CN105875645A - Preparing method of biological agent for preventing and controlling tobacco root black rot - Google Patents
Preparing method of biological agent for preventing and controlling tobacco root black rot Download PDFInfo
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- CN105875645A CN105875645A CN201511020388.2A CN201511020388A CN105875645A CN 105875645 A CN105875645 A CN 105875645A CN 201511020388 A CN201511020388 A CN 201511020388A CN 105875645 A CN105875645 A CN 105875645A
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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
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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
Abstract
The invention discloses a preparing method of a biological agent for preventing and controlling tobacco root black rot. The biological agent is prepared from 10% of rhodopseudomonas palustris, 20% of bacillus subtilis, 10% of bacillus polymyxa, 5% of brevibacillus laterosporus, 15% of bacillus megatherium, 5% of Trichoderma viride, 5% of Trichoderma harzianum, 10% of Issatchenkia orientalis, 10% of streptomyces microflavus, 8% of Thermophilic sporotrichum and 2% of Pythium oligandrum. The preparing method includes the steps that microorganic test tube slant seed culture and microorganic liquid seed are carried out, a fermentation medium is inoculated with liquid seeds, a great number of thalli and metabolite thereof are obtained through liquid fermentation, and the thalli, the metabolite thereof and a carrier are mixed, dried, and smashed into powder. As multiple beneficial bacteria are cultured in the same pot, the biological agent is unique in formula, good in preventing and controlling effect and environmentally friendly. One composite culture medium is shared, and the biological agent has the dual functions of providing nutrition and preventing and controlling diseases, and greatly reduces production cost.
Description
Technical field: the present invention relates to biological pesticide technical field, specifically has black root of tobacco very well
The preparation method of a kind of bacteria agent preventing and treating black root of tobacco of prevention effect.
Background technology: by base take root the mould microbial black root of tobacco of beading be on tobacco leaf production important very
One of fungus diseases, its germ tube intrusion host plant produced by mycelial growth and spore germination causes morbidity,
All can fall ill from seedling to Adult plant, main harm root system.After tobacco seedlings morbidity, children's stem, cotyledon and the tip of a root are sent out
Black rotten, the root system of severe disease strain the most all rots, in black.Cigarette strain is not dead, and it is slow that aerial parts presents growth
Slowly, jaundice, downgrade.Often mix generation with balck shank, cause Nicotiana tabacum L. to lose.Its throughout world mainly produces cigarette
District, once made tobacco leaf production sustain losses severely in states such as the U.S., Canada, Japan, and Main Tobacco Planting district of China is also
All there is generation.The most in recent years, this disease spreads in some cigarette districts, saves in Yunnan Province of China, Guizhou, Hubei etc.
Occurring serious, serious plot sickness rate is up to more than 30%.Shandong, Henan, Anhui, Jilin, Fujian etc. save
Also there is generation, have the trend increasing the weight of harm in recent years.To this, production relies primarily on chemical agent in seedbed
The method spraying the spectrum antibacterial such as thiophanate methyl, thiram with field prevents and treats the outburst of this disease and popular.
But, Nicotiana tabacum L. is leaf with industrial crops, and pesticide residues, pathogen is drug-fast is continuously increased and can not be ignored
Environmental risk, and bring series of problems to Medicated cigarette health, limit the extensive utilization of pesticide.
At present, Biological control has become as the focus of people's research, and the advantage of Biological control is to person poultry safety,
Not polluting environment, use cost is relatively low, and the preventive effect time is long, it is simple to utilize.But domestic Biological control makes
Most of compound bacteria products be all that single bacterium is cultivated, finally remix together blend mixed vaccine product,
Symbiosis after mixed vaccine is manured into soil, alternate situation, understanding inadequate, or after mixing " multiple
Close bacterium " inherently antagonism, so phase between " compound bacteria " after causing prevention effect difference or being manured into soil
Antagonism, cuts one another's throats, causes root system probiotics to reduce mutually, and preventive effect reduces.
Summary of the invention: the purpose of the present invention is primarily directed to the problem existing for above-mentioned prior art, it is provided that
Microorganism fungus kind is used to cultivate, amount reproduction and the microbial inoculum processing method of microorganism and the microbial inoculum that produces, can be right
Black root of tobacco has good inhibiting effect and a prevention effect and a kind of black root of tobacco of preventing and treating of using
Bacteria agent preparation method.
