Bacillus subtilis HF11 and application thereof

Abstract

The invention provides a bacillus subtilis HF11 and application thereof, the preservation number of the bacillus subtilis HF11 is CGMCC No.37401, and the bacillus subtilis HF11 has wide antibacterial activity, shows a remarkable inhibition effect on various important plant pathogens such as fusarium graminearum, fusarium oxysporum, phytophthora nicotianae and alternaria alternata, and can also inhibit generation of vomitoxin. When the bacillus subtilis is applied in the wheat planting period, the wheat growth can be obviously promoted, and the yield and the grain quality are effectively improved. When the bacillus subtilis is used for feeding poultry, not only can the egg laying performance be improved, but also the meat production performance can be improved. The invention provides a new strain resource for developing a multifunctional microbial product integrating green prevention and control of plant diseases, stable yield and quality improvement of crops and quality improvement and efficiency improvement of the breeding industry, and is beneficial to promoting sustainable development of agriculture.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, and more particularly to a strain of Bacillus subtilis HF11 and its applications. Background Technology

[0002] Plant diseases caused by fungi and other pathogens pose a serious challenge to the safety and economic benefits of crop cultivation. They not only reduce crop yield and quality, but the toxins secreted by these pathogens (such as vomitoxin) also pose food safety risks. Chemical pesticides are a traditional method for controlling plant diseases, but they suffer from problems such as increased pathogen resistance, environmental residues, and disruption of the ecological balance. In poultry farming, the problem of feed contamination by mycotoxins, represented by vomitoxin, is particularly prominent. These toxins can directly damage the intestinal health, immune system, and production performance of poultry, leading to a decline in egg and meat production, becoming a key factor restricting farming efficiency. While chemical feed binders can remove mycotoxins, they themselves are prone to leaving residues. Therefore, there is an urgent need to develop environmentally friendly and efficient alternative strategies for both plant cultivation and poultry farming.

[0003] Probiotic preparations, characterized by their environmental friendliness and low likelihood of drug-resistant bacteria development, are currently a hot research topic in the field. Bacillus subtilis, as a widely used representative, can inhibit various pathogens by secreting lipopeptide antibiotics and other substances, and helps improve the rhizosphere microecology. Studies have also found that some Bacillus subtilis strains can improve the intestinal health and bodily functions of poultry. However, Bacillus subtilis strains that simultaneously contribute to plant disease control, plant growth and development, and poultry performance improvement are still rare. Therefore, there is an urgent need to develop new strains with multiple advantages. Summary of the Invention

[0004] To address the aforementioned problems in the prior art, this invention provides a strain of Bacillus subtilis HF11 and its applications.

[0005] The specific solution provided by this invention is as follows: In a first aspect, the present invention provides a strain of Bacillus subtilis ( Bacillus subtilis HF11, the Bacillus subtilis ( Bacillus subtilis The accession number for HF11 is CGMCC No. 37401. It was deposited on January 15, 2026, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 37401. The deposit address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing (Postcode: 100101). The taxonomic name is Bacillus subtilis. Bacillus subtilis The collection name is: HF11.

[0006] In this invention, strain HF11 refers to the Bacillus subtilis (… Bacillus subtilis HF11, this strain was isolated from the rhizosphere soil of reeds on the coast of Yazhou Bay, Sanya City, Hainan Province, and was identified by multiple techniques including morphology, physiology and biochemistry, and genomics.

[0007] Secondly, the present invention provides a microbial inoculant comprising the aforementioned Bacillus subtilis (Bacillus subtilis) Bacillus subtle HF11, or its freeze-dried cells, or its bacterial solution, or the supernatant or resuspension of the bacterial solution after centrifugation.

[0008] According to the present invention, a microbial agent is preferably prepared in which Bacillus subtilis (Bacillus subtilis) is present. Bacillus subtilis HF11 has a viable bacterial count ≥1×10⁻⁶. 9 cfu / g or ≥1×10 8 cfu / ml.

[0009] More preferably, if the microbial agent is in solid form, then the Bacillus subtilis ( Bacillus subtle HF11 has a viable bacterial count ≥1×10⁻⁶. 9 cfu / g.

[0010] More preferably, the microbial agent is in solid form, then the Bacillus subtilis ( Bacillus subtle HF11 has a viable bacterial count ≥1×10⁻⁶. 10 cfu / g.

[0011] More preferably, the microbial agent is in solid form, then the Bacillus subtilis ( Bacillus subtle HF11 has a viable bacterial count ≥ 2 × 10⁻⁶. 10 cfu / g.

[0012] In some specific embodiments, the microbial agent is in solid form, then the Bacillus subtilis ( Bacillus subtilis The viable bacteria content of HF11 was 2.1 × 10⁻⁶. 10 cfu / g.

[0013] More preferably, the carrier of the solid dosage form includes calcium carbonate.

[0014] More preferably, the carrier of the solid dosage form comprises light calcium carbonate.

[0015] More preferably, if the microbial agent is a liquid formulation, then the Bacillus subtilis ( Bacillus subtle HF11 has a viable bacterial count ≥1×10⁻⁶.8 cfu / ml.

[0016] More preferably, the microbial agent is a liquid formulation, then the Bacillus subtilis ( Bacillus subtle HF11 has a viable bacterial count ≥3×10⁻⁶. 8 cfu / ml.

[0017] In some specific embodiments, the microbial agent is a liquid formulation, then the Bacillus subtilis ( Bacillus subtilis The viable bacteria content of HF11 is 3×10⁻⁶. 8 cfu / ml.

[0018] Thirdly, the present invention provides a method for preparing the microbial agent, comprising: culturing the Bacillus subtilis (… Bacillus subtilis HF11.

[0019] According to the preparation method of the microbial agent provided by the present invention, preferably, the Bacillus subtilis ( Bacillus subtilis The culture temperature for HF11 is 28℃~37℃.

[0020] More preferably, the Bacillus subtilis ( Bacillus subtilis The culture temperature for HF11 is 30℃.

