CN103951483A - Biological antibacterial fertilizer for preventing and controlling soil-borne diseases of crops as well as production process and application of biological antibacterial fertilizer - Google Patents
Biological antibacterial fertilizer for preventing and controlling soil-borne diseases of crops as well as production process and application of biological antibacterial fertilizer Download PDFInfo
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The invention discloses a biological antibacterial fertilizer for preventing and controlling soil-borne diseases of crops as well as a production process and an application of the biological antibacterial fertilizer. The production process comprises the steps of culturing to obtain bacillus cereus and pseudomonas choloeaphtis fermentation liquids, and inoculating the two fermentation liquids into cured livestock manure and straw according to a certain proportion to compost and ferment to form the biological antibacterial fertilizer for preventing and controlling soil-borne diseases of crops. The biological antibacterial fertilizer disclosed by the invention can be used for preventing and controlling vegetable botrytis cinerea and increasing the yield of vegetables, safe, pollution-free and easy to realize large-scale production.
Description
Technical field
The invention belongs to microbial organic fertilizer technical field, in particular to a kind of biocontrol bacterial fertilizer and production technique and application that prevents and treats farm crop soil-borne disease.
Background technology
Farm crop soil-borne disease causes huge financial loss to agriculture production every year, so disease control work becomes the task of top priority of current agricultural sciences.At present, the method for preventing and treating of farm crop soil-borne disease is mainly that to utilize disease-resistant strain plant and chemical prevention be main.On the one hand, use the plant of disease-resistant strain can reach cost-effective prevention effect, but wanting to obtain the desirable disease-resistant strain of a strain is not a nothing the matter, and often just there is the even situation such as poor growth of resistant lose in disease-resistant variety after it is promoted several years; On the other hand, chemical prevention because its preventive effect is fast compared with high, speed, antimicrobial spectrum plays very important effect compared with the advantage such as wide in the control of farm crop soil-borne disease, yet it is subject to again the restriction of three factors: the one, and the chemical sterilization pharmacy variety of current high-efficiency low-toxicity is less, great majority are still the medicament that toxicity is larger, with sterilization, are not long-term plan; The 2nd, environmental factors is also not little on the impact of prevention effect, and when envrionment conditions is applicable to pathogenic bacteria intrusion and propagates, its prevention effect often can not be satisfactory; The 3rd, chemical agent easily causes environmental pollution, crisis mankind daily life.As the biological control of one of pest control method, there is under certain conditions good prevention effect, and oneself becomes the focus of current disease control research and development.
In recent years, extensively utilize beneficial microorganism and meta-bolites thereof to suppress the survival activity of pathogenic bacteria both at home and abroad, this control strategy is developed rapidly, Antagonistic Fungi has played very important effect in this field of farm crop soil-borne disease biological control, comprise even mould of bacterium, actinomycetes, yeast, and people still mainly be take antagonistic bacterium as main in application.No matter the bacterium with antagonistic ability is in abiogenous biological control phenomenon or people is applied in the test of disease control, all there is very significantly prophylactic-therapeutic effect, its main advantage is: (l) kind of bacterium and quantity are various, in rhizosphere and soil surface and all existence in a large number of internal layer of plant; (2) antagonistic bacterium is more to the restraining effect mode of pathogenic bacteria, comprises that competition nutrition, parasitism, secretion bacteriocidal substance, inducing plant produce the modes such as resistance, thereby pathogenic bacteria is had a negative impact; (3) have surprising reproduction speed, the growth and breeding speed of general bacterium will be faster than fungi, actinomycetes etc.; (4) bacterium can artificial culture, is convenient to control, in practice easy handling; (5) some bacterium can not only prevent and treat disease, also has the effects such as the plant strain growth of promotion, thereby increases output.
Bio-bacterial manure, refers to the beneficial microorganism of the work that a class contains some amount, and is carried out coordinate plant growth speed, promoted crop yield to increase, improve the particular product of plant rhizosphere or surperficial micro-ecological environment by the biological activity of microorganism.Upper conventional bio-bacterial manure both at home and abroad at present comprises azotobacteria fertilizer, phosphorus decomposing/potassium bacterial manure, mycorhiza bacterial manure, biocontrol bacterial fertilizer etc.Biocontrol bacterial fertilizer as its name suggests, microorganism in bacterial manure mostly is biocontrol bacterium, these Antagonistic Fungis can carry out Suppressing phytopathogens by its growth metabolism process or its meta-bolites, kill pathogenic bacteria even, thereby for the growth of farm crop provides guarantee, some has been identified all has the microorganism of antagonistic action also can promote the fertilizer efficiency of bio-bacterial manure.