The present invention is adopted the technical scheme that for achieving the above object: a kind of life preventing and treating black root of tobacco
The strain component of the preparation method of thing microbial inoculum is: swamp Rhodopseudomonas 10%, bacillus subtilis 20%, many
Viscous bacillus cereus 10%, Brevibacillus laterosporus 5%, bacillus megaterium 15%, Trichoderma viride 5%, Ha Ci
Trichoderma spp. 5%, Issatchenkia orientalis 10%, streptomyces microflavus 10%, sporotrichum thermophile 8%, pythium oligandrum 2%；
Its preparation method is: first carry out microorganism test tube slant seed culture and microbial liquid seed culture, then will
Liquid seeds accesses in fermentation medium, passes through liquid fermentation, it is thus achieved that substantial amounts of thalline and metabolite thereof, will
Thalline and metabolite thereof mix with carrier, and crushed after being dried makes powder.
The invention has the beneficial effects as follows: 1, the present invention is more than ten kinds probioticss to be cultivated with tank, and formula is unique,
Make they symbiosis, alternate, harmonious growth, after being manured into soil, become dominant microflora, collaborative anti-" enemy ", anti-
Effect is substantially.
2, it is to be cultivated with tank, with the cultivation of domestic other products by more than ten kinds probioticss due to this composite bacteria agent capable
Technology is cultivated such as single bacterium, and terminal blended product compares the cost that can save more than ten times.Because this product is to allow
More than ten kinds probioticss are in same fermentation tank, jointly enjoy a kind of complex medium and can harmonious coexistence
Growth, thus reduce production cost.
3, in the production application preventing and treating black root of tobacco, this microbial inoculum has nutrition and the dual work of controlling disease concurrently
With, prevention effect is good, and the preventing and treating persistent period is long, does not results in secondary pollution, low cost, not only environmental protection but also green
Color, nontoxic to cigarette strain and soil, and prevention effect reaches more than 85%.Furthermore, this microbial inoculum contains abundant
Aminoacid that organic carbon source, nitrogen source, phosphorus source and microorganism produce in vital movement, bioactive peptide, auxin,
Gibberellins, heteroauxing, multivitamin etc., produce regulation stimulates the active substance of plant growing simultaneously,
Nicotiana tabacum L. absorb after, in conjunction with self carbon source, nitrogen source promote photosynthetic metabolite, plant by self regulation stalwartness,
Improving cigarette strain anti-adversity ability and drought-resistant ability, the dominant population in soil stably keeps it advantageously simultaneously
Position.
4, the present invention can reach protection environment, improvement soil, improve chemical fertilizer utilization ratio, improve agricultural product number,
Quality, the comprehensive purpose developed eco-agriculture.
5, relevant bacteria species of the present invention obtains from relevant microbial preservation mechanism or other approach, and strain source is clearly.
The strain source brief description of the present invention:
Strain of the present invention, all buys in the common micro-life of China Committee for Culture Collection of Microorganisms
Thing center, and on 01 09th, 2015 and China Committee for Culture Collection of Microorganisms's common micro-organisms
Center has been signed the offer of microorganism hereditary resource and has been utilized agreement (CGMCC/2007/4.00 annex 1) and data
Utilize agreement (CGMCC/2007/4.00 annex 2).
Detailed description of the invention: embodiment 1: the preparation method of a kind of bacteria agent preventing and treating black root of tobacco
Strain component is: swamp Rhodopseudomonas 10%, bacillus subtilis 20%, bacillus polymyxa 10%, side
Spore bacillus brevis 5%, bacillus megaterium 15%, Trichoderma viride 5%, Trichoderma harzianum 5%, her Sa ferment of east
Mother 10%, streptomyces microflavus 10%, sporotrichum thermophile 8%, pythium oligandrum 2%；Its preparation method is: advanced
Row microorganism test tube slant seed culture and microbial liquid seed culture, then liquid seeds is accessed fermentation training
Support in base, pass through liquid fermentation, it is thus achieved that substantial amounts of thalline and metabolite thereof, by thalline and metabolite thereof with
Carrier mixes, and crushed after being dried makes powder.