[0021] Fourthly, the present invention provides the aforementioned Bacillus subtilis ( Bacillus subtilis The application of HF11 or the aforementioned microbial agent in the resistance to plant pathogen infection, wherein the plant pathogen includes Fusarium graminearum (…). Fusarium grasses ), Pyrocystis pyriformis ( Botryosphaeria dothidea ), Tobacco Phytophthora ( Phytophthora Nicotiana ), Alternaria ( Alternaria alternata ), Alternaria appleii ( Alternaria mail ), Hari anthrax bacteria ( Colletotrichum higgindianum Apple anthracnose ( Colletotrichum gloeosporioides Fusarium equisetifolium ( Fusarium horsetail ), Curvularia trofrini Cloverleaf clover ), Corynebacterium cucumeri ( Bryonia spp. Fusarium oxysporum ( Fusarium oxysporum ) and cotton Alternaria ( Alternaria macrospore Any one or more of the following.

[0022] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis The application of HF11 or the aforementioned microbial agent in resisting plant pathogen infection, preferably, the plant pathogen is *Pyrophyllus* (…). Botryosphaeria dothidea), Curvularia trofrini Cloverleaf clover ), Alternaria appleii ( Alternaria mail ), Hari anthrax bacteria ( Colletotrichum higgindianum ) and Fusarium graminearum ( Fusarium gramineae ).

[0023] Compositions containing the aforementioned microbial agents should also be within the scope of protection of this invention.

[0024] Fifthly, the present invention provides the aforementioned Bacillus subtilis ( Bacillus subtilis The application of HF11 or the aforementioned microbial inoculant in the prevention and / or treatment of diseases caused by plant pathogens, including Fusarium graminearum (…). Fusarium gramineae ), Pyrocystis pyriformis ( Botryosphaeria dothidea ), Tobacco Phytophthora ( Phytophthora nicotianae ), Alternaria ( Alternaria alternata ), Alternaria appleii ( Alternaria mail ), Hari anthrax bacteria ( Colletotrichum higgindianum Apple anthracnose ( Colletotrichum gloeosporioides Fusarium equisetifolium ( Fusarium horsetail ), Curvularia trofrini Curved clover ), Corynebacterium cucumeri ( Bryonia spp. Fusarium oxysporum ( Fusarium oxysporum ) and cotton Alternaria ( Alternaria macrospora Any one or more of the following.

[0025] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis The application of HF11 or the aforementioned microbial agent in the prevention and / or treatment of diseases caused by plant pathogens, preferably, the plant pathogen is *Pyrophyllus* (…). Botryosphaeria dothidea ), Curvularia trofrini Cloverleaf clover ), Alternaria appleii ( Alternaria mail ), Hari anthrax bacteria ( Colletotrichum higgindianum ) and Fusarium graminearum ( Fusarium gramineae ).

[0026] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis The application of HF11 or the aforementioned microbial agent in the prevention and / or treatment of diseases caused by plant pathogens, preferably, the diseases include any one or more of wheat scab, corn ear rot, banana wilt, tomato wilt, branch and trunk ring rot, canker, anthracnose, early blight, leaf spot and target spot.

[0027] Sixthly, the present invention provides the aforementioned Bacillus subtilis ( Bacillus subtilisThe use of HF11 or the aforementioned microbial agent in inhibiting the production of vomitoxin and / or degrading vomitoxin.

[0028] In a seventh aspect, the present invention provides the aforementioned Bacillus subtilis ( Bacillus subtilis The application of HF11 or the aforementioned microbial agent in promoting wheat growth, increasing wheat yield and / or improving wheat quality.

[0029] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis The application of HF11 or the aforementioned microbial agent in promoting wheat growth, increasing wheat yield, and / or improving wheat quality, preferably, the improvement of wheat quality includes increasing the protein content of wheat grains.

[0030] Agricultural formulations containing the aforementioned microbial agents should also be within the scope of protection of this invention.

[0031] Preferably, the agricultural formulation is a fungicide or a plant growth regulator.

[0032] Preferably, the formulation of the agricultural preparation is an emulsifiable concentrate, suspension concentrate, wettable powder, water-dispersible granule, or pellet.

[0033] Eighthly, the present invention provides a feed additive comprising the aforementioned microbial agent.

[0034] The feed additive provided by the present invention preferably further includes a feed-grade acceptable carrier.

[0035] According to the present invention, the feed additive is preferably in the form of powder, granules, premix, diluent, microcapsule preparation, block preparation, solution, suspension, emulsifier or gel.

[0036] Compositions containing the aforementioned feed additives should also be within the scope of protection of this invention.

[0037] Feeds containing the aforementioned feed additives should also be within the scope of protection of this invention.

[0038] Ninthly, the present invention provides the Bacillus subtilis ( Bacillus subtilis HF11. The application of the microbial agent or the feed additive in poultry farming. The application is for non-disease treatment purposes.

[0039] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis HF11. The application of the microbial agent or the feed additive in poultry farming, preferably, the poultry includes chickens.

[0040] More preferably, the chicken is a broiler or a laying hen.

[0041] More preferably, the broiler chicken is a chick.

[0042] More preferably, the breed of broiler chicken is a white-feathered broiler chicken.

[0043] More preferably, the laying hen is a laying hen in the late stage of egg production.

[0044] More preferably, the breed of laying hen is the red-feathered laying hen.

[0045] More preferably, the breeding stock of the red-feathered laying hen is Jinghong No. 1.

[0046] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis HF11, the application of the microbial agent or the feed additive in poultry farming, preferably, the Bacillus subtilis ( ) in the feed additive Bacillus subtilis HF11 has a viable bacterial count ≥1×10⁻⁶. 9 cfu / g.

[0047] More preferably, the Bacillus subtilis (Bt) in the feed additive Bacillus subtilis HF11 has a viable bacterial count ≥ 2 × 10⁻⁶. 10 cfu / g.

[0048] In some specific embodiments, the Bacillus subtilis (Bt) in the feed additive Bacillus subtle The viable bacteria content of HF11 was 2.1 × 10⁻⁶. 10 cfu / g.

[0049] In a tenth aspect, the present invention provides the aforementioned Bacillus subtilis ( Bacillus subtilis HF11, the application of the microbial agent or the feed additive in the preparation of poultry feed, wherein the poultry feed is used to achieve any one or more of the following objectives: (1) improving the egg production performance of poultry; (2) improving the egg quality of poultry; (3) improving the growth performance of poultry; (4) improving the meat quality of poultry.

