A bacillus saffron for preventing disease and promoting growth

Abstract

The present invention discloses a disease-preventing and growth-promoting Bacillus safensis. The Bacillus safensis described in the present invention is Bacillus safensis strain H23, which is deposited in the China Center for Type Culture Collection with a deposit number of CCTCCNO:M 20241484 and a deposit date of July 4, 2024. The Bacillus safensis described in the present invention not only has the effect of preventing diseases and promoting growth on peppers, but also has a good antagonistic effect on 8 pathogens of Solanaceae vegetable diseases including Phytophthora capsici. Therefore, the Bacillus safensis described in the present invention or a microbial preparation containing the bacterium can be used to prepare plant biocontrol agents and / or bio-organic fertilizers, reduce the pollution of fertilizers and pesticides brought about in the agricultural production process, and promote the sustainable development of green agricultural production.

Description

Technical Field

[0001] The invention belongs to the technical field of biological control, and in particular relates to a bacillus saffron capable of preventing disease and promoting growth. Background Art

[0002] With the progress of society and the development of science and technology, people's demand and standards for food are constantly increasing. Chemical fertilizers and pesticides have a significant effect on increasing the yield of food crops. However, a series of negative effects have emerged after long-term application of chemical fertilizers and pesticides, such as soil compaction, decreased soil fertility, reduced crop quality, and environmental deterioration. In addition, chemical pesticide residues caused by unreasonable application have led to a decline in food safety, affected human health and safety, and undermined the green and sustainable development principles of agriculture. In recent years, food safety and healthy development have attracted much attention, and many countries have begun to devote themselves to the research of new green, environmentally friendly and efficient fertilizers. Beneficial microorganisms can replace chemical fertilizers and pesticides to a certain extent, reduce the application of chemical fertilizers and pesticides, not only reduce the damage to cultivated land and increase soil organic matter, but also increase crop yields, improve the commodity value of crops, and enhance plant resistance. They are indispensable in modern agricultural production.

[0003] Plant growth-promoting rhizobacteria (PGPR) can produce plant growth regulators, which can promote plant growth and improve stress resistance. Plant growth-promoting rhizobacteria often colonize in the rhizosphere soil of plants, enriching the microbial population in the rhizosphere soil. These microorganisms can not only secrete and metabolize some substances, but also increase the effective content of nitrogen, phosphorus and potassium in the soil through a series of reactions, optimize the nutrient composition of the soil, provide a better growth environment for plants, and thus improve the utilization rate of nutrients by plants. At present, the plant growth-promoting rhizobacteria that have been studied and discovered include Bacillus spp., Pseudomonas spp., Azotobacters spp., Azospirillum spp., Burkholderia spp., Serratia spp., Streptomyces spp., etc. Among them, Bacillus has strong adaptability and is highly expressive in disease prevention and growth promotion.

[0004] Pepper belongs to the genus Capsicum in the Solanaceae family and is an important economic vegetable crop in my country. With the increase in market demand, the planting area continues to expand, and many problems have gradually emerged. Continuous cropping of peppers leads to repeated cropping. Among them, pepper blight (Phytophthora blight) caused by infection of pepper blight (Phytophtora capsici Leon.) is one of the soil-borne diseases that seriously restrict pepper production. In addition, the large-scale use of chemical fertilizers and pesticides not only causes soil compaction and decreased fertility, affecting the growth and development of peppers, but also pollutes the environment, poses potential risks, and harms human health. Therefore, the development of new Bacillus with disease prevention and growth-promoting effects can enrich plant rhizosphere microorganisms, reduce the pollution of chemical fertilizers and pesticides brought about in the agricultural production process, and promote the sustainable development of green agricultural production. Summary of the invention

[0005] In view of the deficiencies in the prior art and actual needs, the present invention provides a new Bacillus saffron.

[0006] Specifically, the present invention adopts the following technical solutions:

[0007] In a first aspect, the present invention provides a Bacillus safensis, wherein the Bacillus safensis is Bacillus safensis strain H23, which is preserved in the China Center for Type Culture Collection, classified and named Bacillus safensis, with a preservation number of CCTCC NO:M 20241484, and a preservation date of July 4, 2024.

