CN113699075B

CN113699075B - Bacillus atrophaeus capable of producing protease and decomposing potassium and application thereof

Info

Publication number: CN113699075B
Application number: CN202111032647.9A
Authority: CN
Inventors: 汪城墙; 丁延芹; 杜秉海; 王欣; 刘凯; 姚良同; 王朝丽
Original assignee: Shandong Agricultural University
Current assignee: Shandong Agricultural University
Priority date: 2021-09-03
Filing date: 2021-09-03
Publication date: 2023-03-28
Anticipated expiration: 2041-09-03
Also published as: CN113699075A

Abstract

The invention provides bacillus atrophaeus producing protease and decomposing potassium and application thereof, belonging to the technical field of microorganisms. The salt-tolerant atrophic bacillus screened by the invention has been preserved in China general microbiological culture Collection center (CGMCC) at 7, 9 and 2021, and the biological preservation number is CGMCC No.22866. The experiment proves that the strain has better salt resistance and simultaneously has the performance of producing protease and potassium, thereby improving the rhizosphere nutrition condition of plants (such as corn) and promoting the growth of the plants, and therefore, the strain has good value of practical application.

Description

Bacillus atrophaeus capable of producing protease and decomposing potassium and application thereof

Technical Field

The invention belongs to the technical field of microorganisms, and particularly relates to bacillus atrophaeus capable of producing protease and decomposing potassium and application thereof.

Background

The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

In recent years, chemical fertilizers play a great role in increasing yield in agricultural production, but pollute soil, harden the soil, reduce soil fertility, reduce quality of agricultural products and other problems. Therefore, it is necessary to develop a novel fertilizer which can reduce the amount of fertilizer and meet the requirement of agricultural sustainable development. Soil microorganisms play an important role in the processes of organic matter decomposition, nutrient circulation, plant nutrient utilization and the like. Plant rhizosphere soil contains a large number of microorganisms, and rhizosphere bacteria are divided into three categories, namely beneficial (2% -5%), harmful (8% -15%) and neutral (80% -90%), according to the action on plants. Plant growth-promoting rhizobacteria (PGPR) refers to a kind of beneficial bacteria that can promote plant growth, prevent and control diseases and increase crop yield and can freely live in soil or be attached to plant rhizosphere. Generally has the capability of fixing nitrogen, dissolving phosphorus, releasing potassium, producing plant hormone, secreting antibiotics and the like or at least one of the capabilities. The PGPR has the biological control effect on harmful pathogenic microorganisms and non-parasitic rhizosphere harmful microorganisms in soil, has the promotion effect on the absorption and utilization of mineral nutrition by plants, and can produce metabolites beneficial to the growth of the plants, thereby promoting the growth and development of the plants.

At present, the reported plant growth-promoting rhizobacteria are mostly concentrated in Bacillus (Bacillus), pseudomonas (Pseudomonas), agrobacterium (Agrobacterium) and ehrlichia (Eriwinia), however, the inventors found that the antagonism of Bacillus atrophaeus is more relevant to research, and the reports on the promotion of corn growth are rare.

Disclosure of Invention

Based on the defects of the prior art, the invention provides bacillus atrophaeus capable of producing protease and decomposing potassium and application thereof. The research shows that the salt-resistant composite fertilizer has better salt resistance and simultaneously has the performance of producing protease and potassium, thereby improving the rhizosphere nutrition condition of plants (such as corn) and promoting the growth of the plants, and therefore, the salt-resistant composite fertilizer has good practical application value.

In order to achieve the technical purpose, the invention relates to the following technical scheme:

in one aspect of the invention, a strain of Bacillus atrophaeus (CNY 01) is provided, which has been deposited in China general microbiological culture Collection center (address: no. 3 of Xilu 1 of Beijing Korean district, north Cheng, china) at 7-9 months in 2021 with the biological preservation number of CGMCC No.22866.

The colony and thallus characteristics of the bacillus atrophaeus CNY01 are as follows: the colony is observed after being cultured on an LB culture medium, and the colony is nearly circular, flat and dry in surface, irregular in edge, milky white and opaque. The thallus is rod-shaped, produces spores and is gram-positive.

