CN114600911B - Compound microbial agent for preventing and treating root rot of fruits and vegetables as well as preparation method and application thereof - Google Patents
Compound microbial agent for preventing and treating root rot of fruits and vegetables as well as preparation method and application thereof Download PDFInfo
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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
- A01N63/28—Streptomyces
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
- A01C1/06—Coating or dressing seed
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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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
- A01N25/08—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 containing solids as carriers or diluents
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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
- A01N25/08—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 containing solids as carriers or diluents
- A01N25/10—Macromolecular compounds
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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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
- A01N63/38—Trichoderma
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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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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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
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
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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
- C12N1/14—Fungi; Culture media therefor
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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
Abstract
The invention discloses a compound microbial agent for preventing and treating root rot of fruits and vegetables and a method for preparing the sameThe preparation method and the application thereof, the compound microbial agent comprises the following steps: streptomyces rochei fermentation liquor, streptomyces fradiae fermentation liquor and Trichoderma harzianum fermentation liquor, wherein rapeseed meal is added in the fermentation process of the Streptomyces fradiae fermentation liquor; the compound microbial agent also comprises sepiolite, talcum powder, trehalose, sodium dodecyl benzene sulfonate, sodium lignosulfonate and polyvinyl alcohol; the effective viable count of the Streptomyces rochei fermentation liquor, the Streptomyces fradiae fermentation liquor and the Trichoderma harzianum fermentation liquor is not less than 1.0 multiplied by 10 8 cfu/g, the three bacterial liquids in the prepared compound microbial agent jointly act synergistically, the synergistic effect is achieved, the effect of preventing and treating the root rot of fruits and vegetables is good, the thallus release rate in the prepared compound microbial agent is high, and the thallus survival time is long.
Description
Technical Field
The invention relates to the field of microbial agents, in particular to a compound microbial agent for preventing and treating root rot of fruits and vegetables, and a preparation method and application thereof.
Background
Root rot is one of the most serious soil-borne diseases in the production of facility fruits and vegetables, particularly the root diseases of fruits and vegetables in cucurbitaceae and solanaceae such as cucumbers and tomatoes are particularly serious, so that the yield loss is generally over 10% -15%, sometimes over 30% -40%, and even the fruits and vegetables are completely harvested. Root rot mainly occurs at the stem base and root of a plant, and can occur in the whole growth period, and vascular tissues at the root of the plant are invaded to cause the plant to die, wherein the root rot caused by fusarium is particularly common. Fusarium oxysporum is reported to be the dominant population causing soil-borne diseases. Taking soybean root rot as an example, besides the pathogen of fusarium oxysporum, there are also phytophthora sojae, pythium ultimum, fusarium solani, fusarium graminearum, rhizoctonia solani and other pathogens. According to statistics, the yield of the soybeans is reduced by more than 30% every year due to root rot, and even the soybeans are completely harvested when the yield is severe. The soybean root rot disease has complex pathogenic species and can be infected compositely, so that the prevention and treatment difficulty is high.
The control of root rot in the prior art mainly comprises the following modes: 1. the method uses the plants of disease-resistant varieties, but the method is not enough to resist various pathogenic bacteria, and the resistance is volatile; 2. chemical control, which takes effect quickly, but causes pathogenic bacteria to generate drug resistance when being used in large quantities, and causes environmental pollution and harm to human health when the drug residue exceeds standard; 3. the biological control agent is safe to crops and human bodies, green and environment-friendly, simple in production process and capable of promoting the growth of the crops. However, the existing microbial bacteria have many problems in biological control application, such as easy inactivation, low release rate, unstable drug effect, short preservation time, easy pollution and degeneration of the strains, etc.
Disclosure of Invention
In view of the above situation, the invention aims to provide a compound microbial agent and a preparation method thereof, which are used for preventing and treating root rot of fruits and vegetables.
