CN111117922A - Bacillus simplex XWY09 and application thereof - Google Patents

Abstract

The invention discloses Bacillus simplex XWY09 and application thereof. The strain XWY09 has a preservation number of CCTCC NO: m2019860 and the strain XWY09 not only have strong organophosphorus dissolving function, but also can well grow in alkaline environment (pH10) and produce various growth promoting substances such as IAA, protease, cellulase, siderophin and the like. Taking corn as an example, the strain XWY09 has obvious plant growth promoting effect, and can increase the stem height of corn seedlings by 29.3%, the root length by 32.84%, the fresh weight of stems by 44%, the fresh weight of roots by 61%, the stem weight by 30.77% and the dry weight of roots by 34.88%. When the strain XWY09 is applied to crop planting, the yield of crops can be increased, the resistance of plants to diseases, drought and the like can be improved, and the strain has important significance for continuously increasing the yield of grains and protecting the ecological environment of farmlands in China.

Description

Bacillus simplex XWY09 and application thereof

Technical Field

The invention relates to agricultural microbiology, in particular to a bacillus simplex XWY09 strain and application thereof.

Background

Plant growth-promoting rhizobacteria (PGPR) refers to a kind of beneficial bacteria that can freely live in soil or attached to plant root system, can promote plant growth and absorption and utilization of mineral nutrition, and can inhibit harmful organisms.

Phosphorus is the second most nutrient element essential for plant growth, and is involved in multiple metabolism of plants, including energy transfer, signal transfer, respiration, macromolecular substance synthesis, and photosynthesis. However, 95% -99% of phosphorus is fixed by calcium ions in soil and phytic acid produced by plants, exists in an insoluble state, a solidified state and a precipitated state, so that the phosphorus is difficult to be absorbed by the plants, and the plants can only absorb soluble phosphorus elements.

The present invention relates to a method for producing phosphorus-solubilizing microorganisms in soil, which are classified into phosphorus-solubilizing bacteria, fungi, and actinomycetes, wherein the phosphorus-solubilizing bacteria include enterobacter (enterobacter sp), Agrobacterium (Agrobacterium sp), Erwinia (Erwinia sp), Pseudomonas (Pseudomonas sp), Serratia (Serratia sp), Flavobacterium (Flavobacterium sp), Bacillus (Bacillus sp), Micrococcus (Micrococcus sp), Azotobacter (azobacter sp), chromobacterium (Chromabacterium sp), Salmonella (Salmonella sp), Bacillus (Salmonella sp), alcaligenes (alcaligenes sp), Bacillus (Escherichia sp), Bacillus coli (Escherichia sp), Bacillus coli (Escherichia sp), and actinomycetes, and the like, and the method for their long storage period, is the most widely applied rhizosphere growth-promoting bacteria at present, but has no application of the simple bacillus XWY09 in plant growth promotion and saline-alkali soil restoration.

Disclosure of Invention

The invention aims to provide a simple bacillus XWY09 strain and application thereof.

In order to achieve the purpose of the invention, in the first aspect, the invention provides an alkali-resistant simple Bacillus XWY09(Bacillus simplex XWY09) separated from soil collected from Xinjiang frontispin, wherein the strain XWY09 not only has strong organophosphorus dissolving function, but also can well grow and produce IAA (indoleacetic acid), protease, cellulase and siderophin under an alkaline environment (pH10.0). The strain is preserved in China center for type culture Collection at the address: wuhan, Wuhan university, zip code 430072, preservation number CCTCC NO: m2019860, date of deposit 2019, 10 months and 29 days.

In a second aspect, the invention provides a bacterial agent comprising said Bacillus simplex XWY 09.

In a third aspect, the invention provides growth promoting substances, such as auxin (IAA) and the like, secreted by the Bacillus simplex XWY 09.

In a fourth aspect, the invention provides enzymes secreted from the bacillus simplex XWY09, including but not limited to cellulases, proteases.

In a fifth aspect, the invention provides a siderophore secreted by said Bacillus simplex XWY09,

in a sixth aspect, the invention provides an agricultural fertilizer, a phosphorus activator, a plant growth promoter, a heavy metal pollution degradation agent or an iron chelating agent prepared from the simple bacillus XWY09 or a microbial inoculum thereof.

