CN115287211A

CN115287211A - Bacillus belgii and application thereof

Info

Publication number: CN115287211A
Application number: CN202210364669.3A
Authority: CN
Inventors: 吴春峰; 孙明宾; 王萌
Original assignee: Yantai Hongyuan Biological Fertilizer Co ltd
Current assignee: Yantai Hongyuan Biological Fertilizer Co ltd
Priority date: 2022-04-08
Filing date: 2022-04-08
Publication date: 2022-11-04
Anticipated expiration: 2042-04-08
Also published as: CN115287211B

Abstract

The invention provides Bacillus belgii and application thereof, and belongs to the technical field of microorganisms. The Bacillus belgii is preserved by the common microorganism center of China Committee for culture Collection of microorganisms with the preservation number as follows: bacillus subtilis FK006 (Bacillus velezensis FK 006) CGMCC No.23980. The Bacillus belgii and the preparation thereof provided by the invention have the functions of promoting the growth of crop plants and broad-spectrum bacteriostasis, such as: cucurbitaceae Colletotrichum (Colletotrichum lagenarium), alternaria solani (Alternaria solani), cladosporium fulvum (Cladosprium fulvum), fusarium solani (haemantonectria haemantococca), botrytis cinerea (Botrytis cinerea), rice sheath blight (Thanatephorus cucumeris), sclerotinia sclerotiorum (scleroderma sclerotiorum). The popularization and the application of the bacterial strain and the preparation are beneficial to reducing the using amount and the residual amount of chemical fertilizers and chemical pesticides in agricultural production, improve the safety of agricultural products and meet the requirements of drug reduction, weight reduction and green production better. Meanwhile, the patent strain and the preparation thereof can be used in the whole growth cycle of crops.

Description

Bacillus belgii and application thereof

Technical Field

The invention belongs to the technical field of microorganisms, and particularly relates to bacillus beiLeisi and application thereof.

Background

In order to improve the agricultural yield, people use a large amount of chemical fertilizers and pesticides during cultivation, so that soil acidification, organic matter deficiency and severe hardening are caused, further soil-borne diseases such as root rot, root rot and withering are caused, and the current situations that the fertilizer consumption is increased year by year, the yield is difficult to increase and pesticide residues are formed. Under the policy of reducing drug and losing weight and simultaneously preserving yield in China, the green sustainable development of agricultural planting can be ensured by researching biological control.

In recent years, a large number of microorganisms have been developed and marketed, such as: the bacillus subtilis, the bacillus polymyxa, the bacillus amyloliquefaciens, the bacillus megaterium, the bacillus pumilus and the like are used for preventing and treating diseases such as bacterial wilt, stem rot, gray mold and the like of crops, do not have the effect of promoting the growth of the crops, and are difficult to overcome the existing soil problems at the same time. And the current market products are mainly spray-dried powder and compound preparations, before spray-drying, thalli are collected centrifugally and then dried, original thalli metabolic active substances in fermentation liquor are lost, and the effect is difficult to see by farmers after use.

Therefore, it is highly desirable to have a microbial strain that solves the above-mentioned problems.

Disclosure of Invention

Aiming at the defects in the prior art, a bacillus with growth promoting and broad-spectrum antibacterial capabilities is obtained by screening a large quantity of strains, the bacillus is identified as bacillus beleisis FK006 through the strains, and the growth promoting effect of the bacillus is extremely remarkable through the test of growth test on rice, wheat and fields. Simultaneously, the laboratory has carried out the treatment of plant pathogens such as: the inhibition rate of the verticillium dahliae, the botrytis cinerea, the fusarium solani, the alternaria solani, the pseudomonas solani, the sclerotinia sclerotiorum and the rice sheath blight disease can reach more than 80 percent, and the resistance is completely superior to the resistance of related resistance strains reported at present.

One of the purposes of the invention is to provide Bacillus belgii FK006, which is preserved in China general microbiological culture Collection center (CGMCC) at 24.02/2022, and is abbreviated as CGMCC;

and (4) storage address: xilu No. 1 Hospital No. 3, beijing, chaoyang, beicheng;

the strain name is as follows: bacillus belgii;

latin name: bacillus velezensis;

the strain number is as follows: FK006;

the preservation number is: CGMCC No.23980.