The physiological and biochemical characteristics of the Bacillus atrophaeus CNY01 are as follows: no oxidase activity, negative; nitrate can be reduced to nitrite and is positive; the generation of the acetyl methyl methanol is realized, and V-P is positive; indole production was negative; mannitol can be used, and the product is positive; xylose is unavailable, and the result is negative; cellobiose can be utilized, and the product is positive; arabinose is unavailable and the result is negative; sorbitol can be used, and the product is positive; maltose can be utilized, and the maltose is positive; lactose is unavailable and is negative; glucose can be used, and the product is positive.

In a second aspect of the present invention, there is provided a fermentation method of the bacillus atrophaeus CNY01, the fermentation method comprising: inoculating the Bacillus atrophaeus CNY01 into a fermentation culture medium for fermentation culture to obtain the bacillus atrophaeus.

In a third aspect of the present invention, a microbial agent is provided, which contains the bacillus atrophaeus CNY01 or its fermentation product or its metabolite.

In a fourth aspect of the present invention, there is provided a microbial fertilizer, wherein the active ingredients of the microbial fertilizer comprise the bacillus atrophaeus CNY01, a fermentation product of the bacillus atrophaeus CNY01, a metabolite of the bacillus atrophaeus CNY01, and/or the microbial agent.

In a fifth aspect of the present invention, the application of the bacillus atrophaeus CNY01 or its fermentation product or its metabolite, microbial agent and/or microbial fertilizer in all or part of the following a) -d) is also within the scope of the present invention:

a) The application of the protease in producing protease;

b) The application in potassium dissolution;

c) The application of the compound in improving the nutrition condition of plant rhizosphere soil;

d) Application in promoting plant growth.

In the above application, the plant may be any one of the following plants:

p1) seed plants;

p2) monocotyledonous plants;

p3) gramineous plants;

p4) corn.

In a sixth aspect of the present invention, there is provided a method for promoting plant growth, which comprises applying the Bacillus atrophaeus CNY01, a fermentation product of the Bacillus atrophaeus CNY01, a metabolite of the Bacillus atrophaeus CNY01, the microbial agent and/or the microbial fertilizer to a plant growth environment.

In the above method, the plant may be any one of the following plants:

p1) seed plants;

p2) monocotyledonous plants;

p3) gramineous plants;

p4) corn.

In the above method, the plant growing environment may include any environment suitable for plant growth, such as soil environment, water environment, plant nutrient solution environment, etc. More preferably, the plant growing environment is a saline environment. Experiments prove that the strain can remarkably promote the growth of corn plants after being applied in a saline corn growth environment (nutrient solution and soil environment), and has the effects of remarkably promoting the height of the corn plants, the physiological plant height and the stem thickness.

The beneficial technical effects of one or more technical schemes are as follows:

according to the technical scheme, the salt-tolerant bacillus atrophaeus producing protease and decomposing potassium is reported for the first time, and the nutrient solution culture and soil culture tests prove that the bacillus atrophaeus can obviously promote the growth of corns, particularly in a salt-containing environment, so that the bacillus atrophaeus has a good practical application value.

Drawings

The accompanying drawings, which are incorporated in and constitute a part of this specification, are included to provide a further understanding of the invention, and are included to illustrate an exemplary embodiment of the invention and not to limit the invention.

FIG. 1 is a microscopic image of Bacillus atrophaeus of example 1 of the present invention.

FIG. 2 is a colony map of LB plate of Bacillus atrophaeus of example 1 of the present invention.

FIG. 3 is a graph showing the growth promoting effect of the Bacillus atrophaeus nutrient solution culture test in example 2 of the present invention.

FIG. 4 is a diagram showing the protease producing ability of Bacillus atrophaeus in example 3 of the present invention.

FIG. 5 is a diagram showing the potassium-solubilizing ability of Bacillus atrophaeus in example 3 of the present invention.

FIG. 6 is a graph showing the effect of different treatments on 28 days of the potting test of Bacillus atrophaeus in example 4 of the present invention.

Detailed Description

It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments in accordance with the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.

In a typical embodiment of the invention, bacillus atrophaeus (CNY 01) is provided, and the strain is preserved in the common microorganism center of the China Committee for culture Collection of microorganisms (address: institute No.1, west way 3, north chen of the rising district in Beijing, china) in 7-9 days in 2021, and the biological preservation number of the strain is CGMCC No.22866.