[0008] The inventors of the present invention screened and studied a variety of strains from pepper rhizosphere soil samples from pepper planting areas in Hainan, and finally obtained Bacillus safensis strain H23, which has a significant blight prevention and control effect on pepper plants and can significantly promote pepper growth. In addition, the Bacillus safensis of the present invention has the effect of antagonizing 8 pathogens of Solanaceae vegetable diseases, so it can be used for the prevention and control of Solanaceae vegetable diseases, especially pepper blight prevention and control, and has a broad-spectrum antagonistic effect.

[0009] In a second aspect, the present invention provides a microbial preparation, which contains the above-mentioned Bacillus sabdariffa, or a fermentation broth of the above-mentioned Bacillus sabdariffa, or a freeze-dried powder of the above-mentioned Bacillus sabdariffa, or an inactivated bacterial cell of the above-mentioned Bacillus sabdariffa, or a lysate of the above-mentioned Bacillus sabdariffa, or an extract of the above-mentioned Bacillus sabdariffa.

[0010] It should be understood that the bacteria generally used for biological control will produce some special products during the growth or fermentation process, such as antibiotics, bacteriocins, protein substances, i.e. other antibacterial substances, etc. Therefore, the protection scope of the present invention should also include the products of the Bacillus sabovii, including but not limited to the fermentation broth of Bacillus sabovii, the lyophilized powder of Bacillus sabovii, the inactivated bacterial cells of Bacillus sabovii, the lysate of Bacillus sabovii and the extract of Bacillus sabovii, etc.

[0011] In a third aspect, the present invention provides the use of the Bacillus saffron or the microbial preparation in preventing and controlling pathogens of Solanaceae plant diseases and / or promoting the growth of Solanaceae plants.

[0012] The shafu bacillus of the present invention has the effect of antagonizing multiple pathogenic bacteria of Solanaceae vegetable diseases, and can also promote the growth of Solanaceae plants. Therefore, the shafu bacillus can be used for the prevention and treatment of Solanaceae vegetable diseases, and for promoting the growth of Solanaceae plants. As mentioned above, the protection scope of the present invention also includes the product of the shafu bacillus, so the shafu bacillus, or the fermentation liquid containing the shafu bacillus, or the freeze-dried powder containing the shafu bacillus, or the inactivated bacterial cell containing the shafu bacillus, or the lysate containing the shafu bacillus, or the extract containing the shafu bacillus can also be used for the prevention and treatment of Solanaceae vegetable diseases, and for promoting the growth of Solanaceae plants.

[0013] In one or more embodiments, the Solanaceae plant is Capsicum annuum.

[0014] The inventors found that the Bacillus sabiotis described in the present invention can significantly promote the growth of peppers, especially the growth of pepper seedlings, including plant height, number of leaves, stem thickness, root length, etc., which are significantly improved compared with the control.

[0015] In one or more embodiments, the pathogens of Solanaceae plant diseases include pepper blight pathogen Phytophthora capsici, pepper anthracnose pathogen Echinops capsici, pepper wilt pathogen Fusarium oxysporum, pepper black spot pathogen Alternaria alternata, pepper root rot pathogen Fusarium spp., pepper white rot pathogen Sclerotium solani, fusarium solani verticillium solani and tomato early blight pathogen Alternaria solani.

[0016] In a fourth aspect, the present invention provides a method for preventing and controlling pathogens of Solanaceae plant diseases or promoting the growth of Solanaceae plants, the method comprising the steps of configuring the Bacillus fusoni or the microbial preparation into a suspension, and watering the suspension around the root base of the Solanaceae plants.

[0017] In a fifth aspect, the present invention provides the use of the Bacillus sabdariffa or the microbial preparation in the preparation of plant biocontrol agents and / or bio-organic fertilizers.

[0018] It should be understood that the plant biocontrol agent of the present invention refers to a composition that uses beneficial microorganisms to kill or reduce the number of pathogenic organisms to control the occurrence and development of plant diseases. The plant biocontrol agent of the present invention comprises the Bacillus sabforniensis or the microbial preparation, which can be in different forms such as solid, liquid, spray, gel or granules. In addition, when the plant biological control preparation of the present invention is prepared into different dosage forms according to needs, corresponding carriers or excipients can be added thereto according to needs, such as solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesive agents, integrators, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, release retardants, etc. Of course, the addition of these carriers or excipients should not affect the original biological control effect of the Bacillus sabdariffa or the microbial preparation.

