A simple bacillus ASE52 and its applications

Abstract

This invention relates to a simple peritrichous bacillus. Peribacillus simplex ASE52 and its applications. This invention relates to a simple aperidobacterium, ASE52, isolated from the seeds of *Leymus chinensis*, which is deposited at the China Center for Type Culture Collection (CCTCC) under accession number M20251159. This bacterium exhibits excellent growth-promoting effects on *Leymus chinensis* seedlings and can be used to prepare microbial agents for application during the seedling stage. It can be applied to the bioremediation of desert grasslands and the planting of *Leymus chinensis*, providing an excellent basic strain for the exploration and application of desert microbial resources.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, specifically to a simple Bacillus peritonis. Peribacillus simplex Application of ASE52 in promoting the growth of *Leymus chinensis* seedlings. Background Technology

[0002] Desertification is an ecological problem caused by the long-term accumulation of multiple factors. Northern and western my country are located in arid and semi-arid climate zones, with annual precipitation mostly below 400 mm, while potential evaporation reaches 2000-3000 mm. This imbalance between water and heat limits the natural recovery capacity of vegetation, constituting the basic conditions for desertification. Human activities, by altering land cover and hydrological conditions, have become the main driving force behind desertification. Under multiple pressures, desertification in Inner Mongolia is becoming increasingly severe. How to effectively improve the desertification situation is a core challenge facing the region's economic development and ecological protection.

[0003] *Elymus nutans* Griseb., a perennial herbaceous plant belonging to the Poaceae family, plays an important role in ecological restoration and grassland animal husbandry development in arid and semi-arid regions of northern my country, possessing both outstanding ecological adaptability and economic value. It is tolerant of extreme low temperatures, able to withstand environments as low as -30 degrees Celsius, with a wintering rate exceeding 90%, and is adapted to temperate to cold-temperate climates. It exhibits significant drought tolerance, able to grow naturally in areas with annual precipitation of 250-400 mm, relying on its deep root system to absorb deep soil moisture and coping with drought through physiological mechanisms such as proline accumulation. In ecological restoration, *Elymus nutans* is a key "pioneer grass species" for desertification control; its well-developed root system and ground cover can reduce wind speed by 30%-50%, reduce soil erosion by 60%-80%, and effectively inhibit the expansion of mobile sand dunes.

[0004] Specific bacterial communities exist on the surface and inside the seeds of *Leymus chinensis*, with *Pseudomonas*, *Bacillus*, and *Pantoea* being the dominant groups. These bacteria significantly influence seed germination and host resistance by secreting auxins, hydrolytic enzymes, and stress-resistance metabolites. Studies have shown that growth-promoting bacteria in the seed sphere and within the seed can decompose seed coat inhibitory substances and regulate plant hormone balance, increasing seed germination rate by 25%-30% and seedling height by 15%-20%. Under saline-alkali or drought stress, stress-resistant bacteria help the host maintain cell membrane stability by enhancing antioxidant enzyme activity and regulating ion transport gene expression, thus maintaining a germination rate of over 60% under stress conditions.

[0005] Studies have found that treatment with these bacteria significantly improved rice seed germination potential, germination index, vigor index, shoot length, and root length, increasing by 12%, 16%, 30%, 28%, and 26%, respectively. In in vitro growth-promoting experiments, rice seedlings grown in insoluble potassium environments showed a 35% increase in underground dry weight and significantly promoted root growth. During the seedling growth stage, root development increased by 54.9%, aboveground plant height increased by 44.7%, and fresh and dry weight increased by 61.8% and 50.0%, respectively. Physiological and biochemical analyses showed that growth-promoting bacteria increased root vigor by 60.7%, chlorophyll content by 32.7%, and promoted the absorption of mineral elements such as nitrogen, phosphorus, and potassium, with nitrogen content increasing by up to 21.3%, thus improving photosynthetic efficiency and nutrient utilization. Other researchers have screened numerous bacteria with growth-promoting and stress-resistance functions and elucidated their mechanisms of action, providing alternative solutions for desert ecological vegetation restoration and sustainable agricultural development. These studies all confirm the growth-promoting potential of bacteria in the seed sphere and within seeds.