The second purpose of the invention is to provide a Bacillus beiLeisi FK006 zymotic fluid, which is characterized in that the Bacillus beiLeisi FK006 is inoculated in a culture medium for culture;

preferably, the culture medium is a bacillus beiLeisi fermentation culture medium, and the bacillus beiLeisi fermentation culture medium comprises a carbon source, a nitrogen source and inorganic salts;

preferably, the carbon source is one or more of molasses, glucose, lactose, corn starch and dextrin;

preferably, the carbon source is molasses;

preferably, the nitrogen source is one or more of soybean meal, defatted soybean meal, chlorella powder, ammonium sulfate and urea;

preferably, the nitrogen source is ammonium sulfate and soybean meal;

preferably, the inorganic salt is one or more of calcium chloride, calcium nitrate, magnesium sulfate, magnesium chloride, corn steep liquor, corn starch, monopotassium phosphate, dipotassium phosphate and diammonium phosphate;

preferably, the inorganic salt is one or a mixture of calcium chloride, magnesium chloride, corn steep liquor powder and diammonium phosphate.

The invention also provides application of the Bacillus beiLeisi fermentation liquor or the Bacillus beiLeisi fermentation liquor in promoting the growth of crops, and particularly, the Bacillus beiLeisi fermentation liquor is prepared to have the effective viable count more than or equal to 1 × 10 ¹⁰ -2*10 ¹⁰ cfu/ml suspension, and adding strain activity protective agent;

the fourth purpose of the invention is to provide an application of Bacillus bleekii or Bacillus bleekii fermentation liquor in inhibiting plant blight and verticillium wilt, and concretely, the Bacillus bleekii fermentation liquor is prepared to have effective viable count more than or equal to 1 x 10 ¹⁰ -2*10 ¹⁰ cfu/ml suspension, and adding strain activity protective agent, wetting penetrant and pH regulator, dipping roots before transplanting crops or flushing with planting water;

preferably, the strain activity protective agent is one or more of glycol, glycerol, sorbitol, erythritol and skim milk powder;

preferably, the strain activity protective agent is a mixture of sorbitol and glycol;

preferably, the wetting penetrating agent is one or a mixture of a plurality of types of nekal BX, tween 80, organic silicon, sodium dodecyl sulfate and azone;

preferably, the wetting penetrant is a mixture of organosilicon and azone;

preferably, the pH regulator is one or more of lactic acid, acetic acid, phosphoric acid, citric acid, ascorbic acid, formic acid and malic acid.

Preferably, the suspension auxiliary agent is one or more of polyacrylamide, gelatin, chitosan, xanthan gum, sodium alginate, sodium carboxymethylcellulose, hydroxyethyl cellulose, hydroxymethyl propyl cellulose, phosphate monoester, hydroxyalkyl starch, hydroxypropyl starch, hydroxymethyl starch and acacia;

preferably, the suspension formulation is a mixture of chitosan and hydroxymethylpropyl cellulose.

The invention provides a Bacillus velezensis FK006 suspension preparation, which has a growth promoting effect on crops and has an obvious growth test effect; has broad-spectrum antibacterial effect, and has remarkable effect in inhibiting fusarium solani, sclerotinia sclerotiorum, botrytis cinerea, cucurbitaceae colletotrichum, cladosporium fulvum, rhizoctonia solani and verticillium dahliae.

Drawings

FIG. 1 is a diagram of a Bacillus belgii resistance screening assay;

FIG. 2 is a single colony diluted and spread by LB medium in physiological identification of Bacillus belgii;

FIG. 3 is a histogram of rice growth promotion test data;

FIG. 4 is a comparison of growth promoting plants of rice;

FIG. 5 is a diagram showing the effect of the Sclerotinia sclerotiorum bacteriostasis test;

FIG. 6 is a graph showing the bacteriostatic effect of Botrytis cinerea;

FIG. 7 is a diagram showing the effect of the Verticillium dahliae bacteriostasis test;

FIG. 8 is a diagram of the effect of the Scutellaria chrysosporium bacteriostasis test;

FIG. 9 is a graph showing the effect of a rice sheath blight bacterial inhibition test;

FIG. 10 is a graph showing the effect of the bacterial inhibition test on Alternaria solani;

FIG. 11 is a diagram showing the effect of the bacteriostatic test on Fusarium solani.

