CN111662849A - Bacillus siamensis and application thereof - Google Patents
Bacillus siamensis and application thereof Download PDFInfo
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- CN111662849A CN111662849A CN202010636819.2A CN202010636819A CN111662849A CN 111662849 A CN111662849 A CN 111662849A CN 202010636819 A CN202010636819 A CN 202010636819A CN 111662849 A CN111662849 A CN 111662849A
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- 241000278457 Bacillus siamensis Species 0.000 title claims abstract description 17
- 241000193830 Bacillus <bacterium> Species 0.000 claims abstract description 65
- 241000255789 Bombyx mori Species 0.000 claims abstract description 56
- 230000002265 prevention Effects 0.000 claims abstract description 9
- 238000004321 preservation Methods 0.000 claims abstract description 6
- 239000001963 growth medium Substances 0.000 claims description 28
- 239000007788 liquid Substances 0.000 claims description 28
- 230000001580 bacterial effect Effects 0.000 claims description 26
- 239000000243 solution Substances 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 238000012258 culturing Methods 0.000 claims description 11
- 240000000249 Morus alba Species 0.000 claims description 9
- 235000008708 Morus alba Nutrition 0.000 claims description 9
- 230000000644 propagated effect Effects 0.000 claims description 8
- 238000010790 dilution Methods 0.000 claims description 6
- 239000012895 dilution Substances 0.000 claims description 6
- 238000002360 preparation method Methods 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 5
- 238000005507 spraying Methods 0.000 claims description 3
- 238000011081 inoculation Methods 0.000 claims description 2
- 241000751139 Beauveria bassiana Species 0.000 abstract description 11
- 241000238675 Periplaneta americana Species 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 5
- 244000005700 microbiome Species 0.000 abstract description 3
- 239000004382 Amylase Substances 0.000 abstract description 2
- 102000013142 Amylases Human genes 0.000 abstract description 2
- 108010065511 Amylases Proteins 0.000 abstract description 2
- 102000004882 Lipase Human genes 0.000 abstract description 2
- 239000004367 Lipase Substances 0.000 abstract description 2
- 108090001060 Lipase Proteins 0.000 abstract description 2
- 108091005804 Peptidases Proteins 0.000 abstract description 2
- 239000004365 Protease Substances 0.000 abstract description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract description 2
- 235000019418 amylase Nutrition 0.000 abstract description 2
- 230000003042 antagnostic effect Effects 0.000 abstract description 2
- 235000019421 lipase Nutrition 0.000 abstract description 2
- 235000019419 proteases Nutrition 0.000 abstract description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 201000010099 disease Diseases 0.000 description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 9
- 239000002609 medium Substances 0.000 description 9
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- 108010080698 Peptones Proteins 0.000 description 6
- 235000019319 peptone Nutrition 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 229920001817 Agar Polymers 0.000 description 5
- 239000008272 agar Substances 0.000 description 5
- 208000015181 infectious disease Diseases 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 230000001954 sterilising effect Effects 0.000 description 5
- 235000015278 beef Nutrition 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical group [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
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- 230000031700 light absorption Effects 0.000 description 3
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- 238000007865 diluting Methods 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 208000031888 Mycoses Diseases 0.000 description 1
- 206010048038 Wound infection Diseases 0.000 description 1
- 238000003975 animal breeding Methods 0.000 description 1
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- 238000011156 evaluation Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
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- 239000000725 suspension Substances 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention provides Siamese Bacillus (Bacillus siamensis) MC2-1, wherein the preservation number is CGMCC No.15974, and the preservation date is 6 months and 20 days in 2018. The Siamese Bacillus MC2-1 Bacillus siamensis CGMCC No.15974 has the activities of protease, amylase and lipase and is separated from the Periplaneta americana alimentary canal. The strain has the function of remarkably antagonizing Beauveria bassiana, is safe to silkworm larvae, and is a silkworm Beauveria bassiana prevention microorganism with application prospect.
