CN111808776B

CN111808776B - Saline-alkali-tolerant air bacillus and preparation method and application of viable bacteria preparation thereof

Info

Publication number: CN111808776B
Application number: CN202010736503.0A
Authority: CN
Inventors: 孙中涛; 辛寒晓; 潘雁鹏; 赵升远; 孙国科; 陈君君; 安良聪
Original assignee: Shandong Zoeticland Biological Technology Co ltd; Dezhou Bahu Biotechnology Co ltd
Current assignee: Shandong Zoeticland Biological Technology Co ltd; Dezhou Bahu Biotechnology Co ltd
Priority date: 2020-07-28
Filing date: 2020-07-28
Publication date: 2022-02-08
Anticipated expiration: 2040-07-28
Also published as: CN111808776A

Abstract

The invention discloses a saline-alkali tolerant bacillus air strain and a preparation method and application of a live bacterial preparation thereof. The invention separates a strain of saline-alkali tolerant air Bacillus (Bacillus aereus) PAPM-14 from corn rhizosphere soil, the strain is preserved in China general microbiological culture Collection center (CGMCC) at 6 and 22 months in 2020, and the preservation number is CGMCC No. 20118. The strain has strong saline-alkali resistance, strong capability of producing IAA, phosphate and potassium, strong antagonistic action on pathogenic bacteria such as corn sheath blight and the like, can be used for producing microbial fertilizers, particularly special microbial fertilizers for saline-alkali soil corns, is used for biologically controlling crop diseases such as corn sheath blight and the like, reduces the occurrence of diseases, and improves the yield and the quality.

Description

Saline-alkali-tolerant air bacillus and preparation method and application of viable bacteria preparation thereof

Technical Field

The invention relates to a bacillus aerosidum with saline-alkali resistance, IAA (International IAA production), phosphate and potassium dissolution and corn sheath blight pathogenic bacterium antagonism effects and application thereof, belonging to the technical field of agricultural biology.

Background

Corn is an annual grass family of plants of the genus zea mays, widely distributed in the united states, china, brazil and other countries, and is an important food crop in the world. The corn sheath blight is one of important diseases for restricting the high-yield and high-quality cultivation of corn, and is caused by rhizoctonia solani, rhizoctonia zeae, rhizoctonia graminearum and other fungi, wherein the diseases caused by the rhizoctonia solani are common. Corn sheath blight commonly occurs all over the country, has a long damage period, and can be attacked from a seedling stage to a later growth stage, but is obvious from a stamina stage to a filling stage, so that leaf sheaths are mainly damaged, and leaves, fruit ears and bracts are secondarily damaged. When the disease is serious, the firm stalks can be damaged, and the yield and the quality of the corn are seriously influenced. In particular to southwest corn planting areas in China, due to high temperature and high humidity, and the main planted corn variety is not resistant to diseases, sheath blight has become the first disease of continuous corn yield reduction.

For a long time, the corn sheath blight disease is mainly controlled chemically, and technical measures such as breeding disease-resistant varieties and optimizing cultivation management are combined. The chemical prevention and treatment mainly comprises a large amount of dimetachlone, validamycin, procymidone, thifluzamide, propiconazole, hexaconazole, tebuconazole, epoxiconazole, difenoconazole-propiconazole and other medicines. Chemical control is simple and efficient, but has many disadvantages. The use of a large amount of chemical pesticides not only causes great harm to human health, but also seriously pollutes the environment, and can cause pathogenic bacteria to generate drug resistance after long-term use. The development of biological control technology and the reduced use of chemical pesticides are inevitable trends in development.

Biological control is a method for preventing and controlling plant diseases and insect pests by using beneficial organisms or metabolites thereof, is characterized by no environmental pollution, safety to people and other organisms, energy conservation and ecological balance maintenance, and is an important means for developing green food and protecting human health. At present, beneficial microorganisms such as trichoderma harzianum, trichoderma viride, bacillus subtilis and the like are used for biological control of corn sheath blight, but the control effect is limited, so that the method cannot be popularized in large area in agricultural production.