The technical scheme for solving the problem is as follows:
a compound microbial agent for preventing and treating fruit and vegetable root rot comprises Streptomyces rochei fermentation liquor, streptomyces fradiae fermentation liquor and Trichoderma harzianum fermentation liquor; rapeseed powder is added in the fermentation process of the streptomyces fradiae fermentation liquid; the compound microbial agent also comprises sepiolite, talcum powder, trehalose, sodium dodecyl benzene sulfonate, sodium lignin sulfonate and polyvinyl alcohol.
Further, according to conventional microorganism culture method, respectively preparing fermentation liquid from Streptomyces rochei and Trichoderma harzianum strain, wherein Streptomyces rochei content is not less than 5 × 10 8 cfu/g, trichoderma harzianum content of not less than 5 × 10 8 cfu/g。
Further, inoculating a seed solution of streptomyces fradiae in an LB slant culture medium, culturing for 28h at the temperature of 30 ℃, transferring to a liquid culture medium, and performing shake fermentation culture for 72h at the temperature of 33 ℃ at 150-250r/min to obtain a streptomyces fradiae fermentation liquid; the content of the streptomyces fradiae is not less than 5 multiplied by 10 8 cfu/g。
Further, the streptomyces fradiae liquid culture medium comprises the following components in percentage by mass: (1) 2.5-4.5% of corn flour, 0.5-1.0% of soybean meal, 5-10% of rapeseed meal, 3-8% of straw powder, 3.5-4.2% of turfy soil, 1.2-1.5% of magnesium iron silicate tailings, 0.54-0.8% of calcium carbonate, 2-3% of cane sugar, 0.04% of ammonium sulfate, 0.01% of monopotassium phosphate and the balance of water, wherein the inoculation amount of streptomyces fradiae is 4%.
Further, the compound microbial agent is prepared by the following method: (1) Uniformly mixing Streptomyces rochei fermentation liquor, streptomyces fradiae fermentation liquor and Trichoderma harzianum fermentation liquor according to the weight ratio of 2-3; (2) Adding sepiolite, talcum powder, trehalose, sodium dodecyl benzene sulfonate, sodium lignin sulfonate and polyvinyl alcohol into the microbial bacterium mixed solution according to the weight ratio, stirring and adsorbing uniformly, pre-dissolving the polyvinyl alcohol in a small amount of hot water at 65-75 ℃, cooling to room temperature, adding into the microbial bacterium mixed solution, and finally supplementing the balance with water to obtain the compound microbial agent.
Furthermore, in the preparation method of the compound microbial agent, the microbial agent mixed solution is 40%, the sepiolite is 8.5%, the talcum powder is 5%, the trehalose is 10%, the sodium dodecyl benzene sulfonate is 2.5%, the sodium lignin sulfonate is 4.5%, the polyvinyl alcohol is 3%, and the balance is water.
The invention also provides a using method of the three composite biological seed coating agents:
the first application method is to coat the composite microbial agent and fruit and vegetable seeds in a weight ratio of 1.
The second use method is that the compound microbial inoculum is used for dipping roots when fruit and vegetable seedlings are transplanted, and the seedlings are transplanted after being soaked and moistened in 100-120 times of diluent of the compound microbial inoculum for preventing and treating fruit and vegetable root rot.
The third using method is that the compound microbial inoculum is used for root irrigation treatment after transplanting fruit and vegetable seedlings, and 30-40mL of root is irrigated in each hole after the compound microbial inoculum is diluted by 300-400 times.
