CN113508821B - Bacillus amyloliquefaciens preparation and application thereof in preventing and treating bacterial diseases - Google Patents
Bacillus amyloliquefaciens preparation and application thereof in preventing and treating bacterial diseases Download PDFInfo
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- CN113508821B CN113508821B CN202110642215.3A CN202110642215A CN113508821B CN 113508821 B CN113508821 B CN 113508821B CN 202110642215 A CN202110642215 A CN 202110642215A CN 113508821 B CN113508821 B CN 113508821B
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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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- 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
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/12—Powders or granules
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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
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
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Abstract
The invention relates to a bacillus amyloliquefaciens preparation and application thereof in preventing and treating bacterial diseases. The invention firstly proposes that the spirulina polysaccharide and the bacillus amyloliquefaciens preparation are mixed and applied, not only does not produce side effect on the bacillus amyloliquefaciens preparation, but also can effectively improve the prevention and treatment effect on bacterial diseases and increase the yield and income.
Description
Technical Field
The invention relates to the technical field of biological control, in particular to a bacillus amyloliquefaciens preparation and application thereof in controlling bacterial diseases.
Background
Bacterial diseases are diseases caused by bacterial pathogen infection, such as soft rot, canker, bacterial wilt and the like. Bacteria invading plants are bacilli, most of which have one or more flagella, can invade through natural orifices (stomata, skin pores, water pores, etc.) and wounds, are spread by running water, rainwater, insects, etc., pass winter in disease residues, seeds, soil, and are prone to attack under high temperature and high humidity conditions.
The mode of preventing and controlling bacterial diseases still mainly adopts disease-resistant breeding and takes chemical prevention and control as assistance. Wherein breeding is limited due to long period and high input cost; chemical control is quick, cheap and efficient, but the existing effective agents for bacterial diseases mainly comprise: copper preparation, zinc preparation, antibiotics, benziothiazolinone and other few pesticides. However, the frequent use of chemical pesticides and the characteristic of quick propagation genetic variation and high suitability of bacteria lead pathogenic bacteria to quickly generate resistance, which affects the control effect of medicaments. The biological control method utilizing the biocontrol bacteria can effectively solve and avoid the resistance problem of the medicament, has high efficiency, no toxicity, no harm, no pollution and no drug resistance, not only meets the requirements of people on green agricultural products, but also provides guarantee for the sustainable development of agriculture, so the research on the biological control of plant bacterial diseases is more and more emphasized.
Bacillus spp is ubiquitous in soil, is one of dominant microbial populations existing in soil and plant micro-ecological systems, has strong capability of resisting external environment pressure due to the generation of endogenous spores, can resist damage caused by drying, heat and ultraviolet radiation in living environments, maintains the self viability unaffected, has very good application prospect due to biological characteristics, and particularly shows strong vitality in biological products using live bacterial preparations. The bacillus has various varieties and huge quantity, can produce various physiologically active substances and metabolites, and has wide industrial application prospect.
Chinese granted patent CN103087951B discloses a strain of Bacillus amyloliquefaciens and application thereof. The strain number of the bacillus amyloliquefaciens is MH71, and the registration number of the bacillus amyloliquefaciens in the common microorganism center of the China Committee for culture Collection of microorganisms is CGMCC No. 6978. Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) MH71 has obvious inhibiting effect on various pathogenic fungi such as botrytis cinerea, fusarium graminearum, brown rot, root rot and anthracnose on fruit and vegetable crops, and pathogenic bacteria such as Chinese cabbage black rot, oyster mushroom brown spot and cucumber angular leaf spot. Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) MH71 has high propagation speed, strong stress resistance and great significance for preventing and treating soil-borne diseases such as cabbage wilt and the like caused by fungi, and can be artificially cultured.
Chinese patent application CN112694998A discloses a Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) ZN-S2, which has a preservation number: CGMCCNO: 18962. the application of the Bacillus amyloliquefaciens ZN-S2 is also disclosed: is used for inhibiting the growth of plant pathogenic bacteria, thereby preventing and controlling plant bacterial diseases.
However, with the development of times and the increase of drug resistance, it has been difficult to achieve good effects of bacillus amyloliquefaciens alone for controlling bacterial diseases. Therefore, the skilled person also tries to combine bacillus amyloliquefaciens with other drugs to improve the control effect. Although the prior art mentions that the bacillus amyloliquefaciens preparation can be mixed with other fungicidal pesticides and plant growth regulators for use, the technical problem to be solved by the technical personnel still needs to be solved by how to ensure that the mixed preparation has no side effect on the bacillus amyloliquefaciens and can improve the control effect.