The bacterial colony and thallus characteristics of the Bacillus atrophaeus CNY01 are as follows: after 24 hours of culture on LB culture medium, the colony appears to be nearly circular, flat and dry in surface, irregular in edge, milky white and opaque. The thallus is rod-shaped, produces spores and is gram-positive.

The physiological and biochemical characteristics of the Bacillus atrophaeus CNY01 are as follows: no oxidase activity, negative; nitrate can be reduced to nitrite and is positive; the generation of the acetyl methyl methanol is realized, and V-P is positive; indole production was negative; mannitol can be used, and the product is positive; xylose is unavailable, and the result is negative; cellobiose can be utilized, and the product is positive; arabinose is unavailable, and the result is negative; sorbitol can be used, and the product is positive; maltose can be utilized, and the maltose is positive; lactose is unavailable and is negative; glucose can be used, and the product is positive.

In another embodiment of the present invention, there is provided a fermentation method of bacillus atrophaeus CNY01, the fermentation method comprising: inoculating the Bacillus atrophaeus CNY01 into a fermentation culture medium for fermentation culture to obtain the bacillus atrophaeus.

The fermentation culture conditions are specifically as follows: culturing for 15-30 h (preferably 16 h) at 30-40 ℃ (preferably 37 ℃), rotating speed: 150-200rpm (preferably 180 rpm).

The fermentation medium is not particularly limited, and a conventional bacterial culture medium, such as an LB culture medium, may be used.

In another embodiment of the present invention, the formulation of the LB medium is: 0.5 percent of yeast extract, 1 percent of peptone, 1 percent of sodium chloride, 1 percent of distilled water, 1000mL, 20min of sterilization at 121 ℃, and pH7.0.

In another embodiment of the present invention, there is provided a microbial agent containing the bacillus atrophaeus CNY01 or a fermentation product thereof or a metabolite thereof.

The term "fermentate" is used to refer to a fermentation product. The corresponding fermentation product may be a liquid obtained from the process of fermentative culture of Bacillus atrophaeus CNY01, and thus, may also be referred to as fermentation broth; the liquid may contain bacteria (bacteria cells), but does not necessarily need to contain bacteria. The liquid preferably contains a metabolite produced by the Bacillus atrophaeus CNY01 of the present invention.

And, in the embodiments of the present invention, the fermentation liquid or culture liquid containing the bacterial cells is centrifuged, filtered, sedimented or other means known in the art to separate the bacterial cells growing in the fermentation liquid or culture liquid from the liquid, and the liquid remaining when the bacterial cells are removed is a "supernatant", and in the present invention, the extracellular metabolite of bacillus atrophaeus CNY01 is contained in the supernatant. In the embodiment of the present invention, the microbial agent may also contain the supernatant.

And, in the embodiments of the present invention, the fermentation liquid or culture liquid containing the bacterial cells is centrifuged, filtered, settled or separated from the liquid by other means known in the art to obtain the bacterial cells, the bacterial cells can be disrupted by ultrasound (e.g., ultrasonic cell disruption in ice bath) or other means known in the art, or further, the disrupted bacterial cells are centrifuged to collect a supernatant, which is referred to as a cell-free extract, and the disrupted bacterial cells or cell-free extract contains intracellular metabolites of Bacillus atrophaeus CNY 01. In the embodiment of the present invention, the microbial inoculum may contain a disrupted microbial cell or a cell-free extract thereof.

And, in the embodiment of the present invention, the microbial inoculum may also be a solid, and more preferably a freeze-dried powder, for the convenience of storage, transportation, improvement of the survival rate of the strain, and the like. Namely, the bacillus atrophaeus CNY01 or the fermentation product thereof or the metabolite thereof is further subjected to freeze drying, and the freeze drying technology (including vacuum freeze drying technology) can be performed by adopting a conventional method, and is not described in detail herein.

In another embodiment of the present invention, there is provided a microbial fertilizer containing, as active ingredients, the bacillus atrophaeus CNY01, a fermentation product of the bacillus atrophaeus CNY01, a metabolite of the bacillus atrophaeus CNY01, and/or the microbial agent.