Bacillus belgii FK006, deposited in China general microbiological culture Collection center (CGMCC) at 24.02/2022, CGMCC, accession number: CGMCC No.23980.

Detailed Description

The present invention will be further illustrated in detail with reference to the following specific examples, which are not intended to limit the present invention but are merely illustrative thereof. The experimental methods used in the following examples are not specifically described, and the materials, reagents and the like used in the following examples are generally commercially available under the usual conditions without specific descriptions.

Example 1 screening and identification of Bacillus

1. Test materials

1.1 test strains

The test strain Bacillus beleisi FK006 is separated from the root system of tomato plants in Taiwan, shandong province, is preserved at-80 ℃ after being successfully separated, and is preserved in the China general microbiological culture Collection center (CGMCC), with the preservation number: CGMCC No.23980

1.2 use of the culture Medium

Screening a culture medium:

LB solid culture medium, proportioning: 10g/L of tryptone, 10g/L of sodium chloride, 5g/L of yeast extract powder, 20g/L of agar powder, 1000Ml of distilled water and 7.0 of ph.

PDA solid medium: 5g/L of potato extract powder, 20g/L of glucose, 20g/L of agar powder and 1000ml of distilled water.

Liquid culture medium, ratio: 10g/L of peptone, 10g/L of sodium chloride, 5g/L of yeast extract powder, 10g/L of glucose, 1000mL of distilled water and 7.0 of ph.

2. Strain screening

2.1 screening method

Adopting a filter paper method, symmetrically placing two strains dipped with different shake flask cultures on a PDA solid flat plate, and then placing filter paper sheets respectively dipped with fusarium solani and alternaria solani in the center of the flat plate; after the paper sheets are firmly stuck, the culture is carried out in an inverted way at the temperature of 28 ℃, and the phenomenon is observed when bacterial colonies grow out.

2.2 screening results

Through a plurality of tests, a strain which has resistance to fusarium solani and alternaria solani at the same time is successfully obtained. The screening results are shown in figure 1, and the fusarium solani and the alternaria solani both have obvious inhibition.

3. Identification of strains

3.1 Strain culture

Selecting strains stored at-80 deg.C, heating in water bath heated to 30 deg.C in advance for dissolving, diluting according to gradient, and sequentially sucking 100 μ L of dilution with dilution degree of 10 ^-4 、10 ^-5 、10 ^-6 The bacterial liquid is put on a solid flat plate and is uniformly coated;

the culture conditions of the constant-temperature incubator are as follows: 24h at 37 ℃;

after the bacterial colony grows out, selecting the single bacterial colony, streaking and separating again, and storing.

3.2 Strain morphology

Description of colony morphology: gram-positive, bacilliform. On an LB culture medium, bacterial colonies are round, semitransparent, colloidal, viscous and smooth and drop-shaped; as shown in fig. 2.

3.3 16S rRNA Gene identification

The 16S rRNA gene sequence of strain FK006 was sequenced using a sequencer ABI3730-XL, aligned with the data in the NCBI BLAST database with a homology of up to 99.80% with Bacillus velezensstrain strain FZB42 1696 ribosomal RNA, complete sequence in the database, so strain FK006 was identified as Bacillus belgii.

Example 2 fermentation culture of bacillus belgii FK 006:

1. strain activation

Inoculating Bacillus velezensis FK006 to LB culture medium plate, culturing at 32 deg.C in incubator for 12-16 hr;

the ratio of LB plate culture medium: 10g/L tryptone, 10g/L sodium chloride, 5g/L yeast extract powder, 20g/L agar powder, 1000ml distilled water and 7.0.

Selecting a ring of relatively large colonies with complete colony morphology, streaking the larger colonies on a slant culture medium, and culturing in a constant-temperature incubator at 32 ℃ for 12-16 hours;

the slant culture medium comprises: 10g/L tryptone, 10g/L sodium chloride, 5g/L yeast extract powder, 20g/L agar powder, 1000ml distilled water and ph7.0.

