CN113396934A - Preparation method of bacillus preparation for promoting crop growth - Google Patents
Preparation method of bacillus preparation for promoting crop growth Download PDFInfo
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- CN113396934A CN113396934A CN202110960580.9A CN202110960580A CN113396934A CN 113396934 A CN113396934 A CN 113396934A CN 202110960580 A CN202110960580 A CN 202110960580A CN 113396934 A CN113396934 A CN 113396934A
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- 238000002360 preparation method Methods 0.000 title claims abstract description 63
- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 46
- 230000012010 growth Effects 0.000 title claims abstract description 43
- 230000001737 promoting effect Effects 0.000 title claims abstract description 34
- 239000000843 powder Substances 0.000 claims abstract description 57
- 241000196252 Ulva Species 0.000 claims abstract description 39
- 238000000855 fermentation Methods 0.000 claims abstract description 29
- 230000004151 fermentation Effects 0.000 claims abstract description 29
- 230000003321 amplification Effects 0.000 claims abstract description 15
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 15
- 241000194107 Bacillus megaterium Species 0.000 claims abstract description 13
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 13
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 13
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims abstract description 12
- 239000005862 Whey Substances 0.000 claims abstract description 8
- 102000007544 Whey Proteins Human genes 0.000 claims abstract description 8
- 108010046377 Whey Proteins Proteins 0.000 claims abstract description 8
- 239000000243 solution Substances 0.000 claims description 32
- 238000002156 mixing Methods 0.000 claims description 16
- 239000012086 standard solution Substances 0.000 claims description 13
- 239000012880 LB liquid culture medium Substances 0.000 claims description 10
- 238000012258 culturing Methods 0.000 claims description 10
- 239000000725 suspension Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 7
- 229910021641 deionized water Inorganic materials 0.000 claims description 7
- 238000009835 boiling Methods 0.000 claims description 5
- 238000001816 cooling Methods 0.000 claims description 5
- 238000004108 freeze drying Methods 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 239000002994 raw material Substances 0.000 abstract description 3
- 230000000813 microbial effect Effects 0.000 abstract description 2
- 244000068988 Glycine max Species 0.000 description 42
- 235000010469 Glycine max Nutrition 0.000 description 42
- 241000196324 Embryophyta Species 0.000 description 27
- 241000894006 Bacteria Species 0.000 description 20
- 241000196253 Ulva prolifera Species 0.000 description 15
- 235000010633 broth Nutrition 0.000 description 10
- 239000002689 soil Substances 0.000 description 7
- 238000001514 detection method Methods 0.000 description 5
- 238000007726 management method Methods 0.000 description 3
- 230000008635 plant growth Effects 0.000 description 3
- 238000002054 transplantation Methods 0.000 description 3
- 208000035240 Disease Resistance Diseases 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 150000002772 monosaccharides Chemical class 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 241000193755 Bacillus cereus Species 0.000 description 1
- 241000195628 Chlorophyta Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000196246 Ulvaceae Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004178 biological nitrogen fixation Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 239000003375 plant hormone Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
Classifications
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- 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
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- 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
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- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- Agronomy & Crop Science (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Pest Control & Pesticides (AREA)
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Abstract
The invention belongs to the technical field of agricultural microbial preparations, and particularly relates to a preparation method of a bacillus preparation for promoting crop growth. According to the invention, enteromorpha is heated and hydrolyzed to prepare enteromorpha extracting solution for amplification to obtain fermentation liquor containing bacillus subtilis, bacillus amyloliquefaciens and bacillus megaterium, and the fermentation liquor is freeze-dried and mixed with a proper amount of whey powder to prepare the bacillus preparation for promoting the growth of crops. The raw materials are easy to obtain, the preparation process is stable, and the prepared bacillus preparation for promoting the growth of crops can effectively promote the growth of crops and is suitable for popularization and application.