[0006] Peribacillus simplex is a Gram-positive, spore-forming bacterium with broad-spectrum plant growth-promoting and biocontrol functions. It secretes indoleacetic acid (IAA), solubilizes phosphorus, and produces siderophores, significantly promoting the growth and nutrient absorption of crops such as wheat, potatoes, and tomatoes. It exhibits strong antagonistic effects against various pathogenic fungi (such as Fusarium), bacteria (such as Xylella fastidiosa), and nematodes, enhancing crop disease resistance by producing antimicrobial lipopeptides, volatile organic compounds (VOCs), and inducing systemic resistance (ISR). It also demonstrates excellent performance in environmental remediation, adsorbing heavy metals (such as cadmium (Cd) and lead (Pb)) and degrading organic pollutants (such as pesticides and crude oil), making it suitable for soil and water bioremediation. Furthermore, this bacterium complies with EU microbial pesticide regulations, possessing commercial potential and representing an important microbial resource for sustainable agriculture as an alternative to chemical pesticides and fertilizers. However, current technologies lack reports on the application of Peribacillus simplex in promoting the growth of Leymus chinensis seedlings, necessitating the screening of targeted strains to meet the ecological restoration needs of desert grasslands.

[0007] CN121379888A discloses *Peribacillus simplex* ZF535, with accession number CGMCC No: 34733. The extracellular polysaccharides of this bacterium, applied as foliar sprays or root irrigation, significantly induce resistance to cucumber angular leaf spot. Strain ZF535 is a *Peribacillus simplex* strain with potential biocontrol application capabilities that induce plant disease resistance, exhibiting good biocontrol effects. CN117356221B discloses a method for promoting the germination of *Leymus chinensis* seeds using soil growth-promoting bacteria, including the following technical solutions: 1) disinfection treatment; 2) seed soaking treatment: soaking disinfected seeds in a suspension of *Bacillus licheniformis*, *Bacillus subtilis*, *Bacillus amyloliquefaciens*, and *Rhizobium rhizobium*; 3) germination. Soaking *Leymus chinensis* seeds with growth-promoting bacteria can significantly improve the germination rate and uniformity of *Leymus chinensis* seeds, and also improve the salt tolerance of *Leymus chinensis* seeds. There are currently no reports of Bacillus simplex being used to assist in promoting the growth of Leymus chinensis seedlings. Summary of the Invention

[0008] To address the shortcomings of existing technologies, this invention provides a simple Bacillus aureus ASE52 isolated from the seeds of Leymus chinensis, which has a significant promoting effect on improving the seedling growth of Leymus chinensis.

[0009] The above strains are Peribacillus simplex The accession number is ASE52, and it is deposited at the China Center for Type Culture Collection (CCTCC) with accession number M20251159.

[0010] The simple perisporidium provided by this invention Peribacillus simplex ASE52 was deposited on May 29, 2025, at the China Center for Type Culture Collection (CCTCC, No. 299 Bayi Road, Wuchang District, Wuhan, Hubei Province, 430072, China), and classified as follows: Peribacillus simplex ASE52, accession number CCTCC No:M20251159.

[0011] Simple Bacillus Peribacillus simplex The 16S rDNA nucleotide sequence of ASE52 is shown in SEQ ID NO:1. The 16S rRNA gene sequence of this strain has 99.87% sequence similarity to that of Peribacillus simplex LMG 11160 (Genbank number: NR_114919).

[0012] The sequence of SEQ ID NO:1 is as follows: CTCCCTGAGGTTAGCGGCGGACGGGTGAGTAACACGTGGGCAACCTGCCTATAAGACTGGGATAACTTCGGGAAACCGGAGCTAATACCGGATACGTTCTTTTCCGCATGAGAGAAGATGGAAAGACGGTTTACGCTGTCACTTATAGATGGGCCCGCGGCGCATTAGCTAGTTGGTGAGGTAATGG CTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAACGAAGAAGGCCTTCGGGTCGTAAAGTTCTGTTGTTAGGGAAGAACA AGTACCAGAGTAACTGCTGGTACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGTTTCCTTAAGTCTGATGTGAAAGCCCACGGCTCAACCGTGGAGGGTCATTGGAAA CTGGGGAACTTGAGTGCAGAAGAGGAAAGTGGAATTCCAAGTGTAGCGGTGAAATGCGTAGAGATTTGGAGGAACACCAGTGGCGAAGGCGACTTTCTGGTCTGTAACTGACACTGAGGCGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAG Simple Bacillus Peribacillus simplex ASE52 is a strain isolated from the seeds of *Leymus chinensis*. This strain is Gram-positive and grows well on R2A solid medium. The colonies are white. The optimal growth temperature is 26℃ and the optimal growth pH is 7.0.