Embodiment 2: the preparation method of described a kind of bacteria agent preventing and treating black root of tobacco is:
1. slant culture: above-mentioned original strain is inoculated in the most respectively in the culture medium of correspondence and lives
Changing, the slant medium of use has the PDA-sucrose cultivation cultivating pythium oligandrum, Trichoderma viride, Trichoderma harzianum
Base cultivation temperature is set under the conditions of 28 DEG C and cultivates 4 days, cultivates sporotrichum thermophile, the PDA-of Issatchenkia orientalis
Dextrose culture-medium cultivation temperature is cultivated 3 days under the conditions of being located at 30 DEG C, and sporotrichum thermophile cultivation temperature controls
Cultivate 3 days, cultivate bacillus subtilis, Paenibacillus polymyxa, Brevibacillus laterosporus, huge for 45 DEG C
Bacillus cereus, the nutrient agar cultivation temperature of swamp Rhodopseudomonas are located at 30 DEG C and cultivate 3 days, training
The Gause I culture medium supporting streptomyces microflavus controls cultivation temperature 30 DEG C cultivation 5 days；
2. seed culture: liquid amplification culture medium has cultivation pythium oligandrum, Trichoderma viride, the PDA-of Trichoderma harzianum
Sucrose medium cultivation temperature is cultivated 2 days in 150r/min shaking table under the conditions of being set in 28 DEG C, cultivates thermophilic side
Spore is mould, the PDA-dextrose culture-medium cultivation temperature of Issatchenkia orientalis is cultivated 1 day under the conditions of being located at 28 DEG C,
Sporotrichum thermophile cultivation temperature controls to cultivate 1 day at 28 DEG C, cultivates the Gause I culture medium of streptomyces microflavus
Cultivation temperature is cultivated 2 days in 150r/min shaking table under the conditions of being set in 28 DEG C, bacillus subtilis, glues class more
Bacillus cereus, Brevibacillus laterosporus, bacillus megaterium, the Nutrient agar of swamp Rhodopseudomonas are cultivated
Base cultivation temperature is cultivated 1 day in 150r/min shaking table under the conditions of being located at 28 DEG C, and seed liquor inoculation is joined by volume
Ratio: swamp Rhodopseudomonas 10%, bacillus subtilis 20%, bacillus polymyxa 10%, the short spore of side spore
Bacillus 15%, bacillus megaterium 15%, Trichoderma viride 5%, Trichoderma harzianum 5%, Issatchenkia orientalis 10%,
Streptomyces microflavus 10%, sporotrichum thermophile 8%, pythium oligandrum 2%；
3. fermentation medium is cultivated: cultured liquid seeds is aseptically received through autoclaved
In fermentation medium, fermentation medium component is: brown sugar 0.6%, glucose 0.3%, corn starch 1.8%,
Bean cake powder 1.5%, magnesium sulfate 0.03%, carbamide 0.12%, potassium dihydrogen phosphate 0.04%, peptone 0.1%, ferment
Female powder 0.05%, sodium carbonate regulation PH to 7.0, liquid microelement 5mL.Condition of culture: inoculum concentration 5-10%,
Cultivation temperature controls at 28-30 DEG C, rotating speed 120-180r/min, ventilation 1: 0.5-0.8, fermentation culture
24-36 hour；
4. prepared by powder: microbial cells fermentation liquid is less than with being crushed to particle diameter with 50% weight ratio after preparing
The sterilizing fertilizer of less than 1 millimeter does carrier with white carbon 3: 1 mass ratio and mixes, and puts into hothouse and is dried
Moisture be less than less than 8%, crushing packing, the solid, powdery bacteria agent of black root of tobacco must be prevented and treated, have
Effect viable count >=5,000,000,000/g.
Embodiment 3: the preparation method of described a kind of bacteria agent preventing and treating black root of tobacco is: microorganism
And metabolite i.e. fermentation end products with 40-75% weight ratio be crushed to particle diameter less than the list of less than 1 millimeter
One or combination carrier such as peat composed of rotten mosses powder, Testa oryzae, Kaolin, sterilizing fertilizer, humic acids, white carbon etc. singly
One carrier or peat composed of rotten mosses powder, Testa oryzae, sterilizing fertilizer and humic acid, Kaolin, white carbon are with 3-5: 1 ratio
Mixing combination carrier, put into hothouse and be dried, moisture be less than less than 8%, crushing packing, be placed in room temperature
The shady and cool place of being dried preserves.
Embodiment 4: the bacteria agent of the preparation method of described a kind of bacteria agent preventing and treating black root of tobacco
Content >=2,000,000,000/g.