[0050] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis HF11. The application of the microbial agent or the feed additive in the preparation of poultry feed, preferably, the poultry are domestic poultry.

[0051] More preferably, the poultry includes chickens.

[0052] More preferably, the chicken is a broiler or a laying hen.

[0053] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilisHF11. The application of the microbial agent or the feed additive in the preparation of poultry feed, preferably, the improvement of poultry egg production performance includes any one or more of the following: increasing the number of eggs produced, increasing the weight of eggs produced, increasing the average weight of eggs, and reducing the feed conversion ratio.

[0054] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis HF11. The application of the microbial agent or the feed additive in the preparation of poultry feed, preferably, the improvement of poultry egg quality includes any one or more of the following: reducing the cholesterol content of poultry eggs, increasing the eggshell strength of poultry eggs, increasing the crude fat content of poultry eggs, and increasing the crude protein content of poultry eggs.

[0055] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis HF11, the application of the microbial agent or the feed additive in the preparation of poultry feed, preferably, the improvement of poultry growth performance includes: promoting weight gain and / or reducing feed conversion ratio.

[0056] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis HF11. The application of the microbial agent or the feed additive in the preparation of poultry feed, preferably, the improvement of poultry meat quality includes any one or more of the following: improving the flavor of poultry meat, improving the nutritional value of poultry meat, improving the water retention of poultry meat, and improving the tenderness of poultry meat.

[0057] The Bacillus subtilis (Bacillus subtilis) provided according to the present invention Bacillus subtilis HF11, the application of the microbial agent or the feed additive in the preparation of poultry feed, preferably, the Bacillus subtilis ( ) in the feed additive Bacillus subtilis HF11 has a viable bacterial count ≥1×10⁻⁶. 9 cfu / g.

[0058] More preferably, the Bacillus subtilis (Bt) in the feed additive Bacillus subtilis HF11 has a viable bacterial count ≥ 2 × 10⁻⁶. 10 cfu / g.

[0059] In some specific embodiments, the Bacillus subtilis (Bt) in the feed additive Bacillus subtle The viable bacteria content of HF11 was 2.1 × 10⁻⁶. 10 cfu / g.

[0060] More preferably, the amount of the feed additive added to the poultry feed is 150 g / ton to 300 g / ton.

[0061] More preferably, the amount of the feed additive added to the poultry feed is 210 g / ton.

[0062] The present invention has the following beneficial effects: The Bacillus subtilis provided by this invention ( Bacillus subtilis HF11 exhibits broad-spectrum antibacterial activity, showing significant inhibitory effects against various important plant pathogens such as *Fusarium graminearum*, *Fusarium oxysporum*, *Phytophthora indicum*, and *Alternaria alternata*, and can also inhibit the production of vomitoxin. Application of this strain during wheat cultivation significantly promotes wheat growth, effectively increasing yield and grain quality. Feeding this strain to poultry improves both egg production and meat production. This invention provides a new strain resource for developing multifunctional microbial products that integrate green control of plant diseases, stable crop yield and quality improvement, and enhanced efficiency in animal husbandry, contributing to the sustainable development of agriculture. Attached Figure Description

[0063] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0064] Figure 1 The results of the confrontation inhibition test between live bacteria of strain HF11 provided in Example 1 of this invention and Fusarium graminearum are shown; A is the control group and B is the treatment group.

[0065] Figure 2 The results of the confrontation inhibition test between the aseptic fermentation supernatant of strain HF11 provided in Example 1 of this invention and Fusarium graminearum are shown; A is the control group and B is the treatment group.

[0066] Figure 3 These are the morphological, Gram staining, and methylene blue staining identification results of strain HF11 provided in Example 1 of the present invention; A is the colony morphology of strain HF11; B is the Gram staining image of strain HF11; C is the methylene blue staining image of strain HF11.

[0067] Figure 4 This is a phylogenetic tree constructed based on the 16S DNA gene sequence of strain HF11, as provided in Example 1 of this invention. Detailed Implementation

[0068] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0069] Unless otherwise specified, the experimental methods involved in the following embodiments are conventional methods in the art. For example, you can refer to the experimental manual in the art or follow the conditions recommended in the manufacturer's instructions.

[0070] Unless otherwise specified, all experimental materials and reagents used in the following examples are commercially available.

[0071] The culture medium formulations used in the following examples are as follows: (1) Endo-KNO3 medium: glucose 28 g / L, KNO3 6 g / L, KH2PO4 1.2 g / L, MgSO4·7H2O 1.2 g / L and Na3C6H5O7·2H2O 0.2 g / L, with 1 mL / L of 1000× trace element stock solution added, 1L of distilled water, and pH adjusted to 7.0 using NaOH solution; (2) Potato agarose (PDA) medium: potato 200 g / L, glucose 20 g / L, agar 15-16 g / L and distilled water 1L, pH adjusted to 7.0; (3) LB liquid medium: yeast extract 5 g / L, NaCl 10 g / L, tryptone 10 g / L (3) LB solid medium: Add 15 g / L agar powder to LB liquid medium; (4) LB solid medium: Add 15 g / L agar powder to LB liquid medium; (5) Fermentation medium: 28 g / L glucose, 6 g / L KNO3, 1.2 g / L KH2PO4, 1.2 g / L MgSO4·7H2O, 0.2 g / L Na3C6H5O7·2H2O, 1 mL / L 1000×SL-10 trace element stock solution and 1 L distilled water, and adjust the pH to 7.0 using NaOH solution. All media need to be prepared at 121℃ (1.034×10⁻⁶ g / L). 5 (Pa) Sterilize with high-pressure steam for 20 minutes before use.

