CN111454860A - Bacillus polymyxa microbial inoculum and preparation method thereof - Google Patents
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Abstract
The invention discloses a bacillus polymyxa microbial inoculum, which comprises the following effective components: bacillus polymyxa; the bacillus polymyxa is preserved in China center for type culture Collection with the preservation number of CCTCC No: the sequence of M2019116, 16srRNA is shown in SEQ ID No. 1; the preparation process comprises the following steps: preparing a culture medium, activating strains, culturing seeds, performing fermentation culture, and preparing a microbial inoculum; the microbial inoculum can effectively prevent and treat citrus canker and reduce the morbidity of the citrus canker.
Description
Technical Field
The invention relates to the technical field of microbial agents, in particular to a bacillus polymyxa microbial agent and a preparation method thereof.
Background
Citrus canker is a disease caused by pseudomonas flavipes, and is frequently found in citrus; the disease damages citrus leaves, branches and tips and fruits, and causes fallen leaves and withered tips when seedlings and saplings are seriously damaged, thereby affecting tree vigor; the fruit victims fall off fruits and the light victims have scabs and are not resistant to storage and rot, so that the commodity value of the fruits is greatly reduced, the pest control cost is increased for fruit growers, and the economic benefit is damaged.
At present, the citrus canker is usually controlled by spraying pesticides on citrus leaves, but on one hand, the pesticides are easy to cause pathogenic bacteria to generate drug resistance, so that the control difficulty is reduced; on the other hand, the environment is polluted.
The bacillus polymyxa is a kind of rhizosphere growth-promoting bacteria with wide hosts, and has control effects on plant diseases caused by fungi, bacteria, nematodes and the like; it can produce a plurality of antibiotics, polymyxin, various hydrolytic enzymes and other antagonistic substances, and the antagonistic substances are one of the important mechanisms of the biological control function of the bacillus polymyxa. Meanwhile, the bacillus polymyxa can induce plants to produce salicylic acid, so that Induced Systemic Resistance (ISR) is triggered; the bacillus polymyxa can also synthesize auxin, cytokinin and exopolysaccharide, and has activities of dissolving phosphorus and fixing nitrogen. If the bacillus polymyxa can be prepared into a microbial inoculum for preventing and treating citrus canker, the bacillus polymyxa can not only effectively prevent pathogenic bacteria, but also prevent and treat environmental pollution.
Therefore, the problem to be solved by the technical personnel in the field is how to provide a bacillus polymyxa microbial inoculum capable of effectively preventing and treating citrus canker and a preparation method thereof.
Disclosure of Invention
In view of the above, the invention provides a bacillus polymyxa microbial inoculum and a preparation method thereof, and the microbial inoculum can effectively prevent and treat citrus canker and reduce the morbidity of the citrus canker.
In order to achieve the purpose, the invention adopts the following technical scheme:
a Bacillus polymyxa inoculant comprising: bacillus polymyxa; the bacillus polymyxa is preserved in China center for type culture Collection with the preservation number of CCTCC No: m2019116, 16srRNA sequence is shown in SEQ ID No. 1.
Compared with other bacilli, the bacillus polymyxa L TPP03 has stronger disease resistance, can effectively promote plant growth and enhance nitrogen fixation capacity of soil, so that the bacillus polymyxa L TPP03 can effectively prevent and treat citrus canker by being prepared into a microbial agent and sprayed on the plants.
The bacillus polymyxa L TPP03 is separated from soil of four-pond forest towns in Xingning district of Nanning city, and has the biological characteristics of (1) growth in a preservation culture medium, small bacterial colony, white or light yellow, round, slightly convex or not convex bacterial surface, neat edge, glossy surface, smoothness, wetness, translucency to nontransparency, (2) growth in a seed slant culture medium, linear and flat bacterial lawn, smooth and semitransparent surface, and (3) growth in a fermentation culture medium, good growth, light yellow, smooth surface, translucency to nontransparency.
