CN111979220A - Carrier molecular chaperone for activating microbial preparation - Google Patents
Carrier molecular chaperone for activating microbial preparation Download PDFInfo
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- CN111979220A CN111979220A CN201910441748.8A CN201910441748A CN111979220A CN 111979220 A CN111979220 A CN 111979220A CN 201910441748 A CN201910441748 A CN 201910441748A CN 111979220 A CN111979220 A CN 111979220A
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- microbial
- bacillus
- activating
- starch
- acid
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- ZLGIYFNHBLSMPS-ATJNOEHPSA-N shellac Chemical compound OCCCCCC(O)C(O)CCCCCCCC(O)=O.C1C23[C@H](C(O)=O)CCC2[C@](C)(CO)[C@@H]1C(C(O)=O)=C[C@@H]3O ZLGIYFNHBLSMPS-ATJNOEHPSA-N 0.000 claims description 2
- 229940113147 shellac Drugs 0.000 claims description 2
- 235000013874 shellac Nutrition 0.000 claims description 2
- 235000010413 sodium alginate Nutrition 0.000 claims description 2
- 239000000661 sodium alginate Substances 0.000 claims description 2
- 229940005550 sodium alginate Drugs 0.000 claims description 2
- 239000000600 sorbitol Substances 0.000 claims description 2
- 229940032147 starch Drugs 0.000 claims description 2
- 239000004575 stone Substances 0.000 claims description 2
- 238000005728 strengthening Methods 0.000 claims description 2
- 239000005720 sucrose Substances 0.000 claims description 2
- 239000000375 suspending agent Substances 0.000 claims description 2
- 239000000811 xylitol Substances 0.000 claims description 2
- 235000010447 xylitol Nutrition 0.000 claims description 2
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 claims description 2
- 229960002675 xylitol Drugs 0.000 claims description 2
- 244000215068 Acacia senegal Species 0.000 claims 1
- 241000192700 Cyanobacteria Species 0.000 claims 1
- 229920000084 Gum arabic Polymers 0.000 claims 1
- 235000010489 acacia gum Nutrition 0.000 claims 1
- 239000000205 acacia gum Substances 0.000 claims 1
- PZZYQPZGQPZBDN-UHFFFAOYSA-N aluminium silicate Chemical compound O=[Al]O[Si](=O)O[Al]=O PZZYQPZGQPZBDN-UHFFFAOYSA-N 0.000 claims 1
- 235000009697 arginine Nutrition 0.000 claims 1
- 239000000560 biocompatible material Substances 0.000 claims 1
- -1 but not limited to Substances 0.000 claims 1
- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical class CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims 1
- 238000009472 formulation Methods 0.000 claims 1
- 235000018977 lysine Nutrition 0.000 claims 1
- 235000006109 methionine Nutrition 0.000 claims 1
- 229940066779 peptones Drugs 0.000 claims 1
- 230000008121 plant development Effects 0.000 claims 1
- 238000007790 scraping Methods 0.000 claims 1
- 238000003860 storage Methods 0.000 abstract description 4
- 238000000034 method Methods 0.000 abstract description 3
- 239000002054 inoculum Substances 0.000 abstract 5
- 239000000575 pesticide Substances 0.000 description 10
- 238000011282 treatment Methods 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000003814 drug Substances 0.000 description 3
- 241000227653 Lycopersicon Species 0.000 description 2
- 238000012271 agricultural production Methods 0.000 description 2
- 230000000443 biocontrol Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 description 2
- 238000004806 packaging method and process Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 241000195493 Cryptophyta Species 0.000 description 1
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 1
- 240000007472 Leucaena leucocephala Species 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 241000589771 Ralstonia solanacearum Species 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- YKTSYUJCYHOUJP-UHFFFAOYSA-N [O--].[Al+3].[Al+3].[O-][Si]([O-])([O-])[O-] Chemical compound [O--].[Al+3].[Al+3].[O-][Si]([O-])([O-])[O-] YKTSYUJCYHOUJP-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000004562 water dispersible granule Substances 0.000 description 1
- 239000004563 wettable powder Substances 0.000 description 1
Classifications
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
-
- 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
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Molecular Biology (AREA)
- Environmental Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- Inorganic Chemistry (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
A carrier chaperone for activating microbial agents. The invention belongs to the technical field of biology, and relates to a method for activating a carrier molecular chaperone of a microbial inoculant to activate the microbial inoculant with short shelf life after long-time storage, so that the microbial inoculant can quickly grow and propagate on a carrier, the microbial inoculant content is increased, a dominant strain is formed, and the technical problem of long-time storage of the microbial inoculant, especially a pseudomonas fluorescens product with weak stress resistance, is solved.