Embodiment 5: the preparation method of described a kind of bacteria agent preventing and treating black root of tobacco is:
1. slant culture: above-mentioned original strain is inoculated in the most respectively in the culture medium of correspondence and lives
Changing, the slant medium used has the PDA-sucrose training cultivating pythium oligandrum, Trichoderma viride, Trichoderma harzianum
Support base cultivation temperature to be set under the conditions of 28 DEG C and cultivate 4 days, cultivate sporotrichum thermophile, Issatchenkia orientalis
PDA-dextrose culture-medium cultivation temperature is cultivated 3 days under the conditions of being located at 30 DEG C, sporotrichum thermophile cultivation temperature control
Make 45 DEG C cultivate 3 days, cultivate bacillus subtilis, Paenibacillus polymyxa, Brevibacillus laterosporus,
Bacillus megaterium, the nutrient agar cultivation temperature of swamp Rhodopseudomonas be located at 30 DEG C cultivate 3 days,
The Gause I culture medium cultivating streptomyces microflavus controls cultivation temperature 30 DEG C cultivation 5 days；
2. seed culture: the strain of slant culture is inoculated in liquid amplification culture medium the most respectively
In, liquid amplification culture medium has cultivation pythium oligandrum, Trichoderma viride, the PDA-sucrose medium of Trichoderma harzianum
Cultivation temperature is cultivated 2 days in 150r/min shaking table under the conditions of being set in 28 DEG C, cultivates sporotrichum thermophile, east
The PDA-dextrose culture-medium cultivation temperature of her Sa yeast is cultivated 1 day under the conditions of being located at 28 DEG C, sporotrichum thermophile
Cultivation temperature controls to cultivate 1 day at 28 DEG C, and the Gause I culture medium culturing temperature cultivating streptomyces microflavus sets
Under the conditions of being scheduled on 28 DEG C in 150r/min shaking table cultivate 2 days, bacillus subtilis, Paenibacillus polymyxa,
Brevibacillus laterosporus, bacillus megaterium, the nutrient agar cultivation temperature of swamp Rhodopseudomonas
Cultivating 1 day in 150r/min shaking table under the conditions of being located at 28 DEG C, seed liquor inoculates proportioning by volume: marsh is red
Pseudomonas 10%, bacillus subtilis 20%, bacillus polymyxa 10%, Brevibacillus laterosporus 15%,
Bacillus megaterium 15%, Trichoderma viride 5%, Trichoderma harzianum 5%, Issatchenkia orientalis 10%, thin yellow strepto-
Bacterium 10%, sporotrichum thermophile 8%, pythium oligandrum 2%；
3. fermentation medium: cultured liquid seeds is aseptically received through autoclaved fermentation
In culture medium, fermentation medium component is: brown sugar 0.6%, glucose 0.3%, corn starch 1.8%, bean cake
Powder 1.5%, magnesium sulfate 0.03%, carbamide 0.12%, potassium dihydrogen phosphate 0.04%, peptone 0.1%, yeast powder
0.05%, sodium carbonate regulation PH to 7.0, liquid microelement 5mL.Condition of culture: inoculum concentration 10%, cultivates
Temperature controls at 28 degree, rotating speed 180r/min, ventilation 1: 0.6, fermentation culture 24 hours；
4. prepared by powder: microbial cells fermentation liquid is less than with being crushed to particle diameter with 50% weight ratio after preparing
The sterilizing fertilizer of less than 1 millimeter does carrier with white carbon 3: 1 mass ratio and mixes, and puts into hothouse and is dried
Moisture be less than less than 8%, crushing packing, the solid, powdery bacteria agent of black root of tobacco must be prevented and treated, have
Effect viable count >=5,000,000,000/g.
5. use: in Sichuan cigarette DIYU on May 4th, 2014 before tobacco transplant, by microbial inoculum by every mu 2
Kilogram consumption mix by 1: 10 with fertilizer or thin sand and soil after spread fertilizer over the fields and plant kind of plant hole is decided at the higher level but not officially announced.Preventing and treating cigarette
The effect of grass roots black rot is 75%, and Nicotiana tabacum L. average yield per mu exceeds than the average yield per mu not using invention formulation
About 15%.