[0072] The pathogens used in the following examples are as follows: (1) Fusarium graminearum ( Fusarium gramineae ): The strain numbers are PH-1, Nanyang 301, Nanyang 1312, Nanyang 609 and Nanyang 20-1; (2) Pyrophyllus ( Botryosphaeria dothidae ): strain number BDH-1; (3) Phytophthora indicum ( Phytophthora nicotianae): The strain number is PNH-1; (4) Alternaria ( Alternaria alternata ): The strain number is AAH-1; (5) Alternaria applea ( Alternaria mail ): strain number AMH-1; (6) Hari anthrax bacteria ( Colletotrichum higgindianum ): The strain number is CHH-1; (7) Apple anthracnose ( Colletotrichum gloeosporioides ): Strain number CGH-1; (8) Fusarium equisetifolium ( Fusarium horsetail ): The strain number is FEH-1; (9) Curvularia trofrini ( Cloverleaf clover ): The strain number is CTH-1; (10) Corynebacterium cucurbita ( Bryonia spp. ): strain number DBH-1; (11) Fusarium oxysporum ( Fusarium oxysporum ): The strain number is FAH-1; (12) Alternaria solani ( Alternaria macrospora The strain number is AMH-1. The above-mentioned pathogen was provided by the research group of Hu Zanmin and Han Fangpu, laboratory technicians at the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences. The public can obtain this biological material from the applicant.

[0073] Example 1 Bacillus subtilis ( Bacillus subtilis Isolation, purification, screening, identification and preservation of HF11 1. Isolation and purification of strains Soil samples were collected from the rhizosphere of reeds along Yazhou Bay in Sanya City, Hainan Province, for bacterial isolation and purification. The specific methods are as follows: Take 5 g of dry soil sample, add 45 ml of sterile water and an appropriate amount of glass beads, shake at 28℃ and 200 rpm for 30 min to obtain a homogeneous soil slurry.

[0074] Take 1 mL of soil homogenate and perform serial dilution with distilled water to obtain a dilution factor of 10. -3 10 -4 and 10 -5 Take 500 μL of each diluted solution and spread it on LB medium (shake left and right to ensure the bacterial solution covers the entire plate, then blow dry), and then incubate at 28°C upside down for 24–48 h.

[0075] Take a 1.5 mL centrifuge tube, add 1 mL of LB liquid medium, pick a single colony into the 1.5 mL centrifuge tube and number it, then inoculate it into an Erlenmeyer flask containing 100 mL of LB liquid medium. Incubate at 28℃ and 200 rpm for 28–48 h to obtain the culture medium for each strain. Adjust the concentration of each strain to be consistent at 1 × 10⁻⁶. 9 CFU / ml.

[0076] 2. Strains screening The strains obtained in the previous step were screened using the plate confrontation method.

[0077] (1) Confrontational inhibition test of live bacteria Fusarium graminearum (Gynostemma pentaphyllum) was dispersed using the agar diffusion method. Fusarium gramineae Activation is achieved by inoculating the fungal disc onto PDA medium. Place the Fusarium graminearum mycelium in the middle of the PDA medium, and place the newly grown mycelium into a fresh PDA medium for later use, thus obtaining the activated Fusarium graminearum.

[0078] The activated Fusarium graminearum was made into mycelial cakes with a diameter of 8 mm and placed in the center of a new PDA medium. A straight line was drawn with the mycelial cake as the center. The culture broth of each strain was inoculated at 1.5 μL at a distance of 25 mm from the mycelial cake as the treatment group. The control group was inoculated with Fusarium graminearum mycelial cake alone. Each treatment was repeated three times and incubated in a constant temperature incubator at 25℃.

[0079] After the control group colonies have fully grown on the plate, the inhibition rate is calculated using the following formula: Inhibition rate (%) = (average diameter of control group colonies - average diameter of treatment group colonies) / average diameter of control group colonies × 100%.

[0080] (2) Screening results Among all strains, strain HF11 showed the highest inhibition rate against Fusarium graminearum, as shown in the results of the live bacteria confrontation inhibition test. Figure 1 As shown in A and B, the antibacterial rate is 64%.

[0081] 3. Standoff inhibition test of aseptic fermentation supernatant of strain HF11 (1) Preparation of sterile fermentation supernatant The culture medium of strain HF11 was centrifuged at 4℃ and 10000 r / min for 10 minutes, the supernatant was collected, and the bacterial cells were removed by filtration through a 0.22 μm filter membrane to obtain the sterile fermentation supernatant of strain HF11.

[0082] (2) Determination of the inhibition rate of aseptic fermentation supernatant against Fusarium graminearum The inhibition rate of aseptic fermentation supernatant of each strain against Fusarium graminearum was determined by the mycelial growth rate method.

[0083] The sterile fermentation supernatant of strain HF11 was uniformly mixed into PDA medium, and the sterile fermentation supernatant was diluted 20 times to obtain fermentation supernatant-PDA medium. Then, the activated Fusarium graminearum was made into mycelial cakes with a diameter of 8 mm and placed in the fermentation supernatant-PDA medium. The PDA medium containing mycelial cakes served as a control group. Each treatment was repeated three times and incubated in a constant temperature incubator at 25°C.

[0084] After the control group colonies have fully grown on the plate, the inhibition rate is calculated using the following formula: Inhibition rate (%) = (average diameter of colonies in the control group - average diameter of colonies in the treatment group) / average diameter of colonies in the control group × 100%.

[0085] (3) Test results The results of the confrontational antibacterial test of the aseptic fermentation supernatant of strain HF11 are as follows: Figure 2 As shown in A and B, the inhibition rate was 77%, indicating that the fermentation supernatant of strain HF11 also had antibacterial activity against Fusarium graminearum.

[0086] 4. Identification of strains The strain HF11 was identified using the methods described in the "Handbook of Systematic Identification of Common Bacteria".

[0087] (1) Morphological identification Strain strain HF11 was inoculated into LB liquid medium and cultured at 37°C for 12 h. Then, it was inoculated into LB liquid medium at an inoculation rate of 4% v / v and cultured at 30°C for 24 h. After that, the bacterial culture was diluted and spread onto LB solid medium. The surface morphology, concavity, size and transparency of the colonies were observed.

[0088] like Figure 3 As shown in A, the colonies of strain HF11 are nearly round, white, flat, and have a moist and opaque surface.

[0089] (2) Gram staining identification Colonies of strain HF11 were selected and Gram-stained.