[0013] The simple Bacillus apocytogenes culture conditions provided by this invention are as follows: culture on R2A solid medium, temperature 26-28℃, pH 7.0-7.2; preferably, the culture temperature is 26-27℃, and the optimal growth pH is 7.0.

[0014] This invention provides a bacterial agent containing Bacillus simplex. Peribacillus simplex ASE52. This inoculant can be used as a liquid inoculant, or it can be prepared as any one of the following: seed soaking solution, inoculant mud, seed coating, or fertilizer.

[0015] This invention provides a product containing Bacillus simplex. Peribacillus simplex ASE52, the total viable count of Bacillus simplex in the product is not less than 10. 6 CFU / mL.

[0016] The above contains simple perisporidum Peribacillus simplex The ASE52 microbial agent can be prepared using conventional techniques and by adding excipients permitted in the field of microbial preparations.

[0017] This invention also provides simple perisporidia. Peribacillus simplex The application of ASE52 or an inoculum containing this strain in the seedling growth of Elymus nutans Griseb. was studied.

[0018] This invention also provides simple perisporidia. Peribacillus simplex ASE52 or an inoculant containing this strain may be used in one or more of the following applications: (1) to improve the germination rate of Leymus chinensis; (2) to improve the height of Leymus chinensis plants; (3) to improve the dry weight of Leymus chinensis on the ground; (4) to improve the dry weight of Leymus chinensis on the ground.

[0019] This invention also provides a microbial inoculant for promoting the growth of *Leymus chinensis* seedlings, which contains *Bacillus simplex*. Peribacillus simplex ASE52 cells or its fermentation product. The fermentation product is the simple Bacillus peritonis. Peribacillus simplex The bacterial cells obtained after fermentation of ASE52.

[0020] The above-mentioned simple Bacillus peritonis Peribacillus simplex Fermentation of ASE52 can be carried out in R2A medium at pH 7.0 and 26°C with shaking. The specific application method is as follows: Bacillus simplex is applied during the seedling stage of *Leymus chinensis*. Peribacillus simplex ASE52 cells.

[0021] Compared with the prior art, the present invention has the following advantages: (1) The present invention provides a simple Bacillus periwinkle ASE52 isolated from the seeds of Leymus chinensis, with the preservation number CCTCC No: M20251159. The strain has a clear source and is adapted to the host characteristics.

[0022] (2) The bacterial cells obtained by fermentation of this strain have a significant promoting effect on the growth of *Leymus chinensis* seedlings and can simultaneously improve key growth indicators such as germination rate, plant height, aboveground dry weight and underground dry weight.

[0023] (3) Simple Bacillus ASE52 can be used to prepare microbial agents for application during the seedling stage of Leymus chinensis. It can be applied to the restoration and expansion of Leymus chinensis communities in desert grasslands, providing excellent basic strains for the exploration and application of desert microbial resources, and has good practical application value. Attached Figure Description

[0024] Figure 1 The morphology of strain Peribacillus simplex ASE52 after 3 days of culture on R2A plates.

[0025] Figure 2 The germination rate of *Leymus chinensis* seeds under different treatments.

[0026] Figure 3 The plant height of *Leymus chinensis* under different treatments.

[0027] Figure 4 The dry weight of drooping, saline-alkali-tolerant grassland under different treatments.

[0028] Figure 5 The understory dry weight of drooping, alkali-tolerant grass under different treatments.

[0029] in, Figures 2 - 5 The study included four treatments: a control group treated with sterile water; a strain LMG 11160 treatment treated with the simple Bacillus peritonis strain LMG 11160; a sterilized strain ASE52 treatment treated by soaking the sterilized strain ASE52 in culture; and a strain ASE52 treatment treated with live ASE52 bacteria. Detailed Implementation

[0030] The following detailed embodiments further illustrate the concept and technical effects of the present invention to provide a full understanding of its purpose, features, and effects. Unless otherwise specified, all methods described are conventional methods, and all materials and reagents are available commercially through publicly available channels. The illustrative embodiments and descriptions of the present invention are for explaining the invention and do not constitute an undue limitation thereof. It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other.