Embodiment 6: the preparation method of described a kind of bacteria agent preventing and treating black root of tobacco is:
1. slant culture: above-mentioned original strain is inoculated in the most respectively in the culture medium of correspondence and lives
Changing, the slant medium used has the PDA-sucrose training cultivating pythium oligandrum, Trichoderma viride, Trichoderma harzianum
Support base cultivation temperature to be set under the conditions of 28 DEG C and cultivate 4 days, cultivate sporotrichum thermophile, Issatchenkia orientalis
PDA-dextrose culture-medium cultivation temperature is cultivated 3 days under the conditions of being located at 30 DEG C, sporotrichum thermophile cultivation temperature control
Make 45 DEG C cultivate 3 days, cultivate bacillus subtilis, Paenibacillus polymyxa, Brevibacillus laterosporus,
Bacillus megaterium, the nutrient agar cultivation temperature of swamp Rhodopseudomonas are located at 30 DEG C and cultivate 3 days,
Gause I is cultivated the culture medium of streptomyces microflavus and is controlled cultivation temperature 30 DEG C cultivation 5 days；
2. seed culture: the strain of slant culture is inoculated in liquid amplification culture medium the most respectively
In, liquid amplification culture medium has cultivation pythium oligandrum, Trichoderma viride, the PDA-sucrose medium of Trichoderma harzianum
Cultivation temperature is cultivated 2 days in 150r/min shaking table under the conditions of being set in 28 DEG C, cultivates sporotrichum thermophile, east
The PDA-dextrose culture-medium cultivation temperature of her Sa yeast is cultivated 1 day under the conditions of being located at 30 DEG C, sporotrichum thermophile
Cultivation temperature controls to cultivate 1 day at 30 DEG C, and the Gause I culture medium culturing temperature cultivating streptomyces microflavus sets
Under the conditions of being scheduled on 30 DEG C in 150r/min shaking table cultivate 2 days, bacillus subtilis, Paenibacillus polymyxa,
Brevibacillus laterosporus, bacillus megaterium, the nutrient agar cultivation temperature of swamp Rhodopseudomonas
Cultivating 1 day in 150r/min shaking table under the conditions of being located at 30 DEG C, seed liquor is inoculated proportioning by volume and is: marsh
Rhodopseudomonas 10%, bacillus subtilis 20%, bacillus polymyxa 10%, Brevibacillus laterosporus 15%,
Bacillus megaterium 15%, Trichoderma viride 5%, Trichoderma harzianum 5%, Issatchenkia orientalis 10%, thin yellow strepto-
Bacterium 10%, sporotrichum thermophile 8%, pythium oligandrum 2%；
3. fermentation medium: cultured liquid seeds is aseptically received through autoclaved fermentation
In culture medium, fermentation medium component is: brown sugar 0.6%, glucose 0.3%, corn starch 1.8%, bean cake
Powder 1.5%, magnesium sulfate 0.03%, carbamide 0.12%, potassium dihydrogen phosphate 0.04%, peptone 0.1%, yeast powder
0.05%, sodium carbonate regulation pH7.0, liquid microelement 5mL；Condition of culture: inoculum concentration 5%, cultivation temperature
Control at 30 DEG C, rotating speed 150r/min, ventilation 1: 0.5, fermentation culture 36 hours；
4. prepared by powder: microbial cells fermentation liquid is less than with being crushed to particle diameter with 60% weight ratio after preparing
The sterilizing fertilizer of less than 1 millimeter does carrier with humic acid 4: 1 mass ratio and mixes, and puts into hothouse and is dried
Moisture be less than less than 8%, crushing packing, the solid, powdery bacteria agent of black root of tobacco must be prevented and treated, have
Effect viable count >=5,000,000,000/g；
5. use: in Henan cigarette DIYU on May 15th, 2014 after tobacco transplant, every mu of this microbial inoculum of use 2
Kilogram dilution 80-100 times of pouring root, every strain 100-300ml.Determine to fill according to soil dry humidity and plant size
Dose is how many, for the purpose of the saturating tobacco plants of thorough tank takes root scope.In the vega using invention formulation,
Root black rot symptom substantially alleviates, and relatively comparison minimizing 80%, Nicotiana tabacum L. average yield per mu ratio does not uses invention formulation
Average yield per mu exceed about 18%.
Embodiment 7: the preparation method field efficacy of described a kind of bacteria agent preventing and treating black root of tobacco
Test is:
Biofungicide prepared by the present invention is applicable to prevent and treat two kinds of the main pathogenic fungi of Tobacco Root maize ear rot.Powdery
Bacteria agent using method: 1, in tobacco seedling transplant before, by microbial inoculum by 2 kilograms every mu consumption with have
Machine is fertile or thin sand and soil mix by 1: 10 after spread fertilizer over the fields and plant kind of plant hole is decided at the higher level but not officially announced.2, after tobacco seedling is transplanted
Before root black rot occurs, every mu of this microbial inoculum of use 80-100 times of pouring root of 2 kilograms of dilutions, every strain 100-300ml.