[0090] like Figure 3 As shown in B, strain HF11 appears blue-purple and rod-shaped after Gram staining, indicating that it is a Gram-positive bacterium.

[0091] (3) Identification by methylene blue staining Colonies of strain HF11 were picked and stained with methylene blue. The morphology of strain HF11 cells under a microscope was observed for both non-spore-forming and spore-forming cells.

[0092] like Figure 3 As shown in C, strain HF11 has spores.

[0093] (4) Physiological and biochemical identification The physiological and biochemical characteristics of strain HF11 were determined by sugar alcohol fermentation test, anaerobic growth test, catalase test, VP test, starch hydrolysis test, gelatin liquefaction test, oxidase test, 7% sodium chloride growth test, propionate test and nitrate reduction test.

[0094] The experimental results showed that strain HF11 could utilize mannitol, D-xylitol, sucrose, maltose, sorbitol, glucose, fructose, and D-sorbitol, but could not utilize lactose and galactose. The starch hydrolysis test, catalase test, gelatin liquefaction test, nitrate reduction test, anaerobic test, and 7% sodium chloride growth test were all positive, while the propionate test was negative.

[0095] The above results indicate that strain HF11 is a facultative anaerobic bacterium, tolerant of high salt, capable of nitrate respiration, and secreting catalase, amylase, and gelatinase.

[0096] (5) Genome identification Single colonies of strain HF11 were inoculated into LB liquid medium and cultured at 30℃ and 200 r / min for 24 h. After centrifugation, the bacterial cells were collected, and DNA of strain HF11 was extracted using a bacterial DNA extraction kit. The 16S DNA gene sequence of strain HF11 was obtained and sequenced as follows:

[0097] The 16S DNA gene sequence (SEQ ID NO.1) obtained by sequencing was compared with the sequence in the GenBank database using BLAST analysis. The sequence of a known strain with high homology was selected as a reference strain. The sequence was analyzed using MEGA5.1 software, and a phylogenetic tree was constructed using the Neighbor-jioning method.

[0098] like Figure 4 As shown, strain HF11 clusters with Bacillus subtilis HF1 in the same lineage and is the most closely related, indicating a high degree of confidence.

[0099] Based on the above identification results, strain HF11 belongs to Bacillus subtilis (Bacillus subtilis). Bacillus subtilis ).

[0100] 5. Preservation of bacterial strains The aforementioned strain HF11 was deposited on January 15, 2026, at the China General Microbiological Culture Collection Center (CGMCC), with accession number CGMCC No. 37401. The deposit address is: Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing (Postcode: 100101). The taxonomic name is Bacillus subtilis. Bacillus subtilis The collection name is: HF11.

[0101] Example 2 Bacillus subtilis ( Bacillus subtilis Determination of the antibacterial spectrum of HF11 1. Activation culture of strain HF11 The preserved strain HF11 was inoculated into a 100 ml Erlenmeyer flask containing 50 ml of LB liquid medium and cultured at 37°C and 200 r / min for 12 h to activate it. The bacterial culture was then collected.

[0102] 2. Confrontation Training Method In this embodiment, *Fusarium graminearum*, *Phytophthora indicum*, *Fusarium oxysporum*, *Alternaria alternata*, *Curvaria trofrini*, *Cervaria cucurbita*, *Alternaria maculata*, *Anthracis harei*, *Alternaria cottoni*, *Anthracis maculata*, and *Fusarium equiseti* were used as pathogens.

[0103] Each activated pathogen was made into an 8 mm mycelial disc and placed in the center of a PDA medium. After new mycelia grew, the mycelial discs were made into 8 mm discs again and placed in a new PDA medium. A straight line was drawn with the mycelial disc as the center.

[0104] The sterile fermentation supernatant of strain HF11 was prepared according to the method in Example 1. 1.5 μL of the sterile fermentation supernatant of strain HF11 was inoculated in a straight line 25 mm from the center of the bacterial cake, with three parallel replicates. PDA medium inoculated only with bacterial cakes of each pathogen served as the corresponding control. After sealing with sealing film, the mixture was incubated at 25°C to observe the antibacterial activity of each pathogen and calculate the inhibition rate according to the method in Example 1.

[0105] 4. Measurement Results Table 1. Results of antibacterial spectrum assay for strain HF11

[0106] As shown in Table 1, strain HF11 exhibits inhibitory activity against 12 pathogens. The top five pathogens with the highest inhibition rates are, in descending order: *Fusarium graminearum*, *Cyclocarya piriformis*, *Cyclocarya trichomoniata*, *Alternaria maculata*, and *Anthracnose harenaeus*. This indicates that strain HF11 has potential control over plant diseases caused by the aforementioned pathogens (such as wheat scab, corn ear rot, banana Panama disease, tomato wilt, leaf spot, branch and trunk canker, cucumber brown spot, and tobacco black shank).

[0107] Example 3 Bacillus subtilis ( Bacillus subtilis Inhibitory effects of HF11 on different strains of Fusarium graminearum 1. Preparation of fermentation agent of strain HF11 The preserved strain HF11 was inoculated into a 100 ml Erlenmeyer flask containing 50 ml of LB liquid medium and incubated at 37°C and 200 r / min for 12 h to activate it.

[0108] The activated bacterial culture was collected and inoculated into a 250 ml shake flask containing 100 ml of fermentation medium at an inoculation rate of 4% v / v. After culturing at 30℃ and 200 r / min for 24 h, the bacterial culture of strain HF11 was obtained, with a bacterial concentration of approximately 1×10⁻⁶. 9 / ml, collect the fermentation broth, centrifuge and retain the supernatant, filter through a 0.22 μm filter membrane to remove the bacterial cells, and dilute 10 times with distilled water to obtain the fermentation preparation of strain HF11.

[0109] 2. Co-culture of Fusarium graminearum and strain HF11 fermentation preparation Different Fusarium graminearum strains (PH-1, Nanyang 301, Nanyang 1312, Nanyang 609 and Nanyang 20-1) were co-cultured with the fermentation preparation of strain HF11 at a ratio of 1:1 (volume ratio) as experimental groups.

[0110] Each Fusarium graminearum strain without strain HF11 was used as the corresponding control group.

