CN101781565B - Multifunctional biological agent containing Trichoderma pseudokiningii - Google Patents

Multifunctional biological agent containing Trichoderma pseudokiningii Download PDF

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CN101781565B
CN101781565B CN2009102309841A CN200910230984A CN101781565B CN 101781565 B CN101781565 B CN 101781565B CN 2009102309841 A CN2009102309841 A CN 2009102309841A CN 200910230984 A CN200910230984 A CN 200910230984A CN 101781565 B CN101781565 B CN 101781565B
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trichoderma pseudokiningii
trichoderma
fermented liquid
pseudokiningii
soil
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CN101781565A (en
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宋保平
陈靠山
尹倩倩
孙斐
李淑梅
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SHANDONG DUOLIDE BIO-TECHNOLOGY Co Ltd
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SHANDONG DUOLIDE BIO-TECHNOLOGY Co Ltd
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Abstract

The invention provides a DLD1 strain of Trichoderma pseudokiningii, which is preserved in the CGMCC, and the number is CGMCC NO.3221. The invention also provides a formula and a preparation method of the multifunctional biological agent for soil melioration and continuous cropping resistance prepared by complex of fermentation liquor of the DLD1 of the Trichoderma pseudokiningii, fulvic acid, zinc sulfate and the like. The biological agent has the functional effects: (1) adopting bio-control microorganism to prevent the harmful flora from infecting plants and killing pathogenic bacteria, inducing the plants to generate disease resistance, and preventing the death of seedling and rot of plant; and (2) adopting the complex synergic effect of the Trichoderma pseudokiningii, fulvic acid and zinc sulfate to induce the root system to grow, realize the radication and soundseedling, and break hardened soil, form crumble structure, and realize cultivation without deep ploughing, water retention and soil moisture conservation, fertilizer damage remission and soil fertility improvement.

Description

A kind of multifunctional biological agent that contains trichoderma pseudokiningii
Technical field
The present invention is the multifunctional biological agent that contains trichoderma pseudokiningii, relates to the prescription and the preparation method of soil improvement and successive planting resistant agent.
Background technology
Good edaphic condition is the prerequisite of crop high yield, but because the weather arid, chemical fertilizer excessively uses, irrational cultivation and management, and reasons such as continuous cropping cause that soil ecology worsens, crop failure has caused enormous economic loss.The deterioration of soil ecology mainly is root system water, vapour, the fertile unbalance and microbial continuous cropping disease of continuous cropping cause of disease that soil compaction causes to the influence of crop.The method of exploring is effectively broken soil compaction, and the control continuous cropping disease has huge economic benefit and social effect.
At present, the method for breaking soil compaction mainly is the machinery deep ploughing and uses soil improvement agent.The machinery deep ploughing is to utilize high-power agricultural machine, and plowed soils thickness reaches more than the 60cm, loose hardened soil, and the reconstruct crumb structure is increase soil fertility, water-retaining effect, this method is pollution-free, does not destroy profitable strain.Shortcoming is that the soil amount of stirring is big, forms uneven ridges easily, influences the land smoothing degree, and labour intensity is big, and cost is high.Use effectively improved soil nodule structure of soil improvement agent, play the effect of ventilation, water-retaining effect.Soil improvement agent comprises synthetic modifying agent and natural modifying agent.SEPIGEL 305, synthetic modifying agent costs such as asphalt emulsion are high, and the time length is short, and ecotope is had disadvantageous effect.Natural modifying agent such as saccharan, humic acid, peat have same improved effect, but cost is low relatively, and the greenization environmental protection has remarkable advantages.