[0019] It should be understood that the bio-organic fertilizer of the present invention refers to a type of fertilizer that has both microbial fertilizer and organic fertilizer effects, which is composed of specific functional microorganisms and mainly animal and plant residues (such as livestock and poultry manure, crop straw, etc.) and harmlessly treated and decomposed organic materials. The bio-organic fertilizer of the present invention comprises the Bacillus fusiformis or the microbial preparation, which can be compounded or fermented with other animal manure or crop straw according to actual needs. It can improve the soil, improve the physical and chemical properties of the soil, enhance the ability of the soil to retain water, fertilizer, and fertilizer, and alleviate the soil compaction caused by the long-term use of chemical fertilizers.

[0020] In one or more embodiments, the plant biocontrol agent or bio-organic fertilizer further comprises other bacterial agents that have a synergistic effect with the Bacillus sabdariffa.

[0021] It should be understood that although the Bacillus sabdariffa described in the present invention has significant control effects and growth-promoting effects on pepper plants; however, it is not excluded that it can also be used in combination with other biocontrol bacteria with similar functions or other pathogen control functions, so that the biocontrol preparation of the present invention has a stronger control effect on pepper pathogens or has a wider control function on other pathogens; or the biological organic fertilizer of the present invention has a stronger growth-promoting effect on peppers.

[0022] In a sixth aspect, the present invention provides a method for preparing a protease, which utilizes the protease-producing property of the Bacillus saffron to prepare the protease.

[0023] It should be understood that the Bacillus safovici described in the present invention has the characteristic of producing protease, and therefore can be used for the extraction of protease to meet the needs of protease fields such as the food industry or the washing industry; it can also be used in agricultural microbial fertilizers to degrade the cell membranes of pathogenic bacteria to control diseases, and to degrade proteins in composting agricultural organic fertilizers to facilitate plant absorption; it can also be used in animal feed to improve the utilization rate of feed by animals.

[0024] In a seventh aspect, the present invention provides a method for preparing cellulase, which utilizes the cellulase-producing property of the Bacillus saffron to prepare cellulase.

[0025] It should be understood that the Bacillus sabdariffa of the present invention has the characteristic of producing cellulase, and can therefore be used for the extraction of cellulase, and has broad application prospects in the food industry and the environmental industry; it can also be used to degrade the cell walls of pathogenic fungi to control diseases, and to decompose cellulose in compost to facilitate plant absorption; it can also be used in feed for livestock and poultry production, overcoming the problem that monogastric animals such as pigs and chickens cannot utilize cellulose.

[0026] The Bacillus sabdariffa of the present invention has a significant antagonistic effect on Phytophthora capsici and other 8 kinds of Solanaceae plant pathogens, and has a significant disease prevention effect on pepper blight, and has a promoting effect on pepper growth. Therefore, the Bacillus sabdariffa of the present invention or a microbial preparation containing the bacterium can be used to prepare plant biocontrol agents and / or bio-organic fertilizers, reduce the pollution of fertilizers and pesticides brought about in the agricultural production process, and promote the sustainable development of green agricultural production. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a diagram showing the colony morphology observation results of Bacillus fusus H23 cultured on a NA plate for 24 hours in Example 1.

[0028] Figure 2 This is a graph showing the inhibitory effect of different strains on pepper phytophthora in Example 1.

[0029] Figure 3 It is the genome colinearity diagram of the Bacillus sp. H23 strain and the comparison strain in Example 1.

[0030] Figure 4 This is a diagram showing the effect of Bacillus saffron H23 on preventing pepper blight in Example 2.

[0031] Figure 5 This is the antagonism graph of Bacillus sp. H23 against 8 tested pathogens in Example 3.

[0032] Figure 6 This is the resistance capacity analysis of Bacillus sp. H23 in Example 4. DETAILED DESCRIPTION

[0033] Bacillus safensis is a type of Bacillus, Gram-positive, thermophilic and heterotrophic. It was first isolated from the surface of spacecraft and assembly facilities at the Jet Propulsion Laboratory in 2006. Since then, Bacillus safensis has been discovered and proven to have multiple functions and broad application prospects.