In a seventh aspect, the invention provides any one of the following applications of the bacillus simplex XWY09 or its microbial inoculum:

1) for promoting plant growth and development and increasing crop yield;

2) the plant stress resistance is improved;

3) dissolving phosphorus;

4) the method is used for repairing heavy metal pollution;

5) the method is used for preparing agricultural fertilizers;

6) used for preparing a phosphorus activator;

7) for the preparation of plant growth promoters;

8) used for preparing heavy metal pollution degradation agent;

9) used for preparing iron chelating agent.

Plants described in the present invention include, but are not limited to, corn, wheat, soybean, cucumber.

The heavy metal comprises iron.

The improvement of the stress resistance of the plants refers to the improvement of the stress resistance of the plants such as disease resistance, drought resistance, heavy metal poison resistance and the like.

In an eighth aspect, the invention provides application of the simple bacillus XWY09 or a microbial inoculum thereof in dissolving insoluble phosphorus.

In the application, the insoluble phosphorus is insoluble organic phosphorus, such as calcium phytate, lecithin and the like.

By the technical scheme, the invention at least has the following advantages and beneficial effects:

the salt-tolerant simple bacillus XWY09 provided by the invention not only has a strong organophosphorus dissolving function, but also can well grow in an alkaline environment (pH10) and generate various growth promoting substances such as IAA, protease, cellulase, siderophin and the like. Taking corn (Zhengdan 958) as an example, the strain XWY09 has obvious effect of promoting the growth of plants, and increases the stem height of corn seedlings by 29.3%, the root length by 32.84%, the fresh stem weight by 44%, the fresh root weight by 61%, the stem weight by 30.77% and the dry root weight by 34.88%. When the strain XWY09 is applied to crop planting, the yield of crops can be increased, the resistance of plants to diseases, drought and the like can be improved, and the strain has important significance for continuously increasing the yield of grains and protecting the ecological environment of farmlands in China.

Drawings

FIG. 1 shows the results of the determination of the phosphorus solubilizing ability of the strain XWY09 in example 2 of the present invention.

FIG. 2 shows the results of the determination of the protease-producing ability of strain XWY09 in example 3 of the present invention.

FIG. 3 shows the results of the cellulase-producing ability of strain XWY09 in example 4 of the present invention.

FIG. 4 shows the results of measurement of the ability of strain XWY09 to produce siderophore according to example 5 of the present invention.

FIG. 5 shows the growth of strain XWY09 in LB liquid medium pH10 in example 7 of the present invention. Wherein, CK: blank medium without inoculation.

FIG. 6 is a graph showing the growth of strain XWY09 under different pH conditions in example 7 of the present invention.

FIG. 7 shows the plant growth promoting effect of strain XWY09 in example 9 of the present invention.

FIG. 8 is a molecular evolutionary tree of the 16S rRNA gene of strain XWY09 in example 1 of the present invention.

Detailed Description

The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are commercially available products.

Example 1 isolation and characterization of Strain XWY09

Collecting soil from Xinjiang frontispon county, adding 10g of soil sample into a triangular flask filled with 100ml of sterile normal saline, standing for 20min, shaking for 30min at 28 ℃ and 200rpm in a shaking table, adding 1ml of sample into 9ml of sterile normal saline, and sequentially diluting for 10 min²，10³，10⁴Taking 100 mu L of the above soil suspension respectively, uniformly coating the soil suspension on an LB culture medium (5 g/L of yeast powder, 10g/L of peptone, 10g/L of NaCl and 15g/L of agar) plate with the pH value of 8.5, culturing for 48h in an incubator at 28 ℃, picking out single bacterial colony of bacteria by using a sterile toothpick, streaking and purifying, and storing for later use.

And (3) identifying the strain XWY09 as Bacillus simplex (Bacillus simplex) by combining the physiological and biochemical characteristics and the molecular biological detection result of the strain. The specific identification results are as follows:

1. molecular biological identification

The strain XWY09 was first identified in molecular biology using primer 27F: 5'-AGAGTTTGATCCTGGTCAGAACGAACGCT-3' and 1492R: 5'-TACGGCTACCTTGTTACGACTTCACCCC-3' amplifying the 16S rRNA gene sequence, sequencing and obtaining the sequence shown in SEQ ID NO: 1 is shown.

The PCR reaction system is as follows: 2 XTaq Mix 12.5. mu.l, primer 27F,1492R 1. mu.l each, DNA template 1. mu.l, ddH₂O9.5. mu.l. The PCR reaction conditions are as follows: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 ℃ for 30s, annealing at 56 ℃ for 30s, extension at 72 ℃ for 1min, and 30 cycles; extension at 72 ℃ for 5 min. Among them, Taq Mix was purchased from TaKaRa.