In another embodiment of the present invention, the microbial fertilizer further contains organic matter, total potassium and total nitrogen for providing nutrients.

In still another embodiment of the present invention, it is within the scope of the present invention to provide the use of the above-mentioned bacillus atrophaeus CNY01 or its fermentation product or its metabolite, microbial agent and/or microbial fertilizer in whole or in part in a) to d) as follows:

a) The application of the protease in producing protease;

b) The application in potassium dissolution;

d) Application in promoting plant growth.

In the above application, the plant may be any one of the following plants:

p1) seed plants;

p2) monocotyledonous plants;

p3) gramineous plants;

p4) maize.

In another embodiment of the present invention, there is provided a method for promoting plant growth, which comprises applying the Bacillus atrophaeus CNY01, a fermentation product of the Bacillus atrophaeus CNY01, a metabolite of the Bacillus atrophaeus CNY01, the microbial agent and/or the microbial fertilizer to a plant growth environment.

In still another embodiment of the present invention, in the above method, the plant may be any one of the following plants:

p1) seed plants;

p2) monocotyledonous plants;

p3) gramineous plants;

p4) maize.

In still another embodiment of the present invention, in the above method, the plant growing environment may include any environment suitable for plant growth, such as soil environment, water environment, plant nutrient solution environment, and the like. More preferably, the plant growing environment is a saline environment. Experiments prove that the strain can remarkably promote the growth of corn plants after being applied in a saline corn growth environment (nutrient solution and soil environment), and has the effects of remarkably promoting the height of the corn plants, the physiological plant height and the stem thickness.

The invention is further illustrated by the following examples, which are not to be construed as limiting the invention thereto. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Example 1 Strain identification

The salt-tolerant bacteria strain obtained by screening can still well grow in a 15% NaCl culture medium, and is determined to be Bacillus atrophaeus (Bacillus atrophaeus) through 16S rDNA and physiological and biochemical index identification. As shown in fig. 1 and 2, the colony and thallus of Bacillus atrophaeus (CNY 01) are characterized by: the colony is observed after being cultured on an LB culture medium, the surface is flat and dry, the edge is irregular, and the colony is milky white and opaque. The thallus is rod-shaped, produces spores and is gram-positive.

The physiological and biochemical characteristics of the Bacillus atrophaeus (CNY 01) are as follows: the strain CNY01 has no oxidase activity and is negative; nitrate can be reduced to nitrite and is positive; the generation of the acetyl methyl methanol is realized, and V-P is positive; indole production was negative; mannitol can be used, and the product is positive; xylose is unavailable, and the result is negative; cellobiose can be utilized, and the product is positive; arabinose is unavailable, and the result is negative; sorbitol can be used, and the product is positive; maltose can be utilized, and the maltose is positive; lactose is unavailable and is negative; glucose can be used, and the product is positive.

The 16S rDNA sequence of the Bacillus atrophaeus CNY01 is as follows: <xnotran> CGGCGGCTGGCTCCATAAAGGTTACCTCACCGACTTTCGGGTGTTACAAACTCTCGTGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCAGCTTCACGCAGTCGAGTTGCAGACTGCGATCCGAACTGAGAACAGATTTGTGGGATTGGCTTAACCTCGCGGTCTCGCTGCCCTTTGTTCTGTCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCACCTTAGAGTGCCCAACTGAATGCTGGCAACTAAGATCAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCACTCTGCCCCCGAAGGGGAAGCCCTATCTCTAGGGGTGTCAGAGGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAGTCTTGCGACCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCAGCACTAAGGGGCGGAAACCCCCTAACACTTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTTACAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCCTCCACATCTCTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTCCCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGAGCCCTTTACGCCCAATAATTCCGGACAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGTGCCGCCCTATTTGAACGGCACTTGTTCTTCCCTAACAACAGAGCTTTACGATCCGAAAACCTTCATCACTCACGCGGCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAGTGTGGCCGATCACCCTCTCAGGTCGGCTACGCATCGTTGCCTTGGTGAGCCATTACCTCACCAACTAGCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCCGAAGCCACCTTTTATGTTTGAACCATGCGGTTCAAACAAGCATCCGGTATTAGCCCCGGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCCGCCGCTAACATCAGGGAGCAAGCTCCCATCTGTCCGCTCGACTTGCATGTATTAGGCACGCCGCCAGCGTTCGTCCTGAGCCAGGTATTCAAACTCTTA (SEQ ID NO. 1). </xnotran>

Biological preservation description:

strain CNY01, taxonomic nomenclature: bacillus atrophaeus (Bacillus atrophaeus) is preserved in China general microbiological culture Collection center (address: no. 3 of Xilu No.1 of Beijing Korean district, north Cheng, china) within 7 months and 9 days of 2021, and the biological preservation number is CGMCC No.22866.