Anti-continuous cropping disease main method has: pyroprocessing, and chemical agent is handled, and microbiobacterial agent is handled.Pyroprocessing is to improve the soil moisture (40-50 ℃, 15 days) through modes such as covering with plastic film to reach the effect of killing pathogenic bacteria.This method can only be carried out the shallow-layer sterilization to the face of land, and deep soil is not had unusual effect.The method that chemical agent is handled, more with trichloronitromethane and monobromethane use, advantage is that effect is remarkable to the strong toxicity of pathogenic soil bacterium.Shortcoming is that environment is caused disadvantageous effect, when killing pathogenic bacteria, the profitable strain in the soil is destroyed, and is unfavorable to plant growth, to the toxic effect of people and animals.Microbial bactericide utilizes the competition between mikrobe through executing probioticses such as biocontrol microorganisms, fixed nitrogen, phosphorus decomposing, potassium decomposing outward, antagonism, and effects such as superparasitism suppress or kill pathogenic bacteria, and protection also promotes plant root growth, increases soil fertility.It is a kind of method of anti-continuous cropping disease of high-efficiency environment friendly.
Harden and the continuous cropping disease of soil is accompanied often, but also do not have a kind of effective means at present, possesses the effect of soil improvement and continuous cropping disease control simultaneously.
Summary of the invention
The object of the invention is to provide a kind of reclamation result of natural soil conditioner and multifunctional biological agent of microbe soil bactericidal agent for preventing and treating continuous cropping disease effect of having combined.Its effect is that (1) utilizes biological and ecological methods to prevent plant disease, pests, and erosion mikrobe blocking-up pernicious bacteria to infect plant and kill pathogenic bacteria, and inducing plant produces disease resistance, mashed of anti-dead seedling; (2) utilize the composite synergistic effect of mikrobe and xanthohumic acid composition, induce root growth, realize strong plantlets and rootage, strengthen resistance against diseases; (3) utilize the composite synergistic effect of mikrobe and xanthohumic acid, zinc sulfate, break soil compaction, form crumb structure, realize exempting from deep ploughing and do, water-retaining effect, alleviate fertilizer damage, increase fertility of land.
To achieve these goals, the invention provides a kind of trichoderma pseudokiningii (Trichoderma pseudokiningii) DLD1 bacterial strain.
Said trichoderma pseudokiningii DLD1 bacterial strain is that the Licheng District corn stalk separates from the Jinan City; The eugonic bacterial strain that utilizes the PDA culture plate to cultivate, screen; Called after trichoderma pseudokiningii (Trichoderma pseudokiningii) DLD1, on August 12nd, 2009 in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) preservation, be numbered CGMCCNO.3221; The address is the Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica.
Trichoderma pseudokiningii (Trichoderma pseudokiningii) DLD1 has following characteristic:
This microbial strains can be in 25 ℃ of-30 ℃ of scopes growth fast, optimum growth temperature is 28 ℃, in being higher than 35 ℃ environment, suppresses growth; In the PDA culture plate, white mycelium, sporophore main shaft secondary branch complex structure has 2-3 and goes back to branch; Bottle metulae portion shrinks obviously, and expand the centre, and the top attenuates, and ultimate bottle stalk prolongs, and base portion does not attenuate, and the conidiogenous cell size is the μ m of (3.00-7.50) μ m * (1.25-2.00), and conidium is short cylindrical shape, and size is the μ m of (2.5-4.5) μ m * (2.0-2.5).
Trichoderma pseudokiningii preparation of fermentation liquid method is following:
At first trichoderma pseudokiningii (Trichoderma pseudokiningii) DLD1 bacterial strain is utilized the dull and stereotyped activation of PDA, after receive in the liquid PDA substratum, cultivate on shaking table the sterilization back, makes seed liquor; Be fermention medium with yam, bran mass again, successively in seeding tank, secondary seed jar and fermentor tank, stir aerobic fermentation; Mixing, embedding, concentrated make the trichoderma pseudokiningii fermented liquid then.
Concrete preparation method is:
The trichoderma pseudokiningii seed culture:
At first the trichoderma pseudokiningii bacterial strain is utilized the dull and stereotyped activation 24h of PDA, after receive in liquid PDA substratum (yam 200g/L, the sucrose 100g/L) triangular flask, cultivate on shaking table the sterilization back, culture condition is 28 ℃, 100rpm, cultivates 36h, obtains seed liquor.