[0034] In the prior art, CN113736711B discloses a strain of Bacillus safensis YZS-C03 and its good effect in the biodegradation of polyethylene plastics, providing a new resource and solution for the biodegradation of polyethylene waste in the environment; CN118165866A discloses a strain of Bacillus safensis AW1, and applies it to the cultivation of Sunshine Rose grapes, which can not only effectively improve the weight of the Sunshine Rose grape bunches and the weight of the grains, but also improve the quality of the Sunshine Rose grapes; CN118064295A discloses a strain of Bacillus safensis V66, which can promote the growth and flowering of rice seeds or peanut seeds; CN117866802A discloses a strain of Bacillus safensis (Bacillus safensis) safensis)CY1, which has a high ability to degrade petroleum hydrocarbons and can degrade more than 99% of petroleum hydrocarbons in diesel, batik wastewater and petrochemical sludge polluted soil in a short period of time; CN115725471B discloses a strain of Bacillus safensis 05-2101, which can effectively maintain the disease index of tobacco plants with bacterial wilt at a low level; CN117701460A discloses a strain of Bacillus safensis AS2505, which can significantly promote the growth of sugarcane seedlings and has good application potential in sugarcane production; CN117701431A discloses a strain of Bacillus safensis PB13, which can significantly promote the accumulation of root dry matter in peanuts, dwarf plants and shorten the flowering period, thereby facilitating the early maturity of peanuts; CN117586921A discloses a strain of Bacillus safensis NHF08, which has a significant effect in preventing and controlling cucumber gray mold and bacterial angular leaf spot, and can also be used for the prevention and treatment of fungal and bacterial diseases of melons; CN117264849A discloses a strain of Bacillus sabforii, which has a good control effect on crop diseases, especially peanut white rot, tomato wilt and potato scab; CN114958665B discloses a strain of Bacillus sabforii S21, which can effectively improve the resistance of melons to infection by Acidovoraxcitrulli, and can also inhibit Peronophythora litchi…

[0035] Although the above-mentioned prior art discloses different applications of Bacillus sabio in agriculture, the application of these strains in disease prevention and growth promotion of peppers has not been found, nor has it been found that these strains can simultaneously have antagonistic effects on multiple pathogens of Solanaceae vegetable diseases.

[0036] The inventors of the present invention screened and studied a variety of strains from pepper rhizosphere soil samples from pepper planting areas in Hainan, and finally obtained Bacillus safensis strain H23, which has a significant disease prevention and control effect on the whole pepper plant and can significantly promote pepper growth. In addition, the Bacillus safensis of the present invention has the effect of antagonizing 8 pathogens of Solanaceae vegetable diseases, so it can be used for the prevention and control of Solanaceae vegetable diseases, especially pepper blight prevention and control, and has a broad-spectrum antagonistic effect.

[0037] It should be understood that in the present invention, Bacillus safensis strain H23, Bacillus safensis strain H23, Bacillus safensis strain H23 or the abbreviated description "H23" are all different names for the same strain, and those skilled in the art will not identify these strains with different names as different strains.

[0038] Example

[0039] The following describes the embodiments of the present invention through specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention.

[0040] Before further describing the specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terms used in the examples of the present invention are for describing specific embodiments rather than for limiting the scope of protection of the present invention.

[0041] When the embodiment gives a numerical range, it should be understood that, unless otherwise specified in the present invention, the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those of ordinary skill in the art to which the present invention belongs.

[0042] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0043] The examples involve the addition amounts, contents and concentrations of various substances, wherein the percentages described therein, unless otherwise specified, are all by mass percentages.

[0044] Example 1: Isolation, screening and identification of Bacillus sabdariffa

[0045] 1. Separation

[0046] 1.1 Sampling: From November 26 to 28, 2021, in Haikou City and Qionghai City, Hainan Province, experimenters collected soil samples from the rhizosphere of pepper planting areas.

[0047] 1.2 Separation: (1) Sample treatment: Take 10g of soil sample that has been sieved through a 20-mesh sieve and put it in 90mL of sterilized deionized water (SDW). Mix it by shaking at 150rpm for 15min and place it in a water bath at 80℃. After 10min, take out the sample and let it stand at room temperature for 10min. Use a pipette to dilute and spread it on a NA medium plate and culture it at 28℃ for 1-2d. Pick out colonies with obvious differences in size, morphology, color, etc., number them and place them on NA medium for culture and purification. Finally, store them in 50% glycerol solution at -20℃ for future use. A total of 30 strains were obtained and preserved in this experiment.

