CN1302710C - Multifunctional biological control bacteria agent and its preparation method - Google Patents
Multifunctional biological control bacteria agent and its preparation method Download PDFInfo
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- CN1302710C CN1302710C CNB2004100210805A CN200410021080A CN1302710C CN 1302710 C CN1302710 C CN 1302710C CN B2004100210805 A CNB2004100210805 A CN B2004100210805A CN 200410021080 A CN200410021080 A CN 200410021080A CN 1302710 C CN1302710 C CN 1302710C
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Abstract
The present invention relates to a multifunctional biocontrol bacterial agent and a preparation method thereof, wherein the multifunctional biocontrol bacterial agent uses the chlamydospore of trichodermaharzianum separated from the rhizosphere soil of crops as main components; a small quantity of mineral nutrient elements are used as a synergistic agent; light calcium carbonate and diatomite are used as a carrier. The bacterial agent is mainly used for preventing and treating multiple soil-borne diseases of crops caused by disease fungus comprising fusarium, decayed fungus, streptoverticillium, rhizoctonia, etc. Meanwhile, the present invention has multiple functions of improving the own resistance of plants and improving productivity. The present invention has the advantages of short periodicity, stable and lasting preventing and treating effects, no residue on crops, no pollution to environment and safety for people and domestic animals.
Description
Technical field
The present invention is a kind of biopesticide of preventing and treating soil-borne disease and preparation method thereof that is used to.
Background technology
Along with the fast development of protection ground cultivated area, the pathogenic bacterial content increases year by year in the soil, and it is more and more heavier to cause disease to take place.With the cucurbits fusarium wilt is example, and the general incidence of disease of past only about 10%, has risen to 15%-20% now, and serious plot reaches more than 50%, even ruins kind, causes great economic loss to the peasant.Produce at present and go up, handle soil as chloropicrin, Celfume and other chemical bactericide etc. to the control of soil-borne disease or based on chemical pesticide.Use chemical pesticide frequently for a long time, heavy dose of, the pesticide resistance of phytopathogen is improved gradually, force dosage to continue to increase, not only increased the control cost, also cause environmental pollution, reduced the edible safety of fruit and vegetable.To this, take measures already in the world, limit the use of some severe toxicity, high-toxic pesticide gradually.China has announced several the chemical pesticides that ban use of in recent years, and advocates energetically and develop, develop the biopesticide to the nontoxic environmentally safe of people, animal.Now, countries in the world are all in the research and development dynamics that strengthens biopesticide.The product of having developed at present registration in the world has the yellow silk of green sticking broom mould (Gliocladium virens) aspergillus (Talaromycesfavus), fluorescent pseudomonas (Pseudomonas fluorescense), bacillus subtilis (Bacillus subtilis) and the mould Trichodex of Ha Shi wood etc.Also there is the patent of some trichoderma biopesticides in China, but really enters Te Like (Trichoderma sp) wetting powder that the large-scale industrial production kind has only the Shandong Province the people live in plenty Industrial Co., Ltd. to produce.Because various technical reasons, as the properties of product instability, effective content is low, can not form effective population in the very fast field planting of crop root after being manured into soil, and makes whole preventive effect not good, and this has restricted the development of biopesticide to a great extent.
The purpose of this invention is to provide a kind of multi-functional biocontrol fungicide of preventing and treating plant soil-borne diseases and preparation method thereof that is used to, overcome the deficiency of prior art, occurrence characteristic according to soil-borne disease, isolate Trichoderma from the root of the crop table, adding multiple mineral nutrient element is adsorbent as synergist with the precipitated calcium carbonate, can improve control efficiency and reach more than 75%, volume increase is more than 15%.