Description
Technical Field
The invention belongs to the technical field of agricultural microbial preparations, and particularly relates to a preparation method of a bacillus preparation for promoting crop growth.
Background
Plant growth-promoting bacteria are a group of microorganisms that are present mainly in the root system of plants. Different plant growth promoting bacteria play different functions of promoting plant growth, including biological nitrogen fixation, dissolving phosphate in soil, improving plant stress resistance, promoting the absorption of trace elements by plants, regulating the secretion of plant hormones and the like. It has been found that various bacteria of the genus bacillus belong to plant growth-promoting bacteria, such as: the bacillus megaterium has a good phosphorus dissolving effect, and the bacillus cereus and the bacillus subtilis can effectively improve the disease resistance of plants.
Currently, various plant growth promoting bacterial preparations are often applied at different stages of the crop growth period to improve crop yield. The existing plant growth-promoting bacteria preparation is prepared by fermenting certain or some plant growth-promoting bacteria by using a special culture medium, and the preparation mode has high cost.
Disclosure of Invention
The invention provides a preparation method of a bacillus preparation for promoting crop growth, aiming at solving the problem of high preparation cost of a plant growth-promoting bacteria preparation in the prior art.
The invention is realized by the following technical scheme:
a preparation method of a bacillus preparation for promoting the growth of crops comprises the following steps: (1) adding enteromorpha powder into sterile deionized water to obtain enteromorpha powder suspension, boiling the enteromorpha powder suspension for 30min to 40min, cooling to room temperature, and adjusting the pH value to 7.0 to 7.5 to obtain an enteromorpha extracting solution; (2) respectively mixing bacillus subtilis powder, bacillus amyloliquefaciens powder and bacillus megaterium powder according to the mass-volume ratio of 1g: inoculating 100mL of the strain in the same LB liquid culture medium, and continuously culturing for 24 hours on a shaking table to obtain a strain amplification solution; (3) OD of strain amplification liquid by LB liquid culture medium600Adjusting the value to 0.6 to obtain a strain standard solution; (4) inoculating the standard strain solution into an enteromorpha extracting solution according to the volume ratio of 1:500 to 1:1000, and continuously culturing for 72h to 96h on a shaking table to obtain a fermentation liquid; (5) and freeze-drying the fermentation liquor, and mixing the fermentation liquor with whey powder according to the mass ratio of 15: 1-20: 1 to prepare the bacillus preparation for promoting the growth of crops.
The invention utilizes enteromorpha extract to prepare a bacillus preparation containing bacillus subtilis, bacillus amyloliquefaciens and bacillus megaterium; the enteromorpha is low in price, so that the amplification culture cost of the three plant growth-promoting bacteria can be greatly reduced.
Preferably, in the step (1), the enteromorpha powder is prepared from enteromorpha powder according to a mass volume ratio of 1g: 50mL to 1g 100mL was added to sterile deionized water.
Preferably, in the step (2), the LB liquid medium inoculated with the Bacillus subtilis powder, the Bacillus amyloliquefaciens powder and the Bacillus megaterium powder is continuously cultured on a shaking table at the conditions of 28-30 ℃ and 90-120 rpm.
Preferably, in the step (4), the enteromorpha extract inoculated with the strain standard solution is continuously cultured on a shaking table at the temperature of 30 ℃ and the rpm of 100.
Preferably, in step (5), the fermentation broth is lyophilized under vacuum at-10 ℃ to-30 ℃.
In the invention, the enteromorpha powder is prepared by crushing enteromorpha. The Enteromorpha prolifera is an algae plant of Ulvaceae and Enteromorpha, has green algae body, is composed of monolayer cells, and is enclosed into a tube shape or a strip shape by adhesion. The enteromorpha prolifera is widely distributed in offshore areas of China, and has strong reproductive capacity and large quantity, so that the raw material cost of the enteromorpha prolifera powder is low. Researches show that the enteromorpha has rich organic matter content.