As a preferable technical scheme of the invention, the number of viable bacteria in the microbial inoculum is 5.0-5.2 × 109cfu/g。
A preparation method of a bacillus polymyxa microbial inoculum comprises the following steps:
1) preparing a culture medium: preparing a preservation culture medium, a seed culture medium and a fermentation culture medium, and sterilizing for later use;
2) activating the strain, namely inoculating L TPP03 strain into a seed slant culture medium for activation to obtain an activated strain;
3) seed culture: selecting the activated strain obtained in the step 2), inoculating the activated strain into a 100-120ml seed culture medium, and culturing to obtain seed bacterial liquid;
4) fermentation culture: pouring the fermentation medium into a fermentation tank, adding a seed bacterium solution with the inoculation amount of 4% of the fermentation medium into the fermentation tank, and performing fermentation culture to obtain a fermentation liquid;
5) preparing a microbial inoculum: adsorbing the fermentation liquor by 15% glucose, and spray drying to obtain wettable powder microbial inoculum.
As a preferred technical scheme of the invention, the preservation medium consists of the following components:
soluble starch 20g, KNO30.5g、NaCl 0.5g、K2HPO4·3H2O 1.0g、FeSO4·7H2O 0.01g、MgSO4·7H20.5g of O, 20g of agar powder and 1L of water.
As a preferred technical scheme of the invention, the seed culture medium consists of the following components:
30g of sucrose, 5g of peptone and FeSO4·7H2O 0.01g,、MgSO4·7H2O 0.5g、KCl 0.5g、K2HPO41g and water 1L;
the seed slant culture medium comprises the following components:
30g of sucrose, 5g of peptone and FeSO4·7H2O 0.01g、MgSO4·7H2O 0.5g、KCl 0.5g、K2HPO41g of agar powder, 20g of agar powder and 1L of water.
As a preferred technical scheme of the invention, the fermentation medium consists of the following components:
5g of molasses liquid, 17g of tryptone, 3g of soybean peptone, 5g of NaCl and K2HPO42.5g and water 1L were 7.2. + -. 0.2.
As a preferred technical scheme of the present invention, in step 2), the conditions for activating the strains are as follows: culturing at 30-37 deg.C for 20-24 hr.
As a preferred embodiment of the present invention, in step 3), the seed culture conditions are: shake culturing at 30-37 deg.C and 110-120 r/min for 40-50 h.
As a preferred technical scheme of the invention, in the step 4), the conditions of fermentation culture are as follows: the rotation speed of the fermentation tank is 110-.
As a preferable technical scheme of the invention, in the step 4), after the seed bacterial liquid is subjected to fermentation culture for 20 hours, an organic silicon defoaming agent with the mass fraction of 0.05 percent is added into a fermentation tank, and the fermentation is continued.
According to the technical scheme, compared with the prior art, the microbial agent with L TPP03 as the main component can effectively prevent and treat citrus canker, improve the disease resistance of citrus and further improve the yield of citrus.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The strain used in the embodiment is bacillus polymyxa, which is preserved in China center for type culture Collection with the preservation number of CCTCC NO: M2019116 and is classified and named as bacillus polymyxa L TPP03(Paenibacillus polymyxa L TPP03), and the nucleotide sequence of 16SrRNA of the bacillus polymyxa L TPP03 is shown as SEQ ID No. 1.
Example 1 screening of Bacillus polymyxa L TPP03
1) Collecting samples: collecting soil (four pond forest land in Xingning district of Nanning city), and placing the soil in a container;
2) diluting and heating in water bath: placing 10g of soil into a 250ml beaker, adding 90ml of sterile water into the beaker, and oscillating to obtain the soil with the concentration of 10-1The soil dilution A of (1); heating the soil diluent A in a water bath kettle at 80 ℃ for 10 min;
3) gradient dilution:
(1) 3 test tubes containing 9m of L sterile water are respectively marked as 10-2、10-3And 10-4；
(2) Shaking the soil dilution A, standing for 30s, and adding the soil dilution A marked with 10m by sucking 1m L with a pipette-2In the test tube, gently and repeatedly blowing and sucking the soil in the test tube for a plurality of times to ensure that the soil is fully and uniformly mixed to obtain a soil diluent B;
(3) similarly, 1m L of soil diluent B is absorbed and added into the soil diluent B with the serial number of 10-3Blowing and sucking the mixture evenly in the test tube to obtain a soil diluent C; obtaining soil diluent D by the same method.