[0111] 3. Determination of DON content After culturing the experimental and control groups at 30℃ for 24 hours, culture medium samples were collected. The DON content was detected by enzyme-linked immunosorbent assay (ELISA) using a deoxynivalenol (DON) assay kit (catalog number RE9147, purchased from Baoruyi Biotechnology Co., Ltd.), and the DON degradation rate was calculated using the formula: DON inhibition rate (%) = .

[0112] 4. Antibacterial test of aseptic fermentation supernatant against different Fusarium graminearum species. The inhibition rate of the aseptic fermentation supernatant of strain HF11 against various Fusarium graminearum strains was determined by the mycelial growth rate method.

[0113] The sterile fermentation supernatant of strain HF11 was prepared according to the method in Example 1. The sterile fermentation supernatant of strain HF11 was uniformly mixed into PDA medium, diluting the supernatant 20-fold to obtain fermentation supernatant-PDA medium. Then, the activated Fusarium graminearum strains were prepared into mycelial discs with a diameter of 8 mm and placed in the fermentation supernatant-PDA medium. PDA medium containing only mycelial discs served as the corresponding control group. Each treatment was repeated three times and incubated in a 25°C incubator.

[0114] After the control group was fully covered with bacteria, the inhibition rate was calculated according to the method in Example 1.

[0115] 5. Measurement Results Table 2. Inhibition of different Fusarium graminearum strains by strain HF11

[0116] Note: * indicates a significant difference between the experimental group and the control group, P < 0.05; ** indicates an extremely significant difference between the experimental group and the control group, P < 0.01; ***P indicates an extremely significant difference between the experimental group and the control group, P < 0.001.

[0117] As shown in Table 2, HF11 had an inhibitory effect on all detected Fusarium graminearum strains and also inhibited the production of DON toxin. Among them, strain HF11 had the strongest inhibitory effect on Fusarium graminearum strain Nanyang 609, with the highest inhibition rate (77.2%) and the highest DON toxin inhibition rate (75%).

[0118] Example 4: Effects of strain HF11 on wheat yield and quality 1. Experimental Methods The wheat variety used in this embodiment is Zhongke 366 (provided by Researcher Liu Zhiyong of the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences; the public can obtain this biological material from the applicant).

[0119] The experimental design employed randomized controlled trials, with a treatment group (T) and a control group (CK). Each group consisted of three replicates, with each replicate having an area of ​​20 square meters.

[0120] For the wheat in the treatment group, fermentation broth of strain HF11 (diluted 100 times, viable cell count 3×10⁻⁶) was used. 8 The fermentation liquid (CFU / ml) was applied once during the seedling stage and then sprayed once during the flowering stage, with each application being 0.5L / plot. For the control group of wheat, the same amount of tap water was applied using the same frequency and method.

[0121] Wheat grains from each group were harvested, wheat yield was tallied, and the protein content in the wheat grains was determined using the Kjeldahl method.

[0122] 2. Experimental Results Table 3. Results of wheat yield and quality determination

[0123] Note: * indicates a significant difference between the treatment group and the control group, P < 0.05.

[0124] As shown in Table 3, compared with the control group, the wheat yield and grain protein content of the treatment group were significantly increased, by 1.63% and 16.18%, respectively. This indicates that the application of strain HF11 can significantly increase wheat yield and improve grain quality.

[0125] Example 5: Effects of strain HF11 as a feed additive on the egg production performance of laying hens. 1. Experimental Methods The laying hen breed used in this embodiment was Jinghong No. 1, with an initial age of 420 days (late laying period). The laying hens were randomly divided into 3 groups, designated as BR group, HF11 group and negative control group, with 100 hens in each group and 3 parallel replicates in each group.

[0126] The laying hens in the BR group were fed a conventional feed supplemented with 240 g / ton (of feed) of BRS feed additive, wherein the BRS feed additive is Bacillus subtilis (… Bacillus subtilisThe spray-dried product of the fermentation broth of strain HF11 (accession number CGMCC No. 23476, disclosed in Chinese invention patent ZL202111555449.0) (light calcium carbonate as carrier, viable bacteria content of 21 billion CFU / g) was used. Laying hens in the HF11 group were fed a conventional feed supplemented with 240 g / ton (of feed) of HF11 feed additive, wherein the HF11 feed additive is the spray-dried product of the fermentation broth of strain HF11 (light calcium carbonate as carrier, viable bacteria content of 21 billion CFU / g). Laying hens in the negative control group were fed a conventional feed. The aforementioned conventional feed was late-stage laying hen feed (purchased from Hebei Yuexing Feed Co., Ltd.).

[0127] The chickens were raised for 49 days with free access to feed. The feed intake and egg production of each group of laying hens were observed and recorded daily. After the rearing period, the number of eggs produced, egg weight, average egg weight, feed consumption, feed conversion ratio and laying rate of each group were counted. The egg quality (cholesterol, eggshell strength, crude fat content and crude protein content) of each group was determined by the Feed Research Institute of the Chinese Academy of Agricultural Sciences.

[0128] 2. Experimental Results Table 4 Results of egg production performance testing of laying hens

[0129] Note: Different lowercase letters indicate significant differences between groups, P < 0.05.

[0130] As shown in Table 4, compared with the BR group and the negative control group, the HF11 group showed significantly higher egg production, egg weight, average egg weight, feed consumption, and egg production rate, and significantly lower feed conversion ratio. Specifically, the egg production increased by 10.53% and 17.51%, respectively; egg weight increased by 17.18% and 28.06%, respectively; average egg weight increased by 6.01% and 8.98%, respectively; feed consumption increased by 1.23% and 0.31%, respectively; egg production rate increased by 10.58% and 17.57%, respectively; and feed conversion ratio decreased by 10.58% and 17.57%, respectively.

[0131] Table 5 Results of egg quality determination from laying hens

[0132] As shown in Table 5, compared with the negative control and the BR group, the cholesterol content in the eggs of the HF11 group was significantly reduced, with decreases of 25.13% and 20.22%, respectively; the eggshell strength was significantly enhanced, with increases of 19.61% and 9.77%, respectively; the fat content decreased by 25.24% and 21.67%, respectively; and the protein content increased by 19.17% and 19.95%, respectively.