[0031] The R2A medium formulation used in the following examples is as follows: starch 0.5 g / L, glucose 0.5 g / L, acid-hydrolyzed casein 0.5 g / L, yeast extract 0.5 g / L, bacteriological peptone 0.5 g / L, sodium pyruvate 0.3 g / L, potassium dihydrogen phosphate 0.05 g / L, magnesium sulfate 0.025 g / L, TES 1.0 g / L, pH 7.2.

[0032] Example 1: Bacillus simplex Peribacillus simplex Separation and screening of ASE52 1. Seed material: The seeds of *Leymus chinensis* were collected from the desert grassland of Jiang'an Sumu in the northern part of Siziwang Banner, Inner Mongolia.

[0033] 2. Sample Pretreatment: First, the seeds were surface-sterilized and verified. 1 g of the preliminarily surface-sterilized seeds were placed in a centrifuge tube containing 9 mL of 0.85% sterile physiological saline and incubated at 180 r / min for 15 min. 200 μL of the supernatant was then spread on a plate and incubated at 28℃ for 3 days. If no bacterial growth was observed on the plate, the surface sterilization was complete, and subsequent processing could proceed. If bacterial growth was observed, the seeds were rinsed for 2 min each with 5% sodium hypochlorite solution and 75% ethanol in a clean bench, followed by rinsing with 0.85% sterile physiological saline. This process was repeated until complete sterilization was achieved. After successful sterilization, the following steps were performed: ① The seeds were placed in a sterilized and dried mortar, and 9 mL of 0.85% sterile physiological saline was added and ground into a homogenate to prepare 10... -1 ① Concentration suspension; ② Place the seeds in a sterile and dried 50 mL centrifuge tube, add 9 mL of 0.85% sterile physiological saline and soak for 1 h.

[0034] 3. Isolation and purification of the bacterial strain: Prepare five sterile 25 mL centrifuge tubes and label them ①-⑤. Dispense 9 mL of 0.85% sterile physiological saline into each tube. Transfer 1 mL from the above seed grinding suspension and soaking solution into tube ① to prepare 10... -2 The concentration suspension was serially diluted to obtain 10. -3 10 -4 10 -5 10 -6 Concentration dilutions. For the initial isolation, all concentration dilutions were plated. For subsequent experiments, intermediate gradients could be selected. 200 μL of each dilution from tubes ③-⑤ was plated onto R2A solid medium, with three replicates. Incubation was carried out at 28°C for 3-4 days. Single colonies with different morphologies were selected and purified using the four-zone streak method until no contaminating bacteria grew. Colony growth was observed and recorded. The final strain obtained was *Peribacillus simpleplex*, designated ASE52. After 3 days of growth on R2A agar, the colonies were white, and the morphology was as shown in the image. Figure 1 .

[0035] Example 2: Identification of Peribacillus simplex ASE52 Total bacterial DNA was extracted using the Chelex-100 extraction method: a suitable amount of single colony was picked and added to 50 μL of resin solution, and the cells were vortexed to mix them thoroughly with the resin; the cells were lysed by incubating in a metal bath at 100℃ for 15 min to release DNA; the cells were centrifuged at 10000 r / min for 10 min, and the supernatant was used as a DNA template and stored at -20℃ for later use.

[0036] The primers used for PCR amplification of the 16S rDNA gene were synthesized by Shanghai Sangon Biotech Co., Ltd., with the sequences: 27F (SEQ ID NO:2): 5′-AGAGTTTGATCCTGGCTCAG-3′; 1492R (SEQ ID NO:3): 5′-TACGGCTACCTTGTTACGACTT-3′. The total PCR reaction volume was 20 μL, including 1 μL DNA template, 10 μL 1×EasyTap PCR SuperMix enzyme, 1.0 μL each of the 27F and 1492R primers, and 7.0 μL sterile, enzyme-free water. The reaction program was as follows: pre-denaturation at 95℃ for 5 min; amplification phase (denaturation at 95℃ for 1 min, annealing at 55℃ for 1 min, extension at 72℃ for 2 min); final extension at 72℃ for 10 min, with multiple PCR cycles.

