CN112322542A - Bacillus amyloliquefaciens and influence thereof on soil nutrient content and enzyme activity - Google Patents
Bacillus amyloliquefaciens and influence thereof on soil nutrient content and enzyme activity Download PDFInfo
- Publication number
- CN112322542A CN112322542A CN202011285194.6A CN202011285194A CN112322542A CN 112322542 A CN112322542 A CN 112322542A CN 202011285194 A CN202011285194 A CN 202011285194A CN 112322542 A CN112322542 A CN 112322542A
- Authority
- CN
- China
- Prior art keywords
- bacillus amyloliquefaciens
- soil
- plbs010
- microbial agent
- content
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention is suitable for the technical field of microbial breeding, and provides a bacillus amyloliquefaciens strain, wherein the bacillus amyloliquefaciens strain is bacillus amyloliquefaciens PLBS010 which is preserved in China general microbiological culture collection center in 2018, 12 and 17 months with the preservation number of CGMCC NO. 16950; the invention also provides application of the bacillus amyloliquefaciens to soil nutrient content and enzyme activity. Therefore, the bacillus amyloliquefaciens PLBS010 has good high-temperature tolerance, acid-base tolerance, salt tolerance and pathogenic fungus inhibition, and a microbial agent containing the bacillus amyloliquefaciens PLBS010 is applied to soil, so that soil nutrient imbalance and soil-borne disease aggravation can be avoided, and stable, healthy and sustainable development of crop growth and agricultural production is promoted.
Description
Technical Field
The invention relates to the technical field of microbial breeding, in particular to bacillus amyloliquefaciens and influence thereof on soil nutrient content and enzyme activity.
Background
The soil is a material foundation for the growth and development of crops and the survival of the crops, and the sustainable utilization of the soil is a precondition for the stable development of agricultural production. In a soil ecosystem, soil enzymes participate in biochemical processes such as decomposition of dead branches and fallen leaves of soil, decomposition and synthesis of various organic substances, nutrient release, circulation and the like, and the soil enzymes have high and low activity and directly or indirectly influence the conversion and circulation rate of various substances in the soil ecosystem. The factors influencing soil enzymes mainly comprise soil microorganisms, soil physical and chemical properties and nutrients, planted crops, fertilization measures and the like, the soil microorganisms and crop root systems are main sources of the soil enzymes, and the activity of the soil microorganisms and the crop root systems is directly influenced by the soil nutrients.
Due to long-term application of chemical fertilizers, soil is hardened, the content of organic matters is reduced, the microbial activity of the soil is reduced, substances are difficult to convert and degrade, soil nutrients are disordered, soil-borne diseases are aggravated, and the like, so that stable, healthy and sustainable development of agricultural production is restricted.
The microbial fertilizer is a novel fertilizer widely applied to production in recent years. The research shows that the microbial fertilizer contains microbial flora, active enzyme, organic matter, multiple trace elements and the like, and has a remarkable effect of promoting the activity of soil enzyme. The microbial fertilizer improves the growth environment and the nutritional condition of crops through microbial activities and related metabolites, stimulates the growth and development of the crops, promotes the conversion of soil nutrients and improves the soil nutrient condition, thereby improving the yield of agricultural products and improving the quality.
The bacillus amyloliquefaciens is used as a microorganism in a microbial fertilizer and has important influence on the content of soil nutrients and the activity of enzyme.
Disclosure of Invention
In view of the above-mentioned drawbacks, the present invention aims to provide a bacillus amyloliquefaciens with good high temperature tolerance, acid-base tolerance, salt tolerance and pathogenic fungus inhibiting property. The microbial agent containing the bacillus amyloliquefaciens PLBS010 is applied to the soil, so that soil nutrient imbalance and soil-borne disease aggravation can be avoided, and the stable, healthy and sustainable development of crop growth and agricultural production can be promoted.
In order to achieve the aim, the invention provides a bacillus amyloliquefaciens, wherein the bacillus amyloliquefaciens is bacillus amyloliquefaciens PLBS010 which is preserved in China general microbiological culture collection center in 2018, 12 and 17 months with the preservation number of CGMCC NO. 16950.
According to the bacillus amyloliquefaciens of the invention, the bacillus amyloliquefaciens is taken from silage.