Description
Technical Field
The invention relates to the technical field of biology, in particular to a Siamese bacillus and application thereof.
Background
Domestic silkworm breeding has a history of more than 5000 years, and is an important model for domestic economic animal breeding, and in recent years, the silkworm cocoon yield of China accounts for about 75% of the global total yield, and the first cocoon silk production country in the world is kept. Beauveria bassiana is one of silkworm fungal diseases, is a disease caused by the fact that Beauveria bassiana (Beauveria bassiana) invades the body of a silkworm through the skin, and if the disease is not properly treated, the prevalence of the Beauveria bassiana in a silkworm chamber can be caused, the yield of silkworm cocoons is seriously influenced, and even the silkworm cocoons are completely harvested. The infection routes of silkworm white muscardine diseases include contact infection and wound infection, wherein the contact infection is the main infection. After the white muscardine silkworm dies, silkworm bodies are softened and then hardened, the silkworm bodies are full of hyphae, and 1-2 days later, the hyphae in the silkworm bodies penetrate through the body wall, white aerial hyphae grow on the body surface, white powdery conidiospores gradually grow on the aerial hyphae and cover the body surface of the whole silkworm body as if the surfaces of the silkworm bodies are full of white hair or white powder, so the silkworm is called white muscardine silkworm. Beauveria bassiana is wet and dry, and in humid environment and rainy season, if the disinfection and disease prevention work is neglected in the silkworm period, the silkworm is easy to suffer from the Beauveria bassiana.
At present, the prevention and control of silkworm white muscardine diseases mainly comprises thorough disinfection, ventilation and mass use of disinfectants, but the method is time-consuming and labor-consuming and is difficult to completely control, once part of silkworms suffer from white muscardine diseases, if the treatment is not timely, the population epidemic is caused, and in the treatment process, if the diseased silkworms are picked up by hands and then the mulberry leaves, silkworm breeding instruments or the mulberry leaves are contacted, new silkworm infection is caused.
Therefore, the research and development of a simple, convenient and low-cost method for preventing and controlling the white muscardine silkworm diseases is a technical problem which needs to be solved urgently by the personnel in the field.
Disclosure of Invention
In view of the above, the invention provides a bacillus siamensis and application thereof.
In order to achieve the purpose, the invention adopts the following technical scheme:
a Siamese Bacillus (Bacillus siamensis) MC2-1 with the preservation number of CGMCC No.15974, which is preserved in the China general microbiological culture Collection center of the China Committee for culture Collection of microorganisms, CGMCC for short, and the address: the microbial research institute of the national academy of sciences No. 3, Xilu No.1, Beijing, Chaoyang, with the preservation date of 2018, 6 months and 20 days, is named after classification: and (3) classification and naming: siamese Bacillus Bacillus siamensis.
The Siamese Bacillus (Bacillus siamensis) MC2-1 provided by the invention has the following biological characteristics:
the bacterial colony cultured by the LB Medium presents typical bacillus characteristics, the bacterial colony is circular or irregular, the diameter of the bacterial colony is 2-4 mm, and the edge is rough. The surface of the bacterial colony is rough and wrinkled and is white and opaque; can grow at 4-54 deg.C, and gram stain is positive.
Further, the Siamese bacillus is applied to prevention and control of silkworm white muscardine.
Further, the application of the Siamese bacillus in prevention and control of silkworm white muscardine comprises the following specific steps: the Siamese bacillus MC2-1 is prepared into Siamese bacillus MC2-1 bacterial liquid which is uniformly sprayed on the surface of a silkworm feeding appliance, the body surface of a silkworm and fed mulberry leaves.
Further, the Siamese bacillus MC2-1 bacterial liquid is uniformly sprayed on the grain surface until the surface is wet and no liquid drips.