The saline-alkali soil is an important land resource in China, and according to the report of Nanjing soil of an agricultural academy, the total area of the saline-alkali soil and saline-alkali obstacle cultivated land in China exceeds 5 hundred million acres, wherein the total area of the saline-alkali soil and the saline-alkali obstacle cultivated land has the agricultural utilization potential of up to 2 hundred million acres, accounts for about 10 percent of the area of the cultivated land in China, and is mainly distributed in northwest, northeast and east coastal areas in China. The corn has strong saline-alkali resistance and is one of crops widely cultivated in saline-alkali soil. The biological prevention and control of the corn sheath blight in the saline-alkali soil requires that the strain has good disease prevention and growth promotion effects, and also requires that the strain has strong saline-alkali resistance and can be planted in saline-alkali soil. Therefore, the screening of the saline-alkali tolerant disease-resistant growth-promoting strain has important significance.

Disclosure of Invention

Aiming at the current situation, the invention provides a strain of bacillus aereus PAPM-14 and a preparation method and application of a live bacterial preparation thereof. The bacillus aerosidus PAPM-14 strain is separated from the corn rhizosphere soil in saline-alkali soil, has a plurality of excellent characteristics of salt and alkali resistance, IAA production, phosphate and potassium dissolution, antagonism of crop pathogenic bacteria (especially corn sheath blight pathogenic bacteria) and the like, and can be used for producing biological fertilizers.

The purpose of the invention is realized by adopting the following technical scheme:

a strain of saline-alkali tolerant air Bacillus (Bacillus aereus) PAPM-14 separated from saline-alkali soil corn rhizosphere soil, wherein the preservation number of the strain in China general microbiological culture Collection center (CGMCC) is CGMCC No. 20118; the preservation address is as follows: the date of preservation is 22 days 6 months 2020. The strain is cultured for 48 hours at 37 ℃ on an NA culture medium, and the bacterial colony is round, raised, moist, wrinkled, opaque and milk-white; the thallus is in a short rod shape.

The invention also discloses a live bacteria preparation taking the air bacillus PAPM-14 as a main effective component.

The preparation method of the bacillus aereus PAPM-14 viable bacteria preparation comprises the following steps: the method is characterized in that after the air bacillus PAPM-14 is subjected to amplification culture, the air bacillus PAPM-14 is inoculated in a fermentation medium (the inoculum size is 2-5 percent) and cultured for 24-48h at the temperature of 35-40 ℃ to obtain the air bacillus PAPM-14 fermentation liquor. Adding 0.01-0.02% (w/w) of Propyl Gallate (PG), 3-5% (w/w) of mineral potassium fulvate, 0.1-0.3% (w/w) of potassium sorbate and 0.02-0.05% (w/w) of carbazone into the air bacillus PAPM-14 fermentation liquor, and stirring uniformly to obtain the viable bacteria preparation.

The formula of the fermentation medium is as follows: 10g of soybean meal, 15g of corn flour, 2g of yeast extract powder, 1.5g of monopotassium phosphate, 0.2g of zinc sulfate, 0.05g of manganese sulfate, 0.05g of complex enzyme preparation and 1000mL of water, wherein the initial pH value is 7.5. The compound enzyme preparation comprises 50% of neutral protease and 50% of cellulase.

The invention also discloses the application of the bacillus aerosidum PAPM-14 or the viable bacteria preparation in the aspects of IAA production, phosphate and potassium dissolution, antagonism of corn sheath blight pathogenic bacteria, corn microsporum pathogenic bacteria, rice bakanae pathogenic bacteria and the like.

The invention has the beneficial effects that:

the bacillus aerosidus PAPM-14 is separated from saline-alkali soil, has strong saline-alkali resistance and high planting rate in the saline-alkali soil, has the effects of producing IAA, dissolving phosphorus and potassium, antagonizing corn sheath blight pathogenic bacteria, corn microsporum pathogenic bacteria and rice bakanae pathogenic bacteria, can be used for producing a special biological fertilizer for the saline-alkali soil, and has the effects of improving soil fertility, promoting crop growth and preventing and treating corn sheath blight.

The production method of the air bacillus PAPM-14 viable bacteria preparation is scientific, the viable bacteria count of the fermentation liquor is high, and the production cost is low. The complex enzyme preparation is added into the fermentation medium, and raw materials such as soybean meal and corn flour are hydrolyzed in the temperature rise process during sterilization, so that the delay period can be shortened, the utilization rate of the raw materials can be improved, and the viable bacteria concentration of the fermentation liquor can be improved.