According to the bacillus amyloliquefaciens, the content of the bacillus amyloliquefaciens in a microbial agent is more than or equal to 2 multiplied by 109cfu/g。
According to the bacillus amyloliquefaciens, the addition amount of the microbial agent in soil is 20-60 kg/hm2。
According to the bacillus amyloliquefaciens, the addition amount of the microbial agent in soil is 37-48 kg/hm2。
According to the bacillus amyloliquefaciens, the invention also provides a method for screening the bacillus amyloliquefaciens, which comprises the following steps of:
step one sample Collection
Pulverizing silage, mixing PBS buffer solution and silage powder, heating in a water bath, and performing gradient dilution on supernatant to obtain bacterial source diluents with different concentrations;
isolation of the strains in step two
Respectively absorbing the bacterial source diluent to inoculate the bacterial source diluent in an LB solid culture medium, and performing static culture to obtain single bacterial colonies;
step three purification of isolate
And (3) selecting the single colony, streaking and inoculating an LB solid culture medium, culturing for 24h, and repeating streaking and inoculating culture for 3 times to obtain the pure-cultured isolate.
The invention aims to provide the bacillus amyloliquefaciens PLBS010 which has better high-temperature tolerance, acid-base tolerance, salt tolerance and pathogenic fungus inhibition. The microbial agent containing the bacillus amyloliquefaciens PLBS010 is applied to the soil, so that the content of organic matters in the soil can be improved to a certain degree, the contents of alkaline hydrolysis nitrogen, available phosphorus and quick-acting potassium in the soil are obviously improved, the activities of soil urease, sucrase and catalase are improved, soil nutrient imbalance and soil-borne disease aggravation are avoided, and the stable, healthy and sustainable development of crop growth and development and agricultural production is promoted.
Drawings
FIG. 1 is a colony morphology of the Bacillus amyloliquefaciens PLBS010 of the invention;
FIG. 2 is a graph showing the growth of the Bacillus amyloliquefaciens PLBS010 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to the accompanying drawings. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The invention provides a bacillus amyloliquefaciens strain, which has the following scientific name: bacillus amyloliquefaciens; the bacillus amyloliquefaciens is bacillus amyloliquefaciens PLBS010 which is preserved in China general microbiological culture Collection center (CGMCC for short, address: No. 3 Xilu No.1 Beichen of the south facing Yang district in Beijing) within 12 and 17 months in 2018, and the preservation number is CGMCC NO. 16950.
The bacillus amyloliquefaciens is taken from silage, and the silage is silage corn or silage corn straw.
The invention provides a screening method of bacillus amyloliquefaciens PLBS010, which comprises the following steps:
step one sample Collection
Pulverizing silage, mixing sterile PBS buffer solution and silage powder at a ratio of 1g silage to 5mL PBS buffer solution, heating in 85 deg.C water bath for 20min, and performing 10-fold serial gradient dilution on 1mL supernatant to obtain 10-1～10-4A series of dilutions of bacterial source at different concentrations.
Isolation of the strains in step two
And (3) absorbing the bacterial source diluent with each dilution degree, inoculating the bacterial source diluent to an LB solid culture medium, and performing static culture at 37 ℃ for 24 hours to obtain a single bacterial colony.
Step three purification of isolate
And (3) selecting the single colony, streaking and inoculating an LB solid culture medium, culturing at 37 ℃ for 24h, and repeating streaking, inoculating and culturing for 3 times to obtain the pure-cultured isolate.
The invention also provides an identification method of the bacillus amyloliquefaciens PLBS010, which comprises morphological identification and gene sequence identification.
Morphological identification
And (3) taking the separated bacterium liquid, diluting the separated bacterium liquid with a proper amount of normal saline, smearing the smear, performing gram staining, and observing the staining condition and morphological characteristics of the bacterium under a microscope. (see FIG. 1)
As can be seen from FIG. 1, the isolated bacteria were milky white, with an initial luster on the surface, and after several days, became wrinkled with raised edges. Gram-positive bacteria are verified through gram staining, the thallus is oval, two ends are purple, and the middle is transparent.
Identification of Gene sequences
The isolated bacterial strain is sent to the company of biological engineering (Shanghai) and Limited for sequencing, and is compared with the 16S database of NCBI according to the obtained 16S r RNA sequence information, and the parameter identification is set to be more than 95. And then selecting the first 30 RNA sequences with the highest identification (if the sequence is insufficient, the RNA sequences are completely taken), performing sequence multiple alignment by using muscle software, and constructing a phylogenetic tree by using FastTree software. The results show that the isolated strain is closest to the published Bacillus amyloliquefaciens Lx-11. Therefore, the isolate strain was classified as bacillus amyloliquefaciens, named bacillus amyloliquefaciens PLBS 010.
