism of Bacillus calfsii ST-1 volatile gases against phytopathogenic fungi; note: a: fusarium oxysporum, Fusarium oxysporum; b: fusarium graminearum of gibberella zeae; c: sweet potato stem rot pathogen Fusarium proliferatu; d: verticillium pyricularis Botryosphaeria dothidea; e: botrytis cinerea; f: sweet potato root rot fungus Fusarium Solani; g: sweet potato vine canker Fusarium bulbigenemum; h: pyri rot pathogen Valsa mali var.

FIG. 4 is the antagonism of a plant pathogenic fungus by a suspension or fermentation broth of Bacillus caldarius ST-1; note: a: botrytis cinerea; b: wheat scab bacteria; c: watermelon fusarium wilt bacteria; d: sweet potato stem rot germs; e: sweet potato black spot pathogen.

FIG. 5 shows the control of the ring spot of pear after harvest by Bacillus karezii ST-1 volatile gas or bacterial suspension or fermentation broth; a: treating gas volatile matters; b: and (4) treating fermentation liquor.

Detailed Description

The present invention will be described in detail below with reference to specific embodiments and examples, and the advantages and various effects of the present invention will be more clearly apparent therefrom. It will be understood by those skilled in the art that these specific embodiments and examples are for the purpose of illustrating the invention and are not to be construed as limiting the invention.

Throughout the specification, unless otherwise specifically noted, terms used herein should be understood as having meanings 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. If there is a conflict, the present specification will control.

Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be obtained by existing methods.

The bacillus karezii ST-1 and its application in the disease control or storage and preservation of bergamot pear 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

Separating and purifying strains: collecting the rhizosphere soil of forest plants in Anshizhou Hefeng county of Hubei province, enriching and culturing, and separating bacteria in the rhizosphere soil by a dilution plate method. Selecting single colonies with different shapes, colors and sizes, streaking and purifying by a 3-5 generation plate, transferring to an LB culture medium inclined plane, and preserving at 4 ℃.

Inoculating the single bacterial colony in LB liquid culture medium, and shake culturing at 30 deg.C and 200rpm for 36-72 hr to obtain fermentation liquid. Uniformly and symmetrically punching two holes at a position 2.0cm away from a central point on a PDA solid plate, inoculating a strain suspension or fermentation liquor, inoculating a bergamot pear rot germ and a bergamot pear ring rot germ 5mm cake at the central point, then culturing for 5-7d at 28 ℃, screening a strain with good antagonistic effect by the presence or absence of an inhibition zone and the size of the inhibition zone, and placing the strain in a glycerol tube for storage at-20 ℃.

2. Identification of the Strain ST-1

(1) Morphological characteristics of the strain ST-1: bacterial colonies of the strain ST-1 on an LB solid plate are irregular circles, thalli are grey white, opaque, rough in surface and sawtooth in edge, and are shown in figure 1; after gram staining, the bacteria are observed under a microscope to be bluish purple shaped like a long rod and are gram-positive bacteria.

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

Physiological and biochemical Properties of Table 1-ST-1

Physiological and biochemical indexes	ST-1	Physiological and biochemical indexes	ST-1
				VP	+	Starch hydrolysis	+
Gram stain	+	Liquefaction of gelatin	+
				Nitrate reduction	+	Glucose	+
Acidified glucose	-	Arabinose	+
				Indoles	-	Maltose	+
Contact enzyme	+	Sucrose	+
				Beta-galactosidase enzyme	+	Mannose	+
Urease	-	Lactose	-

(3) 16S rDNA sequence determination and analysis of the strain: the genome of ST-1 was extracted using a bacterial genome extraction kit, and the 16S rDNA sequence was amplified using bacterial universal primers 27F and 1492R as templates. The amplified product is subjected to 1% agarose gel electrophoresis, the result shows that the size of a PCR product fragment is about 1.5kb, and the PCR product is sent to Shanghai biological engineering Co. BLAST comparison analysis is carried out on the sequencing result at NCBI, the sequence homology of the strain and Bacillus cabrilisii is found to be 99.6 percent, and the strain is determined to be Bacillus calbrialesii by combining the form and the physiological and biochemical characteristics of the strain. The strain is preserved in China center for type culture Collection with the preservation number of CCTCC M2022671.