Drawings

FIG. 1 shows the form of the cells of the Bacillus aereus PAPM-14 strain;

FIG. 2 IAA-producing ability of Bacillus aereus PAPM-14 strain; note: from left to right are: CK (positive control, 50. mu.L 50mg/L IAA added), inoculated with PAPM-14 strain; the result shows that the color turns red after the strain PAPM-14 is inoculated, which indicates that the strain has the capability of producing IAA;

FIG. 3 is a phosphate solubilizing ring of Bacillus aereus PAPM-14 strain on an organophosphorus culture medium, wherein the inner ring is a bacterial colony and the outer ring is a phosphate solubilizing transparent ring;

FIG. 4 is a potassium-solubilizing map of the Bacillus aeruginosus strain PAPM-14, which shows oil-drop colonies grown on silicate medium by the strain PAPM-14.

Detailed Description

The present invention will be further described with reference to the following examples.

Example 1: screening of Bacillus aereus strain PAPM-14

(1) Screening of saline-alkali tolerant strains

Bacillus aereus PAPM-14 was isolated from corn rhizosphere soil. The soil was sampled in the coastal city district of Binzhou, Shandong province in 2018, and was saline-alkali soil. The specific separation method comprises the following steps: the soil sample was mixed well, 5g was weighed and placed in a triangular flask containing 95mL of sterile water and 10 beads, and shaken at 37 ℃ and 180rpm for 30 min. Taking 1mL of soil suspension for 10^-1—10^-7Serial concentration gradient dilutions were made and then 10 taken^-5、10^-6、10^-7Three dilutions were plated onto plates containing selection medium and cultured in an inverted format at 37 ℃ for 2 d. Then, different single colonies were individually picked and streaked onto plates containing selection medium and cultured in an inverted state at 37 ℃ for 2 d. Picking single colony from the plate, transferring to the slant of the storage medium test tube, culturing at 37 deg.C for 2d, growing with thallus Porphyrae, and storing in 4 deg.C refrigerator. And screening 172 saline-alkali tolerant strains.

The formula of the selective culture medium is as follows: 10g of peptone, 5g of yeast extract and composite inorganic salt (NaCl: KCl: MgCl)₂10: 1), 25g of agar, 20g of distilled water 1000mL, pH9.0, and sterilizing at 121 ℃ for 20 min.

The formula of the preservation culture medium is as follows: 10g of peptone, 5g of yeast extract, 10g of NaCl, 20g of agar, 1000mL of distilled water, pH7.5, and sterilizing at 121 ℃ for 20 min.

(2) Screening of IAA (indoleacetic acid) -producing strains

The saline-alkali tolerant strains screened in the test are respectively inoculated into LB liquid culture medium containing L-tryptophan (200mg/L), and shake culture is carried out at 37 ℃ and 180r/min for 4 d. 50. mu.L of the bacterial suspension was dropped on a white ceramic plate, and 50. mu.L of Salkowski colorimetric solution was added thereto. A positive control was prepared by adding 50. mu.L of 50mg/L IAA to the sample. The white ceramic plate was left at room temperature in the dark for 30min, and the red-colored plate showed the ability to secrete IAA. And co-screening 27 strains with IAA production capacity.

The LB liquid culture medium comprises the following components in percentage by weight: 10.0g of peptone, 5.0g of yeast extract, 10.0g of NaCl, 1000mL of distilled water, and pH 7.0.

The formula of the Salkowski colorimetric solution is as follows: 35% HClO₄ 50mL、0.5mol/L FeCl₃ 1mL。

(3) Screening of phosphate solubilizing Strain

And respectively transferring the strains with the IAA production capability to an organophosphorus bacteria culture medium, and determining whether the strains have the phosphate-solubilizing capability. 9 strains with the capability of phosphate solubilizing are selected together.