The PCR amplification product was analyzed by 1% agarose gel electrophoresis, purified and recovered with the purification Kit Gelextraction Kit from OMEGA, and then ligated with the T vector pMD18-T from TaKaRa. Sequencing by using an ABI 3730 DNA sequencer, submitting the gene sequence to GenBank, and recording the number: MN 559034. The sequences were subjected to molecular tree analysis with GEGA5 (fig. 8), and XWY09 was initially judged as bacillus simplex based on the tree.

2. Physiological and biochemical identification

On the basis of molecular biological detection, physiological and biochemical identification is carried out on the strain XWY09, gram staining is firstly carried out on the strain XWY09, and the result shows that the strain is gram positive; inoculating the strain into LB culture medium in a test tube, spreading a layer of culture medium on the surface of the test tube, and adding CO₂Culturing in culture box for 48h, culturing at 4 deg.C and 40 deg.C for 48h, culturing at both temperatures for no growth, activating, inoculating to LB medium with NaCl concentration of 3%, 5% and 7%, culturing at 28 deg.C for 48h to obtain NaCl concentration of 3% and 5%, inoculating to solid culture medium of starch, culturing at 28 deg.C for 48h, staining with iodine solution to obtain colorless transparent ring, culturing at 20 deg.C for 7 days to obtain positive starch hydrolysis, inoculating to gelatin culture medium, culturing at 20 deg.C for 7 days to obtain molten gelatin, culturing at 30 deg.C for 1, 3 and 5 days, inoculating to nitrate liquid culture medium, culturing at 30 deg.C for 1, 3 and 5 days, pouring small amount of culture solution into small hole of white disk, and dropping 1 drop of reagent A (0.5 g of sulfanilic acid dissolved in 10% acetic acid) and distilled acetic acid reagent B (0.150% of acetic acid) to obtain pure acetic acid, and reducing agent B, and culturing at 20-3-52 mL in distilled water to obtain acetic acid reagent B, and reducing agent BRed-based test. In the VP test, the culture medium (about 2ml) is mixed with 40% NaOH, small amount of creatine is added, and the mixture is shaken for 2-5min to obtain a positive VP if the culture medium is red. The test results showed that strain XWY09 was VP negative. Picking single colony with gun head, placing on glass slide, adding one drop of 3% H₂O₂The solution is on the bacterial colony, and bubbles appear, which indicates that the bacterial strain is positive in catalase; inoculating the strain into skimmed milk plate culture medium, culturing at proper temperature for 1, 3, and 5 days, and observing to obtain a transparent circle which is positive, or negative. The casein hydrolysis test showed that strain XWY09 was positive; a piece of filter paper was placed in a clean petri dish, and a 1% aqueous solution of dimethyl-p-phenylene diamine was added dropwise to wet only the filter paper. Selecting a ring of thallus Porphyrae with platinum loop, and spreading on wet filter paper. No red coloration of the applied lawn within 10sec, indicating XWY09 oxidase negative; the results of physiological and biochemical tests of strain XWY09 are shown in Table 1. XWY09 can be judged as simple bacillus according to the evolutionary tree and physiological and biochemical results.

TABLE 1 physio-biochemical characteristics of Bacillus simplex XWY09

Example 2 determination of the phosphorus solubilizing ability of Strain XWY09

The strain XWY09 was first activated on LB medium. The activated strain was inoculated into a phosphate-solubilizing medium (10.00 g of glucose, (NH) shown in appendix A of NY/T1847-2010)₄)₂SO₄0.50g，MnSO₄·7H₂O 0.3g，NaCl 0.3g，KC1 0.30g，FeSO₄·4H₂O 0.036g，MnSO₄·4H₂O0.03 g, distilled water 1000mL, pH 7.0. The organic phosphorus source is calcium phytate, and the addition amount is 2 g. Solid medium: add 1.5% agar powder in proportion). And (3) detecting whether the strain has a phosphorus dissolving function, wherein the strain XWY09 has the capability of dissolving organic phosphorus, and the diameter of a dissolving ring for dissolving the organic phosphorus reaches 1.51cm (figure 1).