2. Shake flask strain preparation

Selecting slant strains or plate strains, inoculating 1-2 rings into shake flask culture medium, shaking table at constant temperature of 32 deg.C, and culturing for 12-16 hr to obtain shake flask seed solution;

liquid shake flask medium composition: 10-50g/L of glucose, 5-20g/L of yeast extract powder, 1-10g/L of calcium chloride, 10-50g/L of peptone, 20-30g/L of soybean meal hydrolysate, 0.01-0.05g/L of biotin and 7.0 of ph, and distilled water is added to 1000mL;

3. first order seed preparation

Inoculating the shake flask seed solution into a primary seed culture medium by the inoculation amount of 2% -5%, culturing at 32 ℃ and 200rpm for 6-12 hours to obtain a primary seed solution;

the first-level seed culture medium comprises the following components: 10-80g/L of glucose, 10g/L of soybean meal powder, 20-30g/L of soybean meal hydrolysate, 0.1-1g/L of calcium chloride, 2-5g/L of ammonium sulfate, 0.1-5g/L of magnesium sulfate, 10-30g/L of corn steep liquor and pH7.4;

4. preparation of fermentation broth

Inoculating the first-stage seed liquid into a fermentation culture medium by using the inoculation amount of 5-10%, culturing for 25-40 hours at the temperature of 32-40 ℃, the rotation speed of 150-250rpm and the air volume of 0.5-3 vvm/min; the ventilation quantity is controlled to be 1-2vvm/min in the early fermentation period, the ventilation quantity is improved to be 2-3vvm/min in the middle fermentation period, and the ventilation quantity is reduced to be 0.5-1vvm/min in the later fermentation period. .

The fermentation medium comprises the following components: 40-150g/L of molasses, 10-30g/L of corn starch, 120-150g/L of soybean meal, 5-10g/L of chlorella powder, 0.1-1g/L of calcium chloride, 10-15g/L of ammonium sulfate, 0.1-5g/L of magnesium sulfate, 0.1-2g/L of manganese sulfate, 5-10g/L of corn steep liquor, 0.5-3g/L of potassium dihydrogen phosphate and 7.2-7.5 of ph; in the fermentation process, molasses diluent is fed in at a ratio of 10L/H, and the content of residual sugar is controlled to be 0.5-0.1% until the fermentation is finished.

The effective viable count of Bacillus beilesensis FK006 in the fermentation liquid is 200-250 x 10 measured by national standard effective viable count measuring method ⁸ cfu/mL。

Example 3 preparation of Bacillus belgii FK006 suspension formulation

A bacillus belgii FK006 fermentation broth was prepared according to the method of example 2;

the preparation method of the suspension preparation comprises the following steps:

step 1. Addition of the strain protective agent of claim 8 to bacillus belgii FK 006: one or more of ethylene glycol, glycerol, sorbitol, erythritol and skimmed milk powder; the amount added is 8% to 30% w/w, most preferably 10% to 20% w/w.

Step 2, adding the wetting penetrant in the claim 9 into the uniform bacterial liquid in the step 1, such as: one or more of nekal BX, tween 80, organosilicon, sodium dodecyl sulfate and azone; the amount added is 0.5 to 3% w/w, most preferably 1 to 2% w/w.

And 3, adding a suspending agent into the uniform bacterial liquid obtained in the step 2, wherein the suspending agent comprises the following components: one or more of polyacrylamide, gelatin, chitosan, xanthan gum, sodium alginate, sodium carboxymethylcellulose, hydroxyethyl cellulose, hydroxymethyl propyl cellulose, phosphate monoester, hydroxyalkyl starch, hydroxypropyl starch, hydroxymethyl starch, and acacia; the amount added is 0.1-1% w/w, most preferably 0.2-0.8% w/w.

And 4, adding a pH regulator into the suspension obtained in the step 3, wherein the pH regulator comprises the following components: one or more of lactic acid, phosphoric acid, acetic acid, citric acid, ascorbic acid, formic acid and malic acid. The addition amount is 1-5% w/w, most preferably 2-4% w/w, to maintain the suspension formulation at a pH in the range of 4-8.