Due to the adoption of the technical scheme, compared with the prior art, the invention has the following advantages;
compared with the prior art, the invention has the following beneficial effects:
1. the streptomyces rochei, streptomyces fradiae and trichoderma harzianum in the prepared compound microbial agent have an inhibiting effect on various root rot pathogenic bacteria, three beneficial bacteria are combined to have a synergistic control effect on the root rot pathogenic bacteria, the proper proportion shows a significant synergistic effect, and the compound microbial agent prepared by utilizing the combined synergistic effect of the beneficial microorganisms can significantly improve the control effect of the compound microbial agent on the root rot of fruits and vegetables. Coating/soaking the seeds/roots of the fruits and vegetables by using a compound microbial agent, so that streptomyces rochei with antagonistic action is inoculated to the seeds/roots, and the streptomyces rochei is metabolized to generate enzymes and antibiotics for preventing pathogenic bacteria from invading, and the propagation of the pathogenic bacteria infected firstly is inhibited; trichoderma harzianum grows around the roots of plants and forms a "protective cover" to prevent infection by root pathogenic fungi; meanwhile, streptomyces fradiae secretes various plant essential growth enzymes, so that the focus is quickly repaired, the immunity is improved, the crop photosynthesis is enhanced, the absorption of elements fixed by soil is promoted, the colonization ability of balanced nutrition at the plant rhizosphere is strong, a community can be quickly formed at the plant rhizosphere, abundant nutrient substances at the rhizosphere are metabolized, the seed decay and the seedling withering are prevented, and the durable prevention and treatment effect is achieved.
2. The rapeseed is added into a liquid culture medium of streptomyces fradiae, a large amount of glucosinolate compounds in the rapeseed are decomposed into active substance isothiocyanate, the control effect of the microbial agent on root rot is further enhanced, and meanwhile, the addition of the rapeseed is beneficial to the improvement of the content of alkaline hydrolysis nitrogen, available phosphorus, quick-acting potassium, total nitrogen and organic matters in soil, so that the abundance and diversity of fungus communities are improved, and the soil microflora develops towards a healthy and stable direction.
2. The compound microbial agent is environment-friendly and safe to use, can remarkably promote germination and propagation of fruit and vegetable seedlings, improves the stress resistance and increases the crop yield. The sepiolite, the talcum powder and the trehalose which are in specific proportion are used for adsorbing the mixed bacteria liquid, the biocompatibility with fungi is good, meanwhile, the composite microbial agent prepared by taking the sodium dodecyl benzene sulfonate and the sodium lignin sulfonate as the surfactants and the polyvinyl alcohol as the film forming agent is good in film forming property and adhesive force, is beneficial to coating seeds or roots, is high in survival rate of bacteria and good in slow release property, remarkably improves the stability of products, is beneficial to storage and transportation, and develops a new way for preventing and treating root rot of fruits and vegetables.
Detailed Description
The technical solution of the present invention is described in detail with reference to the specific embodiments, but it should be emphasized that the present invention is not limited to the specific embodiments; the microbial species used in the examples below can be isolated by themselves or obtained by purchase; the starting materials described in the examples below are, without specific indication, either prepared by conventional methods or commercially available.
Example 1
A. Provides a preparation method of Streptomyces rochei fermentation liquor:
(1) Inoculating activated seed liquid of Streptomyces rochei in NB culture medium according to 1% of inoculation amount, and culturing at 30 deg.C and 180r/min for 24 hr to obtain seed liquid; (2) Inoculating the seed liquid into liquid culture medium according to the inoculation amount of 3%, culturing at 32 deg.C for 48 hr at 200r/min to obtain Streptomyces rochei fermentation liquid with Streptomyces rochei content of not less than 5 × 10 8 cfu/g; the liquid culture medium consists of 8 percent of corn meal, 80 percent of soybean meal, 5 percent of yeast powder, 0.2 percent of magnesium sulfate monohydrate and the balance of water.
B. The preparation method of the trichoderma harzianum fermentation liquor comprises the following steps:
(1) Inoculating the activated Trichoderma harzianum seed solution into an LDA culture medium according to the inoculation amount of 1%, and culturing at 28 ℃ and 150r/min for 24h to obtain the seed solution; (2) Inoculating the seed solution into liquid culture medium according to the inoculation amount of 3%, culturing at 30 deg.C and 180r/min for 72h to obtain Trichoderma harzianum fermentation liquor with Trichoderma harzianum content of not less than 5 × 10 8 cfu/g; the liquid culture medium is prepared by decocting 12% oat flour, 2% sucrose, 0.2% dipotassium hydrogen phosphate, 0.5% magnesium sulfate and the balance of water into paste.