The bacillus subtilis powder, the bacillus amyloliquefaciens powder and the bacillus megaterium powder are respectively dry powders of bacillus subtilis, bacillus amyloliquefaciens and bacillus megaterium, and the three bacteria belong to the bacillus. It has been found in the prior art that the three bacteria belong to plant growth-promoting bacteria.
Compared with the prior art, the invention has the following beneficial effects:
according to the enteromorpha prolifera powder disclosed by the invention, macromolecular organic matters contained in enteromorpha prolifera can be hydrolyzed after heating and hydrolysis, so that an enteromorpha prolifera extracting solution is rich in micromolecular organic matters, and the micromolecular organic matters are more favorable for bacillus subtilis, bacillus amyloliquefaciens and bacillus megatherium to be absorbed and utilized, so that the enteromorpha prolifera extracting solution can be independently used as a culture medium to realize the amplification culture of the three plant growth-promoting bacteria. In the process of preparing the fermentation liquor, monosaccharide substances in the enteromorpha extract are completely consumed by the three plant growth-promoting bacteria, so that the three plant growth-promoting bacteria cannot be rapidly proliferated and play a role after being applied to soil; after the whey powder is added into the bacillus preparation for promoting the growth of crops, the whey powder is rich in lactose, so that monosaccharide can be supplemented for the three plant growth-promoting bacteria, and the quick proliferation of the plant growth-promoting bacteria in soil is promoted. Some polysaccharides in the enteromorpha extract can not be absorbed and utilized by plant growth-promoting bacteria, and after the polysaccharides are applied to soil along with the bacillus preparation for promoting the growth of crops, the disease resistance of the crops can be improved, and the growth of the crops is facilitated.
The raw materials are easy to obtain, the preparation process is stable, and the prepared bacillus preparation for promoting the growth of crops can effectively promote the growth of crops and is suitable for popularization and application.
Detailed Description
The technical solutions of the present invention are described in detail below with reference to specific examples to facilitate understanding of the present invention, but the present invention is not limited thereto.
Example 1: a preparation method of a bacillus preparation for promoting the growth of crops comprises the following steps: (1) mixing enteromorpha powder according to the mass volume ratio of 1g: adding 50mL of the enteromorpha into sterile deionized water to obtain enteromorpha powder suspension, boiling the enteromorpha powder suspension for 30min, cooling to room temperature, and adjusting the pH value to 7.0 to obtain an enteromorpha extracting solution; (2) respectively mixing bacillus subtilis powder, bacillus amyloliquefaciens powder and bacillus megaterium powder according to the mass-volume ratio of 1g: inoculating 100mL of the strain in the same LB liquid culture medium, and continuously culturing for 24h on a shaking table at 28 ℃ and 90rpm to obtain a strain amplification solution; (3) OD of strain amplification liquid by LB liquid culture medium600Adjusting the value to 0.6 to obtain a strain standard solution; (4) inoculating the strain standard solution into the enteromorpha extracting solution according to the volume ratio of 1:500, and continuously culturing for 72h on a shaking table at 30 ℃ and 100rpm to obtain a fermentation solution; (5) and freeze-drying the fermentation liquor at-10 ℃ in vacuum, and mixing the fermentation liquor with whey powder according to the mass ratio of 15:1 to prepare the bacillus preparation for promoting the growth of crops.