The present invention from Guangxi, Yunnan, Henan, Sichuan, the ground such as Guizhou acquire Tobacco Root maize ear rot sample 20
Many parts, according to the separation to its pathogen of the conventional tissue isolation, 6 strain Fusarium spp. bacterial strains and 4 are obtained
Strain base is taken root beading mycete pathogenic strain.Select the strain of test chamber preservation, use growth rate method to measure and protect
Hide the inhibitory action that pathogenic bacterium are grown by strain, its antibacterial percentage rate of statistical analysis, obtain prevention effect good
Several bacterial strains, then determine inoculation ratio according to bacteriostasis size, carry out mixed fungus fermentation cultivation,
Obtain yeast culture thing and include thalline and metabolite thereof, add the particle diameter agriculturally allowing to use less than 1 milli
Following single or the filler such as peat composed of rotten mosses powder, Testa oryzae of combination of rice, Kaolin, sterilizing fertilizer, humic acids,
White carbon mixes, and in mixture, microorganism and metabolite thereof the i.e. percentage by weight of fermentation end products is
40-75%.Put into hothouse and be dried moisture less than less than 8%, crushing packing, make powder, be used for preventing
Control black root of tobacco.Through long-time field test and chemical formulation change determine this bacteria agent production technology and
Application process.Through testing this bacteria agent, black root of tobacco is had stronger inhibitory action.Along with the black corruption of root
The continuous growth of pathogenic bacteria silk, when running into this bacteria agent, root black rot mycelia stops diffusion, and to not having
The local extension of bacterial strain, shows that this bacteria agent has inhibitory action to root black rot.Powdery obtained by the present invention
Bacteria agent respectively reaches 85% and 72% to the suppression ratio of black root of tobacco mycelial growth and spore germination.
In greenhouse pot culture is tested, before inoculation black root of tobacco, 24h uses bacteria agent prevention effect is 83%,
After inoculation, 24h uses the prevention effect of fermentation liquid is 70%；In field test, use powdery bacteria agent
The effect preventing and treating black root of tobacco is 65-80%.This microbial inoculum holding time is 1 year.
Above-described is only some embodiments of the present invention.For the person of ordinary skill of the art,
Without departing from the concept of the premise of the invention, it is also possible to make some deformation and improvement, these broadly fall into
The protection domain of invention.
Claims (4)
1. the preparation method of the bacteria agent preventing and treating black root of tobacco, it is characterised in that: its strain component
For: swamp Rhodopseudomonas 10%, bacillus subtilis 20%, bacillus polymyxa 10%, the short spore of side spore
Bacillus 5%, bacillus megaterium 15%, Trichoderma viride 5%, Trichoderma harzianum 5%, Issatchenkia orientalis 10%,
Streptomyces microflavus 10%, sporotrichum thermophile 8%, pythium oligandrum 2%；Its preparation method is: first carry out microorganism
Test tube slant seed culture and microbial liquid seed culture, then liquid seeds is accessed in fermentation medium,
Pass through liquid fermentation, it is thus achieved that substantial amounts of thalline and metabolite thereof, thalline and metabolite thereof mixed with carrier,
Crushed after being dried makes powder.