In earlier researches of the applicant, the spirulina polysaccharide and the bacillus subtilis preparation are mixed and applied, so that the bacillus subtilis preparation cannot generate side effects on the bacillus subtilis preparation, and the prevention and treatment effects on the clubroot of cruciferae and the yield increase can be effectively improved. However, spirulina polysaccharides have no substantial influence on the activity of bacillus amyloliquefaciens, and therefore, the applicant of the present invention further studies on bacillus amyloliquefaciens and finds that spirulina polysaccharides have a significant meaning for improving the activity thereof, and the present invention is achieved based on the finding.
Disclosure of Invention
Based on the background technology, the technical problem to be solved by the invention is to provide a bacillus amyloliquefaciens preparation and an application thereof in preventing and treating bacterial diseases. In order to realize the purpose of the invention, the following technical scheme is adopted:
the invention relates to an application of a bacillus amyloliquefaciens preparation in preventing and treating bacterial diseases, wherein the bacillus amyloliquefaciens preparation contains bacillus amyloliquefaciens and spirulina polysaccharide. Spirulina polysaccharides (Spirulina polysaccharides) are physiologically active polysaccharides extracted from Spirulina and are nontoxic. Generally colorless or pale yellow powder, soluble in water, insoluble in alcohol and fats. The invention firstly proposes that the spirulina polysaccharide and the bacillus amyloliquefaciens preparation are mixed and applied, so that the side effect on the bacillus amyloliquefaciens preparation can not be generated, the prevention and treatment effect on bacterial diseases can be effectively improved, and the yield and income can be increased.
In a preferred embodiment of the present invention, the weight ratio of bacillus amyloliquefaciens to spirulina polysaccharide is 10: 0.5-2; preferably, the ratio of 10: 0.8-1.2. The preparation of the invention is mixed with bacillus amyloliquefaciens and spirulina polysaccharide according to a specific proportion, and is beneficial to improving the prevention and treatment effect.
In a preferred embodiment of the invention, the bacillus amyloliquefaciens strain has the accession number: bacillus amyloliquefaciens strain CICC10081 with content of 16-22 × 10 8 cfu/g formulation.
In another aspect, the invention relates to a formulation for controlling bacterial diseases, said formulation comprising, by weight, 10: 0.5-2 of bacillus amyloliquefaciens and spirulina polysaccharide; the weight ratio is preferably 10: 0.8-1.2. Preferably, the bacterial disease is bacterial blight of rice.
In a preferred embodiment of the invention, the bacillus amyloliquefaciens formulation is a dry powder formulation.
In a preferred embodiment of the invention, the dry powder formulation is vacuum packed and stored at ambient temperature.
Detailed Description
In order to further understand the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the 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.
Unless otherwise specified, the reagents used in the examples of the present invention are commercially available products, and all of them can be purchased from commercial sources.
Example 1
Taking Bacillus amyloliquefaciens (strain preservation number: CICC 10081) dry powder (content is 8 multiplied by 10) 8 cfu/g)10 parts by weight of the powder and 1 part by weight of spirulina polysaccharide powder are mixed evenly and placed in a vacuum sealing bag to be stored at normal temperature. The number of bacillus amyloliquefaciens in the test sample after 3 months is 7 multiplied by 10 8 cfu/g, mortality was 3.7%. The mortality rate after 3 months storage of the dry powder of bacillus amyloliquefaciens alone was 15%. Therefore, the spirulina polysaccharide has no obvious influence on the activity of the bacillus amyloliquefaciens and also has a certain protection effect.
Example 2:
taking Bacillus amyloliquefaciens dry powder (with the content of 8 multiplied by 10) 8 cfu/g)10 parts by weight and 2 parts by weight of spirulina polysaccharide powder are mixed evenly and put into a vacuum sealing bag for storage at normal temperature. The number of bacillus amyloliquefaciens detected in the test solution after 3 months is 6 multiplied by 10 8 cfu/g, the mortality rate was 10%, thus indicating that an excess of spirulina polysaccharide has a certain effect on the activity of bacillus amyloliquefaciens, but still has a certain protective effect.
Example 3:
taking Bacillus amyloliquefaciens dry powder (with the content of 8 multiplied by 10) 8 cfu/g)10 parts by weight and 0.5 part by weight of spirulina polysaccharide powder are mixed evenly and placed in a vacuum sealing bag to be stored at normal temperature. The number of bacillus amyloliquefaciens in the test sample after 3 months is 7 multiplied by 10 8 cfu/g, mortality was 8.1%, thus indicating that the amount of spirulina polysaccharide has a substantial effect on the protective effect of bacillus amyloliquefaciens.