After the phosphorus dissolving function of the strain XWY09 is determined, the phosphorus dissolving capacity of the strain is quantitatively determined, and the strain is subjected to phosphorus dissolvingActivating the strain on LB liquid culture medium, inoculating the strain into liquid phosphorus-dissolving culture medium and liquid phosphorus-dissolving culture medium with pH of 10.0, shaking for 7 days at 28 ℃, measuring the content of available phosphorus in fermentation supernatant by molybdenum-antimony colorimetry, taking 100 mu L of supernatant, putting the supernatant into a 50ml volumetric flask, diluting the supernatant to about 30ml with water, adding 2 drops of dinitrophenol indicator, dropwise adding 4mol/L NaOH until the solution turns yellow, and then adding 1mol/L H mol₂SO₄1 drop, make the yellow color of the solution just fade away. Adding 5.00ml molybdenum-antimony color-developing resisting agent, adding water to constant volume, and shaking up thoroughly. After standing at room temperature above 15 ℃ for 30min, 200. mu.l of the mixture was added to a 96-well plate, and the absorbance was measured at a wavelength of 882nm using a microplate reader.

The results show that the strain XWY09 has the capacity of dissolving organic phosphorus in a liquid phosphorus-dissolving culture medium and a liquid phosphorus-dissolving culture medium with the pH value of 10.0, and the capacity of dissolving the organic phosphorus reaches 132.65mg/L and 128.58mg/L respectively.

Example 3 determination of the ability of Strain XWY09 to produce protease

Firstly, activating a strain XWY09 on an LB culture medium, inoculating the activated strain on a culture medium for detecting protease (5.00 g of tryptone, 3.00g of yeast extract, 1.00g of glucose, 15.00g of agar, 1000mL of distilled water, pH7.0, and 121 ℃ for 30min in high-pressure sterilization), cooling the sterilized detection culture medium to about 50 ℃, adding skim milk into the culture medium according to the proportion of 10 percent, uniformly mixing, pouring into a culture dish, cooling for later use, culturing at 28 ℃ for 48h, and observing whether a dissolving ring appears, wherein the result shows that the strain XWY09 has good capability of dissolving protein and high yield of protease, and the diameter of the dissolving ring reaches 3.00cm (figure 2). By utilizing the characteristic of high-yield protease of the strain XWY09, the extracted protease can be applied to the protease fields of food industry or washing industry and the like, and can also be used for controlling diseases by degrading pathogenic bacteria cell membranes in agricultural microbial fertilizers and degrading proteins in agricultural wastes in composts.

Example 4 determination of cellulase-producing ability of Strain XWY09

First, the strain XWY09 was activated on LB medium, and the activated strain was inoculated into a medium for detecting cellulose (MgSO4 & 7H)₂O 0.25g，K₂HPO₄0.50g, carboxymethylCellulose 1.88g, agar 15.0g, distilled water 1000mL, pH7.0, 121 ℃ autoclaving for 30 min). Culturing at 28 ℃ for 7 days, after 7 days, adding 5mL of 0.2mg/mL Congo red dye solution into each plate, dyeing for 1h, after discarding the Congo red dye solution, adding 1M NaCl, washing for 1h, discarding the washing solution, observing the generation of a hydrolysis ring around a bacterial colony, wherein the generation of the hydrolysis ring indicates the generation of cellulase. The results show that strain XWY09 has a good capacity to dissolve cellulose with a lysis ring diameter of up to 5.12cm (FIG. 3). The strain XWY09 has high cellulose yield, can degrade the cell wall of pathogenic fungi and control diseases, and can also be used for decomposing cellulose in compost.

EXAMPLE 5 determination of the ability of Strain XWY09 to produce siderophore

Firstly, the strain XWY09 is activated on LB culture medium, and the activated strain is inoculated in a culture medium containing CAS blue detection solution for culture (a certain amount of CAS blue detection solution is added into an iron-deficiency complex culture medium consisting of acid hydrolyzed casein and other inorganic salts to prepare a blue detection plate). If the bacteria produce siderophiles, sequestering the iron (ferric) in the medium, a yellow halo is produced around the colony. The results show that strain XWY09 has a good ability to chelate iron. The halo diameter reached 1.30cm (fig. 4). The strain XWY09 can enrich a part of iron ions to the roots of plants and promote the absorption of the plants to the iron.