The formula of the organophosphorus bacteria culture medium is as follows: glucose 10.0g, (NH)₄)₂SO₄ 0.5g、MgSO₄·7H₂O 0.3g、MnSO₄·4H₂O 0.03g、KCl 0.3g、FeSO₄·7H₂O 0.03g、NaCl 0.3g、CaCO₃5.0g, lecithin 0.2g, agar 1.6%, and 1000mL of distilled water, wherein the pH value is 7.0-7.5. When in preparation, the lecithin is firstly dissolved in absolute ethyl alcohol, water is added into the lecithin after the lecithin is completely dissolved, the lecithin is uniformly mixed, and the mixture is heated until the ethyl alcohol is completely evaporated.

(4) Screening of Potassium-decomposing Strain

Inoculating the strains with the phosphate-solubilizing capability screened in the step to a silicate bacteria culture medium plate by adopting a three-zone streaking method, culturing at the constant temperature of 37 ℃ for 72h, and selecting the strains which can grow on the plate and have larger and thicker bacterial colonies.

The formula of the silicate bacteria culture medium is as follows: sucrose 5g, MgSO₄ 0.5g、CaCO₃ 0.1g、Na₂HPO₄ 2g、FeCl₃0.005g, 1g of glass powder, 1.6 percent of agar and 1000mL of distilled water, and sterilizing at 121 DEG C15min，pH＝7.0。

Through the multiple screening, the growth speed, the IAA producing capacity, the phosphate and potassium dissolving capacity of the strain are comprehensively considered, and the bacterium with strong saline-alkali tolerance, the IAA producing capacity, the phosphate and the potassium dissolving capacity is obtained and is numbered as PAPM-14.

Example 2: identification of Bacillus aereus PAPM-14

(1) Morphological and physiological biochemical characteristics

The morphological characteristics of the PAPM-14 strain are as follows: culturing in NA culture medium at 37 deg.C for 48h to obtain round, raised, moist, wrinkled, opaque, and milky colony; the cells were in the form of short rods, as shown in FIG. 1. The NA culture medium is a nutrient agar culture medium, and the formula of the NA culture medium is as follows: 3g of beef extract, 10g of peptone, 5g of NaCl, 20g of agar, 1000mL of water and pH 7.0.

The physiological and biochemical characteristics of the PAPM-14 strain are as follows: gram-positive bacilli; the test is positive in catalase test, positive in citrate utilization, negative in malonate utilization, positive in nitrate reduction and positive in V-P test; positive starch hydrolysis test, positive D-glucose fermentation test, positive D-mannitol fermentation test, positive lactose fermentation test and positive maltose fermentation test.

(2)16S rDNA sequence analysis

The strain PAPM-14 is inoculated into NB medium and cultured for 24h at 37 ℃ and 180r/min by shaking. The NB culture medium comprises the following components in percentage by weight: 3g of beef extract, 10g of peptone, 5g of NaCl, 1000ml of water and pH 7.0. Collecting thallus, extracting total DNA, and performing PCR amplification of 16S rDNA gene under the guide of universal primers F27: 5'-AGA GTT TGA TCA TGG CTC AG-3' and F27: 5'-AGA GTT TGA TCA TGG CTCAG-3' of prokaryotic 16S rRNA gene by using the total DNA as a template. After the amplification product is separated by 1% agarose gel electrophoresis, the amplification product is recovered by a gel recovery kit and handed over to Shanghai Biotechnology Limited company for sequencing. The 16S rDNA sequence (SEQ No.1) was aligned to sequences in the GenBank database and multiple sequence homology analysis was performed using MEGA 7.0 software.

Through morphological, physiological and biochemical characteristics and 16S rDNA sequence analysis, the strain is Bacillus aereus and is named as Bacillus aereus (Bacillus aereus) PAPM-14. The strain is preserved in China general microbiological culture Collection center (CGMCC) at 6 month and 22 days in 2020, and the preservation number is as follows: CGMCC No. 20118.

Example 3: functional verification of bacillus aereus PAPM-14 strain

(1) Capability of bacillus airoplans PAPM-14 strain to produce IAA

Inoculating Bacillus aereus PAPM-14 into LB liquid culture medium containing L-tryptophan (200mg/L), culturing at 37 deg.C and 180r/min for 4d by shaking. 50 μ L of the bacterial suspension was dropped on a white ceramic plate, and 50 μ L of LSakowski colorimetric solution was added thereto. 50 μ L of 50mg/L IAA-containing colorimetric solution was used as a positive control CK. The white ceramic plate was left at room temperature in the dark for 30min and observed, and the red-colored one indicated that IAA was secreted. After test treatment, the bacterial suspension of the strain PAPM-14 is red in color, which shows that the strain PAPM-14 has stronger capability of secreting IAA (figure 2).