[0133] Similar results were observed in other independent experiments when the HF11 feed additive was added at concentrations of 150 g / ton, 200 g / ton, and 300 g / ton.

[0134] The above results indicate that strain HF11, as a feed additive, can significantly improve the egg production performance and egg quality of laying hens.

[0135] Example 6: Effects of strain HF11 as a feed additive on the growth performance and meat quality of broilers. 1. Experimental Methods The broiler breed used in this embodiment is white-feathered broiler (purchased from Huadu Broiler Technology Co., Ltd.), and the initial age is 0 days.

[0136] Broilers were randomly divided into three groups: BR group, HF11 group and negative control group. Each group had 40 chickens and three replicates were set up. The ratio of roosters to hens in each group was 1:1.

[0137] Broilers in the BR group were fed a conventional diet supplemented with 600 g / ton (of feed) of BRS feed additive, wherein the BRS feed additive is Bacillus subtilis (… Bacillus subtilis The spray-dried product of the fermentation broth of strain HF11 (accession number CGMCC No. 23476, disclosed in Chinese invention patent ZL202111555449.0) (light calcium carbonate as carrier, viable bacteria content of 21 billion CFU / g) was used. Laying hens in the HF11 group were fed a conventional feed supplemented with 600 g / ton (of feed) of HF11 feed additive, wherein the HF11 feed additive is the spray-dried product of the fermentation broth of strain HF11 (light calcium carbonate as carrier, viable bacteria content of 21 billion CFU / g). Laying hens in the negative control group were fed a conventional feed. The aforementioned conventional feed was chick feed (purchased from Hebei Yuexing Feed Co., Ltd.).

[0138] The chickens were raised until 42 days of age, during which time they were fed a restricted diet, with each group receiving a total of 160 kg of feed. After the rearing period, the weight of each group of broilers was measured, and the total weight at 42 days (i.e., the total weight of each group of broilers), average weight of chickens (i.e., the average weight of each group of broilers), feed conversion ratio, and mortality rate were recorded. The amino acid content and physical indicators (drip loss rate, cooking loss, and shear force) of the chicken breast meat of each group of broilers were measured by the Feed Research Institute of the Chinese Academy of Agricultural Sciences.

[0139] 2. Experimental Results Table 6 Results of Broiler Growth Performance Measurement

[0140] Note: Different lowercase letters indicate significant differences between groups, P < 0.05.

[0141] As shown in Table 6, there were no deaths among the three groups of broilers. Compared with the BR group and the negative control group, the HF11 group showed a significant increase in total weight and average weight at 42 days, and a significant decrease in feed conversion ratio. Specifically, the total weight at 42 days increased by 8.41% and 21.84%, respectively, the average weight increased by 8.37% and 22.17%, respectively, and the feed conversion ratio decreased by 8.33% and 18.09%, respectively.

[0142] Table 7. Results of amino acid content determination in broiler breast meat from 42-day-old chickens.

[0143] Note: Different lowercase letters indicate significant differences between groups, P < 0.05.

[0144] As shown in Table 7, compared with the BR group and the negative control group, the HF11 group showed a significant increase in the contents of Lys, Gly, Thr, Arg, Ala, Tyr, and Val in the breast muscle of broilers. Specifically, the contents of Lys increased by 12.01% and 24.52%, respectively; the contents of Gly increased by 37.70% and 45.71%, respectively; the contents of Thr increased by 87.38% and 91.87%, respectively; the contents of Arg increased by 27.81% and 58.96%, respectively; the contents of Ala increased by 19.74% and 37.66%, respectively; the contents of Tyr increased by 247.06% and 244.17%, respectively; and the contents of Val increased by 46.47% and 55.14%, respectively. This indicates that the addition of the HF11 strain resulted in broilers with better flavor and higher nutritional value.

[0145] Table 8. Results of physical property determination of broiler breast meat from 42-day-old chickens.

[0146] Note: Different lowercase letters indicate significant differences between groups, P < 0.05.

[0147] As shown in Table 8, compared with the BR group and the negative control group, the drip loss rate, cooking loss, and shear force of broiler breast meat in the HF11 group were significantly reduced. Specifically, the drip loss rate decreased by 30.73% and 27.91%, respectively; the cooking loss decreased by 11.69% and 2.00%, respectively; and the shear force decreased by 8.72% and 10.19%, respectively. This indicates that the addition of HF11 strain resulted in better water retention and more tender broiler meat.

[0148] Similar results were observed in other independent experiments when the HF11 feed additive was added at concentrations of 150 g / ton, 200 g / ton, and 300 g / ton.

[0149] The above results indicate that strain HF11, as a feed additive, can significantly improve the meat production performance and meat quality of broilers.

[0150] Example 7: Strain HF11 produces abundant fengycin and surfactant and their homologues. 1. Extraction of antimicrobial lipopeptides from strain HF11 The activated bacterial culture of strain HF11 was inoculated at 3% v / v in Endo-KNO3 medium and cultured at 37℃ and 200 r / min for 2 days. The culture was then collected, and its antimicrobial lipopeptides were initially extracted by acid precipitation (6M HCl).

[0151] Take 50 mL of culture medium, centrifuge at 10,000 g for 20 min at 4 °C, and collect the supernatant. Adjust the pH of the supernatant to 2.0 with 6 M HCl and incubate overnight at 4 °C. The next day, centrifuge at 10,000 g for 15 min at 4 °C, discard the supernatant, and retain the precipitate. Add 5 mL of methanol to the precipitate to dissolve it, centrifuge at 10,000 g for 10 min. At this point, lipopeptide compounds are present in the methanol solution. Collect the supernatant and filter it through a 0.22 µm filter membrane for later use.

[0152] 2. Detection of antimicrobial lipopeptides The crude lipopeptide extract in the fermentation broth was identified and its molecular weight was determined using matrix-assisted laser desorption / ionization time-of-flight tandem mass spectrometry (MALDI-TOF / TOF) in reflectance mode.