C. Provides a preparation method of streptomyces fradiae fermentation liquor:
(1) Inoculating the activated seed solution of the streptomyces fradiae into an LB culture medium according to the inoculation amount of 1%, and culturing at 30 ℃ at 180r/min for 28h to obtain the seed solution; (2) Inoculating the seed solution into a liquid culture medium according to the inoculation amount of 4 percent, culturing for 72 hours at the temperature of 33 ℃ and at the speed of 220r/min to obtain streptomyces fradiae fermentation liquor,the content of Streptomyces fradiae is not less than 5 × 10 8 cfu/g; the liquid culture medium consists of 2.5-4.5% of corn flour, 0.5-1.0% of soybean meal, 5-10% of rapeseed meal, 3-8% of straw powder, 3.5-4.2% of turfy soil, 1.2-1.5% of magnesium iron silicate tailings, 0.54-0.8% of calcium carbonate, 2-3% of cane sugar, 0.04% of ammonium sulfate, 0.01% of potassium dihydrogen phosphate and the balance of water.
Example 2
A compound microbial agent for preventing and treating root rot of fruits and vegetables comprises the following components in percentage by weight
40% of microbial mixed solution, 8.5% of sepiolite, 5% of talcum powder, 10% of trehalose, 2.5% of sodium dodecyl benzene sulfonate, 4.5% of sodium lignin sulfonate, 3% of polyvinyl alcohol and the balance of water; the microbial strain mixed liquor is obtained by uniformly mixing Streptomyces rochei fermentation liquor, streptomyces fradiae fermentation liquor and Trichoderma harzianum fermentation liquor according to the weight ratio of 2.
Example 3
A compound microbial agent for preventing and treating root rot of fruits and vegetables comprises the following components in percentage by weight
40% of microbial bacterium mixed solution, 8.5% of sepiolite, 5% of talcum powder, 10% of trehalose, 2.5% of sodium dodecyl benzene sulfonate, 4.5% of sodium lignin sulfonate, 3% of polyvinyl alcohol and the balance of water; the microorganism bacterium mixed solution is obtained by uniformly mixing Streptomyces rochei fermentation liquor, streptomyces fradiae fermentation liquor and Trichoderma harzianum fermentation liquor according to the weight ratio of 5.
Example 4
A compound microbial agent for preventing and treating root rot of fruits and vegetables comprises 40% of microbial agent mixed solution, 8.5% of sepiolite, 5% of talcum powder, 10% of trehalose, 2.5% of sodium dodecyl benzene sulfonate, 4.5% of sodium lignin sulfonate, 3% of polyvinyl alcohol and the balance of water in percentage by weight; the microorganism bacterium mixed solution is obtained by uniformly mixing Streptomyces rochei fermentation liquor, streptomyces fradiae fermentation liquor and Trichoderma harzianum fermentation liquor according to the weight ratio of 3.
Example 5
A preparation method of a compound microbial agent for preventing and treating fruit and vegetable root rot comprises the following steps:
(1) Uniformly mixing Streptomyces rochei fermentation liquor, streptomyces fradiae fermentation liquor and Trichoderma harzianum fermentation liquor according to the weight ratio of the embodiments 2-4 to obtain microorganism bacteria mixed liquor; (2) Adding sepiolite, talcum powder, trehalose, sodium dodecyl benzene sulfonate, sodium lignin sulfonate and polyvinyl alcohol into the microbial bacterium mixed solution according to the weight ratio of the embodiment 2-4, stirring and adsorbing uniformly, pre-dissolving the polyvinyl alcohol with a small amount of hot water at 65-75 ℃, cooling to room temperature, adding the mixture into the microbial bacterium mixed solution, and finally adding water to make up the balance to obtain the compound microbial agent.