The fermentation process of trichoderma pseudokiningii:
Fermention medium is yam, bran mass, and composition is following: wheat bran 40g/L, yam 50g/L, sucrose 20g/L, KH 2PO 40.5g/L, MgSO 47H 2O 0.5g/L, CaCl 20.25g/L.
The condition of fermentation culture is following: pH5-7,28 ℃ of culture temperature, rotating speed 20-50rpm, inoculum size 3%-7%.
Fermentation process is following: by inoculum size bacterial classification is received in the seeding tank, stirred aerobic culture 36h, get into the secondary seed jar then and stir the cultivation 36h that ventilates, stir aerobic culture 96h in the fermentor tank that reinjects; Get into the high-speed shearing machine mixing then, embedding gets into the economic benefits and social benefits thickener again, and under 45 ℃ temperature, dense system trichoderma pseudokiningii culture makes the trichoderma pseudokiningii fermented liquid.Contain Trichoderma silk dry weight 0.1-1.0g in every 100ml trichoderma pseudokiningii fermented liquid of gained, the fermentation culture effect is to contain Trichoderma silk dry weight 0.9-1.0g in every 100ml trichoderma pseudokiningii fermented liquid preferably
Use above-mentioned trichoderma pseudokiningii fermented liquid and soil improvement agent xanthohumic acid, compositions such as zinc sulfate carry out composite, can make the multifunctional biological agent that is used for soil improvement, anti-continuous cropping.
The component of soil improvement according to the invention, anti continuous cropping multifunctional biological agent comprises trichoderma pseudokiningii fermented liquid, xanthohumic acid, zinc sulfate, and each composition weight umber is:
Trichoderma pseudokiningii fermented liquid 30-65
Xanthohumic acid 2-30
Zinc sulfate 0.5-5
Figure G2009102309841D00031
In order to improve effect and to improve the preparation performance, also can add following auxiliary component in soil improvement according to the invention, the anti continuous cropping multifunctional biological agent: vitamins C and/or α naa and/or chlorogenicacid and/or aminosallcylic acid and dextrin etc.
Feed composition and parts by weight thereof in the concrete preparation are following:
Figure G2009102309841D00032
Wherein:
Trichoderma pseudokiningii fermented liquid concentration: contain Trichoderma silk dry weight 0.9-1.0g in every 100ml fermented liquid, other component concentrations are by 100%.
The preparation method of soil improvement according to the invention, anti continuous cropping multifunctional biological agent is:
1. produce the trichoderma pseudokiningii fermented liquid: at first the trichoderma pseudokiningii bacterial strain is utilized the dull and stereotyped activation 24h of PDA, after receive in liquid PDA substratum (yam 200g/L, the sucrose 100g/L) triangular flask, cultivate on shaking table the sterilization back, makes seed liquor; Put seeding tank again into and constantly stir aerobic culture,, get into the secondary seed jar again and stir ventilation; Inject fermentation cylinder for fermentation then, stirring and evenly mixing gets into the high-speed shearing machine mixing again; Embedding gets into the economic benefits and social benefits thickener again, under 45 ℃ temperature, concentrates; Make the trichoderma pseudokiningii fermented liquid, A is subsequent use as microbe composition.(answering aseptic technique in the processing)
2. with xanthohumic acid, zinc sulfate adds in 50-60 ℃ the water fully stirring and dissolving, and chelating progressively is cooled to 30 ± 2 ℃; B is subsequent use as the soil improvement composition.
3. abundant stirring and dissolving in the water of aminosallcylic acid and/or chlorogenicacid and/or α naa and/or vitamins C adding 50-60C progressively is cooled to 30 ± 2 ℃, and C is subsequent use as the plant inducible factor.
4. last with above-mentioned subsequent use microbe composition A, soil improvement composition B, plant inducible factor composition C merging in material-compound tank, mixing, interpolation dextrin composition carry out emulsification processing, and be qualified through detecting, and is preparation finished product of the present invention.(above-mentioned preparation processing technology is except that specifying, all with reference to prior art).