[0037] The PCR products were detected by 1.0% agarose gel electrophoresis. After confirming that the bands were unique and bright, the samples were sent to Shanghai Sangon Biotech Co., Ltd. for sequencing. The sequence is shown in SEQ ID NO:1. The sequence was entered into the NCBI core_nt database (https: / / blast.ncbi.nlm.nih.gov / Blast.cgi) for 16S rRNA sequence alignment. The results showed that the strain had a sequence similarity of 99.87% with Peribacillus simplex LMG 11160 (Genbank number: NR_114919), and it was identified as Peribacillus simplex.

[0038] This strain was deposited on May 29, 2025, at the China Center for Type Culture Collection (CCTCC), located at No. 299 Bayi Road, Wuchang District, Wuhan, Hubei Province, China (Wuhan University). It is classified as Peribacillus simplex ASE52 and has the accession number CCTCC No: M20251159.

[0039] Example 3: Analysis of the growth-promoting effect of Peribacillus simplex ASE52 cells on Leymus chinensis 1. Experimental Methods Preparation of bacterial suspension: Inoculate strain ASE52 into an Erlenmeyer flask containing 200 mL of R2A liquid medium, and incubate with shaking at 180 rpm for 3 days. Measure the OD600 absorbance (at this point, the viable cell count is approximately 10). 6 (CFU / mL). Adjust the absorbance to 1.0. If the absorbance is less than 1.0, centrifuge to remove part of the supernatant, concentrate and retest. If the absorbance is greater than 1.0, centrifuge and dilute with sterile water to meet the standard. After centrifuging to collect the bacterial cells, add the same amount of sterile water as when adjusting OD600=1.0 and resuspend for later use.

[0040] Seed selection and sowing: Select plump, uniformly sized *Leymus chinensis* seeds and soak them in the corresponding bacterial suspension for 30 minutes. Use sterile water as a control. After soaking, remove and air-dry the seeds. Use 7 cm × 7 cm × 9 cm seedling pots. Add 100 g of desert plant germplasm soil collected from Jiang'an Sumu, Siziwang Banner, Inner Mongolia to each pot. Sow 10 seeds of the corresponding treatment into each pot and cover with 30 g of soil. On the day of sowing, water each pot with 1 mL of the corresponding bacterial suspension, while the control group is watered with an equal volume of sterile water. Thereafter, water with 25 mL of sterile water daily. Place the seedling pots on a plant cultivation rack with a light intensity of 4000 lux and a temperature of 25℃.

[0041] Cultivation and growth index determination: Starting from the emergence time of *Leymus chinensis* seedlings, complete plants were collected after 18 days of cultivation. Five plants were selected to determine the germination rate, plant height (measured with a millimeter ruler), above-ground dry weight, and underground dry weight (after drying in a 70℃ oven, weighed using an analytical balance).

[0042] 2. Experimental Results 2.1 Germination rate analysis of *Leymus chinensis* under different treatments Table 1 Germination rate results of *Leymus chinensis* Treatments involving the addition of microorganisms (live and dead bacteria) significantly improved the germination rate of *Leymus chinensis*, with live bacteria treatments (strains LMG11160 and ASE52) showing the best results. Under the ASE52 treatment, the germination rate of *Leymus chinensis* was 84.00%, a 27.27% increase compared to the control group (66.00%), significantly higher than other treatments. Treatments with strains LMG11160 and sterilized strain ASE52 increased the germination rate by 12.12% and 3.03% respectively compared to the control group (Table 1). Figure 2 ).

[0043] 2.2 Analysis of plant height of *Leymus chinensis* under different treatments Five plants of *Leymus chinensis* from four different treatments were selected, and their heights were measured and analyzed using vernier calipers. The results are shown in Table 2.