Example 6
Provides a using method of the compound microbial agent, which comprises the following steps: coating the composite microbial agent and the fruit and vegetable seeds in a weight ratio of 1.
Example 7
Provides a using method of the compound microbial agent, which comprises the following steps: the compound microbial agent is used for root dipping treatment when fruit and vegetable seedlings are transplanted, and the seedlings are transplanted after being soaked and moistened in 100-120 times of diluent of the compound microbial agent for preventing and treating fruit and vegetable root rot.
Example 8
Provides a using method of the compound microbial agent, which comprises the following steps: the compound microbial agent is used for root irrigation treatment after transplanting of fruit and vegetable seedlings, and 30-40mL of root is irrigated in each hole after the compound microbial agent is diluted by 300-400 times.
Comparative example 1
The components and the content of the compound microbial agent for preventing and treating the root rot of the fruits and the vegetables are the same as those in the example 3, except that the talcum powder is replaced by the diatomite, and the rest is unchanged, and the preparation method is the same as that in the example 5.
Comparative example 2
The components and the content of the compound microbial agent for preventing and treating the root rot of the fruits and the vegetables are the same as those in the example 3, except that the 'rapeseed meal' in the liquid culture medium of the streptomyces fradiae fermentation liquid is replaced by the 'soybean meal', and the rest is unchanged, and the preparation method is the same as that in the examples 1 and 5.
Comparative example 3
A composite microbial preparation for preventing and treating root rot of fruit and vegetable has the same components and contents as those in example 3, except that Streptomyces rochei fermentation liquid in microbial mixture is replaced by Streptomyces fradiae fermentation liquid, and the rest is unchanged, and the preparation method is the same as that in example 5.
Comparative example 4
The components and the content of the compound microbial agent for preventing and treating the root rot of the fruits and the vegetables are the same as those in the example 3, except that the streptomyces fradiae fermentation liquor in the microbial bacterium mixed liquor is replaced by the trichoderma harzianum fermentation liquor, and the rest is unchanged, and the preparation method is the same as that in the example 5.
Comparative example 5
A compound microbial agent for preventing and treating root rot of fruits and vegetables has the same components and content as in example 3, except that Trichoderma harzianum fermentation liquor in a microbial bacterium mixed liquor is replaced by Streptomyces rochei fermentation liquor, and the rest is unchanged, and the preparation method is the same as in example 5.
Test example 1
(1) Respectively taking 10g of the compound microbial inoculum prepared in the examples 2-4 and the comparative example 1, putting the compound microbial inoculum into 100mL of sterilized water, pouring sterilized glass beads, fully shaking, uniformly mixing, and detecting the colony of the released living body by adopting a dilution plate counting method (taking the ratio of the amount of the carried bacteria to the amount of the released bacteria as the release rate)
(2) The complex microbial agents prepared in examples 2 to 4 and comparative example 1, which were stored at room temperature, were sampled at 7d, 25d, and 45d, added to sterilized water, applied by gradient dilution, and the number of viable bacteria released from the carrier at different time periods was measured by plate count method, and the survival rate of the cells was measured by the ratio of the number of cells to the initial number of cells ((1) released amount of cells).
The results are shown in Table 1:
| rate of release/%) | 7d survival/%) | 25d survival/%) | 45d survival rate/%) | |
| Example 2 | 72.5 | 88.3 | 62.5 | 49.6 |
| Example 3 | 73.6 | 86.5 | 61.3 | 48.4 |
| Example 4 | 71.7 | 87.2 | 61.8 | 47.9 |
| Comparative example 1 | 40.3 | 86.4 | 55.2 | 32.1 |
As can be seen from Table 1, compared with comparative example 1, the release rate of the bacterial cells in the composite microbial agents prepared in examples 2-4 is up to 72%, and the survival rate of the bacterial cells in the microbial agents is still up to 49% at 45 d.