Example 2 optimization of Bacillus Calleriae Bacillus Calbrialesii ST-1 Medium

The conditions of carbon source, nitrogen source, inorganic salt ion and carbon nitrogen on the growth of ST-1 are researched by adopting a single-factor experiment, the carbon source which has the greatest influence on the benefit of ST-1 is cane sugar, the optimal nitrogen source is yeast extract powder, and then peptone and bran are used, the raw materials with the optimal economic benefit are bran and peptone, and the ratio is 1: 2. the Plackett-burrnan experiment is adopted to determine that sucrose, bran, peptone and potassium dihydrogen phosphate influence the growth of ST-1, the slope climbing experiment is adopted to obtain the approximate central value of the influence factor, the response surface experiment is adopted to obtain that the theoretical value of the spore number reaches the maximum when the sucrose is 13.1g/L, the bran + peptone (1:2) is 17g/L and the potassium dihydrogen phosphate is 2.0g/L, and the maximum is 59.74 x 10 ⁸ CFU/mL, verified by fermentation, the actual value of the number of spores under the condition reaches the maximum of 59.58 x 10 ⁸ CFU/mL, number of spores before optimization with medium 1.85 x 10 ⁸ The CFU/mL ratio is improved by 32.21%.

Example 3 volatile gas produced by Bacillus ST-1 has significant bacteriostatic activity against various pathogenic fungi

1. Cultivation of the Strain

Taking out ST-1 stored in a glycerol tube, subculturing and activating the ST-1, inoculating the ST-1 to an LB solid plate for overnight culture, picking a single colony to an LB liquid culture medium, culturing the colony for 10 to 12 hours at the temperature of 30 ℃ and the speed of 200rpm, centrifuging and collecting thalli, washing the thalli for 3 times by using sterile water, and then resuspending the thalli to the concentration of 1.0 multiplied by 10 by using PBS buffer solution with the pH value of 7.0 of 0.2mol/L ⁸ CFU/mL, aspirate 100. mu.L and spread onto LB plates.

The botrytis cinerea, fusarium graminearum, fusarium oxysporum, rot germs of sweet potato stem base, root rot germs of sweet potato, vine top germs of sweet potato, black spot of sweet potato, rot germs of pear tree and ring rot germs of pear tree are subjected to generation activation on a PDA culture medium and are cultured in the dark at 28 ℃.

The LB culture medium (w/v) comprises the following components: 0.5% of yeast extract powder, 1% of peptone, 0.5% of sodium chloride and 2% of agar, wherein the pH is natural, and the yeast extract powder is sterilized for 20min at 121 ℃.

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

2. Double-layer flat plate method for determining bacteriostatic action of ST-1 volatile gas

Activating ST-1, inoculating into LB liquid culture medium, culturing, and preparing bacterial suspension to concentration of 1.0 x 10 ⁸ CFU/mL, aspirate 100. mu.L and spread onto LB plates for use. A fungal cake was prepared using a 5mm punch, attached to the center of a PDA plate, and then inverted on the ST-1 bacterial plate described above, with the contact area of the two plates sealed with a 3cm plastic wrap. The blank control group was prepared by placing the plates inoculated with the pathogenic fungi upside down on blank LB plates. Each treatment was repeated 4 times, incubated at 28 ℃ and the colony diameter of the pathogenic fungi inhibited for each treatment was tested when the blank control was grown on the plate. And calculating the bacteriostasis rate of each treatment on pathogenic fungi. The specific results are shown in Table 2.

TABLE 2 bacteriostatic rate of ST-1 volatile gas on different fungi

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

The data are subjected to statistical analysis on the bacteriostasis rate by adopting SPSS 26.0 software.

The results show that: volatile gas of the Bacillus cabrialeisi ST-1 strain has an inhibiting 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 base rot disease in the 3 sweet potato diseases is 90.63 percent; the bacteriostasis rates to the pear rot germs and the pear ring rot germs are 92.43 percent and 72.23 percent respectively.

The bacterial strain B.cabrilisii LM-73 (the preservation number is CCTCC NO: M2021686) screened from blueberry rhizosphere at the earlier stage of the patent applicant has the bacteriostatic rate of 76.47% on the pear rot and the inhibitory rate of 32.58% on the pear ring rot, and the bacterial strain disclosed by the invention has the advantages that the bacteriostatic rate of the bacterial strain on the pear rot and the pear ring rot is obviously improved, the bacteriostatic spectrum is wider, the better inhibitory effect on fusarium is particularly realized, and the average bacteriostatic rate on 3 diseases of sweet potatoes reaches more than 60%.