The LB liquid culture medium comprises the following components in percentage by weight: 10.0g of peptone, 5.0g of yeast extract, 10.0g of NaCl and 1000mL of distilled water; sterilizing at 121 deg.C for 20 min; pH 7.0.

(2) Phosphate solubilizing ability of air bacillus PAPM-14 strain

The preparation method comprises the following steps: glucose 10.0g, (NH)₄)₂SO₄ 0.5g、MgSO₄·7H₂O 0.3g、MnSO₄·4H₂O 0.03g、KCl 0.3g、FeSO₄·7H₂O 0.03g、NaCl 0.3g、CaCO₃5.0g, lecithin 0.2g, agar 1.6%, and distilled water 1000mL, and sterilizing at 121 deg.C for 20min at pH 7.0-7.5.

Phosphate solubilizing ability of Bacillus aereus PAPM-14: inoculating Bacillus aereus PAPM-14 strain to an organophosphorus culture medium plate by using a sterilized toothpick, culturing at 37 ℃ for 2-5 days, measuring the diameter of a transparent ring and a colony, and calculating the ratio (HC) of the diameter of the transparent ring to the diameter of the colony. The diameter of a phosphate-solubilizing transparent ring generated by the PAPM-14 on an organophosphorus culture medium is 11.17mm, the diameter of a bacterial colony is 7.02mm, and the HC value is 1.59, which indicates that the strain PAPM-14 has strong capacity of solubilizing organophosphorus (figure 3).

(3) Potassium-dissolving capacity of bacillus aerosidus PAPM-14 strain

Inoculating Bacillus aereus PAPM-14 to a silicate bacteria culture medium plate by using a sterilized toothpick, and culturing for 72h at 37 ℃. Culture results as shown in FIG. 4, the PAPM-14 strain can grow and form colonies on silicate bacteria culture medium, which indicates that it has potassium-dissolving ability. The silicate bacteria culture medium comprises: sucrose 5g, MgSO₄ 0.5g、CaCO₃ 0.1g、Na₂HPO₄2g、FeCl₃0.005g, 1g of glass powder, 1.6% of agar and 1000mL of distilled water, and sterilizing at 121 ℃ for 15min, wherein the pH value is 7.0.

Example 4: preparation of viable bacteria preparation of air bacillus PAPM-14 strain

The preparation method of the live bacillus airlift PAPM-14 preparation comprises the following steps:

1) activating strains: transferring the low-temperature preserved air bacillus PAPM-14 strain to an LB solid culture medium test tube slant, and culturing at 37 ℃ for 48h for activation. The LB solid culture medium comprises the following components in percentage by weight: 5g of yeast powder, 10g of tryptone, 10g of sodium chloride, 20g of agar powder and 1000mL of water, and the pH value is 7.5.

2) Preparing seeds in a triangular flask: scraping the activated air bacillus PAPM-14 lawn by using an inoculating loop, inoculating the air bacillus PAPM-14 lawn in an LB liquid culture medium, and culturing for 48h at 37 ℃. The LB liquid culture medium comprises the following components in percentage by weight: 5g of yeast powder, 10g of tryptone, 10g of sodium chloride and 1000mL of water, and the pH value is 7.5.

3) Preparing strains in a seeding tank: transferring the seeds in the triangular flask into a 10L seeding tank filled with 7L LB liquid culture medium according to the inoculation amount of 2 percent of the volume ratio, culturing for 48h at 37 ℃, stirring at the speed of 200rpm in the whole process, and ventilating for 0-7h at the rate of 4L/min and 7-24h at the rate of 8L/min.