Test example 2
Soybean root rot control test:
the experiments were divided into 10 groups: (1) experimental group: the compound microbial agents prepared in examples 2-4 and comparative examples 1-5 are prepared into slurry (compound microbial agent 2g +120mL of clear water is mixed uniformly); (2) control group: treating with clear water; (3) blank group: treating with clear water; the experimental group and the control group are treated with the pathogen suspension, the blank group is not treated with the pathogen suspension, each treatment is repeated for 4 times, and each time, 10 soybean seedlings are repeated.
(1) Culturing pathogenic bacteria of soybean root rot such as Fusarium oxysporum, phytophthora, pythium ultimum, fusarium solani, fusarium graminearum and Rhizoctonia solani on PDA plate for 3d, taking out bacterial cake with diameter of 1.00cm, transferring to new PDA culture medium, fermenting, and culturing to obtain mixed pathogenic bacteria suspension (each viable bacteria content is 1.0 × 10) 6 cfu·mL -1 )；
(2) Sowing the accelerated soybean seeds, transplanting the seedlings into a nursery after 3-4 true leaves of the seedlings are planted, dipping the root systems of the soybean seedlings with the compound microbial agent slurry in the transplanting process, then planting the seedlings, carrying out mixed pathogenic bacterium suspension root irrigation (12 mL/particle) after 2-3d, managing at normal temperature, and investigating the disease occurrence condition of the plants after 30 d.
The disease investigation method comprises the following steps: adopting soybean root rot classification standard: stage 0: no disease; level 1: slight water-stain-like lesion spots appeared on the stem; and 2, stage: the lesion spots on the stem are expanded, but the plant height is not more than 1/4, and the plant does not wilt; and 3, level: disease part is 1/4 over the whole plant, the disease part extends downwards to root part and is not more than 3/4 of plant height, the stem base is sunken and wilted; and 4, stage 4: the diseased part exceeds the whole plant or spreads to the whole plant, including roots and petioles, the stem base is seriously overflowed, the leaves are withered, and the disease is dead. Disease index = (number of diseased plants at each level × corresponding level)/(total number of investigated plants × maximum number of levels) × 100; control effect (%) = (blank disease index-treatment disease index)/blank disease index × 100, and the results are shown in table 2:
| TABLE 2 | Index of disease condition | Preventive effect/%) |
| Example 2 | 9 | 88.7 |
| Example 3 | 10 | 87.5 |
| Example 4 | 8 | 90.0 |
| Comparative example 1 | 22 | 72.5 |
| Comparative example 2 | 12 | 85.0 |
| Comparative example 3 | 23 | 71.3 |
| Comparative example 4 | 29 | 63.8 |
| Comparative example 5 | 28 | 65.0 |
| Control group | 80 | - |
| Blank group | 0 | - |
As can be seen from the results in Table 2, the disease index of the compound microbial inoculant provided by the invention on soybean root rot caused by various pathogenic bacteria is 8-10, the control effect can be up to 88.7%, compared with comparative example 1, the compound microbial inoculant provided by the invention has a good control effect on soybean root rot, and compared with comparative example 3-5, the compound microbial inoculant provided by the invention has a stronger control effect when being used in combination with Streptomyces rochei, streptomyces fradiae and Trichoderma harzianum. In addition, the comparison of the growth conditions of soybean plants in the blank group shows that the soybean plants in the examples 2-4 grow vigorously and have tough root systems, and the soybean plants in the comparative example 2 are short, small and easy to pour.
While the invention has been described in further detail with reference to specific embodiments thereof, it is not intended that the invention be limited to the specific embodiments thereof; for those skilled in the art to which the present invention pertains and related arts, the extension, operation method, and data replacement shall fall within the scope of the present invention based on the technical idea of the present invention.