Example 2 nutrient solution culture growth promotion test of Bacillus atrophaeus CNY01 on corn

The preparation method of the bacterial liquid comprises the following steps: transferring the strain preserved on the inclined plane into a test tube filled with 5mL of LB liquid culture medium, and culturing in a constant temperature shaking table for 12h (temperature: 37 ℃, rotating speed: 180 rpm); inoculating the activated bacterial liquid into 50mL LB liquid culture medium according to the inoculation amount of 1%, placing into a shaking table for culturing for 16h (the temperature is 37 ℃, the rotating speed is 180 rpm), and preparing the bacterial liquid.

Wherein, LB liquid culture medium: yeast extract, 0.5%, peptone, 1%, sodium chloride, 1%, distilled water, 1000mL. Sterilizing at 121 deg.C for 20min, and pH7.0.

Soaking Zhengdan 958 semen Maydis in 75% ethanol for 5min, and shaking the container sufficiently to remove the coating of semen Maydis with ethanol. After pouring out 75% alcohol, soaking semen Maydis seed with 1% sodium hypochlorite for 10min, and chargingAnd oscillating the containers, and finally cleaning the corn seeds with sterile water for 1min each time until the sterile water added into the containers has no obvious color, pouring the sterile water off, and reserving the corn seeds for later use. Setting four groups of different treatments of sterile NaCl-free, sterile 100mM NaCl and sterile 100mM NaCl, respectively, and setting partial corn seeds at 10 deg.C according to experimental requirements ⁶ The cfu/mL bacterial solution is dipped and placed in a sterilized culture medium for later use. Illumination conditions of the illumination incubator: 16h, 25 ℃ and 60% of illumination intensity; dark conditions: the illumination intensity is 0 at 25 ℃ for 8 h. When the growth promotion effect of the corn is measured, the corn kernels are placed on kraft paper and are sequentially arranged at positions where lines are drawn in advance, the end close to the embryo is close to the kraft paper, the position of the radicle is lower, the position of the endosperm is upper, the corn seeds are wrapped by the kraft paper and tied by a string, and the corn seeds are prevented from falling off from the kraft paper. The corn seeds are uniformly distributed, the kraft paper roll is placed in a transparent culture bottle filled with nutrient solution, the cover is covered, the interference of other strains is avoided, and the corn seed is placed in an illumination incubator for culture. On day 5, the cap was opened and the height (longest leaf length of selected maize plant), root length (main root length selected) and fresh weight (weighed after removal of corn kernels) of the maize was measured on day 8.

As shown in fig. 3, at day 8, the sterile NaCl group increased 3.03% and the sterile NaCl group decreased 55.07% and 37.64%, respectively, in the height of the corn plants compared to the sterile NaCl-free group (both reaching a very significant difference, p < 0.01). In the aspect of corn root length, compared with the sterile NaCl-free group, the sterile NaCl-free group is increased by 12.11%, and the sterile NaCl-containing group are respectively reduced by 60.09% (reaching the extremely significant difference, p is less than 0.01) and 50.62% (reaching the significant difference, p is less than 0.05). In the aspect of the total fresh weight of the corn, compared with the sterile NaCl-free group, the sterile NaCl-free group is increased by 16.89%, and the sterile NaCl-containing group are respectively reduced by 38.4% (reaching the extremely significant difference, p is less than 0.01) and 21.04% (reaching the significant difference, p is less than 0.05). Under the condition of NaCl, the height, the root length and the fresh weight of the corn added with the strain CNY01 are respectively improved by 38.80 percent (reaching extremely obvious difference, p is less than 0.01), 23.73 percent and 28.19 percent (reaching extremely obvious difference, p is less than 0.01). The data obtained on the 8 th day are analyzed to obtain: the strain CNY01 shows growth promoting effect on corn under NaCl-free condition or NaCl-containing condition.