Technical scheme content of the present invention is:
Biocontrol microorganisms of the present invention is the Trichoderma that is taken from crop rhizosphere, be transferred to after being purified and make female microbial inoculum in the test tube slant, and shaken cultivation is a liquid spawn in triangular flask, makes spore suspension through producing a jar fermentation again, and the liquid fermentation medium composition that is adopted is:
Potato 150-200 gram, glucose 15-20 gram,
Potassium dihydrogen phosphate 1-2 gram, magnesium sulfate 0.5-1 gram,
Add water to 1000 milliliters;
Aforesaid liquid medium adjust pH is 6-7, and following 30 minutes of 121 ℃ of sterilization times, strain fermentation are with 250 milliliters of triangular flasks, and fermentation temperature 28-30 ℃, fermentation time is 3-5 days; Producing a jar fermentation temperature is 25-30 ℃, and fermentation time is 5-9 days, makes the Trichoderma spore suspension.
Multi-functional biocontrol fungicide constituent is by weight percentage:
Trichoderma spore suspension (chlamydospore content 1 * 10
8It is individual/ml) 15-20,
Precipitated calcium carbonate 20-30, diatomite 40-53,
Potassium dihydrogen phosphate 3-5, calcium sulphate 1-2,
Magnesium sulfate 0.5-1, molybdic acid 0.5-1,
Boric acid 0.1-0.4, zinc sulphate 0.1-0.4,
Manganese sulphate 0.1-0.4, ferrous sulfate 0.1-0.4;
Earlier carrier precipitated calcium carbonate and diatomite are mixed by weight percentages of components, again the Trichoderma spore suspension is fully adsorbed with carrier, successively add mineral nutrient element potassium dihydrogen phosphate, magnesium sulfate, calcium sulphate, molybdic acid, boric acid, manganese sulphate, ferrous sulfate, zinc sulphate again, fully mix, make multi-functional biocontrol fungicide.
The production and processing technology flow process of multi-functional biocontrol fungicide is seen Figure of description 1.
Advantage of the present invention is: the antagonistic strain Trichoderma (Trichodermaharzianum) that separates from the root of the crop table is the soil saprophytic bacteria, after using, can be in the rapid field planting of root of the crop table, by causing the disintegration of pathogen mycelium at short notice with pathogen competition for nutrients and hyperparasitism.Its chlamydospore has the ability of poor environments such as opposing high temperature, drying and nutritional deficiency, belong to nature by putting again after the artificial culture, and make it in the natural microorganisms fauna, further to multiply, be formed with the benign cycle of beneficial microorganism in soil, help keeping the natural equilibrium of the little ecology of rhizosphere, reach Sustainable Control, to the crop noresidue disease, environmentally safe is to the person poultry safety.
The present invention utilizes crop, pathogeny bacterium and the Trichoderma physiological reaction difference to Different Nutrient Elements, add an amount of nutritive element as synergistic factor, these mineral nutrient elements all are that plant growing is necessary, can improve plant resistance to environment stress on the one hand, strengthen biocontrol effect, can promote plant growing on the other hand, improve the output and the quality of product, have the dual-use function of disease-preventing and yield-increasing.Cost is low, and high efficiency is used conveniently, utilizes preparation of the present invention to prevent and treat soil-borne disease, can obtain being higher than the control efficiency of chemical pesticide.
Description of drawings
Fig. 1 is a technological process of production schematic diagram of the present invention.
Embodiment
Embodiment 1
Get root of the crop table soil 10 grams, add 100 milliliters of sterile waters, after vibration, be diluted to 10000 times of liquid, get 1 milliliter of dilution and make flat board with 9 milliliters of PDA medium that add streptomycin sulphates (1000 milliliters of potato 200 grams, glucose 20 grams, agar 20 grams, water), cultivated 48 hours down at 26 ℃, after separation obtains Trichoderma (Trichoderma harzianum), be purified, be transferred to and make female microbial inoculum in the test tube slant.
The liquid fermentation medium prescription that the second class inoculum fermentation is adopted is:
Potato 200 grams, glucose 20 grams, potassium dihydrogen phosphate 2 grams, magnesium sulfate 1 restrain, add water to 000 milliliter.