In order to verify the strain properties of the bacillus amyloliquefaciens PLBS010 disclosed by the invention, strain property tests are carried out, wherein the strain property tests comprise a growth curve of a strain, a high-temperature tolerance test, an acid-base tolerance test, a salt tolerance test and a pathogenic fungus inhibition test.
Growth curve of the Strain
After the isolated bacterial strains are activated, the isolated bacterial strains are respectively inoculated into a sterilized LB liquid culture medium according to the inoculation amount of 5 percent and are placed into a shaking table for culture at the temperature of 30 ℃. And measuring the absorbance value of the bacterial liquid by using a spectrophotometer every 4 hours, and drawing a strain growth curve. (see FIG. 2)
As can be seen from FIG. 2, Bacillus amyloliquefaciens PLBS010 grows rapidly in LB liquid medium; the growth is rapid within 0-26h, which can be regarded as logarithmic growth phase, the concentration is maintained at a higher level and reaches a stable state within 26-40h, and the growth speed gradually decreases after 40 h.
High temperature resistance test
Culturing the isolated strain with liquid LB culture medium for 48h, centrifuging to obtain supernatant, diluting with aseptic technique, placing in water bath at 37, 55, 65, 80 and 100 deg.C for 30min, cooling with flowing water, culturing at room temperature without water bath as control, and calculating the survival rate of the strain after water bath by plate counting method.
See Table 1
As can be seen from Table 1, the survival rate of the isolated strain at 80 ℃ is higher than 85%, which indicates that the Bacillus amyloliquefaciens PLBS010 has strong high-temperature tolerance.
TABLE 1 survival Rate of Bacillus amyloliquefaciens at different bath temperatures%
Acid-base resistance test
The isolated bacterial strains were inoculated into liquid LB medium at pH 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 and 10.0, respectively, to give an initial concentration of the isolated bacterial strains of about 1X 106cfu/mL, cultured in a shaker at 37 ℃ and then plated on a solid medium for plate counting, and the survival rate was calculated. See Table 2
As shown in Table 2, the survival rate of the isolated strain is higher than 99% under the condition that the pH value is 6.0-8.0, and the survival rate of the isolated strain is higher than 85% under the conditions that the pH value is 3.0-5.0 and the pH value is 9.0-10.0, which indicates that the Bacillus amyloliquefaciens PLBS010 has wider acid-base tolerance.
TABLE 2 survival Rate of Bacillus amyloliquefaciens in different pH environments%
Salt resistance test
Respectively inoculating the separated bacterial strains into liquid LB culture media with NaCl mass fractions of 1%, 3%, 5%, 7%, 10%, 15% and 20%, culturing at 37 ℃, coating on a solid culture medium, counting plates, and calculating the survival rate. See Table 3
As can be seen from Table 3, the survival rates of the isolated bacterial strains are higher than 98% when the mass fraction of NaCl is 1-5%, and higher than 70% when the mass fraction of NaCl is 7-15%, which indicates that the salt tolerance of the Bacillus amyloliquefaciens PLBS010 is good.
TABLE 3 survival Rate of Bacillus amyloliquefaciens at different salt concentrations%
Test for inhibiting pathogenic fungi
Preparing PDA solid culture medium by pouring method, inoculating pathogenic fungi such as Rhizoctonia solani, Fusarium oxysporum, Rhizopus nigricans, Penicillium, Aspergillus viridis, Rhizopus, Alternaria rugosa, Botrytis cinerea and Penicillium citrinum, adding 0.2mL of isolated bacterium liquid into each solid culture medium, culturing at 28 deg.C for 48h, and measuring colony diameter and inhibition rate. See Table 4
As can be seen from Table 4, the isolated bacterial strains have strong inhibition effects on pathogenic fungi such as fusarium oxysporum, rhizopus nigricans, penicillium, aspergillus viridis, alternaria, botrytis cinerea, penicillium citrinum and the like, the inhibition rates are all over 70 percent, and the inhibition effects on pathogenic fungi such as rhizoctonia solani and rhizopus are also over 60 percent; the bacillus amyloliquefaciens PLBS010 can inhibit various pathogenic fungi and has broad-spectrum antifungal capability.