EXAMPLE 3 protease production and Potassium lysis test in Strain CNY01

The protease and potassium-releasing ability of Bacillus atrophaeus CNY01 was measured, and the results are shown in fig. 4 and fig. 5. The test result shows that the strain CNY01 obtained by screening has the functions of producing protease, decomposing potassium and the like.

EXAMPLE 4 Strain CNY01 Pot culture test

In this embodiment, four groups are provided: a treatment group in which neither bacteria nor NaCl is added and sterile water is added; treatment groups to which bacteria were added but no NaCl was added; treatment group with no added bacteria but with 100mM NaCl; the treatment group to which 100mM NaCl was added was also added. The effect of applying the inoculum under different NaCl conditions on the growth of maize seedlings can be compared by this test. Selecting corn seeds with uniform size and disinfected to plant, thinning the corn seeds when two leaves are grown, performing NaCl treatment two days after thinning, and setting two treatment groups: one group was irrigated with 200mL of the prepared 100mM NaCl solution and the other group with an equal amount of tap water. 200ml 10 was carried out two days after NaCl treatment ⁶ The cfu/mL bacterial liquid is used for treating corn plants, and initial data are determined and recorded. In the test period, the plant height, the physiological plant height (the maximum distance of the straightened crop) and the stem thickness of the plant are measured every 7 days, and seedlings are harvested in 28 days and agronomic character data (including the dry and fresh weight on the ground and the dry and fresh weight under the ground) are measured.

After the strain CNY01 is applied, the plant height, the physiological plant height and the stem thickness are measured every 7 days, and the data such as the dry and fresh weight on the ground, the dry and fresh weight under the ground and the like are measured when the corn seedlings grow to 28 days. As can be seen from FIG. 6, the effect of different NaCl treatment conditions on plant height, physiological plant height and stem thickness of plants in different periods is obtained. Under the condition of no salt, the strain CNY01 shows inhibition effect on the height, physiological plant height and stem thickness of corn plants from the 21 st day; under the condition of salt, the strain CNY01 shows the growth promoting effect on corn, and the strains are respectively increased by 13.55%, 14.13%, 15.51% and 10.84% in the strains on days 7, 14, 21 and 28 compared with the sterile group (the significant difference is achieved, and p is less than 0.05); on the aspect of physiological plant height, the number of the florigen groups on days 7, 14, 21 and 28 is respectively increased by 7.72%, 10.56%, 11.63% and 10.47% (reaching significant difference, p is less than 0.05); in terms of stem thickness, there was a promoting effect on day 7, and there were 12.95%, 4.71% and 7.56% increases in the germ-free group (significant differences were achieved, p < 0.05) over the germ-free group on days 14, 21, 28.

It should be noted that the above-mentioned embodiments are only preferred embodiments of the present invention, and the present invention is not limited thereto, and although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications and equivalents can be made in the technical solutions described in the foregoing embodiments, or equivalents thereof. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention. Although the present invention has been described with reference to the specific embodiments, it should be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention.

SEQUENCE LISTING

<110> Shandong university of agriculture

<120> Bacillus atrophaeus producing protease and decomposing potassium and application thereof