3. Detection of ST-1 volatile antibacterial components by headspace solid-phase microextraction combined with GC-MS

Step 1: activating Bacillus cabrialisii ST-1, selecting a single colony, inoculating the single colony into an LB culture medium with the content of 50mL (250 mL in a bottle), culturing at 30 ℃ and 200r/min for 24 hours to serve as a seed solution, sucking the seed solution into the LB liquid fermentation culture medium according to the inoculation amount with the final concentration of 5%, then placing the seed solution into a headspace sampling bottle, and placing the headspace bottle into a constant-temperature shaking table with the temperature of 30 ℃ and 200r/min for culturing for 60 hours.

Step 2: and (3) placing the headspace sample injection bottle at 40 ℃ for balancing for 1h, and placing the micro-extraction head in a gas chromatography sample injection port for activating at 230 ℃ for 10 min. And (4) after the headspace bottle is balanced, inserting the activated micro-extraction head into the headspace bottle, and performing solid-phase extraction on volatile components collected in the headspace bottle. And (3) performing solid-phase extraction by using a micro-extraction head of divinylbenzene/carbon molecular sieve/polydimethylsiloxane.

And step 3: and inserting the extraction head into a headspace sample injection bottle for extraction for 20min, quickly 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.

And (3) gas condition: the column was an HP-5MS (30mm 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 5 min. The temperature of the column box is 250 ℃, the carrier gas is high-purity He (more than or equal to 99.99 percent), the pre-column pressure is 7.93psi, the flow rate of the carrier gas is 1mL/min, and the sample injection amount is 1 mu L. Mass spectrum conditions: the ion source is an EI source, the temperature of the ion source is 230 ℃, the temperature of the injection port is 250 ℃, the temperature of the quadrupole rod is 150 ℃, the electron energy is 70eV, the emission current is 34.6Ma, and the solvent delay is 6 min. After detection, the samples were identified by NIST (National Institute of Standards and technology) and fractions with similarity of more than 80% were selected for component analysis.

TABLE 3-ST-1 volatile gas composition

-: no bacteriostatic effect; +: 10-20% of antibacterial effect; + 21-50% of antibacterial effect; the bacteria inhibition effect is +51 to 90 percent; ND: without data

4. Plate confrontation method for determining antibacterial effect of fermentation liquor

And (3) selecting various plant pathogenic fungi hyphae, and growing on the PDA culture medium for 5-10 days for later use. Punching the cultured fungus with a puncher to prepare 5mm fungus cake, and connecting to the center of a PDA plate. Two holes are uniformly and symmetrically drilled on a PDA solid plate 2.0cm away from the central point, and strain suspension or fermentation liquor is inoculated and placed in the dark at the temperature of 28 ℃ for cultivation for 5-7 d. Each treatment was repeated 4 times and the diameter of colonies with inhibited pathogenic fungi was tested for each treatment.

Example 4 application of Bacillus Calleriae Bacillus cabrialeisi ST-1 in prevention and treatment of ring rot after bergamot pear harvest

1. Prevention and treatment of pear ring spot after picking by ST-1 fermentation liquor

Step 1: selecting Korla bergamot pears with weight close to health, disinfecting, cleaning, wiping for later use, and punching 3 inoculation holes with the depth of 3mm at the neck of the bergamot pear at equal intervals by using an aseptic puncher.

Step 2: a sterile puncher is used for preparing the ring rot of bergamot pear fungus cakes, and the size of the ring rot of bergamot pear fungus cakes is 5 mm.

And step 3: 50 mu L of ST-1 bacterial suspension is taken to be placed in three inoculation holes at the neck of the bergamot pear, equal volume of sterile deionized water is used as a blank control, and 6 bacteria in each group are parallel.

And 4, step 4: group A of bergamot pears: no perforation, no inoculation of pathogenic bacteria, group B bergamot pears: punching, not inoculating pathogenic bacteria, and preparing a group C bergamot pear: punching and inoculating a pathogenic bacteria cake without fermentation liquor, and carrying out D group of bergamot pears: and punching holes and inoculating pathogenic bacteria cakes and inoculating fermentation liquor, wherein 6 bacteria cakes are parallel in each group. The treated bergamot pears are put into a polyethylene film self-sealing bag to be sealed and stored at the temperature of 25 ℃ and the HR of 60-80 percent. And observing the morbidity of the bergamot pears in the control group and the experimental group, photographing and recording the data of each treatment group.