4) Fermentation culture: inoculating the seed tank strain into 500L fermentation tank with inoculum size of 2% by volume. The fermentation tank is filled with 350L of fermentation medium, and cultured for 24-48h at 37 ℃ to obtain the fermentation liquor of air bacillus PAPM-14. The whole stirring speed is 180-200rpm, the ventilation rate is 180-200L/min for 0-8h, and the ventilation rate is 300-400L/min.After the fermentation is finished, the number of viable bacteria of the air bacillus PAPM-14 in the fermentation liquor is (2-3) multiplied by 10¹⁰cfu/ml。

The formula of the fermentation medium in the step 4) is as follows: 10g of soybean meal, 15g of corn flour, 2g of yeast extract powder, 1.5g of monopotassium phosphate, 0.2g of zinc sulfate, 0.05g of manganese sulfate, 0.05g of complex enzyme preparation and 1000mL of water, wherein the initial pH value is 7.5. The compound enzyme preparation comprises 50% of neutral protease and 50% of cellulase. The neutral protease and the cellulase are both produced by Taian Xindeli biological technology limited company, and the product specification is 10 ten thousand U/g. The unit definition and detection method of the activity of the neutral protease execute the national standard GB/T23527-2009 protease preparation, and the unit definition and detection method of the activity of the cellulase execute the light industry standard QB 2583-2003 cellulase preparation of the people's republic of China.

The preparation method of the fermentation medium in the step 4) comprises the following steps: weighing raw materials according to the formula, dissolving in water, heating to 40-50 deg.C, maintaining the temperature for 0.5-1h, heating to 121 deg.C, maintaining the temperature for 15-30min, and sterilizing.

5) Preparation of the viable bacteria preparation: adding 0.01% (w/w) of Propyl Gallate (PG), 3% (w/w) of mineral potassium fulvate, 0.1% (w/w) of potassium sorbate and 0.02% (w/w) of carbazone into the fermentation liquor of the bacillus aereus PAPM-14 in the step 4), and uniformly stirring to obtain a viable bacteria preparation, wherein the viable bacteria content is preferably (1.0-2.0) multiplied by 10¹⁰cfu/g. The mineral source potassium fulvate is produced by Shanxi Guangyuton science and technology Limited, and the fulvic acid content of the mineral source potassium fulvate is 45%.

Example 5: antagonistic action of Bacillus aereus PAPM-14 on pathogenic fungi

Adopting a confrontation culture method. The center of the PDA plate was inoculated with a cake of pathogenic fungi of 7mm in diameter. The pathogenic fungi are respectively: the pathogenic bacteria of the corn sheath blight disease, namely rhizoctonia solani; the pathogen of corn small leaf spot-Bipolaris maydis; fusarium moniliforme, the pathogenic bacterium of bakanae disease of rice. Uniformly dibbling 3-5 PAPM-14 strains on a circumference which takes the center of a pathogenic fungus cake as the center of a circle and has the radius of 3cm, taking the treatment without inoculating the PAPM-14 strains as a control, culturing at 37 ℃ for 7-8d, and determining the diameter or the radius of a fungus colony. The bacteriostatic activity is expressed by the inhibition rate, and the calculation formula is as follows: the bacteriostatic rate is [ (R-R)/R ] × 100%. In the formula: r is the radius of pathogenic bacteria colony on the control plate; r is the colony radius of pathogenic bacteria at the position where the PAPM-14 strain and the pathogenic bacteria face each other. As shown in Table 1, the Bacillus aerosidus PAPM-14 strain has different degrees of antagonism on three pathogenic fungi tested, wherein the antagonism on the pathogenic bacteria of the maize sheath blight disease is the best.

TABLE 1 inhibitory Effect of Bacillus aereus PAPM-14 on plant pathogens

SEQUENCE LISTING

<110> Texas eight tiger Biotechnology Ltd, Shandong Zongtian Biotechnology Ltd

<120> saline-alkali tolerant air bacillus and preparation method and application of viable bacteria preparation thereof

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<170> PatentIn version 3.3

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<213> 16S rDNA sequence of Bacillus aereus (Bacillus aereus) PAPM-14