Grey mold Pseudomonas Deuteromycotina Staphlosporonites.Botrytis cinerea conidiophore is elongated, grey black, and size is (960～1200) μ m * (16～22) μ m, irregular tree-shaped branch, the apical cell of branch enlarges into spherical, on it raw many stigmas, on stigma raw conidium, likeness in form grape spike.Conidium unit cell is colourless, ellipse or oval, and size is (8～14) μ m * (6～9) μ m, the raw gray of spore collection.Sclerotium brown, out-of-shape, size is (2～4) mm * (1～3) mm[1].Botrytis cinerea can infect 200 various plants, belongs to fungal disease, the flower of various plants, really, leaf, stem all can fall ill.This sclerotium is survived the winter the summer on soil or invalid body, and germ is low temperature resistant, at 7-20 ℃ of a large amount of spore that produces, seedling stage temperature of shed 15-23 ℃.The low light level, relative humidity is more than 90% or seedling is surperficial while having moisture film susceptible disease.Florescence is the most susceptible, borrows air-flow, and irrigation and farming operation are invaded from wound, old and feeble organ.
The at present domestic agricultural chemicals to gray mold control is mainly chemical products, mainly contains: the mould prestige of second, procymidone, RP-26019, dimetachlone, phonetic mould amine, jingganmycin etc.In view of there is problem of environmental pollution in chemical agent, therefore study the suitable Novel biocontrol bacterial manure of preventing and treating gray mold, this long term growth for agriculture production seems particularly important.
Summary of the invention
First object of the present invention is to provide a kind of biocontrol bacterial fertilizer of preventing and treating farm crop soil-borne disease, and the Antagonistic Fungi in this bacterial manure has good inhibition to botrytis cinerea, can prevent and treat vegetable botrytis cinerea disease, improves vegetable crop.In addition, another object of the present invention is to provide a kind of production technique and application of above-mentioned biocontrol bacterial fertilizer.
The object of the present invention is achieved like this:
Prevent and treat a biocontrol bacterial fertilizer for farm crop soil-borne disease, wherein this biocontrol bacterial fertilizer, by every gram, contains and is greater than 1.0 * 10
8individual biocontrol microorganisms, described biocontrol microorganisms is comprised of bacillus cereus and Pseudomonas chlororaphis.
A production technique for above-mentioned biocontrol bacterial fertilizer, comprises the steps:
(1) bacillus cereus, Pseudomonas chlororaphis are inoculated into respectively in nutrient solution and carry out liquid state fermentation, obtain viable count and be greater than 1.0 * 10
9the bacillus cereus fermentation liquid of individual/ml, Pseudomonas chlororaphis fermented liquid, standby;
(2) fresh feces of livestock and poultry is evenly mixed by the weight ratio of 1:2-3 with stalk particle, compost 10-20 days to ight soil become thoroughly decomposed, stalk rots, sterilising treatment, forms solid fermentation matrix;
(3) by bacillus cereus fermentation liquid, Pseudomonas chlororaphis fermented liquid step (1) Suo Shu, according to the inoculum size of 20-40L/ ton, 50-100L/ ton, be inoculated in the described solid fermentation matrix of step (2) and carry out solid fermentation successively, leavening temperature is 25-32 ℃, in fermenting process, turning is 3-5 time, fermentation 5-8 days, obtains biocontrol bacterial fertilizer.
The production technique of the above-mentioned biocontrol bacterial fertilizer of the present invention, wherein in step (1): the consisting of of described nutrient solution: peptone 10g/L, extractum carnis 5g/L, NaCl 5g/L, pH7.0; The condition of the liquid state fermentation of described bacillus cereus is: under aeration-agitation in 28 ℃-32 ℃ fermentation 30-60h; The condition of the liquid state fermentation of described Pseudomonas chlororaphis is: under aeration-agitation in 25 ℃-30 ℃ fermentation 20-36h; Stalk particle described in step (2) is one or more particles after crushed in corn stalk, straw, paddy stalk, Rape Straw and rice straw.