Description
Technical Field
The present invention belongs to the field of biological pesticide and biochemical technology.
Background
The pesticide plays an important role in controlling agricultural pests, protecting agricultural production safety, reducing labor intensity and the like, but with the wide application of chemical pesticides, environmental problems and video safety problems caused by chemical pesticide residues are increasingly concerned. The search for new agricultural measures meeting the agricultural sustainable development needs becomes a hot spot of the current agricultural production research. The biocontrol bacteria and metabolites thereof can not only inhibit the growth of various pathogenic bacteria, but also promote the growth of plants and improve the yield of crops, and the development of biocontrol strains and effective antibacterial active ingredients thereof become the focus of research and competition at present.
The fatal defects of microbial pesticides are sensitivity to external conditions under natural conditions, easy death and short shelf life. The problems of short shelf life, easy death in nature, easy influence by environment and the like of microbial pesticides can be solved by improving the dosage form. In order to prolong the storage period, many microbial pesticides are optimized through fermentation to culture organisms with strong stress resistance such as spores, chlamydospores and the like, and then are prepared into wettable powder, microcapsule granules (ZL 201210419235.5), water dispersible granules and the like. The method increases the resistance of the microbial pesticide under natural conditions to a certain extent, but the shelf life of some microbial bacteria is still difficult to prolong because the microbial bacteria do not generate structures with stronger stress resistance such as spores or chlamydospores, for example, the shelf life of the pseudomonas fluorescens microbial inoculum widely used in the world is very short, and the national standard of the pseudomonas fluorescens shelf life established in our country is 6 months.
Disclosure of Invention
The carrier molecular chaperone for activating microbial preparation is prepared with one or several kinds of polysaccharide, protein, amino acid, proteinase, nucleic acid, fatty acid or their derivative and one or several kinds of mineral matters, and is used in activating the microbial activity of microbial preparation, increasing the microbial content, strengthening the effect of microbial preparation in regulating plant growth and preventing and controlling plant diseases and pests in water.
The molecular chaperone for activating microbial preparation is prepared by combining one or more of polysaccharide or protein or amino acid or protease or nucleic acid or fatty acid or their derivatives with one or more mineral carriers, and is characterized by comprising, but not limited to, milk powder, carboxymethyl starch, glyceryl monostearate, lysine, arginine, glutamic acid, aluminum silicate, methionine, S-adenosylmethionine, aspartic acid, glycine, methionine, histidine, polypeptide, peptone, aluminosilicate, trehalose, sucrose, sorbose, melezitose, glycerol, oligosaccharide, mannose, maltose, lactose, chitosan, lactic acid, calcium lactate, butyric acid, arabinose, xylose, ribose, rhamnose, magnesium silicate, glucose, mannitol, glycerol, xylitol, erythritol, threitol, sorbitol and raffinose, dextrin, dextran and soluble starch, more preferably mineral and chitosan.
The microbial preparation is characterized in that: the microbial preparation is prepared by adsorbing fermented microbial bacteria with an adsorbing material to prepare a capsule core, wrapping the capsule core with a wall material and directly preparing the capsule core into a suspending agent or preparing microcapsule powder and granules by drying.
The microorganism includes, but is not limited to, Bacillus subtilis, Pseudomonas fluorescens, Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacillus methylotrophicus, Bacillus cereus, Paenibacillus polymyxa, Bacillus hypothermia, Streptomyces hygroscopicus, Trichoderma, Verticillium, Bacillus firmus, Bacillus thuringiensis, Beauveria bassiana, Paecilomyces lilacinus, and the like, and more preferably Pseudomonas fluorescens.
The adsorbing material includes but is not limited to activated carbon, white carbon black, diatomite, medical stone, kaolin, bran shell, starch, porous starch, soluble starch, macroporous adsorbent and the like or the combination of the materials. The wall material is a material with good biocompatibility and comprises but is not limited to beta-cyclodextrin, shellac, gelatin, acacia, dextrin, starch, sodium alginate, calcium alginate and the like.