Beneficial effect:
Trichoderma pseudokiningii of the present invention (Trichoderma pseudokiningii) DLD1 is a kind of plant biological control strain with multiple disease-resistant mechanism, to the person poultry harmless, does not produce environmental pollution, and growing ability is extremely strong.This bacterium can breed at crop root fast, forms the protection structure, the blocking-up pathogen infection.Simultaneously can pass through hyperparasitism, secrete various bacteriocidal substances and cell wall lysis enzyme, play the effect of killing pathogenic bacteria.In addition, trichoderma pseudokiningii can be induced and made the deposits yields disease resistance and promote plant-growth.Realize the crop strong plantlets and rootage, effectively reduced mashed of dead seedling.
Xanthohumic acid of the present invention belongs to humic-acid kind, can promote soil granular to form, and makes loosing soil, has good adsorption property.Thereby protect fertile, steady fertilizer, water conservation, ventilative, overcome the phenomenon that hardens of soil, lay the first stone for increasing soil fertility.Humic acids can also promote root growth and development to realize strong sprout through promoting the crop respiration.Humic acids is a kind of natural soil conditioner of high-efficiency environment friendly, and ME is simple, and is cheap.
Zinc sulfate of the present invention is a kind of trace element supplement.Because the plantation of the continuous cropping of crop, constantly absorb the specific trace element in the soil and can not get replenishing, especially zinc element is the easiest lacks in the continuous cropping continuous cropping soil, and additional zinc sulfate helps to alleviate the underproduction that continuous cropping causes.
Aminosallcylic acid, chlorogenicacid, the α naa, vitamins C helps promoting plant growth, disease resistance enhancemen as plant inducible factor C.
Above-mentioned each component rationally composite can produce synergistic function.
Biotechnological formulation of the present invention has soil improvement and anti-continuous cropping disease double effects, has solved soil ecology deterioration causing soil compaction and continuous cropping disease comprehensively.
The concrete method of use of the present invention can be irritated root, watered, sprays, dips in root with water.
Biotechnological formulation of the present invention has soil improvement and anti-continuous cropping disease double effects, has solved soil ecology deterioration causing soil compaction and continuous cropping disease more comprehensively.
Embodiment:
Pharmaceutical formulation embodiment:
Example 1, soil improvement of the present invention, anti continuous cropping multifunctional biological agent are formed by the preparation of raw material of following (part) by weight ratio:
Trichoderma pseudokiningii fermented liquid 42, xanthohumic acid 15, zinc sulfate 4, vitamins C 7, α naa 0.8, chlorogenicacid 10, aminosallcylic acid 7, dextrin 8, add water to 100 parts.
Example 2, soil improvement of the present invention, anti continuous cropping multifunctional biological agent are formed by the preparation of raw material of following (part) by weight ratio:
Trichoderma pseudokiningii fermented liquid 40, xanthohumic acid 10, zinc sulfate 3, vitamins C 6, α naa 0.7, chlorogenicacid 9, aminosallcylic acid 6, dextrin 7, add water to 100 parts.
Example 3, soil improvement of the present invention, anti continuous cropping multifunctional biological agent are formed by the preparation of raw material of following (part) by weight ratio:
Trichoderma pseudokiningii fermented liquid 45, xanthohumic acid 20, zinc sulfate 5, vitamins C 7, α naa 0.8, chlorogenicacid 9, aminosallcylic acid 6, dextrin 8, add water to 100 parts.
The preparation method of soil improvement of the present invention, anti continuous cropping multifunctional biological agent:
1. the preparation method of trichoderma pseudokiningii fermentation: (answering aseptic technique in the processing)
At first the trichoderma pseudokiningii bacterial strain is utilized the dull and stereotyped activation 24h of PDA, after receive in liquid PDA substratum (yam 200g/L, the white sugar 100g/L) triangular flask, the sterilization back under the 100rpm condition, is cultivated 36h at 28 ℃, obtains seed liquor; , receive in the seeding tank by 3% inoculum size, stir aerobic culture 36h, get into the secondary seed jar again and stir the cultivation 36h that ventilates, inject fermentor tank then and stir aerobic culture 96h; Fermention medium is potato, bran mass (wheat bran 40g/L, yam 50g/L, sucrose 20g/L, KH 2PO 40.5g/L, MgSO 47H 2O 0.5g/L, CaCl 20.25g/L, pH5-7), 28 ℃ of culture temperature, rotating speed 20-50rpm.Get into the high-speed shearing machine mixing again, embedding gets into the wooden mould culture of economic benefits and social benefits thickener dense system under 45 ℃ temperature then; A is subsequent use as microbe composition.