4) Coating a flat plate:
respectively sucking A, B, C, D100 ul of soil diluent from low concentration, correspondingly coating the soil diluent on plates which are marked with A, B, C, D and are filled with culture medium, coating 2 plates on each diluent, uniformly coating the dilution by using a coater, and then placing the plates in an environment at 37 ℃ for constant-temperature culture for 20 hours; stopping culturing until bacterial colony grows out of the plate;
5) and (3) streak inoculation: selecting a flat plate with a growing bacterial colony for plate scribing, firstly, carrying out parallel scribing on 3 strips on one side of a plate culture medium, then, rotating a culture dish by an angle of 40 degrees, burning off residues on an inoculating loop, cooling, then, carrying out parallel scribing for the 2 nd time through a 1 st scribing part, carrying out the 3 rd time parallel scribing and the 4 th time parallel scribing through a 2 nd time parallel scribing part by the same method, placing the flat plate after scribing in an incubator for continuous culture until the bacterial colony grows out at the scribing part, and stopping culture;
6) gram staining microscopy:
(1) tabletting: selecting the bacterial colony obtained in the step 5), drying and fixing the bacterial colony on a glass slide to obtain a smear;
(2) primary dyeing: dripping ammonium oxalate crystal violet dye solution on the smear obtained in the step (1), dyeing for 0.5min, pouring out the dye solution, and washing with running water until the purple color fades;
(3) mordant dyeing: washing residual water on the smear obtained in the step (2) by using iodine solution, then covering the coated surface by using the iodine solution with the same concentration for 0.5min, and washing by using running water;
(4) and (3) decoloring: removing residual water of the smear obtained in the step (3), dripping 95% alcohol on the smear, decoloring for 15s, and immediately washing with running water;
(5) counterdyeing: dripping a safranine staining solution on the smear obtained in the step (4), staining for 3min, washing with water, and then blotting with absorbent paper;
(6) microscopic examination: under oil-lens observation, the nucleus was stained blue and the cytoplasm was stained pink.
7) Collecting the strain to obtain the bacillus polymyxa L TPP 03.
Example 2 identification of Bacillus polymyxa L TPP03
The identification of the strain comprises morphological observation, physiological and biochemical characteristic test and 16SrRNA gene sequence determination and analysis of the strain of bacillus polymyxa L TPP03, and the classification position of the strain is determined according to the identification result, the strain is identified to be bacillus polymyxa L TPP03(Paenibacillus polymyxa L TPP03), and the detection result of the physiological and biochemical characteristic is shown in tables 1-2;
TABLE 1 physiological and biochemical characteristics of strain L TPP 03-enzyme activity, carbon source assimilation
+: positive reaction; -: negative reaction; w.weak positive reaction
TABLE 2 Biochemical and physiological Properties of Strain L TPP 03-production of acid Using carbon Source
+: positive reaction; -: negative reaction;
example 3
A preparation method of a bacillus polymyxa microbial inoculum comprises the following steps:
1) preparing a culture medium: preparing a preservation culture medium, a seed culture medium and a fermentation culture medium, and sterilizing for later use;
2) activating strain, namely inoculating L TPP03 strain into a seed slant culture medium for activation, and culturing at 37 ℃ for 20h to obtain activated strain;
3) seed culture: selecting the activated strain obtained in the step 2), inoculating the activated strain into 120ml of seed culture medium, and performing shake culture at 37 ℃ and 110r/min for 40h to obtain seed bacterial liquid;
4) fermentation culture: pouring the fermentation medium into a fermentation tank, adding seed bacteria liquid with the inoculation amount of 4% of the fermentation medium into the fermentation tank, performing fermentation culture at the rotation speed of 110r/min and the ventilation amount of 1:1 at the temperature of 37 ℃ for 20 hours, adding an organic silicon defoaming agent with the mass fraction of 0.05% into the fermentation tank according to the foaming condition, and continuing to perform fermentation for 50 hours to obtain fermentation liquid;
5) preparing a microbial inoculum, namely adsorbing fermentation liquor by using 15 percent of glucose, and preparing a wettable powder microbial inoculum after spray drying, wherein the viable count in the microbial inoculum is 5.0 × 109cfu/g。
Wherein, the preservation culture medium comprises the following components:
soluble starch 20g, KNO30.5g、NaCl 0.5g、K2HPO4·3H2O 1.0g、FeSO4·7H2O 0.01g、MgSO4·7H20.5g of O, 20g of agar powder and 1L of water.