Preparation method is: will be cut into the fritter of 1 centimeter square behind the peeling potatoes, adding 1000 ml waters boiled 30 minutes, with gauze elimination potato, add water again and mend 1000 milliliters, add glucose 20 gram, potassium dihydrogen phosphate 2 grams, magnesium sulfate 1 gram then and make it to dissolve fully, this liquid nutrient medium adjust pH is 6.5,121 ℃ of sterilization times are following 30 minutes, strain fermentation is 5 days with 250 milliliters of triangular flasks, 28 ℃ of fermentation temperatures, fermentation time.It is identical with the second class inoculum fermentation process to produce jar medium and sterilizing methods thereof that fermentation is adopted, and connecing the bacterium amount is 5%, and fermentation temperature is 26 ℃, fermentation time 9 days.Make the Trichoderma spore suspension.
By weight percentages of components carrier diatomite 42 grams and precipitated calcium carbonate 30 grams are mixed, then with Trichoderma spore suspension (the chlamydospore content 1 * 10 that makes
8Individual/ml) 20 milliliters fully adsorb with carrier diatomite and precipitated calcium carbonate, again with mineral nutrient element potassium dihydrogen phosphate 5 grams, calcium sulphate 1 gram, magnesium sulfate 0.5 gram, molybdic acid 1 gram, boric acid 0.2 gram, manganese sulphate 0.1 gram, ferrous sulfate 0.1 gram, zinc sulphate 0.1 gram, fully mix, make multi-functional biocontrol fungicide.
Multi-functional biocontrol fungicide and 500-600 times of nutrition soil are mixed, the dress feeding block seedlings raising, the control soil-borne diseases of vegetable, control efficiency reaches more than 75%.
Embodiment 2
Get root of the crop table soil 10 grams, add 100 milliliters of sterile waters, after vibration, be diluted to 10000 times of liquid, get 1 milliliter of dilution and make flat board with 9 milliliters of PDA medium that add streptomycin sulphates (1000 milliliters of potato 200 grams, glucose 20 grams, agar 20 grams, water), cultivated 48 hours down at 26 ℃, after separation obtains Trichoderma (Trichoderma harzianum), be purified, be transferred to and make female microbial inoculum in the test tube slant.
The liquid fermentation medium prescription that the second class inoculum fermentation is adopted is:
Potato 200 grams, glucose 20 grams, potassium dihydrogen phosphate 1 gram, magnesium sulfate 0.5 restrain, add water to 1000 milliliters.
Preparation method is: will be cut into the fritter of 1 centimeter square behind the peeling potatoes, adding 1000 ml waters boiled 30 minutes, with gauze elimination potato, add water and mend 1000 milliliters, add glucose 20 gram, potassium dihydrogen phosphate 1 gram, magnesium sulfate 0.5 gram then and make it to dissolve fully, this liquid nutrient medium adjust pH is 7,121 ℃ of sterilization times are following 30 minutes, strain fermentation is 5 days with 250 milliliters of triangular flasks, 28 ℃ of fermentation temperatures, fermentation time.It is identical with the second class inoculum fermentation process to produce jar medium and sterilizing methods thereof that fermentation is adopted, and connecing the bacterium amount is 5%, and fermentation temperature is 28 ℃, and fermentation time is 7 days.Make the Trichoderma spore suspension
By weight percentages of components carrier diatomite 48 grams and precipitated calcium carbonate 25 grams are mixed, then with Trichoderma spore suspension (the chlamydospore content 1 * 10 that makes
8Individual/ml) 20 grams fully adsorb with carrier diatomite and precipitated calcium carbonate, again with mineral nutrient element potassium dihydrogen phosphate 3.5 grams, magnesium sulfate 1 gram, calcium sulphate 1 gram, molybdic acid 0.6 gram, boric acid 0.3 gram, manganese sulphate 0.2 gram, ferrous sulfate 0.2 gram, zinc sulphate 0.2 gram, fully mix, make multi-functional biocontrol fungicide.