Further, the preparation method of the Siamese bacillus MC2-1 bacterial liquid comprises the following steps:
(1) selecting a standard solid LB culture medium, adjusting the pH value to 5.5-8.5, after a plate culture dish is cooled to room temperature, inoculating a strain Siamese bacillus MC2-1, and culturing for 1-3 days at the temperature of 25-35 ℃ to obtain a propagated Siamese bacillus MC2-1 colony;
(2) selecting a standard liquid LB culture medium, adjusting the pH value to 5.5-8.5, inoculating the expanded and propagated Siamese bacillus MC2-1 colony obtained in the step (1), and culturing for 1-3 days on a shaking table with the rotating speed of 220rpm under the condition that the temperature is 25-35 ℃ to obtain a culture solution;
(3) and (3) standing the culture solution obtained in the step (2) to normal temperature, and then adding water for dilution to obtain Siamese bacillus MC2-1 bacterial solution.
Further, the pH regulator used for regulating the pH value is NaOH solution.
Further, the inoculation ratio in the step (2) is as follows: inoculating 1cm of standard liquid LB culture medium per 1-1.5L2And (2) expanding the propagated Siamese bacillus MC2-1 colony.
Further, the volume ratio of the culture solution to water in the step (3) is 1: 100-10000.
The invention has the beneficial effects that: the Siamese Bacillus MC2-1 Bacillus siamensis CGMCC No.15974 has the activities of protease, amylase and lipase and is separated from the Periplaneta americana Periplaneta americana digestive tract. The strain has the function of remarkably antagonizing Beauveria bassiana, is safe to silkworm larvae, and is a silkworm Beauveria bassiana prevention microorganism with application prospect.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The periplaneta americana is a strain raised in a laboratory for the pathogenic organisms and the insect-borne diseases of the first college of medical science in Shandong, and is continuously subcultured in the laboratory for 15 years. When Siamese Bacillus siemensis is separated, firstly the body surface of the periplaneta americana adult is rinsed 3 times by 75% alcohol, 2 minutes each time, and then is rinsed 3 times by sterile water and then is dissected to obtain a complete periplaneta americana digestive tract.
1. Sterilization
The experimental devices such as the culture dish, the centrifuge tube, the test tube, the gun head and the like and the sterile water are sterilized by high pressure for 30 minutes at the temperature of 121 ℃ under the pressure of 0.1 MPa.
2. Grinding
The periplaneta americana digestive tract is cut into pieces and then put into a sterile centrifuge tube, 1ml of sterile water is added, and a grinding rod is used for full grinding.
3. Gradient dilution
Taking 1ml of the ground juice, and placing into a test tube containing 9ml of sterile water to obtain 10-1Diluted bacterial liquid, from 10-1Taking 1ml of the diluted bacterial liquid, and putting the diluted bacterial liquid into a test tube filled with 9ml of sterile water to obtain 10-2Diluting the bacterial liquid, and repeating the steps to obtain 10-3、10-4、10-5、10-6、10-7And (4) diluted bacteria liquid.
4. Preparation of culture Medium
A medium commonly used for isolation of bacteria is beef extract peptone medium (NA medium), and LB medium commonly used for purification.
NA medium: 3g of beef extract, 10g of peptone, 5g of NaCl, 18g of agar and 1000ml of water, wherein the pH value is 7.0-7.2, and the beef extract is sterilized at 121 ℃ for 20 min.
LB culture medium: 10g of peptone; 5g of yeast extract; 10g of sodium chloride; 15g of agar; 1L of distilled water; sterilizing at 121 deg.C for 20min at pH 7.0.
Pour plate, pour about 15ml of medium per petri dish, lay flat and stand, wait to cool for use.
5. Coating of
Diluting the bacterial liquid 10-4、10-5、10-6、10-7The plates were coated separately, 3 dishes per gradient and marked on the bottom of the dish.
6. Culturing
The plates were inverted and incubated in an incubator at 28 ℃ for 3-5 days.
7. Purification of bacteria
Picking single colony growing on the NA culture medium by using a picking needle under the aseptic condition to perform streak culture on an LB culture medium; the single colony after purification can be obtained after 3-4 times of purification.