The seed culture medium consists of the following components:
30g of sucrose, 5g of peptone and FeSO4·7H2O 0.01g,、MgSO4·7H2O 0.5g、KCl 0.5g,、K2HPO41g of agar powder and 1L g of water, and 20g of agar powder is added into the slant culture medium.
The seed slant culture medium comprises the following components:
30g of sucrose, 5g of peptone and FeSO4·7H2O 0.01g、MgSO4·7H2O 0.5g、KCl 0.5g、K2HPO41g of agar powder, 20g of agar powder and 1L of water.
The fermentation medium consists of the following components:
5g of molasses liquid, 17g of tryptone, 3g of soybean peptone, 5g of NaCl and K2HPO42.5g of 1L with water to be 7.2 +/-0.2.
Example 4
A preparation method of a bacillus polymyxa microbial inoculum comprises the following steps:
1) preparing a culture medium: preparing a preservation culture medium, a seed culture medium and a fermentation culture medium, and sterilizing for later use;
2) activating strain, namely inoculating L TPP03 strain into a seed slant culture medium for activation, and culturing at 33 ℃ for 22h to obtain activated strain;
3) seed culture: selecting the activated strain obtained in the step 2), inoculating the activated strain into 110ml of seed culture medium, and performing shake culture at 33 ℃ and 110r/min for 45 hours to obtain seed bacterial liquid;
4) fermentation culture: pouring the fermentation medium into a fermentation tank, adding seed bacteria liquid with the inoculation amount of 4% of the fermentation medium into the fermentation tank, performing fermentation culture at the rotation speed of 130r/min and the ventilation amount of 1:2 at the temperature of 33 ℃ for 20 hours, adding an organic silicon defoaming agent with the mass fraction of 0.05% into the fermentation tank according to the foaming condition, and continuing to perform fermentation for 60 hours to obtain fermentation liquid;
5) preparing a microbial inoculum, namely adsorbing fermentation liquor by using 15 percent of glucose, and preparing a wettable powder microbial inoculum after spray drying, wherein the viable count in the microbial inoculum is 5.12 × 109cfu/g。
Wherein, the preservation culture medium comprises the following components:
soluble starch 20g, KNO30.5g、NaCl 0.5g、K2HPO4·3H2O 1.0g、FeSO4·7H2O 0.01g、MgSO4·7H20.5g of O, agar powder 20 and water 1L.
The seed culture medium consists of the following components:
30g of sucrose, 5g of peptone and FeSO4·7H2O 0.01g,、MgSO4·7H2O 0.5g、KCl 0.5g,、K2HPO41g and water 1L;
the seed slant culture medium comprises the following components:
30g of sucrose, 5g of peptone and FeSO4·7H2O 0.01g、MgSO4·7H2O 0.5g、KCl 0.5g、K2HPO41g of agar powder, 20g of agar powder and 1L of water.
The fermentation medium consists of the following components:
5g of molasses liquid, 17g of tryptone, 3g of soybean peptone, 5g of NaCl and K2HPO42.5g and water 1L were 7.2. + -. 0.2.
Example 5
A preparation method of a bacillus polymyxa microbial inoculum comprises the following steps:
1) preparing a culture medium: preparing a preservation culture medium, a seed culture medium and a fermentation culture medium, and sterilizing for later use;
2) activating the strain, namely inoculating L TPP03 strain into a seed slant culture medium to activate for 24 hours at 30 ℃ to obtain activated strain;
3) seed culture: selecting the activated strain obtained in the step 2), inoculating the activated strain into 100ml of seed culture medium, and performing shake culture at 30 ℃ and 115r/min for 50h to obtain seed bacterial liquid;
4) fermentation culture: pouring the fermentation medium into a fermentation tank, adding seed bacteria liquid with the inoculation amount of 4% of the fermentation medium into the fermentation tank, performing fermentation culture at the rotation speed of 120r/min and the ventilation amount of 1:1.5 at the temperature of 30 ℃ for 20 hours, adding an organic silicon defoaming agent with the mass fraction of 0.05% into the fermentation tank according to the foaming condition, and continuing to perform fermentation for 52 hours to obtain fermentation liquid;
5) preparing a microbial inoculum, namely adsorbing fermentation liquor by using 15 percent of glucose, and preparing a wettable powder microbial inoculum after spray drying, wherein the viable count in the microbial inoculum is 5.2 × 109cfu/g。
Wherein, the preservation culture medium comprises the following components:
soluble starch 20g, KNO30.5g、NaCl 0.5g、K2HPO4·3H2O 1.0g、FeSO4·7H2O 0.01g、MgSO4·7H20.5g of O, agar powder 20 and water 1L.