Example 4: evaluation of Effect of spray treatment of Stem and leaf of Bacillus amyloliquefaciens preparation on controlling bacterial leaf blight of rice
(1) Preparation of potted rice seedlings: sterilizing the surface of the rice seeds, and then placing the rice seeds in an artificial climate box at the temperature of 30 ℃ for sterile water soaking and germination acceleration for 3 d; simultaneously sterilizing the cultivation soil by high pressure steam (121 ℃, 110KPa) for 50min, placing in an incubator at 25 ℃ for 24h, then repeating sterilization once, cooling the soil, then potting, sowing the germinated rice seeds in pots with the length and width of 20cm multiplied by 15cm, sowing 20 seeds in each pot, and supplying the rice seedlings for test when the rice seedlings grow to 4-leaf stage.
(2) And (3) inoculation treatment: when the rice seedlings grow to the 4-leaf stage, inoculating bacterial blight original bacteria of rice, and keeping moisture for 24 h. The preparations described in examples 1 to 3 and a control Bacillus amyloliquefaciens dry powder preparation were diluted 1000 times with sterile water and then spray treated (2 times with 3d intervals) with the rice plants described in step (1) at room temperature of 30 ℃ and relative humidity of about 90%, and were subjected to moist culture. When the area of the diseased leaves in the blank control group is about 1/2%, the diseased leaf area of the leaves is measured, and the control effect is calculated, wherein the control effect is (control lesion length-treatment lesion length)/control lesion length multiplied by 100%.
(3) The results of the experiment are shown in table 1. Experimental results show that the control effect of the bacillus amyloliquefaciens preparation on bacterial diseases is obviously better than that of a blank control group and a positive control group, wherein the preparation of the embodiment 1 has the best effect.
TABLE 1 prevention and treatment of bacterial diseases with Bacillus amyloliquefaciens preparation
The foregoing describes preferred embodiments of the present invention, but is not intended to limit the invention thereto. Modifications and variations of the embodiments disclosed herein may be made by those skilled in the art without departing from the scope and spirit of the invention.
Claims (4)
1. The application of the bacillus amyloliquefaciens preparation in the prevention and treatment of bacterial diseases is characterized in that the bacillus amyloliquefaciens preparation contains bacillus amyloliquefaciens and spirulina polysaccharide, and the weight ratio of the bacillus amyloliquefaciens to the spirulina polysaccharide is (10): 0.5-2; the bacillus amyloliquefaciens is a strain preservation number: bacillus amyloliquefaciens strain CICC10081, its content is 8X 10 8 cfu/g。
2. The use of claim 1, wherein the weight ratio of bacillus amyloliquefaciens to spirulina polysaccharide is 10: 0.8-1.2.
3. The use according to claim 1 or 2, wherein the bacillus amyloliquefaciens formulation is a dry powder formulation.
4. The use according to claim 3, wherein the dry powder formulation is vacuum packed and stored at ambient temperature.
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| CN113508821B true CN113508821B (en) | 2022-07-26 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| EP1306008A1 (en) * | 2001-10-24 | 2003-05-02 | Natural Bio Technology Co. | Composition for treatment against mosquito larvae and process for its preparation |
| CN105707072B (en) * | 2014-12-03 | 2018-08-21 | 中国科学院大连化学物理研究所 | A kind of spirulina polysaccharide and its application |
| CN105294870B (en) * | 2015-06-09 | 2017-07-25 | 深圳海王药业有限公司 | A kind of spirulina polysaccharide and preparation method thereof |
| WO2017205800A1 (en) * | 2016-05-27 | 2017-11-30 | Agro Research International, LLC | Biopesticide composition for use preventing or minimizing plant disease |
| EP3565413A1 (en) * | 2017-01-03 | 2019-11-13 | Université de Mons | Activators of plant metabolic changes |
| CN107375982B (en) * | 2017-07-17 | 2020-07-03 | 王婧宁 | Air freshener suitable for northern houses |
| CN108300676A (en) * | 2018-02-10 | 2018-07-20 | 厦门昶青生物科技有限公司 | A kind of red seaweed polysaccharide composite bacteria agent and its preparation method and application |
| CN110106163B (en) * | 2019-06-19 | 2021-02-19 | 西南大学 | Application and method of fucoidin and hydrolyzed oligosaccharide thereof in preparation of probiotic protective agent |
| CN111248196B (en) * | 2020-03-26 | 2021-04-23 | 南京工业大学 | Cell membrane bionic agricultural microbial agent microcapsule and preparation method thereof |
| CN112694998A (en) * | 2021-01-19 | 2021-04-23 | 浙江农林大学 | Biocontrol plant endophytic bacterium ZN-S2 and application thereof |
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