Example 2
Research on culture conditions of Siam bacillus MC2-1
1. Temperature of
Inoculating Siamese bacillus into LB culture medium with pH 7.0, respectively at 15 deg.C, 25 deg.C, 30 deg.C, 35 deg.C and 40 deg.C, and rotating at 22 deg.CShaking culture at 0r/min, sampling at 24h to determine light absorption value (OD) at 600nm600). See Table 1 below for a comparison of the OD of Siamese Bacillus cultures at various temperature conditions600The value can be known that the optimum culture temperature of Siamese Bacillus Bacillus Siamensis MC2-1 is 35 ℃, and the more suitable growth range is between 30 ℃ and 35 ℃.
TABLE 1 Effect of temperature on growth of Siamese Bacillus
| Temperature of | OD600 |
| 15℃ | 0.99±0.04de |
| 20℃ | 1.11±0.12d |
| 25℃ | 1.38±0.07c |
| 30℃ | 1.52±0.04ab |
| 35℃ | 1.77±0.09a |
| 40℃ | 1.26±0.06cd |
pH value
Inoculating Siamese bacillus into LB culture medium with pH of 5.5, 6, 6.5, 7, 7.5, 8 and 8.5 respectively, and shaking table (rotating speed is220r/min at 25 ℃ and a sample was taken at 24h to determine the light absorption at a wavelength of 600nm (OD 600). See Table 2 for comparison of OD of Siamese Bacillus cultures at various pH conditions600It is known that Siamese Bacillus grew better in the pH range of 5.5-8.5, with the best growth at pH 7.0.
TABLE 2 influence of pH on growth of Siamese Bacillus
3. Culture medium
Siamese bacillus is respectively inoculated in LB culture medium, TSB culture medium and NA culture medium with pH 7.0, cultured in a shaking table (the rotating speed is 220r/min, the temperature is 25 ℃), and sampled at 24h to determine the light absorption value (OD600) at the wavelength of 600 nm. As can be seen from Table 3, both LB and TSB were suitable for Siamese Bacillus growth in all three media.
TABLE 3 Effect of culture Medium on growth of Siamese Bacillus
| Culture medium | OD600 |
| LB | 1.82±0.07a |
| TSB | 1.61±0.14a |
| NA | 1.44±0.12b |
Note: LB culture medium: 10g of peptone; 5g of yeast extract; 10g of sodium chloride; 15g of agar; 1L of distilled water; sterilizing at 121 deg.C for 20 min.
TSB medium: tryptone 15 g; 5g of soybean peptone; 5g of sodium chloride; 15g of agar; 1L of distilled water; sterilizing at 121 deg.C for 20 min.
NA medium: 3g of beef extract, 10g of peptone, 5g of NaCl, 18g of agar and 1000ml of water, and sterilizing at 121 ℃ for 20 min.
Example 3
The preparation method of the Siamese bacillus MC2-1 bacterial liquid specifically comprises the following steps:
(1) selecting a standard solid LB culture medium, adjusting the pH value to 5.5, inoculating a strain Siamese bacillus MC2-1 after a plate culture dish is cooled to room temperature, and culturing for 1 day at the temperature of 25 ℃ to obtain a propagated Siamese bacillus MC2-1 colony;
(2) selecting standard liquid LB culture medium, adjusting pH to 5.5, inoculating 1cm of standard liquid LB culture medium per 1L2Culturing the Siamese bacillus MC2-1 colony expanded in the step (1) on a shaking table at the rotating speed of 220rpm for 1 day at the temperature of 25 ℃ to obtain a culture solution;
(3) and (3) standing the culture solution obtained in the step (2) to normal temperature, and then adding water for dilution, wherein the volume ratio of the culture solution to the water is 1:100, so as to obtain Siamese bacillus MC2-1 bacterial solution.