The inventor finds unexpectedly when screening suppresses the Antagonistic Fungi of botrytis cinerea growth, and the composite bacteria liquid that contains bacillus cereus CICC21687 and Pseudomonas chlororaphis CICC20676 has restraining effect extremely significantly to botrytis cinerea.Based on this discovery, the inventor obtains the fermented liquid of bacillus cereus, Pseudomonas chlororaphis by cultivation, and these two kinds of fermented liquids are accessed to compost fermentation in the feces of livestock and poultry of slaking and stalk by a certain percentage, formed the biocontrol bacterial fertilizer that control farm crop soil passes botrytis cinerea disease.The biocontrol bacterial fertilizer that the present invention makes, wherein in every gram of fertilizer, contained effective Antagonistic Fungi number (cfu) is greater than 1.0 * 10
8individual, organic content (in butt) is greater than 25%, and moisture content is lower than 30%, and other index meets biological organic fertilizer national sector standard.Therefore, the invention provides the application of above-mentioned biocontrol bacterial fertilizer, that is: the application of above-mentioned biocontrol bacterial fertilizer in control farm crop soil-borne disease, described soil-borne disease is that botrytis cinerea is sick.
Bio-bacterial manure of the present invention is as a kind of biological product, and it is compared with chemical fertilizer has following several feature: (1) is spoiled soil structure, free from environmental pollution not, and to person poultry harmless; (2) improve soil fertility, fertilizer efficiency is lasting; (3) can prevent and treat soil-borne disease, promote the growth of farm crop, increase output, improve the quality of agricultural-food; (4) bio-bacterial manure raw material is feces of livestock and poultry and stalk the like waste, is easy to obtain, and turn waste into wealth, and the equipment that forms a complete production network is less demanding, and cost is lower.
Embodiment
Be below specific embodiment of the present invention, it should be noted that, various embodiments of the present invention bacillus cereus used be bacillus cereus (
bacillus cereus) CICC21687, Pseudomonas chlororaphis used be Pseudomonas chlororaphis (
pseudomonas chlororaphis) CICC20676, all buy in Chinese industrial microbial strains preservation administrative center (China Center of Industrial Culture Collection, CICC, is positioned at No. 6 building of No. 24, Road, Jiuxianqiao, Chaoyang District, Beijing City institute).
the inhibition test research of embodiment 1 different strains to botrytis cinerea
The preparation of PDA substratum: take peeling potato 200g, 30min is boiled in stripping and slicing, then gets juice by filtered through gauze, add again 20.0g glucose and 20.0g agar, after dissolving, supply water to 1000mL, divide the triangular flask that installs to 10 150mL, 121 ℃ of sterilizing 30min.
Under aseptic condition, get the PDA substratum that 20mL is cooled to 45 ℃ of left and right after sterilizing and add culture dish, make PDA flat board.Place after 24h, dull and stereotyped central authorities inoculation pathogenic bacteria botrytis cinerea mycelia, cultivate 4d for 22 ℃.In the botrytis cinerea plate of having grown, with diameter 1 cm punch tool, at colony edge, cut bacterium colony nahlock, standby.
Getting respectively bacillus cereus fermentation liquid, the Pseudomonas chlororaphis fermented liquid according to embodiment 2 techniques, prepared, is 1 * 10 with sterilized water dilution fermented liquid to cell concentration
5individual/ml
-1diluent, what wherein thalline counting process adopted is micro-direct-counting method.To placing the Antagonistic Fungi fermentation diluent that adds 100 μ L in the PDA flat board of l days (in composite bacteria liquid bacillus cereus with the quantity of Pseudomonas chlororaphis than being 1:10), with spreader, smoothen, each processes 4 repetitions.The botrytis cinerea bacterium piece cutting is seeded in to the flat board central authorities that coat, has the one side contact of mycelia dull and stereotyped.Cultivate after 4d for 25 ℃, with ruler " right-angled intersection method ", measure botrytis cinerea colony diameter (unit is cm), result is got its mean value.Separately get bacillus cereus fermentation liquid, Pseudomonas chlororaphis fermented liquid prepared by embodiment 1 step (3), (4), the centrifugal 30min of 4000r/min, supernatant liquor is through 0.22 μ L membrane filtration, and filtering thalline, obtains aseptic fermented supernatant fluid, in contrast.Be calculated as follows bacteriostasis rate: bacteriostasis rate=(D
contrast-D
process)/D
contrast* 1O0%.
According to the test-results of table 1, can find out, Pseudomonas chlororaphis or bacillus cereus are not remarkable to the inhibition of botrytis cinerea separately.Yet, use and compare with single bacterium, the composite bacteria liquid that these two kinds of bacterial strains form but shown to the very significant inhibition of botrytis cinerea (
p< 0.01).