[0153] The structure of lipopeptide molecules in the fermentation broth was identified using ultra-high performance liquid chromatography-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The HPLC parameters were as follows: mobile phase A was 0.1% HPLC-grade formic acid aqueous solution, and mobile phase B was HPLC-grade methanol; the elution gradient was: 0.1 min, 70% mobile phase B; 0.1 min–2.0 min, 70% mobile phase B; 2.0 min–8.0 min, 70%–100% mobile phase B; 8.0 min–10 min, 100% mobile phase B; 10.1 min, 70% mobile phase B; 10.1 min–13 min, 70% mobile phase B; the flow rate was 0.3 mL / min; and the chromatographic column used was a Waters BEH C18 (50 mm × 2.1 mm, 1.7 µm particle). The mass spectrometry method was as follows: positive ion mode was used for analysis. MS tuning parameters were: degassing rate 500 L / h, degassing temperature 500℃, cone gas flow rate 50 L / h, source temperature 120℃, capillary voltage 3.0 kV, and cone voltage 30 V. Data acquisition time was 0.5 sec, and the mass-to-charge ratio range was 100 m / z to 1,200 m / z. The collision voltage was set to 25 V and fine-tuned according to actual conditions. Mass spectrometry results were analyzed in accordance with standard mass spectra of fengycin and surfactant and references (Confirmation of the Structure of Surfactin by Mass Spectrometry (https: / / doi.org / 10.1080 / 00021369.1969.10859524); Purification and structural characterization of fengycin homologues produced by Bacillus subtilis LSFM-05grown on raw glycerol (DOI: 10.1007 / s10295-011-0980-1)).

[0154] 3. Test Results Table 9. Molecular identification results of Fengycin and its homologues

[0155] Note: * indicates that the fatty acid chain has one double bond; m / z refers to the ratio of the mass of the charged particle to its charge, [M+H] + Refers to the protonated molecular ion, [M+Na]. + It refers to the sodium adduct ion in the molecule.

[0156] Table 10 Molecular identification results of Surfactin and its homologues

[0157] Note: m / z refers to the ratio of the mass of a charged particle to its charge, [M+H] + Refers to the protonated molecular ion, [M+Na]. + It refers to the sodium adduct ion in the molecule.

[0158] As shown in Tables 9 and 10, strain HF11 produces abundant homologues of cytokinin and surfactant. Surfactants help strain HF11 colonize, move, and form biofilms on the host surface, disrupting the integrity of pathogen cell membranes and creating favorable conditions for the penetration and killing of cytokinin homologues, thus endowing strain HF11 with broad antibacterial activity.

[0159] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A strain of Bacillus subtilis ( Bacillus subtilis HF11, characterized in that, The Bacillus subtilis ( Bacillus subtilis The accession number of HF11 is CGMCC No. 37401.

2. A microbial inoculant, characterized in that, Contains the Bacillus subtilis of claim 1 ( Bacillus subtilis HF11, or its freeze-dried cells, or its bacterial solution, or the supernatant or resuspension of the bacterial solution after centrifugation.

3. The method for preparing the microbial inoculant according to claim 2, characterized in that, include: Cultivating the Bacillus subtilis of claim 1 ( Bacillus subtilis HF11.

4. The Bacillus subtilis strain described in claim 1 ( Bacillus subtilis The application of the microbial agent according to HF11 or claim 2 in resisting plant pathogen infection is characterized in that... The plant pathogens include Fusarium graminearum (… Fusarium graminearum ), Pyrocystis pyriformis ( Botryosphaeria dothidea ), Tobacco Phytophthora ( Phytophthora nicotianae ), Alternaria ( Alternaria alternate ), Alternaria appleii ( Alternaria mail ), Hari anthrax bacteria ( Colletotrichum higgindianum Apple anthracnose ( Colletotrichum gloeosporioides Fusarium equisetifolium ( Fusarium equiseti ), Curvularia trofrini Curvularia trifolii ), Corynebacterium cucumeri ( Didymella bryoniae Fusarium oxysporum ( Fusarium oxysporum ) and cotton Alternaria ( Alternaria macrospora Any one or more of the following.

5. The Bacillus subtilis strain described in claim 1 ( Bacillus subtilis The application of the microbial agent as described in HF11 or claim 2 in the prevention and / or treatment of diseases caused by plant pathogens, characterized in that... The plant pathogens include Fusarium graminearum (… Fusarium graminearum ), Pyrocystis pyriformis ( Botryosphaeria dothidea ), Tobacco Phytophthora ( Phytophthora nicotianae ), Alternaria ( Alternaria alternate ), Alternaria appleii ( Alternaria mail ), Hari anthrax bacteria ( Colletotrichum higgindianum Apple anthracnose ( Colletotrichum gloeosporioides Fusarium equisetifolium ( Fusarium equiseti ), Curvularia trofrini Curvularia trifolii ), Corynebacterium cucumeri ( Didymella bryoniae Fusarium oxysporum ( Fusarium oxysporum ) and cotton Alternaria ( Alternaria macrospora Any one or more of the following; Preferably, the disease includes any one or more of the following: wheat scab, corn ear rot, banana wilt, tomato wilt, branch and trunk ring rot, canker, anthracnose, early blight, leaf spot, and target spot.

6. The Bacillus subtilis of claim 1 ( Bacillus subtilis The use of the microbial agent of HF11 or claim 2 in inhibiting the production of vomitoxin and / or degrading vomitoxin.

7. The Bacillus subtilis strain according to claim 1 ( Bacillus subtilis The application of the microbial agent described in HF11 or claim 2 in promoting wheat growth, increasing wheat yield and / or improving wheat quality.

8. A feed additive, characterized in that, It includes the microbial agent as described in claim 2.

9. The Bacillus subtilis of claim 1 ( Bacillus subtilis HF11, the application of the microbial agent of claim 2 or the feed additive of claim 8 in poultry farming.

10. The Bacillus subtilis strain according to claim 1 ( Bacillus subtilis HF11, the application of the microbial agent of claim 2 or the feed additive of claim 8 in the preparation of poultry feed, characterized in that, The poultry feed is used to achieve one or more of the following purposes: (1) Improve the egg production performance of poultry; (2) Improve the quality of poultry eggs; (3) Improve the growth performance of poultry; (4) Improve the quality of poultry meat.

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