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ggggcggggt gctatacatg caagtcgagc ggaccgacgg gagcttgctc ccttaggtca 60

gcggcggacg ggtgagtaac acgtgggtaa cctgcctgta agactgggat aactccggga 120

aaccggggct aataccggat gcttgattga accgcatggt tcaatcataa aaggtggctt 180

ttagctacca cttacagatg gacccgcggc gcattagcta gttggtgagg taacggctca 240

ccaaggcgac gatgcgtagc cgacctgaga gggtgatcgg ccacactggg actgaaacac 300

ggcccagact cctacgggag gcagcagtag ggaatcttcc gcaatggacg aaagtctgac 360

ggaacaacgc cgcgtgagtg atgaaggttt tcggatcgta aaactctgtt gttagggaag 420

aacaagtacc gttcgaatag ggcggcacct tgacggtacc taaccagaaa gccacggcta 480

actacgtgcc agcagccgcg gtaatacgta ggtggcaagc gttgtccgga attattgggc 540

gtaaagcgcg cgcaggcggt ttcttaagtc tgatgtgaaa gcccccggct caaccgggga 600

gggtcattgg aaactgggga acttgagtgc agaagaggag agtggaattc cacgtgtagc 660

ggtgaaatgc gtagagatgt ggaggaacac cagtggcgaa ggcgactctc tggtctgtaa 720

ctgacgctga ggcgcgaaag cgtggggagc gaacaggatt agataccctg gtagtccacg 780

ccgtaaacga tgagtgctaa gtgttagagg gtttccgccc tttagtgctg cagcaaacgc 840

attaagcact ccgcctgggg agtacggtcg caagactgaa actcaaagga attgacgggg 900

gcccgcacaa gcggtggagc atgtggttta attcgaagca acgcgaagaa ccttaccagg 960

tcttgacatc ctctgacaac cctagagata gggcttcccc ttcgggggca gagtgacagg 1020

tggtgcatgg ttgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg 1080

caacccttga tcttagttgc cagcattcag ttgggcactc taaggtgact gccggtgaca 1140

aaccggagga aggtggggat gacgtcaaat catcatgccc cttatgacct gggctacaca 1200

cgtgctacaa tgggcagaac aaagggcagc gaagccgcga ggctaagcca atcccacaaa 1260

tctgttctca gttcggatcg cagtctgcaa ctcgactgcg tgaagctgga atcgctagta 1320

atcgcggatc agcatgccgc ggtgaatacg ttcccgggcc ttgtacacac cgcccgtcac 1380

accacgagag tttgtaacac ccgaagtcgg tgaggtaacc ttttggagcc agccgccgaa 1440

ggggatagt 1449

Claims

1. A saline-alkali tolerant Bacillus air strain is characterized by being a Bacillus air (Bacillus aereus) strain PAPM-14, wherein the preservation number of the strain is CGMCC No. 20118.

2. The use of the saline-alkali tolerant air bacillus strain of claim 1 for producing IAA, solubilizing phosphorus and potassium, and antagonizing maize sheath blight pathogenic bacteria, maize microspott pathogenic bacteria, and rice bakanae pathogenic bacteria.

3. A live bacterial preparation comprising the Bacillus air-tolerant saline-alkali-tolerant bacterial strain of claim 1 as the main active ingredient.

4. The method for preparing the viable bacteria preparation according to claim 3, wherein the Bacillus aereus strain PAPM-14 is inoculated in a fermentation medium after being subjected to expanded culture, and cultured for 24-48h at 35-40 ℃ to obtain a Bacillus aereus PAPM-14 fermentation liquor; further preparing into viable bacteria preparation.

5. The method of preparing the viable bacteria preparation according to claim 4, wherein the fermentation medium is: 10g of soybean meal, 15g of corn flour, 2g of yeast extract powder, 1.5g of monopotassium phosphate, 0.2g of zinc sulfate, 0.05g of manganese sulfate, 0.05g of complex enzyme preparation and 1000mL of water, wherein the initial pH value is 7.5; the complex enzyme preparation comprises the following components: 50% of neutral protease and 50% of cellulase according to the mass ratio.

6. The method for preparing the viable bacteria preparation according to claim 4 or 5, wherein 0.01-0.02% of propyl gallate, 3-5% of mineral source fulvic acid potassium, 0.1-0.3% of potassium sorbate and 0.02-0.05% of kaempferol are added into the fermentation liquor of the bacillus aero-bacillus PAPM-14 according to the mass ratio, and the mixture is stirred uniformly to obtain the viable bacteria preparation.