[0048] 2. Screening: The plate confrontation method was used for screening. The above-preserved strains were inoculated on NB liquid culture medium and cultured at 28°C for 24 hours. SDW was used to adjust the OD 600 =1.0 as the test bacterial solution for later use.

[0049] The indicator fungus Phytophthora capsici Leonian LT1534 strain (provided by the team of Professor Chen Qinghe of the College of Tropical Agriculture and Forestry, Hainan University) was placed on potato culture medium (PDA) for activation. An indicator bacterial block was taken with an 8mm puncher and placed in the center of the PDA culture (90mm culture dish). 0.5μL of the test bacterial solution was inoculated at 20mm from the center on all sides. The same amount of SDW was inoculated as a control. Each test strain was repeated three times. The culture was placed at 28°C until the control indicator bacteria grew all over the culture dish. The diameter of the indicator colony was measured and the inhibition rate was calculated as (control colony diameter-treatment colony diameter) / control colony diameter × 100%. A strain with strong antibacterial ability was obtained, numbered H23. The NA culture chart (24h) of H23 is shown as follows Figure 1 As shown, the antibacterial effect is Figure 2 As shown in Table 1, the inhibition rate of strain H23 against Phytophthora capsici was 81.2%.

[0050] Table 1 Inhibitory effect of strain H23 on pepper phytophthora

[0051] deal with Phytophthora growth diameter / mm Inhibition rate / % Comparison 7.80±0.06a / H19 7.53±0.15a 3.5 H23 1.42±0.19b 81.2

[0052] Note: Different lowercase letters indicate significant differences among treatments (P<0.05)

[0053] 3 Identification: The total DNA of H23 was extracted using the FastPure Bacteria DNA Isolation Mini Kit (Nanjing Novogene Biotechnology Co., Ltd.). Guangdong Meige Gene Technology Co., Ltd. performed whole genome sequencing, and the sequenced sequences were compared for average nucleic acid similarity. Average nucleic acid similarity (ANI) is one of the most powerful measurement indicators for identifying the closeness of bacterial genome relationships. The average value of the comparison of all orthologous protein sequences based on genome quality inspection reflects the evolutionary distance of genome quality inspection. When ANI>95%, it means that the two genomes belong to the same species. The target genome and the reference genome were compared using MUMmer software (Version 3.23).

[0054] The results showed that the ANI of the H23 genome and the reference genome Bacillus safensis strain PgKB20 reached 98.9947%, which was greater than 95% (Table 2), and the colinear relationship between the genomes was determined with the reference genome ( Figure 3 ), so H23 was identified as Bacillus safensis and named Bacillus safensis strain H23.

[0055] Table 2: Average nucleic acid similarity analysis results of H23

[0056]

[0057] Example 2: Evaluation of resistance of Bacillus saffron H23 to pepper blight

[0058] 2.1 Preparation of pepper seedlings: Seedlings of drooping pepper CB81 (Capsicum baccatum L., provided by the pepper research group of Hainan University) at the 4-5 leaf stage were used as experimental materials. Pepper seeds were sterilized with 75% ethanol solution for 1 min, then with 2% sodium hypochlorite for 3 min, and then washed with sterilized deionized water (SDW) for 5-6 times. After germination at room temperature, the seeds were sown in 50-hole seedling trays and placed in the seedling room with normal water and fertilizer pipes until the peppers grew 4-5 true leaves.

[0059] 2.2 Preparation of Phytophthora liquid: The stored Phytophthora was propagated with PDA medium. After the Phytophthora grew all over the plate, it was cultured in light for 6 days. The sporangia were scraped off and placed in SDW in a 4°C refrigerator for 40 minutes. Then it was taken out and placed in a greenhouse. The Phytophthora liquid was filtered through 3 layers of gauze and the concentration was adjusted to 1.0×10 by hemocytometer counting method. 5 CFU / mL.