The inhibition comparison of table 1 different strains to botrytis cinerea
| Antagonistic Fungi | Bacteriostasis rate (%) |
| Pseudomonas chlororaphis | 36.70±6.08 |
| Bacillus cereus | 15.52±5.96 |
| Composite bacteria liquid (Pseudomonas chlororaphis+bacillus cereus) | 92.17±10.31 ★★■■ |
With Pseudomonas chlororaphis comparison,
★ ★ p< 0.01; With the comparison of lactose streptomyces microflavus,
■ ■ p< 0.01.
The inventor finds in test, why the mixed bacteria liquid of Pseudomonas chlororaphis and bacillus cereus can bring into play the synergy that suppresses botrytis cinerea growth, possible cause is that two kinds of Antagonistic Fungis have the relation of the symbiosis of being similar to, the breeding that the metabolite of bacillus cereus is Pseudomonas chlororaphis provides some necessary somatomedins, thereby promote Pseudomonas chlororaphis Fast-propagation also to produce a large amount of antagonistic substances, then the effect by this class material causes the botrytis cinerea can not normal growth.The concrete mechanism of action, contriver can do further further investigation in follow-up test.In addition, this test is just studied for the antagonism of botrytis cinerea, as for the prevention effect that other soil is passed to pathogenic bacterium disease, also awaits further research.
the preparation of embodiment 2 biocontrol bacterial fertilizers
(1) preparation of liquid seed culture medium: take peptone 10g, extractum carnis 5g, NaCl 5g, adds purified water stirring and evenly mixing, supplies water to 1000mL, adjusts pH=7.0, divides the triangular flask that installs to 10 150mL, 121 ℃ of sterilizing 30min.
(2) preparation of liquid fermentation medium: take peptone 1kg, extractum carnis 0.5 kg, NaCl 0.5kg, adds purified water stirring and evenly mixing, supplies water to 100L, adjusts pH=7.0,121 ℃ of sterilizing 30min.
(3) bacillus cereus is accessed in the liquid nutrient medium in above-mentioned triangular flask, in 30 ℃, 180r/min shaking table shaking culture 30h, form seed liquor; Seed liquor is inoculated into and in the liquid nutrient medium in fermentor tank, carries out fermentative production by the volume ratio of 1:500, the groundwork parameter of fermentor tank is: 30 ℃ of temperature, stirring velocity 200r/min, pH7.0, dissolved oxygen 30%, tank pressure 0.04MPa, stops fermentation after 48 hours, obtain viable count and be greater than 9.6 * 10
9the bacillus cereus fermentation liquid of individual/ml.
(4) Pseudomonas chlororaphis is accessed in the liquid nutrient medium in above-mentioned triangular flask, in 28 ℃, 150r/min shaking table shaking culture 20h, form seed liquor; Seed liquor is inoculated into and in the liquid nutrient medium in fermentor tank, carries out fermentative production by the volume ratio of 1:500, the groundwork parameter of fermentor tank is: 28 ℃ of temperature, stirring velocity 150r/min, pH7.0, dissolved oxygen 30%, tank pressure 0.04MPa, stops fermentation after 25 hours, obtain viable count and be greater than 2.1 * 10
9the Pseudomonas chlororaphis fermented liquid of individual/ml.
(5) straw is ground into particle, getting fresh pig dung evenly mixes by the weight ratio of 1:2.5 with straw particle, regulating water content is 32% left and right, pile highly 1.2 meters, when heap temperature rise turning during to 78-80 ℃, and add water and take that to keep compost water content be 30-33%, after turning 2-3 time, ight soil becomes thoroughly decomposed, stalk rots, and sterilising treatment forms solid fermentation matrix;
(6) be cooled in the solid fermentation matrix of 25-32 ℃, the bacillus cereus fermentation liquid of preparing according to the standard access step (3) of matrix 30L per ton, the Pseudomonas chlororaphis fermented liquid of simultaneously preparing according to the standard access step (4) of matrix 90L per ton, mix, pile high 0.50 meter, the turning while reaching 32 ℃ of heap temperature, ferment 5 days, is no more than that under the condition of 35 ℃, the water content of fertilizer is evaporated to must biocontrol bacterial fertilizer below 30% in temperature.The biocontrol bacterial fertilizer that the present invention makes, wherein in every gram of fertilizer, contained living bacteria count (cfu) is greater than 1.0 * 10
8individual, organic content (in butt) is greater than 25%, and moisture content is lower than 30%, and other index meets biological organic fertilizer national sector standard.