The stable and uniformly dispersed Bacillus belgii FK006 suspension preparation is prepared by the steps, the active ingredients can be uniformly dispersed in the root system or the leaf surface of the plant, the surface tension of water on the leaf surface can be obviously reduced, and the product efficacy is greatly improved.

Example 4 growth promoting action of Bacillus belgii FK006

1. Growth promotion test for rice

Products were prepared according to examples 1-3 and were prepared as solutions with a dilution factor of 300 and 500, respectively, and clear water was used as a control (ck).

And (3) selecting rice seeds with full grains, wrapping the rice seeds with gauze sterilized in advance, soaking the rice seeds into the diluted solution respectively for 4 hours, and uniformly placing the rice seeds into a flat culture dish with the bottom paved with 2-3 layers of filter paper, wherein each culture dish contains 30 grains. After the placement is finished, the container is placed in a constant-temperature artificial climate box at 28 ℃, and the illumination is set for 15 hours and the darkness is set for 9 hours.

The germination was observed daily until the plates were full and no new seeds germinated, and the culture was stopped. Taking out each seed, measuring the root length and the bud length respectively, and carrying out data analysis.

2. Test for inhibiting plant pathogenic bacteria

2.1 test materials

Bacillus belgii FK006 suspension formulation, sclerotinia sclerotiorum, botrytis cinerea, cladosporium fulvum, thanatephorus cumieri, alternaria solani, verticillium dahliae, fusarium solani Haematonecta Haematococcu, PDA medium, sterile plastic plate, tweezers, punch, coating rod, pipette gun, triangular flask PDA solid medium: 5g/L of potato soaking powder, 20g/L of glucose, 20g/L of agar powder and 1000ml of distilled water.

2.2 test methods

1ml of the Bacillus belgii FK006 suspension preparation is added into 99ml of sterile water to be diluted by 100 times, thus obtaining the mother solution used in the test. Absorbing mother liquor with different volumes, adding into sterilized PDA culture medium (culture medium temperature is 50 ℃), mixing, pouring into 9cm culture dish, condensing, inoculating fungus cake with diameter of 6mm, placing into 28 ℃ incubator, culturing, and taking PDA plate inoculated with fungus cake without adding bacterial liquid as control. 3 times of each treatment, after culturing for 7d, measuring the colony diameter by a cross method, and calculating inhibition effect inhibition rate (%) = (control colony diameter-treatment colony diameter)/(control colony diameter-6 mm) × 100% according to the following formula;

3. test results

3.1 growth promoting Effect of Rice

Compared with the control group, the test group and the control group are shown in table 1 and figure 2, and the test data clearly shows that the Bacillus belgii FK006 diluent has the effect of obviously promoting the germination and growth of plants, and the maximum yield of the root length and the stem length are respectively 28.9% and 27.9% of that of the ck control group. The picture shows that the rhizomes of plants using the Bacillus belgii FK006 suspending agent are obviously stronger than those of a control group, and the influence of the strains on the growth of crops is further verified.

3.2 test effects on inhibition of plant pathogenic fungi

The data of the inhibition rates of the Bacillus belgii FK006 against different pathogenic bacteria are shown in Table 2, and it is obvious from the test data that the inhibition rates of the Bacillus belgii FK006 suspension formulation against Sclerotinia sclerotiorum, botrytis cinerea, cladosporium fulvum, rhizoctonia solani Thanatephorus cumaris, alternaria solani, verticillium dahliae, fusarium solani Haematococcus 7 plant pathogenic bacteria are 97.4%, 61.2%, 71.2%, 89%, 58.1%, 75% and 42.8%, respectively.