[0060] 2.3 Seedling inoculation test: The pepper seedlings were transferred to 8×8 cm seedling cups and placed in an artificial incubator (26±2°C, 14h light / 10h dark) for 2 days. The roots were injured with a knife 2 cm away from the roots of the pepper seedlings, and 10 mL of Bacillus subtilis H23 solution was injected for each strain. The SDW treatment was used as a control. After 3 days of incubation in the same conditions, the Phytophthora was inoculated. 5 mL of pathogen solution was injected into the injured roots and then placed in an artificial incubator for culturing. The disease incidence of the experimental seedlings was observed and recorded. Ten seedlings were treated in each group, and repeated three times. After the disease stabilized (21 days), the incidence rate was calculated. The incidence rate (%) = number of diseased plants / total number of plants investigated × 100%, and the control effect (%) = (control incidence rate - treatment incidence rate) / control incidence rate × 100%. The test results are shown in Tables 3 and Figure 4 As shown in the figure, the control effect of Bacillus saffron H23 on pepper blight was 75.4%. Figure 4 The graphs are the results of the control and treatment groups on the 21st day.

[0061] Table 3 Effect of Bacillus saffron H23 on the control of pepper plant blight

[0062] deal with Incidence (%) Control effect (%) Control (SDW) 96.7±2.8 / H23 23.3±5.8* 75.4

[0063] Note: * indicates significant difference between the two treatments (P<0.05)

[0064] Example 3: Evaluation of the antagonism of Bacillus sabdariffa H23 against plant pathogens of the Solanaceae family

[0065] The plate confrontation method was used to detect the antagonistic effect of Bacillus sabdariffa H23 on various pathogenic fungi of Solanaceae vegetables, including Phytophthora capsici Leonian, the pathogen of pepper anthracnose, Colletotrichum capsici, the pathogen of pepper wilt, Fusarium oxysporu, the pathogen of pepper black spot, Alternaria alternate, the pathogen of pepper root rot, Fusarium spp., the pathogen of pepper white rot, Sclerotium rolfsii Sacc., the pathogen of pepper white rot, Fusarium verticillioide, and Alternaria solani (Ell. et Mart.) Sorauer, the pathogen of tomato early blight. The above pathogenic fungi strains were provided by Hunan Academy of Agricultural Sciences. The above pathogenic fungi were activated on PDA medium, punched with an 8mm puncher and transferred to new PDA medium. The bacterial liquid of Bacillus fusiformis H23 was cultured in NB medium and the concentration was adjusted to OD after centrifugation. 600 =1.0(ca.1×10 8 CFU / mL), inoculate 0.5 μL around the pathogen block 2 mm away, and inoculate with sterile distilled water as a blank control. Cultivate at 28°C until the control fills the entire culture dish, take photos and record the experimental results. Repeat the experiment three times.

[0066] The results are as follows Figure 5 As shown, Bacillus shaforniensis H23 had a strong antagonistic effect on the eight pathogens tested.

[0067] Example 4: Functional properties of Bacillus sabdariffa H23

[0068] 4.1 Protease detection: Pour about 10mL of protease culture medium into a 90mm culture dish. After cooling to room temperature, use a 7mm diameter hole puncher to punch a hole in the middle of the culture medium. Then, add 5μL of H23 bacterial solution to the hole, incubate at 30℃ for 72h, and observe the formation of transparent circles in the culture medium. The appearance of transparent circles indicates that the strain has the ability to hydrolyze proteases, otherwise it does not. The experimental treatment is set up with 3 replicates.

[0069] 4.2 Cellulose detection: Pour about 10mL of cellulose culture medium into a 90mm culture dish. After cooling to room temperature, use a 7mm diameter hole puncher to punch a hole in the middle of the culture medium. Then, inoculate 5μL of H23 bacterial solution into the hole and culture it at 30℃. After 48h, first add 1mg / mL Congo red solution to soak for 1h, and then add 1mo1 / L NaCl to soak for 1h. Observe the formation of transparent circles in the culture medium. The appearance of transparent circles indicates that the strain has a certain hydrolysis effect on cellulose, otherwise it has no effect. The experimental treatment is set up with 3 replicates.

[0070] 4.3 Siderophore detection: Pour about 10 mL of siderophore culture medium into a 90 mm culture dish. After cooling to room temperature, use a 7 mm diameter puncher to punch a hole in the middle of the culture medium. Then inoculate the H23 bacterial solution into the hole and culture it at 30°C. After 5 days, observe the color change in the culture medium. If the color of the culture medium changes from blue to orange, it means that the strain can hydrolyze siderophore, otherwise it cannot. Set up 3 replicates for the experimental treatment.