EXAMPLE 6 determination of IAA-producing ability of Strain XWY09

The strain XWY09 was first activated on LB liquid medium, and the activated strain was inoculated in an amount of 1% to DF medium (peptone 5.00g, yeast extract 1.50g, beef extract 1.50g, NaCl 5.00g, tryptophan 0.50g, distilled water 1000mL, pH7.0, autoclaved at 121 ℃ for 30min) at pH 10.0. Shaking at 28 deg.C for 7 days, taking out the fermentation broth after 7 days, centrifuging at 12000rpm for 5min, and determining IAA content in the fermentation broth by Salkowkin colorimetry. The results showed that strain XWY09 produced IAA in an amount of 8.62 mg/L. Further analysis confirmed IAA synthesis by HPLC, strain culture for 7 days, centrifugation at 12000rpm for 5min, taking 30mL of supernatant, extracting with two volumes of ethyl acetate in a constant temperature oscillator for 3 times, combining the extracts, distilling with ethyl acetate under reduced pressure, and distilling with 5mL of methanolDissolving in alcohol, diluting to desired volume, and filtering with 0.22um filter membrane. Reference literature (Liancuifei, Jiangzhi, Lihuci, Luxiouyun, chaulmoogra and Maping) utilizes high performance liquid chromatography to screen plant hormone producing bacteria [ J]North china agro-scientific newspaper 2006 (02): 66-69; high performance liquid chromatography shear wavelength method for determining endogenous hormone [ J ] in folium Artemisiae Argyi]Modern agricultural technology, 2007 (03): 9-11) for sample detection. A detection instrument: waters2998 high performance liquid chromatography, column: agilert Zorbax SB-C₁₈250mm × 4.6mm, 5 um. Mobile phase, methanol: acetonitrile: 0.6% aqueous glacial acetic acid (50: 5: 45 by volume). Sample introduction amount: 20 μ l. The flow rate was 0.8 mL/min. Column temperature: and (4) room temperature. Detection wavelength: 255 nm. The detection result shows that the yield of IAA is 8.71mg/L, which is slightly higher than the detection result of the colorimetric method.

Example 7 determination of alkali resistance of Strain XWY09

Firstly, the strain XWY09 was activated on LB liquid medium, the activated strain was inoculated in an amount of 1% to LB liquid medium of pH8.5, pH9.5, pH10.0, pH10.5, respectively, and simultaneously inoculated to normal LB liquid medium as a control, and the control was shaken at 28 ℃ for 48 hours to measure the OD₆₀₀The value is obtained. The results showed that OD of strain XWY09 in the medium at pH10.0₆₀₀No significant difference in values compared to the control occurred (fig. 6), indicating that strain XWY09 can grow well in an environment of pH10.0 (fig. 5).

Example 8 preparation of microbial inoculum

1. Strain activation

The strain XWY09 was first activated on LB medium. The Bacillus simplex XWY09 was inoculated into LB liquid medium and cultured at 28 ℃ for 12 hours.

2. Cultivation of seed liquid

The seed culture medium comprises the following components: 15g/L of peptone, 8g/L of yeast extract, 12g/L of glucose, 10g/L of NaCl and 7.2 of pH value. Inoculating the activated strain XWY09 into a seed liquid culture medium with the inoculation amount of 1.2%, and performing shake culture at 28 ℃ on a shaking table at the rotating speed of 200rpm for 12 h.

3. Fermenting in a fermentation tank

The fermentation medium comprises the following components:18g/L of soybean meal, 25g/L of molasses, 6g/L of corn meal, 2.2g/L of yeast extract, 2.4g/L of peptone, 3.2g/L of glucose and KH₂PO₄0.6g/L，MgSO₄。7H₂O 0.05g/L，MnSO₄·7H₂O 0.018/L，CaCl₂4g/L, 1.2g/L of polyether defoamer and 7.5 of pH value. The cultured seed liquid OD₆₀₀When the inoculation amount reaches 5.00, the mixture is inoculated into a fermentation tank according to 5 percent of the inoculation amount, the stirring speed is 180rpm at 28 ℃, and the ventilation quantity is adjusted to ensure that the dissolved oxygen is more than 50 and the tank pressure is 0.05 MPa. The bacterial amount can reach 10 after 28h fermentation¹⁰cfu/ml, stopping fermentation when the spore amount reaches more than 95%, cooling and discharging to obtain 10 of Bacillus simplex XWY09¹⁰The cfu/mL bacterial liquid optimizes the formula of the fermentation medium, so that the yield of the IAA is improved to 22.2mg/L from 8.71 mg/L. Preparing solid or liquid microbial inoculum according to the requirements. Directly diluting and filling the liquid microbial inoculum with zymogen liquid; the solid microbial inoculum is prepared by centrifuging, spray drying, diluting to the required concentration, and packaging.