The plant diseases and insect pests include, but are not limited to, diseases caused by plant-harmful microorganisms such as nematodes, viruses, fungi or bacteria and insect pests caused by insects such as chewing mouthparts, piercing-sucking mouthparts, filing-sucking mouthparts, scraping-sucking mouthparts and feeding-sucking mouthparts. The water body pests are characterized by comprising but not limited to blue algae, dinoflagellate, moss and the like which pollute a water body.
The invention has the innovation point that the molecular chaperone of the carrier for activating the microbial preparation is creatively prepared, so that the microbial preparation with short shelf life can be stored for a long time, and the molecular chaperone is mixed with the microbial preparation before or during the use of the microbial preparation to activate the microbial bacteria by the molecular chaperone, so that the microbial bacteria can rapidly grow and reproduce on the molecular chaperone to form a dominant strain, and the technical problem of long-term storage of the microbial preparation, especially the pseudomonas fluorescens product with weak stress resistance is solved.
The invention solves the technical problem that the biological pesticide can still stably exert the effect after being stored for a long time through the carrier molecular chaperone provided for the live bacteria biological pesticide.
Detailed description of the preferred embodiments
The present invention is further described in terms of the following examples, which are included to provide further understanding to those skilled in the art. Any modifications and alterations made without departing from the spirit of the invention are within the scope of the invention.
Embodiment one
1. Preparation of microbial agent
Selecting pseudomonas fluorescens to be inoculated into an LB culture medium, culturing for 72 hours at the temperature of 28 ℃ and at the speed of 150 r/min, preparing microcapsule powder according to the method described in the embodiment I of the viable organism pesticide microcapsule (ZL 201210419235.5), packaging into a sealed container, and storing at normal temperature.
Taking 10 ml of 0.5% chitosan water (purchased from Du Tepu biotechnology Co., Ltd.), adding 10 g aluminosilicate, mixing well to obtain carrier molecular chaperone for activating microbial preparation, packaging in a sealed container, and storing at room temperature.
When the pseudomonas fluorescens microcapsule powder is stored for 1 week, 1 month, 3 months, 6 months, 12 months, 18 months and 24 months, 30 g of the pseudomonas fluorescens microcapsule powder is uniformly mixed with 10 g of carrier molecular chaperones, and the bacteria content is measured after the pseudomonas fluorescens microcapsule powder is placed for 24 hours at normal temperature. The results are shown in Table 1.
TABLE 1 Effect of Carrier chaperones on Pseudomonas fluorescens microcapsule powder content
The result shows that the number of viable bacteria of the pseudomonas fluorescens microbial microcapsule is obviously reduced after the pseudomonas fluorescens microbial microcapsule is stored for 1 year at normal temperature. The carrier molecular chaperone is added, the pseudomonas fluorescens microbial agent still keeps relatively stable after being stored for 24 months at normal temperature, and the effect of activating the microbial agent by the carrier molecular chaperone is very obvious.
Embodiment two
30 g of pseudomonas fluorescens microbial microcapsule powder stored for 12 months is uniformly mixed with 10 g of carrier molecular chaperone, and the mixture is added into 10L of water after 24 hours, and is uniformly stirred to prepare liquid medicine. Selecting tomato plants to be planted in a greenhouse, inoculating ralstonia solanacearum, designing a test according to a random block group, and carrying out root irrigation treatment on each seedling in 100 ml of liquid medicine 24 hours after inoculation, wherein 50 plants are treated each time and repeated for 4 times. The experiment was divided into 3 treatments: treating 1, diluting the microcapsule powder by 500 times of liquid medicine; and (3) treatment 2: diluting the microcapsule powder and carrier molecular chaperone by 500 times to obtain medicinal liquid; and (3) treatment: clear water control; the results are shown in the table, the seedlings of the control plot are seriously attacked by the bacterial wilt, the microcapsule powder has certain effect on controlling the bacterial wilt, and the combination of the microcapsule powder and the carrier molecular chaperone has the most obvious effect on controlling the bacterial wilt.
TABLE 2 Pseudomonas fluorescens effect on control of tomato bacterial wilt
Note that1: the trial was designed as a random panel, with 4 replicates per 50 replicates, and the different letters followed by the data in each column of the table indicate significant differences at the 5% level between treatments.