The seed culture medium consists of the following components:
30g of sucrose, 5g of peptone and FeSO4·7H2O 0.01g、MgSO4·7H2O 0.5g、KCl 0.5g、K2HPO41g and water 1L;
the seed slant culture medium comprises the following components:
30g of sucrose, 5g of peptone and FeSO4·7H2O 0.01g、MgSO4·7H2O 0.5g、KCl 0.5g、K2HPO41g of agar powder, 20g of agar powder and 1L of water.
The fermentation medium consists of the following components:
5g of molasses liquid, 17g of tryptone, 3g of soybean peptone, 5g of NaCl, 42.5g of K2HPO42 and 1L of water are 7.2 +/-0.2.
Example 6 prevention and treatment Effect of 6L TPP03 microbial inoculum on Wako canker disease
(1) Reagent for testing
50 billion spores/gram L TPP03 wettable powder and 100 billion spores/gram Bacillus subtilis wettable powder (Deqiang biology) prepared in example 3.
(2) Design of experiments
The test is designed to be 5 treatments, and repeated for 4 times, wherein L TPP03 bacterium wettable powder is set to have 3 concentrations which are respectively 500 times, 750 times and 1000 times of water, a contrast reagent (bacillus subtilis Deqiang organism) is diluted 1000 times of water, clear water is used as a blank contrast, and when the young sprout of the Or is 3-5cm, the wettable powder is sprayed for the first time, and is sprayed for 3 times at intervals of 14 days.
(4) Test method
The test site is arranged in the Wuwoo citrus base of Lingwu in Wuming area of Nanning, and the tree age is 4 years. Each treatment cell is 2 Wo citrus trees, the procedure is repeated for 4 times, the trees are randomly arranged in blocks, 1 fruit tree is arranged between each treatment cell, and protection rows are arranged around the treatment cells. The old leaves of the citrus reticulata blanco have ulcer disease spots and are uniform in disease occurrence.
Each cell was investigated for 2 citrus plants, each plant was investigated for 5 directions in north and south, and each direction was investigated for all leaves on 2 shoots. The leaf grading method comprises the following steps: level 0: no disease; level 1: 1-5 lesions per leaf; and 3, level: 6-10 lesions per leaf; and 5, stage: each leaf lesion is 11-15; and 7, stage: each leaf lesion is 16-20; and 9, stage: each leaf lesion is more than 21;
(4) calculation method
The prevention and treatment effect is investigated 14 days after the last spraying, the classification standard executes the national standard of the people's republic of China, "test criteria of pesticide field efficacy" (II), and the prevention and treatment effect is calculated according to the disease index.
The drug effect calculation method comprises the following steps:
(5) test results
TABLE 3 prevention and treatment effects of 3L TPP03 bacterial agent on citrus canker
As can be seen from Table 1, the treatments were sprayed 3 times at a spraying interval of 14 days, and the leaf spot investigation was carried out 14 days after the third treatment, compared with the control of clear water, the control effects of the L TPP03 fungicide 500-fold liquid on the Wai-gan ulcer were stable, the control effects were 71.49% -73.58%, and were significantly higher than the control agent, and the control effect of the control Bacillus subtilis (Deqiang) was 63.00%.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other. The device disclosed by the embodiment corresponds to the method disclosed by the embodiment, so that the description is simple, and the relevant points can be referred to the method part for description.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Sequence listing
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cggcccagac tcctacggga ggcagcagta gggaatcttc cgcaatgggc gaaagcctga 360
cggagcaacg ccgcgtgagt gatgaaggtt ttcggatcgt aaagctctgt tgccagggaa 420
gaacgctttg tagagtaact gctcttgaag tgacggtacc tgagaagaaa gccccggcta 480
actacgtgcc agcagccgcg gtaatacgta gggggcaagc gttgtccgga attattgggc 540
gtaaagcgcg cgcaggcggc tctttaagtc tggtgtttaa tcccgaggct caacttcggg 600
tcgcactgga aactggggag cttgagtgca gaagaggaga gtggaattcc acgtgtagcg 660
gtgaaatgcg tagagatgtg gaggaacacc agtggcgaag gcgactctct gggctgtaac 720
tgacgctgag gcgcgaaagc gtggggagca aacaggatta gataccctgg tagtccacgc 780
cgtaaacgat gaatgctagg tgttaggggt ttcgataccc ttggtgccga agttaacaca 840
ttaagcattc cgcctgggga gtacggtcgc aagactgaaa ctcaaaggaa ttgacgggga 900
cccgcacaag cagtggagta tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt 960
cttgacatcc ctttgaccgg tctagagata gatctttcct tcgggacaga ggagacaggt 1020
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acgtactaca atggccggta caacgggaag cgaaggagcg atctggagcc aatcctagaa 1260
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aagt 1444
Claims (10)
1. The bacillus polymyxa microbial inoculum is characterized by comprising effective components of bacillus polymyxa, wherein the bacillus polymyxa is preserved in China center for type culture collection with the preservation number of CCTCC No. M2019116 and named L TPP03, and the 16srRNA sequence is shown in SEQ ID No. 1.