Example 4
The preparation method of the Siamese bacillus MC2-1 bacterial liquid specifically comprises the following steps:
(1) selecting a standard solid LB culture medium, adjusting the pH value to 8.5, inoculating a strain Siamese bacillus MC2-1 after a plate culture dish is cooled to room temperature, and culturing for 3 days at the temperature of 35 ℃ to obtain a propagated Siamese bacillus MC2-1 colony;
(2) selecting standard liquid LB culture medium, adjusting pH to 8.5, inoculating 1cm of standard liquid LB culture medium per 1.5L2Culturing the Siamese bacillus MC2-1 colony expanded in the step (1) on a shaking table at the rotating speed of 220rpm for 3 days at the temperature of 35 ℃ to obtain a culture solution;
(3) and (3) standing the culture solution obtained in the step (2) to normal temperature, and then adding water for dilution, wherein the volume ratio of the culture solution to the water is 1:1000, so as to obtain Siamese bacillus MC2-1 bacterial solution.
Example 5
The preparation method of the Siamese bacillus MC2-1 bacterial liquid specifically comprises the following steps:
(1) selecting a standard solid LB culture medium, adjusting the pH value to 7, inoculating a strain Siamese bacillus MC2-1 after a plate culture dish is cooled to room temperature, and culturing for 2 days at the temperature of 30 ℃ to obtain a Siamese bacillus MC2-1 bacterial colony after propagation;
(2) selecting standard liquid LB culture medium, adjusting pH to 7, inoculating 1cm of standard liquid LB culture medium per 1L2Culturing the Siamese bacillus MC2-1 colony expanded in the step (1) on a shaking table at the rotating speed of 220rpm for 2 days at the temperature of 30 ℃ to obtain a culture solution;
(3) and (3) standing the culture solution obtained in the step (2) to normal temperature, adding water for dilution, wherein the volume ratio of the culture solution to the water is 1:10000, and obtaining Siamese bacillus MC2-1 bacterial liquid.
Example 6
Test of prevention and treatment effect of Siamese bacillus MC2-1 on white muscardine disease of silkworm
(1) Spraying conidia of Beauveria bassiana on the surface of 5-year-old silkworm body, wherein the concentration is 1010The white muscardine fungus spore suspension is sprayed on silkworm bodies to infect silkworms, and the test is carried out after the silkworm bodies are slightly aired;
the silkworms sprayed with sterile water were used as controls, and 3 boxes of silkworms were treated in each of the treatment group and the control group.
Respectively spraying the Siamese bacillus MC2-1 bacterial liquid obtained in the embodiment 3-5 on the surfaces of mulberry leaves, and airing for later use; control was made with leaves sprayed with sterile water.
Feeding infected silkworms and control silkworms with Siamese bacillus MC2-1 bacterial solution treated mulberry leaves and control mulberry leaves respectively until the silkworms and the control silkworms are raised to a mounting position,
counting the incidence of silkworm white muscardine of the test group and the control group; the safety of Siamese bacillus MC2-1 to silkworms is measured, 5-year-old silkworms are fed by the mulberry leaves sprayed with Siamese bacillus MC2-1 fermentation liquor, the cage death rate of the silkworms is examined, the cocoon yield of box seeds, the cocoon number of hundreds of kilograms of mulberry leaves and the number of kilograms of cocoon particles are examined, and the safety of Siamese bacillus MC2-1 to the silkworms is evaluated.
TABLE 4 safety evaluation of Bombyx mori by Siamese Bacillus MC2-1
Note: the letters following the numbers in the same column represent no significant difference between the data.
Research shows that Siamese bacillus is very safe to 5-year-old silkworms, and biological indexes and cocoon yield of the Siamese bacillus have no obvious difference compared with those of a control, and the Siamese bacillus can be used for preventing and controlling white muscardine diseases of the silkworms as shown in a table 4.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (6)
1. A Siamese Bacillus is characterized in that the Siamese Bacillus is Bacillus siamensis MC2-1, the preservation number is CGMCC No.15974, and the preservation date is 2018, 6 and 20 days.