2. with xanthohumic acid, zinc sulfate adds in 50-60 ℃ the water fully stirring and dissolving, and chelating progressively is cooled to 30 ± 2 ℃; B is subsequent use as the soil improvement composition.
3. with aminosallcylic acid, chlorogenicacid, α naa, vitamins C add in 50-60 ℃ the water fully stirring and dissolving, progressively are cooled to 30 ± 2 ℃, and C is subsequent use as the plant inducible factor.
4. last with above-mentioned subsequent use microbe composition A, soil improvement composition B, plant inducible factor composition C merging in material-compound tank, mixing, interpolation dextrin composition also carry out emulsification, again through detecting the qualified finished product of the present invention that is;
Concrete application example:
Instance 1:
The prescription reference formulation embodiment 1 of (1) soil improvement, anti continuous cropping multifunctional biological agent.
(2) method of use: Qufu City Dong local strawberry booth carries out in Shandong.Use the present invention to dilute 60 times earlier and spray ground, underground in conjunction with plough under wholely.After the strawberry field planting, use the present invention to dilute 300-400 and doubly spray ground, logotype 3-4 time sprayed 10-15 days at interval.Every 666.7m 2The each 2-3 liter of consumption.Strawberry cultivars is beautiful king No. 1.
In adjacent plot not use the present invention as contrast.Every sub-district area is 100m 2, three repetitions.Other management are all with conventional.
Upgrowth situation to the root of seedling phase, stem, leaf is investigated.
Test-results is seen table 1:
Table 1 the present invention is to the influence of strawberry seedling phase root, stem, leaf
Figure G2009102309841D00061
Can know by table 1, use the present invention after, average individual plant root system number, root length, capillary radical amount have increased by 33.3%, 13.6% and 54.1% respectively; Average stem rugosity has increased by 0.5 times; Blade quantity, the speed of growth, effectively leaf age has improved 55.6%, 22.2% and 33.3% respectively.Explain and use the present invention can improve edatope, promote the strawberry strong plantlets and rootage, the effect highly significant.
Instance 2
The prescription reference formulation embodiment 1 of (1) soil improvement, anti continuous cropping multifunctional biological agent.
(2) method of use: carry out in Jinxiang County, Shandong, this regional cotton continuous cropping was planted in more than 5 years, and the soil compaction that the unreasonable use of chemical fertilizer causes is serious, and soil-borne disease is in vogue.Trial crops is a cotton.Use the present invention to dilute 60 times and spray ground and fertilizer and mix and execute, underground in conjunction with plough under wholely.Every 666.7m 2Consumption 2-3 liter.
In adjacent plot not use the present invention as contrast.Every sub-district area is 100m 2, three repetitions.Other management are all with conventional.
Carry out the investigation of sprout term disease control effect; Be respectively after the field planting 10 days, 20 days, 30 days, 40 days, 45 days; To this stage by sprout term diseases such as the samping off that causes because of soil-borne disease, cold spell in later spring, temperature, ground temperature, high humidity, damping-off, root rot, red rot, carry out investigation statistics.
Test-results is seen table 2:
Table 2 the present invention is to the influence of cotton in seedling stage resistance against diseases
Figure G2009102309841D00071
Can know by table 2, used the present invention after, the seedling of dying of illness; The seedling of catching an illness has obtained control effectively in whole seedling stage (10 days-45 days), has kept higher healthy seedling rate, shows that this invention can be prone to susceptible seedling stage in cotton; Improve edatope, effectively control the generation of soil-borne disease.The continuous cropping disease that causes for continuous cropping for many years has significant preventive effect.