TABLE 4 bacteriostatic effect of Bacillus amyloliquefaciens on pathogenic fungi
The bacillus amyloliquefaciens PLBS010 has good high-temperature tolerance, acid-base tolerance, salt tolerance and pathogenic fungus inhibition, so the bacillus amyloliquefaciens PLBS010 is applied to soil, and in order to verify the influence of the bacillus amyloliquefaciens PLBS010 on the soil nutrient content and the enzyme activity, the influence is verified.
First, test materials
The contents of the nutrient substances in the test soil are respectively as follows: 27.6g/kg of organic matter, 1.25g/kg of total nitrogen, 0.71g/kg of total phosphorus, 13.2g/kg of total potassium, 110.4mg/kg of alkaline hydrolysis nitrogen, 64.2mg/kg of available phosphorus, 182.6mg/kg of quick effect and 5.75 of pH; the strains in the test soil are respectively as follows: bacterium 0.11X 107cfu/g, actinomycete 0.96X 106cfu/g, fungus 1.60X 104cfu/g。
Planting corns in test soil, wherein the row spacing is 55-75 cm, the plant spacing is 35-50 cm, and the nitrogen application amount is 105kg/hm2. The content of Bacillus amyloliquefaciens PLBS010 in the microbial agent is more than or equal to 2 multiplied by 109cfu/g。
Second, design and process of experiment
The test soil is divided into 4 experimental groups, wherein one control group and three experimental groups are adopted, the addition amount of the microbial agent in the soil of the control group is 0, and the addition amount of the microbial agent in the soil of the experimental groups is 20-60 kg/hm2。
The microbial agent is applied by a hole application method along with the base fertilizer 3d before transplanting.
Third, items and methods of measurement
Soil nutrient determination
Collecting the middle ridge bodies of two corn plants by a sampler in a 5-point sampling method at 30, 45 and 60 days after the corn seedlings emerge respectively, taking a plough layer soil sample with the depth of 0-20 cm as about 1kg, uniformly mixing the soil samples by taking a cell as a unit, packaging the mixture by using sterile self-sealing bags, naturally drying the mixture in the absence of direct sunlight, grinding and removing impurities and residual roots of the corn, and determining the soil nutrients after sieving.
The organic matter adopts a hydration thermogravimetric potassium chromate oxidation colorimetric method; alkaline hydrolysis nitrogen is subjected to alkali de-spreading; the effective phosphorus adopts a sodium bicarbonate method; the quick-acting potassium is obtained by flame photometry.
Soil enzyme activity assay
And (3) collecting 0-20 cm soil layers of each experimental group by a 5-point method, removing impurities and stones, mixing and preparing samples, and screening the samples by a 1mm sieve for later use after the samples are naturally air-dried.
The catalase activity is determined by potassium permanganate titration method, and the result shows that 0.02mol/L KMnO is consumed by 1g of soil4Expressed in volume (mL); the soil urease activity is measured by adopting a phenol-sodium hypochlorite colorimetric method, and the result is that NH is contained in 1g of soil after 24 hours4 +-mass of N (mg); the sucrase is measured by a 3, 5-dinitrosalicylic acid colorimetric method, and the enzyme activity is expressed by the mass (mg) of glucose in 1g of soil after 24 hours.
In order to further verify the influence of the bacillus amyloliquefaciens PLBS010 on the soil nutrient content and the enzyme activity, the bacillus amyloliquefaciens PLBS010 is applied to the soil by the method, the following examples are set, and the soil nutrient content and the enzyme activity in each example are measured.
The addition amount of the microbial agent in each example is shown in Table 5; the results of the soil indexes in the examples are shown in tables 6 and 7.
Comparative example
The addition amount of the microbial agent in the soil is 0.
After the soil nutrient content and the enzyme activity are measured, the content of organic matters in the soil is 26.63g/kg, the content of alkaline hydrolysis nitrogen is 138.25mg/kg, the content of available phosphorus is 76.06mg/kg, and the content of quick-acting potassium is 220.17 mg/kg; the soil was found to have urease activity of 3.06 mg/g.d, catalase activity of 2.26 mg/g.h, and invertase activity of 42.64 mg/g.d.