2. The use of bacillus siamensis according to claim 1 in prevention and control of silkworm white muscardine.
3. The application of bacillus siamensis in prevention and control of silkworm white muscardine according to claim 2, which comprises the following steps: and preparing Siamese bacillus MC2-1 into Siamese bacillus MC2-1 bacterial liquid, and uniformly spraying the Siamese bacillus MC2-1 bacterial liquid onto the surface of a silkworm feeding appliance, the body surface of a silkworm and fed mulberry leaves.
4. The application of bacillus siamensis in prevention and control of silkworm white muscardine according to claim 3, wherein the preparation method of the bacillus siamensis MC2-1 bacterial liquid comprises the following steps:
(1) selecting a standard solid LB culture medium, adjusting the pH value to 5.5-8.5, after a plate culture dish is cooled to room temperature, inoculating a strain Siamese bacillus MC2-1, and culturing for 1-3 days at the temperature of 25-35 ℃ to obtain a propagated Siamese bacillus MC2-1 colony;
(2) selecting a standard liquid LB culture medium, adjusting the pH value to 5.5-8.5, inoculating the expanded and propagated Siamese bacillus MC2-1 colony obtained in the step (1), and culturing for 1-3 days on a shaking table with the rotating speed of 220rpm under the condition that the temperature is 25-35 ℃ to obtain a culture solution;
(3) and (3) standing the culture solution obtained in the step (2) to normal temperature, and then adding water for dilution to obtain Siamese bacillus MC2-1 bacterial solution.
5. The use of bacillus siamensis MC2-1 as claimed in claim 4 for preventing grain mildew, wherein the inoculation ratio in step (2) is: inoculating 1cm of standard liquid LB culture medium per 1-1.5L2And (2) expanding the propagated Siamese bacillus MC2-1 colony.
6. The use of Bacillus siamensis MC2-1 in preventing grain mildew as claimed in claim 4, wherein the volume ratio of the culture solution to water in step (3) is 1: 100-10000.
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| CN112410261A (en) * | 2020-12-03 | 2021-02-26 | 云南省烟草公司昆明市公司 | Bacillus siamensis MC2-1 and application thereof |
| CN116064280A (en) * | 2022-08-12 | 2023-05-05 | 大连工业大学 | Siamese bacillus and application thereof |
| CN116064279A (en) * | 2022-08-12 | 2023-05-05 | 大连工业大学 | Siamese bacillus and application thereof in low-salt fermentation of food |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA3039308A1 (en) * | 2016-10-07 | 2018-04-12 | Idemitsu Kosan Co., Ltd. | Method for culturing bacillus bacterium, and method for producing useful substance |
| CN109266570A (en) * | 2018-08-21 | 2019-01-25 | 山东省花生研究所 | A kind of microorganism formulation of antagonism aspergillus flavus growth and application |
-
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA3039308A1 (en) * | 2016-10-07 | 2018-04-12 | Idemitsu Kosan Co., Ltd. | Method for culturing bacillus bacterium, and method for producing useful substance |
| CN109266570A (en) * | 2018-08-21 | 2019-01-25 | 山东省花生研究所 | A kind of microorganism formulation of antagonism aspergillus flavus growth and application |
Non-Patent Citations (1)
| Title |
|---|
| 林志楷等: "暹罗芽孢杆菌研究进展", 《亚热带植物科学》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112410261A (en) * | 2020-12-03 | 2021-02-26 | 云南省烟草公司昆明市公司 | Bacillus siamensis MC2-1 and application thereof |
| CN116064280A (en) * | 2022-08-12 | 2023-05-05 | 大连工业大学 | Siamese bacillus and application thereof |
| CN116064279A (en) * | 2022-08-12 | 2023-05-05 | 大连工业大学 | Siamese bacillus and application thereof in low-salt fermentation of food |
| CN116064279B (en) * | 2022-08-12 | 2024-04-02 | 大连工业大学 | Siamese bacillus and application thereof in low-salt fermentation of food |
| CN116064280B (en) * | 2022-08-12 | 2024-05-07 | 大连工业大学 | Siamese bacillus and application thereof |
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