Detection and application test: (1) in the step (4) of this example, fermentation broths cultured continuously for 24h and 72h were used for OD detection600The value is obtained. (2) Selecting 10 soybean seedlings with the height of 15cm to 18cm, transplanting each soybean seedling from a seedling tray into a planting pot filled with 200g of sterilized soil, and randomly selectingTaking 5 transplanted soybean seedlings as a test group, and taking the other 5 transplanted soybean seedlings as a control group; the bacillus preparation for promoting the growth of crops prepared in the embodiment is prepared by mixing the following components in a mass-volume ratio of 1g: dissolving 100mL of the bacillus preparation solution in water to prepare a bacillus preparation solution, and irrigating 30mL of the bacillus preparation solution to the soybean seedlings of the test group once every 2 days; the soybean seedlings of the control group are watered only, and the other crop management methods are the same as those of the soybean seedlings of the test group. And respectively taking the plant heights and the dry weights of the test group soybean seedlings and the control group soybean seedlings transplanted on the 0 th day as standard values, and calculating the average growth rates of the plant heights and the dry weights of the test group soybean seedlings and the control group soybean seedlings transplanted on the 7 th day, the 14 th day and the 28 th day.
And (3) test results: (1) in this example, in step (4), OD of fermentation broth after 24 hours of continuous culture was measured600OD of fermentation broth with value of 0.14, continuously cultured for 72h600The value was 0.68.
(2) The average growth rate of each parameter of the test group soybean seedlings and the control group soybean seedlings at the 7 th, 14 th and 28 th days of transplantation is as follows:
the above results illustrate that: in the embodiment, the enteromorpha extract can effectively promote the amplification of three plant growth-promoting bacteria in the strain standard solution; the bacillus preparation for promoting the growth of crops prepared by the embodiment can also effectively promote the growth of soybean seedlings.
Example 2: a preparation method of a bacillus preparation for promoting the growth of crops comprises the following steps: (1) mixing enteromorpha powder according to the mass volume ratio of 1g: adding 100mL of the enteromorpha prolifera powder into sterile deionized water to obtain enteromorpha prolifera powder suspension, boiling the enteromorpha prolifera powder suspension for 40min, cooling to room temperature, and adjusting the pH value to 7.5 to obtain an enteromorpha prolifera extracting solution; (2) respectively mixing bacillus subtilis powder, bacillus amyloliquefaciens powder and bacillus megaterium powder according to the mass-volume ratio of 1g: inoculating 100mL of the strain in the same LB liquid culture medium, and continuously culturing for 24h on a shaker at 30 ℃ and 120rpm to obtain a strain amplification solution; (3) strain expansion by LB liquid culture mediumOD of liquid increase600Adjusting the value to 0.6 to obtain a strain standard solution; (4) inoculating the strain standard solution into the enteromorpha extracting solution according to the volume ratio of 1:1000, and continuously culturing for 96h on a shaking table at 30 ℃ and 100rpm to obtain a fermentation solution; (5) and freeze-drying the fermentation liquor at the temperature of minus 30 ℃ in vacuum, and mixing the fermentation liquor with whey powder according to the mass ratio of 20:1 to prepare the bacillus preparation for promoting the growth of crops.
Detection and application test: (1) in the step (4) of this example, fermentation broths of 24h and 96h were continuously cultured for OD detection600The value is obtained. (2) Selecting 10 soybean seedlings with the height of 15cm to 18cm, transplanting each soybean seedling from a seedling tray into a planting pot filled with 200g of sterilized soil, randomly selecting 5 transplanted soybean seedlings as a test group, and taking the other 5 transplanted soybean seedlings as a control group; the bacillus preparation for promoting the growth of crops prepared in the embodiment is prepared by mixing the following components in a mass-volume ratio of 1g: dissolving 100mL of the bacillus preparation solution in water to prepare a bacillus preparation solution, and irrigating 30mL of the bacillus preparation solution to the soybean seedlings of the test group once every 2 days; the soybean seedlings of the control group are watered only, and the other crop management methods are the same as those of the soybean seedlings of the test group. And respectively taking the plant heights and the dry weights of the test group soybean seedlings and the control group soybean seedlings transplanted on the 0 th day as standard values, and calculating the average growth rates of the plant heights and the dry weights of the test group soybean seedlings and the control group soybean seedlings transplanted on the 7 th day, the 14 th day and the 28 th day.