TABLE 4-ST-1 fermentation liquid for preventing and treating disease of pear

The experimental results show that: the disease incidence rates of the bergamot pears in the blank control groups inoculated with the ring rot fungus Botryosphaeria dothidea are 100% at 120h, 144h and 168h, the ring rot disease rates of the bergamot pears in the experimental group inoculated with the ST-1 fermentation liquor are 20%, 20% and 26.7%, and the control effects are all more than 70%, as shown in Table 4. The ST-1 bacterial suspension or the fermentation liquid can effectively inhibit the occurrence of the bergamot pear ring rot.

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

Step 1: example 4 step 1.

And 2, step: example 4 step 2.

And step 3: group A of bergamot pears: no perforation and no inoculation of pathogenic bacteria; b group of bergamot pears: punching without inoculating pathogenic bacteria; group C bergamot pear: punching and inoculating pathogenic bacteria cake, and simultaneously placing 3 blank LB flat plates into a polyethylene film self-sealing bag for sealing; d, group of bergamot pears: perforating and inoculating pathogenic bacteria cake, and coating 3 pieces with 10 pieces ⁸ The LB plate of CFU/mL Bacillus cabrialeisi ST-1 was placed in a polyethylene film bag and sealed, each group of 6 parallel bergamot pears. Storing at 25 deg.C and HR 60% -80%.

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

TABLE 5-ST-1 volatile gas effect on preventing and treating disease of bergamot pear

The experimental results show that: the incidence rates of the bergamot pears in 120h and 144h of the positive control group inoculated with the ring rot germs of the pear trees are 93.3 and 100 percent respectively, the incidence rates of the ring rot pears in the experimental group inoculated with the ST-1 are 26.7 percent and 46.6 percent, and the control effects are both more than 50 percent, which is shown in Table 5. Shows that the volatile gas generated by ST-1 can inhibit the occurrence of the bergamot pear ring rot.

In conclusion, the volatile gas and the thallus fermentation liquid generated by the strain Bacillus cabrialeii ST-1 have broad-spectrum resistance to plant pathogenic fungi, have obvious bacteriostatic action on the pear rot and the pear ring rot, have the bacteriostatic rates of 92.43 percent and 72.23 percent respectively, and have wide application prospect in the aspects of preventing, controlling, storing and refreshing the pear diseases.

Meanwhile, the strain has strong bacteriostatic activity on fusarium, and can also be used for preventing and treating cotton wilt and sweet potato diseases. The strain has stable performance, simple preparation method of bacterial suspension, low cost, high viable count and great comprehensive development potential, can be used for preparing functional biocontrol agents and microbial agents for fruit preservation, and has wide application prospect.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Finally, it should also be 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. Therefore, it is intended that the appended claims be interpreted as including 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 changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.

Sequence listing

<110> Hubei university of industry

<120> bacillus carinii ST-1 and application thereof in prevention and control of pear diseases or storage and preservation

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gaggagcgaa agcgtgggga gcgaacagga ttagataccc tggtagtcca cgccgtaaac 780

gatgagtgct aagtgttagg gggtttccgc cccttagtgc tgcagctaac gcattaagca 840

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agtca 1445

Claims

1. The Bacillus calmette-guerin ST-1 is characterized in that the Bacillus calmette-guerin ST-1 is classified and named as Bacillus calbrialesiiST-1, and the preservation number is as follows: CCTCC NO: m2022671.

2. A fermentation inoculum, comprising:

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

3. The fermentation inoculant according to claim 2, wherein the bacterial suspension is prepared by a process comprising:

4. The fermentation inoculant 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. The fermentation inoculant according to claim 3, wherein the formula of the culture medium is: 13.1g/L of sucrose, 17.0g/L of bran + peptone (1:2) and 2.0g/L of potassium dihydrogen phosphate.

6. A volatile gas prepared from the bacterial suspension of claim 2 or 3, comprising: and (3) coating the bacterial suspension on an LB solid flat plate, placing the bacterial suspension in a polyethylene film bag for sealing, standing at the temperature of 25 ℃ and in the HR 60-80%, and detecting by adopting headspace solid phase microextraction and GC-MS.

7. Use of the bacillus carinii ST-1 of claim 1, the fermentation inoculant of claims 2-5 or the volatile gas of claim 6 for disease control or storage and freshness preservation of bergamot pear.

8. The use according to claim 7, characterized in that the pathogenic fungi of the bergamot pear disease comprise the species Pyrus pyrifolia Valsa mali var. pyri and Pyrus pyrifolia Botryosphaeria dothidea.

9. Use of the bacillus kadsii ST-1 of claim 1 or the fermentation inoculant of claims 2-5 or the volatile gas of claim 6 for the control of apple rot, cotton wilt or sweet potato disease.