Example 1
The addition amount of the microbial inoculum in the soil is 37kg/hm2。
After the soil nutrient content and the enzyme activity are measured, the content of organic matters in the soil is 29.44g/kg, the content of alkaline hydrolysis nitrogen is 183.24mg/kg, the content of available phosphorus is 94.61mg/kg, and the content of quick-acting potassium is 305.23 mg/kg; the soil was found to have urease activity of 4.92 mg/g.d, catalase activity of 3.40 mg/g.h, and invertase activity of 82.35 mg/g.d.
Example 2
The addition amount of the microbial inoculum in the soil is 44kg/hm2。
After the soil nutrient content and the enzyme activity are measured, the content of organic matters in the soil is 30.15g/kg, the content of alkaline hydrolysis nitrogen is 180.55mg/kg, the content of available phosphorus is 93.75mg/kg, and the content of quick-acting potassium is 312.01 mg/kg; the soil was found to have urease activity of 5.10 mg/g.d, catalase activity of 3.09 mg/g.h, and invertase activity of 80.49 mg/g.d.
Example 3
The addition amount of the microbial inoculum in the soil is 48kg/hm 2.
After the soil nutrient content and the enzyme activity are measured, the content of organic matters in the soil is 29.33g/kg, the content of alkaline hydrolysis nitrogen is 179.64mg/kg, the content of available phosphorus is 90.44mg/kg, and the content of quick-acting potassium is 294.72 mg/kg; the soil was found to have urease activity of 5.07 mg/g.d, catalase activity of 3.15 mg/g.h, and invertase activity of 81.67 mg/g.d.
Because the implementation processes of the embodiments are the same, the specific processes of other embodiments are not listed, and only the three embodiments with better effects are listed.
TABLE 5 addition amount of microbial inoculum in soil of each example kg/hm2
TABLE 6 nutrient content in soil of each example
TABLE 7 soil enzyme Activity of the examples
The embodiment shows that the microbial agent containing the bacillus amyloliquefaciens PLBS010 is applied to the soil, so that the content of organic matters in the soil can be improved to a certain degree, and the contents of alkaline hydrolysis nitrogen, available phosphorus and quick-acting potassium in the soil are obviously improved; the bacillus amyloliquefaciens PLBS010 has a certain effect on improvement of soil organic matters and quick-acting nitrogen, phosphorus and potassium, and can activate soil nutrients, prevent soil hardening, inhibit residual germs in soil, promote growth and development of plant seeds and seedlings and enhance root activity.
The above examples show that the application of the microbial agent containing the bacillus amyloliquefaciens PLBS010 in the soil can improve the activities of soil urease, sucrase and catalase, and the generated high-activity decomposition enzyme decomposes organic matters in the growth process of the bacillus amyloliquefaciens PLBS010, and the generated metabolic products such as organic acid, enzyme, physiological active matters and the like are easily absorbed by animals, plants and microorganisms. The bacillus amyloliquefaciens PLBS010 can accelerate the conversion and circulation rate of various substances in a soil ecosystem and promote the growth and development of crops.
In addition, the above examples show that the addition amount of the microbial agent containing Bacillus amyloliquefaciens PLBS010 in the soil is 37-48 kg/hm2In the process, the soil nutrient content and the soil enzyme activity are the best, so that the optimal addition amount of the microbial agent containing the bacillus amyloliquefaciens PLBS010 in the soil is 37-48 kg/hm2。
In conclusion, the bacillus amyloliquefaciens PLBS010 provided by the invention has better high-temperature tolerance, acid-base tolerance, salt tolerance and pathogenic fungus inhibition. The microbial agent containing the bacillus amyloliquefaciens PLBS010 is applied to the soil, so that the content of organic matters in the soil can be improved to a certain degree, the contents of alkaline hydrolysis nitrogen, available phosphorus and quick-acting potassium in the soil are obviously improved, the activities of soil urease, sucrase and catalase are improved, soil nutrient imbalance and soil-borne disease aggravation are avoided, and the stable, healthy and sustainable development of crop growth and development and agricultural production is promoted.
The present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof, and it should be understood that various changes and modifications can be effected therein by one skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (6)
1. The bacillus amyloliquefaciens is characterized in that the bacillus amyloliquefaciens is bacillus amyloliquefaciens PLBS010 and is preserved in China general microbiological culture collection center (CGMCC) for 12-17 th in 2018 with the preservation number of CGMCC NO. 16950.