And (3) test results: (1) in this example, in step (4), OD of fermentation broth after 24 hours of continuous culture was measured600OD of fermentation broth with value of 0.10, continuously cultured for 96h600The value was 0.72.
(2) The average growth rate of each parameter of the test group soybean seedlings and the control group soybean seedlings at the 7 th, 14 th and 28 th days of transplantation is as follows:
the above results illustrate that: in the embodiment, the enteromorpha extract can effectively promote the amplification of three plant growth-promoting bacteria in the strain standard solution; the bacillus preparation for promoting the growth of crops prepared by the embodiment can also effectively promote the growth of soybean seedlings.
Example 3: a preparation method of a bacillus preparation for promoting the growth of crops comprises the following steps: (1) mixing enteromorpha powder according to the mass volume ratio of 1g: adding 80mL of the enteromorpha prolifera powder into sterile deionized water to obtain enteromorpha prolifera powder suspension, boiling the enteromorpha prolifera powder suspension for 35min, cooling to room temperature, and adjusting the pH value to 7.2 to obtain an enteromorpha prolifera extracting solution; (2) respectively mixing bacillus subtilis powder, bacillus amyloliquefaciens powder and bacillus megaterium powder according to the mass-volume ratio of 1g: inoculating 100mL of the strain in the same LB liquid culture medium, and continuously culturing for 24h on a shaking table at 29 ℃ and 105rpm to obtain a strain amplification solution; (3) OD of strain amplification liquid by LB liquid culture medium600Adjusting the value to 0.6 to obtain a strain standard solution; (4) inoculating the strain standard solution into the enteromorpha extracting solution according to the volume ratio of 1:700, and continuously culturing for 84h on a shaking table at 30 ℃ and 100rpm to obtain a fermentation solution; (5) and freeze-drying the fermentation liquor at the temperature of-20 ℃ in vacuum, and mixing the fermentation liquor with whey powder according to the mass ratio of 17:1 to prepare the bacillus preparation for promoting the growth of crops.
Detection and application test: (1) taking the fermentation broth continuously cultured for 24h and 84h in the step (4) of the example to detect OD600The value is obtained. (2) Selecting 10 soybean seedlings with the height of 15cm to 18cm, transplanting each soybean seedling from a seedling tray into a planting pot filled with 200g of sterilized soil, randomly selecting 5 transplanted soybean seedlings as a test group, and taking the other 5 transplanted soybean seedlings as a control group; the bacillus preparation for promoting the growth of crops prepared in the embodiment is prepared by mixing the following components in a mass-volume ratio of 1g: dissolving 100mL of the bacillus preparation solution in water to prepare a bacillus preparation solution, and irrigating 30mL of the bacillus preparation solution to the soybean seedlings of the test group once every 2 days; the soybean seedlings of the control group are watered only, and the other crop management methods are the same as those of the soybean seedlings of the test group. And respectively taking the plant heights and the dry weights of the test group soybean seedlings and the control group soybean seedlings transplanted on the 0 th day as standard values, and calculating the average growth rates of the plant heights and the dry weights of the test group soybean seedlings and the control group soybean seedlings transplanted on the 7 th day, the 14 th day and the 28 th day.
And (3) test results: (1) in this example, in step (4), OD of fermentation broth after 24 hours of continuous culture was measured600OD of fermentation broth with value of 0.11, continuously cultured for 84h600The value was 0.71.
(2) The average growth rate of each parameter of the test group soybean seedlings and the control group soybean seedlings at the 7 th, 14 th and 28 th days of transplantation is as follows:
the above results illustrate that: in the embodiment, the enteromorpha extract can effectively promote the amplification of three plant growth-promoting bacteria in the strain standard solution; the bacillus preparation for promoting the growth of crops prepared by the embodiment can also effectively promote the growth of soybean seedlings.
In the above embodiments, the best mode of the present invention has been described, and it is apparent that many changes can be made under the inventive concept of the present invention. It should be noted here that any changes made under the inventive concept of the present invention shall fall within the protective scope of the present invention.