[0071] The test results are as follows Figure 6 As shown, compared with the control, transparent halos appeared around the protease medium and cellulase medium of H23 after inoculation, indicating that H23 has the ability to hydrolyze proteases and cellulases but does not produce siderophores.

[0072] Example 5: Effect of Bacillus saffron on the growth promotion of pepper

[0073] 5.1 Preparation of pepper seedlings: Pepper (Capsicum annuum L., CA181) seeds were sterilized with 75% ethanol solution for 1 min, then with 2% sodium hypochlorite for 3 min, and then washed 5-6 times with sterilized deionized water (SDW). After the seeds were germinated at room temperature, they were sown in 50-hole seedling trays and placed in the seedling room with normal water and fertilizer management.

[0074] 5.2 Seedling stage growth promotion test: When the pepper seedlings grow to two leaves and one heart, select healthy and uniformly growing seedlings and transplant them into seedling bags, and apply 5mL / plant of Bacillus fusus H23 bacterial solution around the rhizosphere, inoculate once every ten days, for a total of 3 times, and replace the bacterial solution with SDW for the control treatment. Move the treated seedlings to the seedling room (26℃-28℃, 14h light / 10h dark) for normal water and fertilizer management, and measure the following growth indicators after 30 days: plant height, stem thickness, number of leaves, and root length. Fresh weight and dry weight measurement: Separate the aboveground and underground parts of each treated plant, weigh the fresh weight of the aboveground and underground parts of the plant, and then dry them in an oven at 85℃ to constant weight, and weigh the dry weight of the aboveground and underground parts.

[0075] After applying the H23 bacterial solution, compared with the control group, all growth indicators of H23 were significantly higher than those of the control group (Table 4), indicating that H23 has a promoting effect on the growth of peppers.

[0076] Table 4H23 growth-promoting effect on pepper pot experiment

[0077]

[0078] Note: * indicates significant difference between the two treatments (P<0.05)

[0079] In summary, the Bacillus sabio H23 discovered in the present invention has a significant antagonistic effect on Capsici Phytophthora and other 8 Solanaceae plant pathogens, has a significant disease prevention effect on pepper blight, and has a promoting effect on pepper growth.

[0080] Although the content of the present invention has been described in detail through the above preferred embodiments, it should be appreciated that the above description should not be considered as a limitation of the present invention. After reading the above content, it will be apparent to those skilled in the art that various modifications and substitutions of the present invention will occur. Therefore, the protection scope of the present invention should be limited by the appended claims.

Claims

1. A Bacillus sabdariffa ( Bacillus safensis ) strain H23, characterized in that The deposit number of the Bacillus sabdariffa strain H23 is CCTCC NO: M 20241484.

2. A microbial preparation, characterized in that: The microbial preparation contains the Bacillus sp. strain H23 according to claim 1.

3. Use of the Bacillus subtilis strain H23 according to claim 1 or the microbial preparation according to claim 2 in preventing and controlling pathogens of Solanaceae plant diseases and / or promoting the growth of Solanaceae plants; the Solanaceae plant is pepper, and the pathogens of Solanaceae plant diseases are the pathogen of pepper blight, Phytophthora capsici, the pathogen of pepper wilt, Fusarium oxysporum, the pathogen of pepper black spot, Fusarium spp., the pathogen of pepper root rot, Sclerotium solani, Fusarium solani and Alternaria solani of tomato early blight.

4. A method for preventing and controlling pathogens of Solanaceae plant diseases or promoting the growth of Solanaceae plants, characterized in that: The method comprises the steps of configuring the Bacillus subtilis strain H23 described in claim 1 or the microbial preparation described in claim 2 into a suspension, and the step of irrigating or spraying the suspension around the root base of a Solanaceae plant, wherein the Solanaceae plant is pepper, and the pathogens of Solanaceae plant diseases are the pathogen of pepper blight, Phytophthora capsici, the pathogen of pepper wilt, Fusarium oxysporum, the pathogen of pepper black spot, Fusarium spp., the pathogen of pepper root rot, Sclerotium solani, the pathogen of pepper white rot, Fusarium solani and Alternaria solani, the pathogen of tomato early blight.

5. Use of the Bacillus sabdariffa strain H23 according to claim 1 or the microbial preparation according to claim 2 in the preparation of plant biocontrol agents and / or bio-organic fertilizers.

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