<130>

<160> 1

<170> PatentIn version 3.3

<210> 1

<211> 1473

<212> DNA

<213> Artificial sequence

<400> 1

cggcggctgg ctccataaag gttacctcac cgactttcgg gtgttacaaa ctctcgtggt 60

gtgacgggcg gtgtgtacaa ggcccgggaa cgtattcacc gcggcatgct gatccgcgat 120

tactagcgat tccagcttca cgcagtcgag ttgcagactg cgatccgaac tgagaacaga 180

tttgtgggat tggcttaacc tcgcggtctc gctgcccttt gttctgtcca ttgtagcacg 240

tgtgtagccc aggtcataag gggcatgatg atttgacgtc atccccacct tcctccggtt 300

tgtcaccggc agtcacctta gagtgcccaa ctgaatgctg gcaactaaga tcaagggttg 360

cgctcgttgc gggacttaac ccaacatctc acgacacgag ctgacgacaa ccatgcacca 420

cctgtcactc tgcccccgaa ggggaagccc tatctctagg ggtgtcagag gatgtcaaga 480

cctggtaagg ttcttcgcgt tgcttcgaat taaaccacat gctccaccgc ttgtgcgggc 540

ccccgtcaat tcctttgagt ttcagtcttg cgaccgtact ccccaggcgg agtgcttaat 600

gcgttagctg cagcactaag gggcggaaac cccctaacac ttagcactca tcgtttacgg 660

cgtggactac cagggtatct aatcctgttc gctccccacg ctttcgctcc tcagcgtcag 720

ttacagacca gagagtcgcc ttcgccactg gtgttcctcc acatctctac gcatttcacc 780

gctacacgtg gaattccact ctcctcttct gcactcaagt tccccagttt ccaatgaccc 840

tccccggttg agccgggggc tttcacatca gacttaagaa accgcctgcg agccctttac 900

gcccaataat tccggacaac gcttgccacc tacgtattac cgcggctgct ggcacgtagt 960

tagccgtggc tttctggtta ggtaccgtca aggtgccgcc ctatttgaac ggcacttgtt 1020

cttccctaac aacagagctt tacgatccga aaaccttcat cactcacgcg gcgttgctcc 1080

gtcagacttt cgtccattgc ggaagattcc ctactgctgc ctcccgtagg agtctgggcc 1140

gtgtctcagt cccagtgtgg ccgatcaccc tctcaggtcg gctacgcatc gttgccttgg 1200

tgagccatta cctcaccaac tagctaatgc gccgcgggtc catctgtaag tggtagccga 1260

agccaccttt tatgtttgaa ccatgcggtt caaacaagca tccggtatta gccccggttt 1320

cccggagtta tcccagtctt acaggcaggt tacccacgtg ttactcaccc gtccgccgct 1380

aacatcaggg agcaagctcc catctgtccg ctcgacttgc atgtattagg cacgccgcca 1440

gcgttcgtcc tgagccaggt attcaaactc tta 1473

Claims

1. Bacillus atrophaeus strainBacillus atrophaeus) CNY01, which has been preserved in China general microbiological culture Collection center (CGMCC) on 7/9/2021 with the biological preservation number of CGMCC No.22866.

2. The fermentation process of bacillus atrophaeus CNY01 of claim 1, characterized in that it comprises: and inoculating the Bacillus atrophaeus CNY01 into a fermentation culture medium for fermentation culture.

3. The fermentation process according to claim 2, wherein the fermentation culture conditions are in particular: culturing at 30-40 ℃ for 15-30h; rotating speed: 150-200rpm.

4. A fermentation process according to claim 3, wherein the fermentation culture conditions are in particular: culturing at 37 deg.C for 16h; rotating speed: 180 rpm.

5. A microbial inoculant containing bacillus atrophaeus CNY01 according to claim 1 or a fermentation product of bacillus atrophaeus CNY01 obtained by the method according to claim 2.

6. A microbial fertilizer, characterized in that the active ingredients of the microbial fertilizer comprise the Bacillus atrophaeus CNY01 of claim 1, the fermentation product of the Bacillus atrophaeus CNY01 obtained by the method of claim 2, or the microbial agent of claim 5.

7. The microbial fertilizer according to claim 6, further comprising organic matter, total potassium and total nitrogen.

8. Use of the bacillus atrophaeus CNY01 of claim 1 or a bacillus atrophaeus CNY01 fermentation obtained by the method of claim 2, the microbial agent of claim 5 or the microbial fertilizer of claim 6 in any one or more of the following a) -d):

a) The application of the protease in producing protease;

b) The application in potassium dissolution;

d) The application in promoting the growth of plants;

the plant is corn.

9. A method for promoting plant growth, comprising applying to a plant growth environment the bacillus atrophaeus CNY01 of claim 1, a bacillus atrophaeus CNY01 fermentation obtained by the method of claim 2, the microbial agent of claim 5 and/or the microbial fertilizer of claim 6 or 7;

the plant is corn;

the plant growing environment is a salt-containing environment.