Claims (8)
1. The carrier molecular chaperone for activating microbial preparation is prepared with one or several kinds of polysaccharide, protein, amino acid, proteinase, nucleic acid, fatty acid and their derivatives and one or several kinds of mineral matters, and is used in activating the microbial activity of microbial preparation or increasing the microbial content, strengthening the effect of microbial preparation in regulating plant growth and development and preventing and controlling plant diseases and pests in water.
2. The chaperone for a carrier for activating microbial agents according to claim 1, prepared by combining one or more of polysaccharides or proteins or amino acids or proteases or nucleic acids or fatty acids or their derivatives with one or more mineral carriers including, but not limited to, milk powder, carboxymethyl starch, glyceryl monostearate, lysine, arginine, glutamic acid, aluminium silicate, methionine, S-adenosylmethionine, aspartic acid, glycine, methionine, histidine, polypeptides, peptones, aluminosilicates, trehalose, sucrose, sorbose, melezitose, glycerol, oligosaccharides, mannose, maltose, lactose, chitosan, lactic acid, calcium lactate, butyric acid, arabinose, xylose, ribose, rhamnose, magnesium silicate, glucose, mannitol, glycerol, xylitol, erythritol, threitol, sorbitol and raffinose, dextrin, dextran and soluble starch, and more preferably a combination of minerals and chitosan.
3. The microbial formulation of claim 1, wherein: the microbial preparation is prepared by adsorbing fermented microbial bacteria with an adsorbing material to prepare a capsule core, wrapping the capsule core with a wall material and directly preparing the capsule core into a suspending agent or preparing microcapsule powder and granules by drying.
4. A plant pest according to claim 1 including, but not limited to, plant-damaging micro-organisms such as nematodes, viruses, fungi or bacteria infested to cause disease and insect pests from chewing mouthparts, piercing-sucking mouthparts, rasping-sucking mouthparts, scraping mouthparts and feeding mouthparts.
5. The water body pest of claim 1, including but not limited to cyanobacteria, dinoflagellates, lichens, and the like that contaminate water.
6. The microbial organism of claim 3, wherein: the microorganism includes, but is not limited to, Bacillus subtilis, Pseudomonas fluorescens, Bacillus pumilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacillus methylotrophicus, Bacillus cereus, Paenibacillus polymyxa, Bacillus hypothermia, Streptomyces hygroscopicus, Trichoderma, Verticillium, Bacillus firmus, Bacillus thuringiensis, Beauveria bassiana, Paecilomyces lilacinus, and the like, and more preferably Pseudomonas fluorescens.
7. The microcapsule according to claim 3, wherein the wall material is a biocompatible material selected from the group consisting of beta-cyclodextrin, shellac, gelatin, gum arabic, dextrin, starch, sodium alginate, calcium alginate, etc.
8. The adsorbent material of claim 3, wherein: the adsorbing material includes but is not limited to activated carbon, white carbon black, diatomite, medical stone, kaolin, bran shell, starch, porous starch, soluble starch, macroporous adsorbent and the like or the combination of the materials.
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| CN201910441748.8A CN111979220A (en) | 2019-05-24 | 2019-05-24 | Carrier molecular chaperone for activating microbial preparation |
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| CN201910441748.8A CN111979220A (en) | 2019-05-24 | 2019-05-24 | Carrier molecular chaperone for activating microbial preparation |
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| CN111979220A true CN111979220A (en) | 2020-11-24 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112206242A (en) * | 2020-08-05 | 2021-01-12 | 内蒙古农业大学 | Fungal microcapsules for killing animal parasite eggs and preparation method and application thereof |
| CN112514918A (en) * | 2020-12-23 | 2021-03-19 | 湖南恒圣创新建筑科技有限公司 | Embedding body composite insecticidal probiotic agent and preparation method and application thereof |
| CN115011525A (en) * | 2022-06-28 | 2022-09-06 | 中国农业大学 | A compound microbial inoculant for inhibiting bacterial wilt disease of tomato and preparation method of enhanced compost |
| CN119191895A (en) * | 2024-11-27 | 2024-12-27 | 山东蓝驼农业技术开发有限公司 | Microbial preparation for promoting corn growth and preparation method thereof |
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