Example 9 plant growth promotion test for Strain XWY09

The method comprises the steps of selecting plump and healthy corn (Zhengdan 958) seeds, sterilizing, cleaning, soaking, and then putting into a biochemical incubator at 28 ℃ for culture and germination acceleration for about 24 hours. Selecting single colony from a flat plate of a test strain, activating the single colony in an LB liquid culture medium, inoculating the activated bacterial liquid into a DF culture medium according to the bacterial inoculation amount of 1%, shaking at 28 ℃, 150rpm for 72 hours, and diluting the bacterial liquid to 10⁵cfu/mL. The corn seedlings were tested by the method described in example 7 of CN201010511686.2, and the stem height, root length, fresh stem weight, fresh root weight, dry stem weight and dry root weight were measured.

The results show that strain XWY09 has a good growth promoting effect (FIG. 7). Can increase the stem height of corn seedling by 29.3%, the root length by 32.84%, the fresh weight of stem by 44%, the fresh weight of root by 61%, the stem weight by 30.77% and the dry weight by 34.88% (Table 2).

TABLE 2 growth promoting effect of Strain XWY09 on maize

Note: the significant differences are represented by P < 0.05 and P < 0.01.

Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Sequence listing

<110> Zhang family, Kokou Gen-Ji, Multi-ecological agriculture technology Co Ltd

<120> simple bacillus XWY09 and application thereof

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<170>SIPOSequenceListing 1.0

<210>1

<211>785

<212>DNA

<213> Bacillus simplex (Bacillus simplex)

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tatactgcag tcgagcgaat cgatgggagc ttgctccctg agattagcgg cggacgggtg 60

agtaacacgt gggcaacctg cctataagac tgggataact tcgggaaacc ggagctaata 120

ccggatacgt tcttttctcg catgagagaa gatggaaaga cggattacgc tgtcacttat 180

agatgggccc gcggcgcatt agctagttgg tgaggtaatg gctcaccaag gcgacgatgc 240

gtacccgacc tgagagggtg atcggccaca ctgggactga gacacggccc agactcctac 300

gggaggcagc agtagggaat cttccgcaat ggacgaaagt ctgacggagc aacgccgcgt 360

gaacgaagaa ggccttcggg tcgtaaagtt ctgttgttag ggaagaacaa gtaccagagt 420

aactgctggt accttgacgg tacctaacca gaaagccacg gctaactacg tgccagcagc 480

cgcggtaata cgtaggtggc aagcgttgtc cggaattatt gggcgtaaag cgcgcgcagg 540

tggttcctta agtctgatgt gaaagcccac ggctcaaccg tggagggtca ttggaaactg 600

gggaacttga gtgcagaaga ggaaagtgga attccaagtg tagcggtgaa atgcgtagag 660

atttggagga acaccagtgg cgaaggcgac tttctggtct gtaactgaca ctgaggcgcg 720

aaagcgtggg ggagcaaaca ggattagata ccctggtagt ccacgccgta aacgatgagt 780

gctag 785

Claims (10)

1. Simple Bacillus (Bacillus simplex) XWY09 with the preservation number of CCTCC NO: m2019860.

2. A bacterial agent comprising Bacillus simplex XWY09 according to claim 1.

3. A growth-promoting substance secreted by bacillus simplex XWY09 of claim 1.

4. An enzyme secreted from the Bacillus simplex XWY09 of claim 1, wherein the enzyme comprises a cellulase or a protease.

5. An agricultural fertilizer, a phosphorus activator, a plant growth promoter, a heavy metal pollution degradation agent or an iron chelator prepared from the Bacillus simplex XWY09 of claim 1 or the microbial agent of claim 2.

6. The use of the bacillus simplex XWY09 of claim 1 or the microbial inoculum of claim 2 in any one of the following applications:

1) for promoting plant growth and development and increasing crop yield;

2) the plant stress resistance is improved;

3) dissolving phosphorus;

4) the method is used for repairing heavy metal pollution;

5) the method is used for preparing agricultural fertilizers;

6) used for preparing a phosphorus activator;

7) for the preparation of plant growth promoters;

8) used for preparing heavy metal pollution degradation agent;

9) used for preparing iron chelating agent.

7. Use according to claim 6, wherein the plant comprises maize, wheat, soybean, cucumber.

8. Use according to claim 6 or 7, wherein the heavy metal comprises iron.

9. Use of the Bacillus simplex XWY09 of claim 1 or the microbial inoculum of claim 2 for solubilizing poorly soluble phosphorus.

10. The use according to claim 9, wherein said sparingly soluble phosphorus is a sparingly soluble organophosphorus.

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