Instance 3
The prescription reference formulation embodiment 1 of (1) soil improvement, anti continuous cropping multifunctional biological agent.
(2) method of use: university student's thing chemistry carries out the soil improvement test with molecular biology research institute experimental plot in Shandong.With experimental plot soil as experimental subjects.Using the present invention to dilute 60 times and spray soil, is control group to spray zero(ppm) water.。Sampling, measuring method are with reference to State Standard of the People's Republic of China's soil environment quality standard (GB15618-1995).Measure control group and soil organic matter content, water holding capacity, the unit weight of using experimental group of the present invention respectively.
Table 3 the present invention is to the influence of the soil organism, water cut, unit weight
Can know by table 3, use the present invention after, the soil organism, water holding capacity, the soil weight all are significantly increased.The result shows that the present invention can effectively improve the soil particle degree, breaks soil compaction, increases soil fertility and water holding capacity.Has tangible reclamation result.
Instance 4
The prescription reference formulation embodiment 1 of (1) soil improvement, anti continuous cropping multifunctional biological agent.
(2) method of use: university student's thing chemistry carries out the soil parameters detection with molecular biology research institute experimental plot in Shandong.Soil adopts experimental plot soil.Use the present invention to dilute 60 times and spray soil.Potted plant cucumber seedling is used zero(ppm) water as contrast.Totally 100 basins, every basin 3 strains.Other management are all with conventional.
Using back 0d respectively, 3d, 6d, 9d, 12d, 24d takes a sample to seedlings root.Method is with reference to Li Chunxi chief editor " biostatistics " the 4th edition (Science Press).Each sampling spot triplicate.Measure plant disease-resistant relevant enzyme vigor, polyphenol oxides enzyme (PPO), superoxide-dismutase (SOD), px (POD), beta-1,3-glucanase content (β-1,3-Gluconase).Method is with reference to Zhang Zhiliang chief editor " plant physiology test direction " third edition (Higher Education Publishing House).
Table 4 the present invention is to the influence (unit: U/mg) of disease-resistant relevant enzyme vigor in the root system
Figure G2009102309841D00081
Can know by table 4, use the present invention after, the vigor of several kinds of disease-resistant key enzymes in the root system of plant improved rapidly in 3 days, these enzymes can effectively suppress, kill the soil-borne disease that infects root system in the soil.Enzyme activity all is significantly higher than control group in after using 24 days.Can effectively resist soil-borne disease of crop at this product of Physiology and biochemistry level proof.

Claims (7)

1. a multifunctional biological agent that contains trichoderma pseudokiningii is characterized in that the component of said biotechnological formulation comprises trichoderma pseudokiningii fermented liquid, xanthohumic acid, zinc sulfate;
Each composition weight umber is:
Trichoderma pseudokiningii fermented liquid 30-65 part
Xanthohumic acid 2-30 part
Zinc sulfate 0.5-5 part;
Wherein, The bacterial strain of trichoderma pseudokiningii fermented liquid is trichoderma pseudokiningii (Trichoderma pseudokiningii) DLD1 bacterial strain; This bacterial strain in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) preservation, is numbered CGMCC NO.3221; Contain Trichoderma silk dry weight 0.9-1.0g in every 100ml trichoderma pseudokiningii fermented liquid.
2. biotechnological formulation as claimed in claim 1,, it is characterized in that the component of said biotechnological formulation also comprises vitamins C, α naa, chlorogenicacid, aminosallcylic acid and dextrin;
Each composition weight umber is:
Wherein, the bacterial strain of trichoderma pseudokiningii fermented liquid is trichoderma pseudokiningii (Trichoderma pseudokiningii) DLD1 bacterial strain; Contain Trichoderma silk dry weight 0.9-1.0g in every 100ml trichoderma pseudokiningii fermented liquid.
3. biotechnological formulation as claimed in claim 2 is characterized in that each composition weight umber of said biotechnological formulation is:
Figure FSB00000744509900012
Figure FSB00000744509900021
Wherein, the bacterial strain of trichoderma pseudokiningii fermented liquid is trichoderma pseudokiningii (Trichoderma pseudokiningii) DLD1 bacterial strain; Contain Trichoderma silk dry weight 0.9-1.0g in every 100ml trichoderma pseudokiningii fermented liquid.