The preparation method of a kind of bacteria agent preventing and treating black root of tobacco the most according to claim 1,
It is characterized in that: the preparation method of described bacteria agent is:
1. slant culture: above-mentioned original strain is inoculated in the most respectively in the culture medium of correspondence and lives
Changing, the slant medium of use has the PDA-sucrose cultivation cultivating pythium oligandrum, Trichoderma viride, Trichoderma harzianum
Base cultivation temperature is set under the conditions of 28 DEG C and cultivates 4 days, cultivates sporotrichum thermophile, the PDA-of Issatchenkia orientalis
Dextrose culture-medium cultivation temperature is cultivated 3 days under the conditions of being located at 30 DEG C, and sporotrichum thermophile cultivation temperature controls
Cultivate 3 days, cultivate bacillus subtilis, Paenibacillus polymyxa, Brevibacillus laterosporus, huge for 45 DEG C
Bacillus cereus, the nutrient agar cultivation temperature of swamp Rhodopseudomonas are located at 30 DEG C and cultivate 3 days, training
The Gause I culture medium supporting streptomyces microflavus controls cultivation temperature 30 DEG C cultivation 5 days；
2. seed culture: liquid amplification culture medium has cultivation pythium oligandrum, Trichoderma viride, the PDA-of Trichoderma harzianum
Sucrose medium cultivation temperature is cultivated 2 days in 150r/min shaking table under the conditions of being set in 28 DEG C, cultivates thermophilic side
Spore is mould, the PDA-dextrose culture-medium cultivation temperature of Issatchenkia orientalis is cultivated 1 day under the conditions of being located at 28 DEG C,
Sporotrichum thermophile cultivation temperature controls to cultivate 1 day at 28 DEG C, cultivates the Gause I culture medium of streptomyces microflavus
Cultivation temperature is cultivated 2 days in 150r/min shaking table under the conditions of being set in 28 DEG C, bacillus subtilis, glues class more
Bacillus cereus, Brevibacillus laterosporus, bacillus megaterium, the Nutrient agar of swamp Rhodopseudomonas are cultivated
Base cultivation temperature is cultivated 1 day in 150r/min shaking table under the conditions of being located at 28 DEG C, and seed liquor inoculation is joined by volume
Ratio: swamp Rhodopseudomonas 10%, bacillus subtilis 20%, bacillus polymyxa 10%, the short spore of side spore
Bacillus 15%, bacillus megaterium 15%, Trichoderma viride 5%, Trichoderma harzianum 5%, Issatchenkia orientalis 10%,
Streptomyces microflavus 10%, sporotrichum thermophile 8%, pythium oligandrum 2%；
3. fermentation medium is cultivated: cultured liquid seeds is aseptically received through autoclaved
In fermentation medium, fermentation medium component is: brown sugar 0.6%, glucose 0.3%, corn starch 1.8%,
Bean cake powder 1.5%, magnesium sulfate 0.03%, carbamide 0.12%, potassium dihydrogen phosphate 0.04%, peptone 0.1%, ferment
Female powder 0.05%, sodium carbonate regulation PH to 7.0, liquid microelement 5mL.Condition of culture: inoculum concentration 5-10%,
Cultivation temperature controls at 28-30 DEG C, rotating speed 120-180r/min, ventilation 1: 0.5-0.8, fermentation culture
24-36 hour；
4. prepared by powder: microbial cells fermentation liquid is less than with being crushed to particle diameter with 50% weight ratio after preparing
The sterilizing fertilizer of less than 1 millimeter does carrier with white carbon 3: 1 mass ratio and mixes, and puts into hothouse and is dried
Moisture be less than less than 8%, crushing packing, the solid, powdery bacteria agent of black root of tobacco must be prevented and treated, have
Effect viable count >=5,000,000,000/g.
The preparation side of a kind of bacteria agent preventing and treating black root of tobacco the most according to claim 1 and 2
Method, is characterized in that: described microorganism and metabolite thereof i.e. fermentation end products is with 40-75% weight ratio and powder
It is broken to the particle diameter single or carrier such as peat composed of rotten mosses powder, Testa oryzae, Kaolin, the sterilizing of combination less than less than 1 millimeter
The single carriers such as fertilizer, humic acids, white carbon or peat composed of rotten mosses powder, Testa oryzae, sterilizing fertilizer and humic acid,
The combination carrier that Kaolin, white carbon mix with 3-5: 1 ratio, puts into hothouse and is dried, and moisture is less than
Less than 8%, crushing packing, it is placed in the room temperature cool place place of being dried and preserves.