Multi-functional biocontrol fungicide and 500-600 times of nutrition soil are mixed, and the dress feeding block seedlings raising is prevented and treated melon soil-borne disease, and control efficiency reaches more than 70%.
Embodiment 3
Get root of the crop table soil 10 grams, add 100 milliliters of sterile waters, after vibration, be diluted to 10000 times of liquid, get 1 milliliter of dilution and make flat board with 9 milliliters of PDA medium that add streptomycin sulphates (1000 milliliters of potato 200 grams, glucose 20g, agar 20g, water), cultivated 48 hours down at 26 ℃, after separation obtains Trichoderma (Trichoderma harzianum), be purified, be transferred to and make female microbial inoculum in the test tube slant.
The liquid fermentation medium prescription that the second class inoculum fermentation is adopted is:
Potato 150 grams, glucose 15 grams, potassium dihydrogen phosphate 1 gram, magnesium sulfate 0.5 restrain, add water to 1000 milliliters
Preparation method is: will be cut into the fritter of 1 centimeter square behind the peeling potatoes, adding 1000 ml waters boiled 30 minutes, with gauze elimination potato, add water and mend 1000 milliliters, add glucose 20 gram, potassium dihydrogen phosphate 1 gram, magnesium sulfate 0.5 gram then and make it to dissolve fully, this liquid nutrient medium adjust pH is 6,121 ℃ of sterilization times are following 30 minutes, strain fermentation is 5 days with 250 milliliters of triangular flasks, 28 ℃ of fermentation temperatures, fermentation time.It is identical with the second class inoculum fermentation process to produce jar medium and sterilizing methods thereof that fermentation is adopted, and connecing the bacterium amount is 5%, and fermentation temperature is 30 ℃, fermentation time 5 days.
By weight percentages of components carrier diatomite 51 grams and precipitated calcium carbonate 20 grams are mixed, then with Trichoderma spore suspension (the chlamydospore content 1 * 10 that makes
8Individual/ml) 20 milliliters fully adsorb with carrier diatomite and precipitated calcium carbonate, again with mineral nutrient element potassium dihydrogen phosphate 4 grams, magnesium sulfate 0.5 gram, calcium sulphate 2 grams, molybdic acid 0.5 gram, boric acid 0.3 gram, manganese sulphate 0.2 gram, ferrous sulfate 0.2 gram, zinc sulphate 0.3 gram, fully mix, make multi-functional biocontrol fungicide.
Multi-functional biocontrol fungicide and 500-600 times of nutrition soil are mixed, 50 grams of spreading manuer in holes, the control of maize soil-borne disease, control efficiency reaches more than 80%.
Claims (2)
1. multi-functional biocontrol fungicide is characterized in that contained various composition and percentage by weight are as follows:
Trichoderma spore suspension (chlamydospore content 1 * 10
8It is individual/ml) 15-20,
Precipitated calcium carbonate 20-30, diatomite 40-53,
Potassium dihydrogen phosphate 3-5, calcium sulphate 1-2,
Magnesium sulfate 0.5-1, molybdic acid 0.5-1,
Boric acid 0.1-0.4, zinc sulphate 0.1-0.4,
Manganese sulphate 0.1-0.4, ferrous sulfate 0.1-0.4.
2. the preparation method of a multi-functional biocontrol fungicide as claimed in claim 1, it is characterized in that: utilize the Trichoderma that is separated to from the root of the crop table, adopting liquid fermentation method to make spore suspension is main component, earlier carrier precipitated calcium carbonate and diatomite are mixed by weight percentages of components, again the Trichoderma spore suspension is fully adsorbed with carrier, successively add mineral nutrient element potassium dihydrogen phosphate, magnesium sulfate, calcium sulphate, molybdic acid, boric acid, manganese sulphate, ferrous sulfate, zinc sulphate again, fully mix, make;
Wherein the used medium component of liquid fermentation is: potato 150-200 gram, and glucose 15-20 gram, potassium dihydrogen phosphate 1-2 gram, magnesium sulfate 0.5-1 gram adds water and becomes 1000 ml solns.The pH value of culture fluid is 6-7, following 30 minutes of 121 ℃ of sterilization times, and fermentation temperature 25-30 ℃, fermentation time is 5-9 days, forms chlamydospore and gets final product to detect mycelia more than 90%.
