CN113508821A - 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 PDF

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CN113508821A
CN113508821A CN202110642215.3A CN202110642215A CN113508821A CN 113508821 A CN113508821 A CN 113508821A CN 202110642215 A CN202110642215 A CN 202110642215A CN 113508821 A CN113508821 A CN 113508821A
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bacillus amyloliquefaciens
preparation
bacterial diseases
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preventing
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CN113508821B (en
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仇俊涛
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Yunnan Xingyao Biological Products Co ltd
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    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION 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
    • A01N63/00Biocides, 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
    • A01N63/20Bacteria; Substances produced thereby or obtained therefrom
    • A01N63/22Bacillus
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION 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
    • A01N25/00Biocides, 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
    • A01N25/12Powders or granules
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION 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
    • A01N43/00Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
    • A01N43/02Biocides, 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
    • A01N43/04Biocides, 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
    • A01N43/14Biocides, 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
    • A01N43/16Biocides, 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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  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Dentistry (AREA)
  • Plant Pathology (AREA)
  • Pest Control & Pesticides (AREA)
  • Agronomy & Crop Science (AREA)
  • Environmental Sciences (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Virology (AREA)
  • Toxicology (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention relates to a bacillus amyloliquefaciens preparation and an 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

Bacillus amyloliquefaciens preparation and application thereof in preventing and treating bacterial diseases
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, ulcer, bacterial wilt and the like. Bacteria which attack plants are bacillary bacteria, most of which have one to several flagella, can invade through natural orifices (stomata, skin pores, water pores, and the like) and wounds, are spread by running water, rainwater, insects, and the like, live through winter in disease residues, seeds, and soil, and are easy to cause diseases 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. But the frequent use of chemical pesticides and the characteristics of quick propagation genetic variation and high fitness of bacteria lead pathogenic bacteria to quickly generate resistance, and influence 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 with the 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 staff in the field is still how to ensure that the mixed preparation has no side effect on 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. Usually 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 to 2; preferably 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 × 108cfu/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 present 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 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 involved in the examples of the present invention are all commercially available products, and all of them are commercially available.
Example 1
Taking Bacillus amyloliquefaciens (strain preservation number: CICC 10081) dry powder (content is 8 x 10)8cfu/g)10 parts by weight and 1 part 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 in the test sample after 3 months is 7 multiplied by 108cfu/g, mortality was 3.7%. The mortality rate after 3 months storage of the bacillus amyloliquefaciens dry powder alone was 15%. Therefore, the spirulina polysaccharide has no obvious influence on the activity of the bacillus amyloliquefaciens and also has a certain protective effect.
Example 2:
taking Bacillus amyloliquefaciens dry powder (with the content of 8 multiplied by 10)8cfu/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 in the test sample after 3 months is 6 multiplied by 108cfu/g, the mortality rate is 10%, thereby showing that the excessive spirulina polysaccharide has certain influence on the activity of the bacillus amyloliquefaciens, but still has certain protection effect.
Example 3:
taking Bacillus amyloliquefaciens dry powder (with the content of 8 multiplied by 10)8cfu/g)10 parts by weight and 0.5 part 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 in the test sample after 3 months is 7 multiplied by 108cfu/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 preparation described in examples 1-3 and a control Bacillus amyloliquefaciens dry powder preparation were diluted 1000 times with sterile water and then spray treated (2 times with 3d intervals) to the rice plants described in step (1) at room temperature of 30 ℃ and relative humidity of about 90%, and then subjected to moist culture. When the area of the diseased leaves in the blank control group is about 1/2%, the disease 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
Figure BDA0003108398240000041
Figure BDA0003108398240000051
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 (6)

1. The application of a bacillus amyloliquefaciens preparation in preventing and treating bacterial diseases is disclosed, wherein the bacillus amyloliquefaciens preparation contains bacillus amyloliquefaciens and spirulina polysaccharide.
2. The use of claim 1, wherein the weight ratio of bacillus amyloliquefaciens to spirulina polysaccharide is 10: 0.5 to 2; preferably 10: 0.8-1.2.
3. The use according to claim 2, wherein the bacillus amyloliquefaciens strain is deposited under the accession number: bacillus amyloliquefaciens strain CICC10081 with content of 16-22 × 108cfu/g formulation.
4. A Bacillus amyloliquefaciens preparation for controlling bacterial diseases, which comprises the following components in a weight ratio of 10: 0.5-2 of bacillus amyloliquefaciens and spirulina polysaccharide; the weight ratio is preferably 10: 0.8-1.2.
5. The formulation of claim 4, wherein the Bacillus amyloliquefaciens formulation is a dry powder formulation.
6. The formulation of claim 5, wherein the dry powder formulation is vacuum packaged and stored at ambient temperature.
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