Claims (7)
1. The compound microbial agent for preventing and treating the root rot of fruits and vegetables is characterized by comprising Streptomyces rochei fermentation liquor, streptomyces fradiae fermentation liquor and Trichoderma harzianum fermentation liquor, wherein rapeseed meal is added in the fermentation process of the Streptomyces fradiae fermentation liquor; the compound microbial agent also comprises sepiolite, talcum powder, trehalose, sodium dodecyl benzene sulfonate, sodium lignosulfonate and polyvinyl alcohol;
the compound microbial agent is prepared by the following method: (1) Uniformly mixing Streptomyces rochei fermentation liquor, streptomyces fradiae fermentation liquor and Trichoderma harzianum fermentation liquor according to the weight ratio of 2-3; (2) Adding sepiolite, talcum powder, trehalose, sodium dodecyl benzene sulfonate, sodium lignin sulfonate and polyvinyl alcohol into the microbial bacterium mixed solution according to the weight ratio, stirring and adsorbing uniformly, pre-dissolving the polyvinyl alcohol in a small amount of 65-75 ℃ hot water, cooling to room temperature, adding into the microbial bacterium mixed solution, and finally supplementing the balance with water to obtain a compound microbial agent;
the preparation method of the compound microbial agent comprises 40% of microbial mixed solution, 8.5% of sepiolite, 5% of talcum powder, 10% of trehalose, 2.5% of sodium dodecyl benzene sulfonate, 4.5% of sodium lignosulfonate, 3% of polyvinyl alcohol and the balance of water.
2. The compound microbial agent for preventing and treating root rot of fruits and vegetables as claimed in claim 1, wherein the Streptomyces rochei and Trichoderma harzianum strains are prepared into fermentation liquid respectively according to conventional microbial culture method, and the content of Streptomyces rochei is not less than 5 x 10 8 cfu/g, trichoderma harzianum content not less than 5X 10 8 cfu/g。
3. The compound microbial agent for preventing and treating root rot of fruits and vegetables according to claim 1, wherein seed liquid of streptomyces fradiae is inoculated in an LB slant culture medium, cultured for 28 hours at 30 ℃, transferred to a liquid culture medium, and subjected to shake fermentation culture for 72 hours at 33 ℃ at 150-250r/min to obtain streptomyces fradiae fermentation liquid; the content of the streptomyces fradiae is not less than 5 multiplied by 10 8 cfu/g。
4. The compound microbial agent for preventing and treating fruit and vegetable root rot according to claim 1, wherein the streptomyces fradiae liquid culture medium consists of the following components in percentage by mass: (1) 2.5-4.5% of corn flour, 0.5-1.0% of soybean meal, 5-10% of rapeseed meal, 3-8% of straw powder, 3.5-4.2% of turfy soil, 1.2-1.5% of magnesium iron silicate tailings, 0.54-0.8% of calcium carbonate, 2-3% of cane sugar, 0.04% of ammonium sulfate, 0.01% of monopotassium phosphate and the balance of water, wherein the inoculation amount of streptomyces fradiae is 4%.
5. The use method of the compound microbial inoculant for preventing and treating the root rot of fruits and vegetables as claimed in any one of claims 1 to 4, wherein the compound microbial inoculant and the fruit and vegetable seeds are coated in a weight ratio of 1.
6. The use method of the compound microbial inoculant for preventing and treating the root rot of fruits and vegetables as claimed in any one of claims 1 to 4, wherein the compound microbial inoculant is used for root dipping treatment during fruit and vegetable seedling transplantation, and seedlings are transplanted after being soaked and moistened in a diluent 100-120 times of the compound microbial inoculant for preventing and treating the root rot of fruits and vegetables.
7. The use method of the compound microbial inoculant for preventing and treating the root rot of fruits and vegetables as claimed in any one of claims 1 to 4, wherein the compound microbial inoculant is used for root irrigation treatment after transplanting of fruit and vegetable seedlings, and 30-40mL of roots are irrigated in each hole after the compound microbial inoculant is diluted by 300-400 times.
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