[0044] Table 2. Measurement results of the height of *Leymus chinensis* plants (cm) Treatment with live ASE52 strain significantly promoted the height of *Leymus chinensis*, with an average height of 11.98 cm, a significant increase of 40.28% compared to the control group (8.54 cm), demonstrating superior efficacy compared to other treatments. Treatment with sterilized ASE52 resulted in an average height of 9.36 cm, an increase of 9.60% compared to the control group, but its growth-promoting effect was significantly weaker than that of live strain treatment. Treatment with LMG11160 resulted in an average height of 10.34 cm, an increase of 21.08% compared to the control group, showing a weaker promoting effect compared to ASE52, but a better effect compared to inactivated ASE52. Figure 3 ).

[0045] 2.3 Analysis of surface dry weight of drooping, alkali-tolerant grassland under different treatments Table 3. Dry weight measurement results (g) of drooping, alkali-tolerant grassland. The mean aboveground dry weight of the three microbial treatment groups was significantly higher than that of the control group (mean 0.017 g), verifying the growth-promoting effect of microbial inoculation. The live ASE52 strain treatment showed the most significant effect, with a mean aboveground dry weight of 0.033 g, an increase of 94.12% compared to the control group. The LMG11160 treatment (mean 0.025 g) and the sterilized ASE52 treatment (mean 0.021 g) also showed growth-promoting effects, but these were weaker than the live ASE52 treatment, indicating that the growth-promoting effect of the ASE52 strain depends on its live bacterial activity (Table 3). Figure 4 ).

[0046] 2.4 Analysis of the underground dry weight of drooping, wheat-laden grass under different treatments Table 4. Dry weight measurement results (g) of drooping, alkali-tolerant grass. Underground dry weight is an important indicator reflecting the root growth status of *Leymus chinensis*. A higher value indicates a more developed root system, stronger water and nutrient absorption capacity, and better drought and stress resistance. Table 4 shows the results of underground dry weight measurements. The mean underground dry weight of all three microbial treatment groups was significantly higher than the control group (mean 0.014 g), indicating that microbial inoculation also significantly promoted the root growth of *Leymus chinensis*. Among them, the treatment with live bacteria strain ASE52 showed the best effect, with a mean underground dry weight of 0.031 g, far exceeding the other treatment groups, further confirming the significant growth-promoting effect of this strain on the overall growth of *Leymus chinensis* (Table 4). Figure 5 ).

[0047] The results of this embodiment indicate that live Peribacillus simplex ASE52 cells can significantly promote the germination rate, plant height, aboveground dry weight, and underground dry weight of *Leymus chinensis* seedlings at a concentration of 10%. 6 The fact that this strain can exert a significant effect at a concentration of CFU / mL indicates that it has a prominent promoting effect on the seedling growth of *Leymus chinensis*.

[0048] Although the present invention has been described in detail with general description and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention are within the scope of protection claimed by the present invention.

Claims

1. A simple Bacillus peritonis, characterized in that, It is Peribacillus simplex The accession number is ASE52, and it is deposited at the China Center for Type Culture Collection (CCTCC) with accession number M20251159.

2. A microbial agent, characterized in that, Containing the contents of claim 1 Peribacillus simplex ASE52.

3. A product comprising the simple Bacillus peritonis or its fermentation product as described in claim 1, characterized in that, The product is any one of the following: seed soaking solution, bacterial mud, seed coating, or fertilizer.

4. The product as described in claim 3, characterized in that, The total viable count of Bacillus simplex in the product is not less than 10. 6 CFU / g or 10 6 CFU / mL.

5. The application of the simple Bacillus pertussis as described in claim 1 or the inoculum as described in claim 2 in improving the growth performance of Leymus chinensis.

6. The application as described in claim 5, characterized in that, The application described is for promoting the growth of *Leymus chinensis* seedlings during the seedling stage.

7. The application according to claim 5, characterized in that, The fermentation culture conditions for Bacillus simplex ASE52 were as follows: cultured on R2A medium at a temperature of 26-28℃ and a pH of 7.0-7.

2.

8. The application according to claim 5, characterized in that, The application is one or more of the following: Application in improving the germination rate of Leymus chinensis; Application in improving the height of *Leymus chinensis* plants; Application in improving the dry weight of drooping, alkali-tolerant grasslands; Application in improving the dry weight of drooping, alkali-tolerant grasses.

9. The application according to claim 8, characterized in that, The total viable count of the simple Bacillus aureus is not less than 10. 6 CFU / g or 10 6 CFU / mL.

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