Claims (5)
1. A preparation method of a bacillus preparation for promoting the growth of crops is characterized by comprising the following steps: (1) adding enteromorpha powder into sterile deionized water to obtain enteromorpha powder suspension, boiling the enteromorpha powder suspension for 30min to 40min, cooling to room temperature, and adjusting the pH value to 7.0 to 7.5 to obtain an enteromorpha extracting solution; (2) respectively mixing bacillus subtilis powder, bacillus amyloliquefaciens powder and bacillus megaterium powder according to the mass-volume ratio of 1g: inoculating 100mL of the strain in the same LB liquid culture medium, and continuously culturing for 24 hours on a shaking table to obtain a strain amplification solution; (3) OD of strain amplification liquid by LB liquid culture medium600Adjusting the value to 0.6 to obtain a strain standard solution; (4) inoculating the standard strain solution into an enteromorpha extracting solution according to the volume ratio of 1:500 to 1:1000, and continuously culturing for 72h to 96h on a shaking table to obtain a fermentation liquid; (5) and freeze-drying the fermentation liquor, and mixing the fermentation liquor with whey powder according to the mass ratio of 15: 1-20: 1 to prepare the bacillus preparation for promoting the growth of crops.
2. The method for preparing a bacillus preparation for promoting crop growth according to claim 1, wherein the bacillus preparation comprises: in the step (1), the enteromorpha powder is prepared from enteromorpha powder according to a mass volume ratio of 1g: 50mL to 1g 100mL was added to sterile deionized water.
3. The method for preparing a bacillus preparation for promoting crop growth according to claim 1, wherein the bacillus preparation comprises: in the step (2), LB liquid culture medium inoculated with bacillus subtilis powder, bacillus amyloliquefaciens powder and bacillus megaterium powder is continuously cultured on a shaking table at the conditions of 28-30 ℃ and 90-120 rpm.
4. The method for preparing a bacillus preparation for promoting crop growth according to claim 1, wherein the bacillus preparation comprises: in the step (4), the enteromorpha extract inoculated with the strain standard solution is continuously cultured on a shaking table at the temperature of 30 ℃ and the speed of 100 rpm.
5. The method for preparing a bacillus preparation for promoting crop growth according to claim 1, wherein the bacillus preparation comprises: in the step (5), the fermentation liquor is freeze-dried under vacuum at the temperature of between 10 ℃ below zero and 30 ℃ below zero.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113980834A (en) * | 2021-09-23 | 2022-01-28 | 中化农业（临沂）研发中心有限公司 | Bacillus subtilis and application thereof in fertilizer |
| CN115088718A (en) * | 2022-07-18 | 2022-09-23 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Application of enteromorpha polysaccharide in promoting formation of bacillus biofilm |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113980834A (en) * | 2021-09-23 | 2022-01-28 | 中化农业（临沂）研发中心有限公司 | Bacillus subtilis and application thereof in fertilizer |
| CN113980834B (en) * | 2021-09-23 | 2022-08-30 | 中化农业（临沂）研发中心有限公司 | Bacillus subtilis and application thereof in fertilizer |
| CN115088718A (en) * | 2022-07-18 | 2022-09-23 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Application of enteromorpha polysaccharide in promoting formation of bacillus biofilm |
| CN115088718B (en) * | 2022-07-18 | 2024-02-09 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Application of enteromorpha polysaccharide in promoting formation of bacillus biomembrane |
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| CN113396934B (en) | 2021-11-02 |
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Denomination of invention: A method for preparing a Bacillus preparation that promotes crop growth Effective date of registration: 20231227 Granted publication date: 20211102 Pledgee: Rizhao Bank Co.,Ltd. Weifang Hanting Branch Pledgor: Shandong De Ming Xing Biotechnology Co.,Ltd. Registration number: Y2023980074731 |