2. The bacillus polymyxa microbial agent as claimed in claim 1, wherein the number of viable bacteria in the microbial agent is 5.0-5.2 × 109cfu/g。
3. A preparation method of a bacillus polymyxa microbial inoculum is characterized by comprising the following preparation processes:
1) preparing a culture medium: preparing a preservation culture medium, a seed culture medium and a fermentation culture medium, and sterilizing for later use;
2) activating the strain, namely inoculating L TPP03 strain into a seed slant culture medium for activation to obtain an activated strain;
3) seed culture: selecting the activated strain obtained in the step 2), inoculating the activated strain into a 100-120ml seed culture medium, and culturing to obtain seed bacterial liquid;
4) fermentation culture: pouring the fermentation medium into a fermentation tank, sterilizing, cooling, adding seed bacteria liquid with 4% inoculation amount of the fermentation medium into the fermentation tank, and performing fermentation culture to obtain fermentation liquid;
5) preparing a microbial inoculum: adsorbing the fermentation liquor by 15% glucose, and spray drying to obtain wettable powder microbial inoculum.
4. The method for preparing a bacillus polymyxa inoculant according to claim 3, wherein the preservation medium comprises the following components:
soluble starch 20g, KNO30.5g、NaCl 0.5g、K2HPO4·3H2O 1.0g、FeSO4·7H2O 0.01g、MgSO4·7H20.5g of O, 20g of agar powder and 1L of water.
5. The method for preparing a bacillus polymyxa inoculant according to claim 3, wherein the seed medium comprises the following components:
30g of sucrose, 5g of peptone and FeSO4·7H2O 0.01g,、MgSO4·7H2O 0.5g、KCl 0.5g、K2HPO41g and water 1L;
the seed slant culture medium comprises the following components:
30g of sucrose, 5g of peptone and FeSO4·7H2O 0.01g、MgSO4·7H2O 0.5g、KCl 0.5g、K2HPO41g of agar powder, 20g of agar powder and 1L of water.
6. The method for preparing a bacillus polymyxa inoculant according to claim 3, wherein the fermentation medium comprises the following components:
5g of molasses liquid, 17g of tryptone, 3g of soybean peptone, 5g of NaCl and K2HPO42.5g and water 1L, pH 7.2. + -. 0.2.
7. The method for preparing a bacillus polymyxa inoculant according to claim 3, wherein in step 2), the conditions for activating the inoculant are as follows: culturing at 30-37 deg.C for 20-24 hr.
8. The method for preparing a bacillus polymyxa inoculant according to claim 3, wherein in step 3), the seed culture conditions are as follows: shake culturing at 30-37 deg.C and 110-120 r/min for 40-50 h.
9. The method for preparing a bacillus polymyxa inoculant according to claim 3, wherein in step 4), the fermentation culture conditions are as follows: the rotation speed of the fermentation tank is 110-.
10. The method for preparing a bacillus polymyxa inoculant according to any one of claims 3-9, wherein in step 4), after the seed inoculant is subjected to fermentation culture for 20 hours, an organic silicon defoamer with the mass fraction of 0.05% is added into a fermentation tank, and the fermentation is continued.
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