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| CNB2004100210805A CN1302710C (en) | 2004-01-18 | 2004-01-18 | Multifunctional biological control bacteria agent and its preparation method |
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| CNB2004100210805A CN1302710C (en) | 2004-01-18 | 2004-01-18 | Multifunctional biological control bacteria agent and its preparation method |
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| CN1557164A CN1557164A (en) | 2004-12-29 |
| CN1302710C true CN1302710C (en) | 2007-03-07 |
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Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100381055C (en) * | 2006-04-07 | 2008-04-16 | 浙江大学 | Preparation method and purpose of zinc-carried silicate antimicrobial agent |
| CN100466909C (en) * | 2006-07-31 | 2009-03-11 | 云南生物制药有限公司 | Production of trichodermin agricultural chemicals |
| CN102219602B (en) * | 2011-04-22 | 2013-10-16 | 浙江科技学院 | Novel biocontrol and plant vigor complexing agent and preparation method thereof |
| CN102499265B (en) * | 2011-11-10 | 2013-08-28 | 宁夏农林科学院 | Trichoderma hamatum biological agent for controlling soil borne disease for greenhouse vegetable cultivation |
| CN102845478A (en) * | 2012-08-16 | 2013-01-02 | 上海交通大学 | Trichoderma harzianum chlamydospore granules and preparation method thereof |
| CN102870822B (en) * | 2012-10-29 | 2015-01-14 | 黄永 | Aqueous dispersible granules of trichoderma asperellum chlamydospores biopesticide and preparation of aqueous dispersible granules |
| CN106045599A (en) * | 2016-05-31 | 2016-10-26 | 博白县那林镇菌兴家庭农场 | Efficient disease-preventing auricularia polytricha culture medium and preparation method thereof |
| CN105837346A (en) * | 2016-05-31 | 2016-08-10 | 博白县那林镇菌兴家庭农场 | Disease prevention medium for cultivating auricularia polytricha and method for preparing disease prevention medium |
| CN106083277A (en) * | 2016-05-31 | 2016-11-09 | 博白县那林镇菌兴家庭农场 | A kind of Auricularia polytricha (Mout) Sacc. cultivation matrix and preparation method thereof |
| CN106631433A (en) * | 2016-12-12 | 2017-05-10 | 新昌县奥而特农业科技有限公司 | Pesticide preparation for plant protection and growth regulation |
| TR2021013538A2 (en) * | 2021-08-27 | 2021-09-21 | Atatuerk Ueniversitesi Rektoerluegue Bilimsel Arastirma Projeleri Bap Koordinasyon Birimi | A MICROBIAL FERTILIZER AND BIOPESTICIDE FORMULATION WITH SOLID CARRIER |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5068105A (en) * | 1989-03-13 | 1991-11-26 | W. R. Grace & Co.-Conn. | Fungal formulation for biocontrol of soilborne plant pathogens |
| WO1997016974A1 (en) * | 1995-11-07 | 1997-05-15 | Universidad De Salamanca | Liquid formulation based on strains of the stringy fungi trichoderma harzianum and trichoderma viride |
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2004
- 2004-01-18 CN CNB2004100210805A patent/CN1302710C/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5068105A (en) * | 1989-03-13 | 1991-11-26 | W. R. Grace & Co.-Conn. | Fungal formulation for biocontrol of soilborne plant pathogens |
| WO1997016974A1 (en) * | 1995-11-07 | 1997-05-15 | Universidad De Salamanca | Liquid formulation based on strains of the stringy fungi trichoderma harzianum and trichoderma viride |
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