the experimental study of embodiment 3 biocontrol bacterial fertilizer control graw mold of tomato
Soil property is consistent, fertility is evenly medium to select two, and gray mold falls ill tomato planting field heavier, be grouped into experimental plot and contrast field, and the tomato that these two ground are planted is handled as follows respectively:
Experimental plot (0.5 mu): use embodiment 2 biocontrol bacterial fertilizer 100kg for every mu;
Contrast field (0.5 mu): use embodiment 2 sterilizing biocontrol bacterial fertilizer 100kg in contrast for every mu.
The sampling survey state of an illness before and after test (after testing, the time of sampling is latter 2 weeks of fertilising).Each experimental plot adopts 5 point samplings, every some the investigation whole blades of 4 strain and fruit, and the percentage that every blade accounts for leaf area by scab carries out severity Scaling record, calculates disease index.Fruit gathers each investigation result by grade scale, calculates disease index.
Disease blade investigation grade scale: 0 grade: without scab; 1 grade: single blade has 3 of scabs; 3 grades: single blade has scab 4-6; 5 grades: single blade has scab 7-10; 7 grades: single blade has 11～20 of scabs, part is intensive in flakes; 9 grades: single blade has scab, account for leaf area more than 1/4.
Disease fruit investigation grade scale: 0 grade: anosis class; L level: residual petal morbidity or column cap morbidity; 3 grades: sepal rots or column cap morbidity spreads to areola portion; 5 grades: there is the spot of infiltration in areola portion without mould layer; 7 grades: there is mould layer in areola portion but does not expand to other positions; 9 grades: mould layer expands to other positions of fruit.
Blade disease index=[∑ (the sick number of sheets * relative level numerical value at different levels)/(investigating the total number of sheets * 9)] * 100
Fruit disease index=[∑ (sick fruit number * relative level numerical value at different levels)/(the total fruit of investigation number * 9)] * 100
By table 2,3 test-results, can be found out, by field test biocontrol bacterial fertilizer of the present invention, can significantly reduce tomato leaf and fruit gray mold disease index, graw mold of tomato is had to good prevention effect.
The impact of table 2 biocontrol bacterial fertilizer on tomato leaf gray mold disease index
| Group | Investigation strain number | Blade (before fertilising) | Blade (after fertilising) |
| Experimental plot | 20 | 8.06 | 0.49 |
| Contrast field | 20 | 8.14 | 7.72 |
The impact of table 3 biocontrol bacterial fertilizer on tamato fruit gray mold disease index
| Group | Investigation strain number | Fruit (before fertilising) | Fruit (after fertilising) |
| Experimental plot | 20 | 6.81 | 0.54 |
| Contrast field | 20 | 6.69 | 6.47 |
Claims (7)
1. a biocontrol bacterial fertilizer of preventing and treating farm crop soil-borne disease, is characterized in that, this biocontrol bacterial fertilizer, by every gram, contains and is greater than 1.0 * 10
8individual biocontrol microorganisms, described biocontrol microorganisms is comprised of bacillus cereus and Pseudomonas chlororaphis.
2. a production technique for biocontrol bacterial fertilizer according to claim 1, is characterized in that comprising the steps:
(1) bacillus cereus, Pseudomonas chlororaphis are inoculated into respectively in nutrient solution and carry out liquid state fermentation, obtain viable count and be greater than 1.0 * 10
9the bacillus cereus fermentation liquid of individual/ml, Pseudomonas chlororaphis fermented liquid, standby;
(2) fresh feces of livestock and poultry is evenly mixed by the weight ratio of 1:2-3 with stalk particle, compost 10-20 days to ight soil become thoroughly decomposed, stalk rots, sterilising treatment, forms solid fermentation matrix;
(3) by bacillus cereus fermentation liquid, Pseudomonas chlororaphis fermented liquid step (1) Suo Shu, according to the inoculum size of 20-40L/ ton, 50-100L/ ton, be inoculated in the described solid fermentation matrix of step (2) and carry out solid fermentation successively, leavening temperature is 25-32 ℃, in fermenting process, turning is 3-5 time, fermentation 5-8 days, obtains biocontrol bacterial fertilizer.