2. The bacillus amyloliquefaciens according to claim 1, wherein the bacillus amyloliquefaciens is taken from silage.
3. The Bacillus amyloliquefaciens according to claim 1, wherein the content of the Bacillus amyloliquefaciens in a microbial agent is more than or equal to 2 x 109cfu/g。
4. The bacillus amyloliquefaciens according to claim 3, wherein the addition amount of the microbial agent in soil is 20-60 kg/hm2。
5. The Bacillus amyloliquefaciens according to claim 4, wherein the addition amount of the microbial agent in soil is 37-48 kg/hm2。
6. A method of screening the Bacillus amyloliquefaciens strain of claim 1, comprising the steps of:
step one sample Collection
Pulverizing silage, mixing PBS buffer solution and silage powder, heating in a water bath, and performing gradient dilution on supernatant to obtain bacterial source diluents with different concentrations;
isolation of the strains in step two
Respectively absorbing the bacterial source diluent to inoculate the bacterial source diluent in an LB solid culture medium, and performing static culture to obtain single bacterial colonies;
step three purification of isolate
And (3) selecting the single colony, streaking and inoculating an LB solid culture medium, culturing for 24h, and repeating streaking and inoculating culture for 3 times to obtain the pure-cultured isolate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011285194.6A CN112322542A (en) | 2020-11-17 | 2020-11-17 | Bacillus amyloliquefaciens and influence thereof on soil nutrient content and enzyme activity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011285194.6A CN112322542A (en) | 2020-11-17 | 2020-11-17 | Bacillus amyloliquefaciens and influence thereof on soil nutrient content and enzyme activity |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112322542A true CN112322542A (en) | 2021-02-05 |
Family
ID=74320853
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011285194.6A Pending CN112322542A (en) | 2020-11-17 | 2020-11-17 | Bacillus amyloliquefaciens and influence thereof on soil nutrient content and enzyme activity |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112322542A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114436698A (en) * | 2022-02-17 | 2022-05-06 | 青岛力力惠生物科技股份有限公司 | Liquid compound microbial fertilizer and application thereof in agricultural production |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110274673A1 (en) * | 2010-05-04 | 2011-11-10 | Novozymes Biologicals, Inc. | Bacillus amyloliquefaciens Strain |
| CN103275907A (en) * | 2013-06-18 | 2013-09-04 | 武汉科前动物生物制品有限责任公司 | Bacillus amyloliquefacien and preparation method and application thereof |
| CN107267423A (en) * | 2017-07-28 | 2017-10-20 | 中国农业科学院果树研究所 | The bacillus amyloliquefaciens L 1 of antagonism the operatic circle disease and its application |
| KR101837695B1 (en) * | 2016-10-10 | 2018-03-13 | 전남대학교 산학협력단 | Novel Bacillus amyloliquepaciens Y1 having Control Activity against Plant Pathogenic Fungi |
| CN108658679A (en) * | 2018-07-23 | 2018-10-16 | 湖南泰谷生态工程有限公司 | It is a kind of to be used to prevent organic fertilizer of early blight of tomato and preparation method thereof |
| CN108707560A (en) * | 2018-05-23 | 2018-10-26 | 福建省农业科学院农业工程技术研究所 | A kind of microbial liquid bacterial manure and its preparation method and application |
| CN110791456A (en) * | 2019-11-26 | 2020-02-14 | 黄河三角洲京博化工研究院有限公司 | Salt-tolerant growth-promoting bacillus and application thereof |
| EP3659440A1 (en) * | 2018-11-28 | 2020-06-03 | Bio-Iliberis Research and Development S.L. | Plant growth promoting microorganism and enzymes for soil biogenic cycles |
-
2020
- 2020-11-17 CN CN202011285194.6A patent/CN112322542A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110274673A1 (en) * | 2010-05-04 | 2011-11-10 | Novozymes Biologicals, Inc. | Bacillus amyloliquefaciens Strain |
| CN103275907A (en) * | 2013-06-18 | 2013-09-04 | 武汉科前动物生物制品有限责任公司 | Bacillus amyloliquefacien and preparation method and application thereof |
| KR101837695B1 (en) * | 2016-10-10 | 2018-03-13 | 전남대학교 산학협력단 | Novel Bacillus amyloliquepaciens Y1 having Control Activity against Plant Pathogenic Fungi |
| CN107267423A (en) * | 2017-07-28 | 2017-10-20 | 中国农业科学院果树研究所 | The bacillus amyloliquefaciens L 1 of antagonism the operatic circle disease and its application |