4. biotechnological formulation as claimed in claim 2 is characterized in that each composition weight umber of said biotechnological formulation is:
Figure FSB00000744509900022
Wherein, the bacterial strain of trichoderma pseudokiningii fermented liquid is trichoderma pseudokiningii (Trichoderma pseudokiningii) DLD1 bacterial strain; Contain Trichoderma silk dry weight 0.9-1.0g in every 100ml trichoderma pseudokiningii fermented liquid.
5. according to claim 1 or claim 2 the preparation method of biotechnological formulation is characterized in that said trichoderma pseudokiningii preparation of fermentation liquid method is following:
At first trichoderma pseudokiningii (Trichoderma pseudokiningii) DLD1 bacterial strain is utilized the dull and stereotyped activation of PDA, after receive in the liquid PDA substratum, cultivate on shaking table the sterilization back, makes seed liquor; Be fermention medium with potato, bran mass then, successively in seeding tank, secondary seed jar and fermentor tank, stir aerobic fermentation; Mixing, embedding, concentrated make the trichoderma pseudokiningii fermented liquid then.
6. according to claim 1 or claim 2 the preparation method of biotechnological formulation is characterized in that the concrete preparation method of said trichoderma pseudokiningii fermented liquid is following:
The trichoderma pseudokiningii seed culture:
At first the trichoderma pseudokiningii bacterial strain is utilized the dull and stereotyped activation 24h of PDA, after receive liquid PDA substratum: yam 200g/L, among the sucrose 100g/L, cultivate on shaking table the sterilization back, culture condition is 28 ℃, 100r/min, cultivates 36h, obtains seed liquor;
The fermentation process of trichoderma pseudokiningii:
Fermention medium is potato, bran mass, and composition is following: wheat bran 40g/L, yam 50g/L, sucrose 20g/L, KH 2PO 40.5g/L, MgSO 47H 2O 0.5g/L, CaCl 20.25g/L;
The condition of fermentation culture is following: pH5-7,28 ℃ of culture temperature, rotating speed 20-50rpm, inoculum size 3%-7%;
Fermentation process is following: by inoculum size bacterial classification is received in the seeding tank, stirred aerobic culture 36h, get into secondary seed jar stir culture 36h then, stir aerobic culture 96h in the fermentor tank that reinjects; Get into the high-speed shearing machine mixing then, embedding gets into the economic benefits and social benefits thickener again, and under 45 ℃ temperature, the wooden mould culture of dense system makes the trichoderma pseudokiningii fermented liquid.
7. like the preparation method of claim 2,3 or 4 described biotechnological formulations, it is characterized in that concrete steps are following:
1. produce the trichoderma pseudokiningii fermented liquid, A is subsequent use as microbe composition;
2. with xanthohumic acid, zinc sulfate adds in 50-60 ℃ the water fully stirring and dissolving, and chelating progressively is cooled to 30 ± 2 ℃; B is subsequent use as the soil improvement composition;
3. aminosallcylic acid and/or chlorogenicacid and/or α naa and/or vitamins C add in 50-60 ℃ the water fully stirring and dissolving, progressively are cooled to 30 ± 2 ℃, and C is subsequent use as the plant inducible factor;
4. last with above-mentioned subsequent use microbe composition A, soil improvement composition B, plant inducible factor composition C merging in material-compound tank, mixing, interpolation dextrin composition carry out emulsification processing, and be qualified through detecting, and is the biotechnological formulation finished product.
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CN110386844A (en) * 2019-07-25 2019-10-29 王晓雯 A kind of fulvic acid microbial bacterial agent and preparation method thereof
CN111559933A (en) * 2020-04-20 2020-08-21 杨新正 Special fertilizer for continuous cropping prevention cotton and preparation method thereof

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CN1422828A (en) * 2002-12-20 2003-06-11 黎保文 Microbial compound liquid fertilizer and method for preparing the same
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