The inhibition of Bacillus belezii FK006 against different pathogenic bacteria is shown in FIGS. 5-11, wherein FIG. 5 is a diagram of the inhibition effect of Sclerotinia sclerotiorum, and the colony diameter of the left control group is 83mm and has grown over the whole plate; the diameter of the colony of the right test group is 8mm, which is not much different from the initial stage of inoculation and does not spread around. FIG. 6 is a diagram showing the bacteriostatic effect of Botrytis cinerea, the colony diameter of the control group on the left side is 73mm, the edge is rough, the surface has wrinkles, and the growth trend is continued; the diameter of the bacterial colony of the right test group is 32mm, the growth speed is far less than that of the control group, and the inhibition effect is obvious. FIG. 7 is a graph of the bacteriostatic effect of Cladosporium fulvum in the Cladosporium fulvum, the surface of the colony in the left control group is blackened and may already produce spores, and the colony in the right test group is not sporulated although the color of the colony is lighter and is inhibited, so that the colony cannot grow in an enlarged manner. FIG. 8 is a graph showing the effect of inhibiting Rhizoctonia solani on rice, showing that the colony of the left control group becomes black as a whole, has a wrinkle diameter of 88mm, and the growth rate of the colony of the right test group, which has a diameter of 15mm, is much lower than that of the control group. FIG. 9 is a graph showing the inhibitory effect of Alternaria solani, the colonies of the left control group are dispersed, and the division of hyphae and spores is obvious; and hypha at the edge of the colony of the right test group grows into the culture medium, the center of the colony grows convexly, but the overall diameter is far smaller than that of the control group. FIG. 10 is a graph showing the inhibitory effect of Verticillium dahlia on Verticillium dahlia, wherein the colonies in the two groups are similar in morphology, the color of the center is dark, the color of the edge is light, and the diameter of the left control group is 66mm which is far larger than that of the right control group which is 21mm. FIG. 11 is a graph showing the inhibition effect of Fusarium solani Haematonnectria Haematococcus, wherein the two groups of colonies have regular and white edges, the diameter of the colony of the left control group is 83mm, the diameter of the colony of the right control group is 50mm, and the difference between the diameters of the two groups is the smallest, but the inhibition effect can still be seen. From the above data, it can be seen that Bacillus belgii FK006 has a wide inhibitory activity against phytopathogens.

TABLE 1 growth promoting effect of B.bailii on rice

TABLE 2 inhibition of different pathogenic bacteria by Bacillus belgii

Sequence listing

<110> tobacco platform Hongyuan biofertilizer Co Ltd

<120> Bacillus belgii and application thereof

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 1454

<212> DNA

<213> Bacillus belgii (Bacillus velezensis)