The preparation side of a kind of bacteria agent preventing and treating black root of tobacco the most according to claim 1 and 2
Method, is characterized in that: described bacteria agent content >=2,000,000,000/g.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201511020388.2A CN105875645A (en) | 2015-12-23 | 2015-12-23 | Preparing method of biological agent for preventing and controlling tobacco root black rot |
Applications Claiming Priority (1)
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| CN106431661A (en) * | 2016-09-28 | 2017-02-22 | 西南大学 | Preparation method and application of Pythium oligandrum microbial fertilizer |
| CN107646878A (en) * | 2017-10-11 | 2018-02-02 | 杨建设 | Prevent and treat microbial inoculum of tobacco mosaic virus (TMV) and preparation method thereof |
| CN107668073A (en) * | 2017-10-10 | 2018-02-09 | 云南农业大学 | A kind of bacillus polymyxa biological agent and preparation method for preventing and treating plant root rot |
| CN108841752A (en) * | 2018-07-06 | 2018-11-20 | 四川农业大学 | The application of bacillus megaterium BM22 and its gemma liquid preparation in prevention and treatment cyclamen black root |
| CN109135762A (en) * | 2018-10-30 | 2019-01-04 | 上海理工大学 | A kind of microbial-bacterial fertilizer and preparation method thereof for promoting tangerine orange to grow |
| CN109526986A (en) * | 2018-11-22 | 2019-03-29 | 云南省烟草公司临沧市公司 | A kind of tobacco diseases prevention biological prevention and control agent and preparation method thereof |
| CN110150094A (en) * | 2019-06-14 | 2019-08-23 | 甘肃绿能农业科技股份有限公司 | A kind of disease and insect resistance Radix Angelicae Sinensis seedling medium |
| CN111139186A (en) * | 2019-12-28 | 2020-05-12 | 河南省农业科学院植物保护研究所 | Trichoderma viride with disease prevention and growth promotion functions and application thereof |
| CN111500644A (en) * | 2020-04-28 | 2020-08-07 | 中国农业科学院烟草研究所 | Preparation method of composite bacteria fermentation product for preventing and treating phytophthora root rot of plants, composite bacteria fermentation product and application of composite bacteria fermentation product |
| CN112195138A (en) * | 2020-11-06 | 2021-01-08 | 河南省农业科学院植物营养与资源环境研究所 | Biological agent for livestock and poultry manure composting and preparation method thereof |
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Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106431661A (en) * | 2016-09-28 | 2017-02-22 | 西南大学 | Preparation method and application of Pythium oligandrum microbial fertilizer |
| CN107668073A (en) * | 2017-10-10 | 2018-02-09 | 云南农业大学 | A kind of bacillus polymyxa biological agent and preparation method for preventing and treating plant root rot |
| CN107646878A (en) * | 2017-10-11 | 2018-02-02 | 杨建设 | Prevent and treat microbial inoculum of tobacco mosaic virus (TMV) and preparation method thereof |
| CN108841752B (en) * | 2018-07-06 | 2020-08-18 | 四川农业大学 | Bacillus megaterium BM22 and application of spore liquid preparation thereof in preventing and treating cyclamen persicum radices |
| CN108841752A (en) * | 2018-07-06 | 2018-11-20 | 四川农业大学 | The application of bacillus megaterium BM22 and its gemma liquid preparation in prevention and treatment cyclamen black root |
| CN109135762A (en) * | 2018-10-30 | 2019-01-04 | 上海理工大学 | A kind of microbial-bacterial fertilizer and preparation method thereof for promoting tangerine orange to grow |
| CN109526986A (en) * | 2018-11-22 | 2019-03-29 | 云南省烟草公司临沧市公司 | A kind of tobacco diseases prevention biological prevention and control agent and preparation method thereof |
| CN110150094A (en) * | 2019-06-14 | 2019-08-23 | 甘肃绿能农业科技股份有限公司 | A kind of disease and insect resistance Radix Angelicae Sinensis seedling medium |
| CN111139186A (en) * | 2019-12-28 | 2020-05-12 | 河南省农业科学院植物保护研究所 | Trichoderma viride with disease prevention and growth promotion functions and application thereof |
| CN111139186B (en) * | 2019-12-28 | 2022-07-15 | 河南省农业科学院植物保护研究所 | Trichoderma viride with disease prevention and growth promotion functions and application thereof |
| CN111500644A (en) * | 2020-04-28 | 2020-08-07 | 中国农业科学院烟草研究所 | Preparation method of composite bacteria fermentation product for preventing and treating phytophthora root rot of plants, composite bacteria fermentation product and application of composite bacteria fermentation product |
| CN111500644B (en) * | 2020-04-28 | 2022-02-08 | 中国农业科学院烟草研究所 | Preparation method of composite bacteria fermentation product for preventing and treating phytophthora root rot of plants, composite bacteria fermentation product and application of composite bacteria fermentation product |
| CN112195138A (en) * | 2020-11-06 | 2021-01-08 | 河南省农业科学院植物营养与资源环境研究所 | Biological agent for livestock and poultry manure composting and preparation method thereof |
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