3. the production technique of biocontrol bacterial fertilizer according to claim 1, is characterized in that, the consisting of of the nutrient solution described in step (1): peptone 10g/L, extractum carnis 5g/L, NaCl 5g/L, pH7.0.
4. the production technique of biocontrol bacterial fertilizer according to claim 1, is characterized in that, described in step (1), the condition of the liquid state fermentation of bacillus cereus is: under aeration-agitation in 28 ℃-32 ℃ fermentation 30-60h.
5. the production technique of biocontrol bacterial fertilizer according to claim 1, is characterized in that, described in step (1), the condition of the liquid state fermentation of Pseudomonas chlororaphis is: under aeration-agitation in 25 ℃-30 ℃ fermentation 20-36h.
6. the production technique of biocontrol bacterial fertilizer according to claim 1, is characterized in that, the stalk particle described in step (2) is one or more particles after crushed in corn stalk, straw, paddy stalk, Rape Straw and rice straw.
7. the application of biocontrol bacterial fertilizer claimed in claim 1 in control farm crop soil-borne disease, described soil-borne disease is that botrytis cinerea is sick.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673719A (en) * | 2015-02-15 | 2015-06-03 | 山东省科学院生物研究所 | Biocontrol strain BCJB01 and application thereof |
| CN105052623A (en) * | 2015-07-15 | 2015-11-18 | 广西宏华生物实业股份有限公司 | Method for controlling soil-borne diseases |
| CN106497831A (en) * | 2016-10-21 | 2017-03-15 | 广州市林业和园林科学研究院 | A kind of preventing and treating Phytophthora nicotianae disease composite bacteria agent capable and its preparation method and application |
| CN110066194A (en) * | 2019-04-02 | 2019-07-30 | 漳州三炬生物技术有限公司 | Novel more bacterium composite microbe fertilizers of one kind and its preparation method and application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101007746A (en) * | 2007-01-11 | 2007-08-01 | 扬州市森大肥业有限公司 | Process for preparing anti-second-stubble efficient biologic-organic fertilizer |
| CN101143800A (en) * | 2007-09-05 | 2008-03-19 | 韩大山 | Stubble resisting and continuous and alternate cropping fertilizer and producing method thereof |
| US20120045427A1 (en) * | 2009-09-18 | 2012-02-23 | Jiangsu New Ground Bio-Fertilizer Engineering Center Co., Ltd. | Antagonistic bacteria for preventing and eliminating the bacterial wilt of continuous cropping tobacco and their microbial organic fertilizer |
| CN103524253A (en) * | 2013-10-31 | 2014-01-22 | 河北科技师范学院 | Seedling culturing matrix, preparing method and application of matrix |
-
2014
- 2014-05-18 CN CN201410207914.5A patent/CN103951483B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101007746A (en) * | 2007-01-11 | 2007-08-01 | 扬州市森大肥业有限公司 | Process for preparing anti-second-stubble efficient biologic-organic fertilizer |
| CN101143800A (en) * | 2007-09-05 | 2008-03-19 | 韩大山 | Stubble resisting and continuous and alternate cropping fertilizer and producing method thereof |
| US20120045427A1 (en) * | 2009-09-18 | 2012-02-23 | Jiangsu New Ground Bio-Fertilizer Engineering Center Co., Ltd. | Antagonistic bacteria for preventing and eliminating the bacterial wilt of continuous cropping tobacco and their microbial organic fertilizer |
| CN103524253A (en) * | 2013-10-31 | 2014-01-22 | 河北科技师范学院 | Seedling culturing matrix, preparing method and application of matrix |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104673719A (en) * | 2015-02-15 | 2015-06-03 | 山东省科学院生物研究所 | Biocontrol strain BCJB01 and application thereof |
| CN105052623A (en) * | 2015-07-15 | 2015-11-18 | 广西宏华生物实业股份有限公司 | Method for controlling soil-borne diseases |
| CN106497831A (en) * | 2016-10-21 | 2017-03-15 | 广州市林业和园林科学研究院 | A kind of preventing and treating Phytophthora nicotianae disease composite bacteria agent capable and its preparation method and application |
| CN106497831B (en) * | 2016-10-21 | 2019-10-25 | 广州市林业和园林科学研究院 | A kind of prevention and treatment Phytophthora nicotianae disease composite bacteria agent and its preparation method and application |
| CN110066194A (en) * | 2019-04-02 | 2019-07-30 | 漳州三炬生物技术有限公司 | Novel more bacterium composite microbe fertilizers of one kind and its preparation method and application |
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