<400> 1

ttcggcggct ggctccataa aggttacctc accgacttcg ggtgttacaa actctcgtgg 60

tgtgacgggc ggtgtgtaca aggcccggga acgtattcac cgcggcatgc tgatccgcga 120

ttactagcga ttccagcttc acgcagtcga gttgcagact gcgatccgaa ctgagaacag 180

atttgtggga ttggcttaac ctcgcggttt cgctgccctt tgttctgtcc attgtagcac 240

gtgtgtagcc caggtcataa ggggcatgat gatttgacgt catccccacc ttcctccggt 300

ttgtcaccgg cagtcacctt agagtgccca actgaatgct ggcaactaag atcaagggtt 360

gcgctcgttg cgggacttaa cccaacatct cacgacacga gctgacgaca accatgcacc 420

acctgtcact ctgcccccga aggggacgtc ctatctctag gattgtcaga ggatgtcaag 480

acctggtaag gttcttcgcg ttgcttcgaa ttaaaccaca tgctccaccg cttgtgcggg 540

cccccgtcaa ttcctttgag tttcagtctt gcgaccgtac tccccaggcg gagtgcttaa 600

tgcgttagct gcagcactaa ggggcggaaa ccccctaaca cttagcactc atcgtttacg 660

gcgtggacta ccagggtatc taatcctgtt cgctccccac gctttcgctc ctcagcgtca 720

gttacagacc agagagtcgc cttcgccact ggtgttcctc cacatctcta cgcatttcac 780

cgctacacgt ggaattccac tctcctcttc tgcactcaag ttccccagtt tccaatgacc 840

ctccccggtt gagccggggg ctttcacatc agacttaaga aaccgcctgc gagcccttta 900

cgcccaataa ttccggacaa cgcttgccac ctacgtatta ccgcggctgc tggcacgtag 960

ttagccgtgg ctttctggtt aggtaccgtc aaggtgccgc cctatttgaa cggcacttgt 1020

tcttccctaa caacagagct ttacgatccg aaaaccttca tcactcacgc ggcgttgctc 1080

cgtcagactt tcgtccattg cggaagattc cctactgctg cctcccgtag gagtctgggc 1140

cgtgtctcag tcccagtgtg gccgatcacc ctctcaggtc ggctacgcat cgtcgccttg 1200

gtgagccgtt acctcaccaa ctagctaatg cgccgcgggt ccatctgtaa gtggtagccg 1260

aagccacctt ttatgtctga accatgcggt tcagacaacc atccggtatt agccccggtt 1320

tcccggagtt atcccagtct tacaggcagg ttacccacgt gttactcacc cgtccgccgc 1380

taacatcagg gagcaagctc ccatctgtcc gctcgacttg catgtattag gcacgccgcc 1440

agcgttcgtc ctga 1454

Claims

1. Bacillus beleisi, which is deposited in China general microbiological culture Collection center (CGMCC) in 2022, 24.02.month, and is abbreviated as CGMCC;

the preservation address is as follows: xilu No. 1 Hospital No. 3, beijing, chaoyang, north;

the strain name: bacillus belgii;

latin name: bacillus velezensis;

the strain number is as follows: FK006;

the preservation number is: CGMCC No.23980.

2. A bacillus belgii broth according to claim 1, wherein the bacillus belgii broth is obtained by inoculating bacillus belgii FK006 according to claim 1 in a culture medium.

3. The Bacillus belgii fermentation broth of claim 2, wherein the medium is a Bacillus belgii fermentation medium comprising a carbon source, a nitrogen source, and inorganic salts;

the carbon source is one or a mixture of molasses, glucose, lactose, corn starch and dextrin;

the nitrogen source is one or more of bean pulp, defatted bean flour, soybean protein, ammonium sulfate and urea;

the inorganic salt is one or more of calcium chloride, calcium nitrate, magnesium sulfate, magnesium chloride, corn steep liquor powder, potassium dihydrogen phosphate, dipotassium hydrogen phosphate and diammonium phosphate.

4. A Bacillus belgii broth according to claim 3, wherein the carbon source is molasses;

the nitrogen source is ammonium sulfate and soybean meal;

the inorganic salt is one or more of calcium chloride, magnesium chloride, corn starch and diammonium phosphate.

5. Use of a bacillus beijerinckii according to claim 1 or a bacillus beijerinckii fermentation broth according to claim 2 for promoting the growth of crops.

6. Use of a B.bailii fermentation broth for promoting growth of crops as claimed in claim 5, wherein the B.bailii fermentation broth of claim 2 is formulated to have an effective viable count of 1 to 10 ¹⁰ -2*10 ¹⁰ Suspension of cfu/ml, and addition of viable bacteriaA sexual protective agent.

7. Use of the Bacillus belgii according to claim 1 or the Bacillus belgii fermentation broth according to claim 2 for inhibiting diseases such as blight and verticillium wilt in plants.

8. The use of a Bacillus beijerinckii fermentation broth according to claim 7 for inhibiting diseases such as verticillium wilt in plants, wherein the Bacillus beijerinckii fermentation broth according to claim 2 is formulated to have an effective viable count of not less than 1 x 10 ¹⁰ -2*10 ¹⁰ cfu/ml suspension preparation, bacterial strain activity protective agent, wetting penetrating agent and pH regulator are added, and the suspension preparation is dipped in roots or is applied along with planting water before the crops are transplanted.

9. The use according to claim 6 or 8, characterized in that the strain activity protective agent is one or more of glycol, glycerol, sorbitol, erythritol and skimmed milk powder;

the suspension auxiliary agent is one or more of polyacrylamide, gelatin, chitosan, xanthan gum, sodium alginate, sodium carboxymethylcellulose, hydroxyethyl cellulose, hydroxymethyl propyl cellulose, phosphate monoester, hydroxyalkyl starch, hydroxypropyl starch, hydroxymethyl starch and Arabic gum.

10. The use of a bacillus belgii suspension formulation for the inhibition of blight and verticillium wilt in plants as claimed in claim 8, wherein said wetting penetrant is one or more of powdered broccoli BX, tween 80, silicones, sodium lauryl sulfate, azone;

the pH regulator is one or more of lactic acid, acetic acid, phosphoric acid, citric acid, ascorbic acid, formic acid and malic acid.