| CN108707560A (en) * | 2018-05-23 | 2018-10-26 | 福建省农业科学院农业工程技术研究所 | A kind of microbial liquid bacterial manure and its preparation method and application |
| CN108658679A (en) * | 2018-07-23 | 2018-10-16 | 湖南泰谷生态工程有限公司 | It is a kind of to be used to prevent organic fertilizer of early blight of tomato and preparation method thereof |
| EP3659440A1 (en) * | 2018-11-28 | 2020-06-03 | Bio-Iliberis Research and Development S.L. | Plant growth promoting microorganism and enzymes for soil biogenic cycles |
| CN110791456A (en) * | 2019-11-26 | 2020-02-14 | 黄河三角洲京博化工研究院有限公司 | Salt-tolerant growth-promoting bacillus and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 高亮等: "腐植酸生物菌肥对保护地次生盐渍化土壤改良效果研究", 《腐殖酸》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114436698A (en) * | 2022-02-17 | 2022-05-06 | 青岛力力惠生物科技股份有限公司 | Liquid compound microbial fertilizer and application thereof in agricultural production |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2021185021A1 (en) | Microbial agent yf beneficial to corn growth in saline-alkali soil and application thereof | |
| CN105936881B (en) | One kind is for alignic thermophilic sugared bacillus and its application method of degrading | |
| CN103539535A (en) | Active biological matrix product specially used for culture of cucumber seedlings | |
| CN108192838B (en) | Bacillus amyloliquefaciens with dual functions of inorganic phosphorus degradation and disease prevention | |
| CN104818233A (en) | Bacillus vallismortis and functional vegetable seedling raising biological matrix prepared from bacillus vallismortis | |
| CN104726378B (en) | The method for improving salt stress turfgrass defence enzyme activity using Salt-tolerant microbial agent is strengthened | |
| CN111172052B (en) | Bacillus P75 for improving soil fertility of sandstone and application thereof | |
| CN107142210A (en) | A kind of compound method of hard stalk fermentation microbial inoculum | |
| CN116396888A (en) | Microbial composite microbial agent suitable for livestock and poultry manure and straw fermentation and preparation method thereof | |
| CN102816719B (en) | Growth-promoting rhizobacteria SXH-2 and application thereof | |
| CN102174428A (en) | Antagonistic bacteria for preventing bacterial wilt of potatoes and microorganism organic fertilizer thereof | |
| CN109136142B (en) | Streptomyces alboflavus, method for preparing biocontrol microbial inoculum by using same and application thereof | |
| CN113881599B (en) | Bacillus subtilis and tomato compound microbial fertilizer, preparation method and application thereof | |
| CN109456915B (en) | Bacillus safensis strain X3 and application thereof | |
| CN108048360B (en) | Bacillus subtilis with dual functions of degrading organic phosphorus and preventing diseases | |
| CN107628894A (en) | Composite bacteria agent increase soil fertility and its preparation method and application | |
| CN110791459B (en) | Bacillus subtilis for preventing and controlling continuous cropping lily soil-borne blight and application thereof | |
| CN110229757B (en) | Trichoderma citrinoviride JS84 capable of effectively promoting crop growth and bio-organic fertilizer developed by trichoderma citrinoviride JS84 | |
| CN112322542A (en) | Bacillus amyloliquefaciens and influence thereof on soil nutrient content and enzyme activity | |
| CN104498413B (en) | Functional vegetable seedling biological matrix containing bacillus subtilis G10 and preparation method thereof | |
| CN114874953B (en) | Peanut rhizosphere biocontrol bacterium-Bacillus beleisi SW-1 and application thereof | |
| CN116354766A (en) | Carbon-based bio-organic fertilizer for promoting vegetable growth and improving soil and preparation method thereof | |
| CN115959931A (en) | Method for preparing organic fertilizer by deep fermentation of livestock and poultry manure and straws | |
| CN112391311A (en) | Bacillus subtilis and influence thereof on soil nutrient content and enzyme activity | |
| CN113481111B (en) | Efficient biological straw fermentation inoculant and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20210205 |
|
| RJ01 | Rejection of invention patent application after publication |