CN1294816C - Prepn process of corn stalk trichoderma as biological antiseptic - Google Patents
Prepn process of corn stalk trichoderma as biological antiseptic Download PDFInfo
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- CN1294816C CN1294816C CNB2005100129121A CN200510012912A CN1294816C CN 1294816 C CN1294816 C CN 1294816C CN B2005100129121 A CNB2005100129121 A CN B2005100129121A CN 200510012912 A CN200510012912 A CN 200510012912A CN 1294816 C CN1294816 C CN 1294816C
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Abstract
The present invention relates to a preparation process of a trichoderma biological germicide of corn stalks, which belongs to the technical field of an agricultural germicide and is used for solving the problem of preparing biological germicide with corn stalks. The present invention uses corn stalk powder as raw material and manufactured through the steps of raw material processing, liquid fermentation, solid fermentation, germicide preparation, etc., wherein during the step of solid fermentation, pulverized corn stalk powder of 8-10 grams, (NH4) 2SO4 of 0.12-0.14 gram, MgSO4 of 0.002-0.004 gram and KH2PO4 of 0.004-0.005 gram are taken; the inorganic salt are dissolved with water of 200 ml, added with the corn stalk powder, accessed with culture fluid after agitation, sterilization and cooling and manufactured into finished goods after cultivation and air drying. The germicide of the present invention does not generate pollution to human, animals and ecological environment, has good control efficiency to fungal soil-borne diseases of crops and has the functions of growth promotion and yield increase; in addition, a large number of corn stalks can be used for reducing environment pollution; the present invention is an ideal biological pesticide for developing high-efficiency, green and ecological agriculture.
Description
Technical field
The present invention relates to a kind of agricultural biological bactericide, particularly is the preparation method of the trichoderma as biological antiseptic of raw material production with the maize straw, belongs to agricultural microorganism bactericide technical field.
Background technology
Domestic greatly an agricultural production, straw behind the annual results crops can be up to more than one hundred million tons, the wherein direct and indirect also not enough half in field, some is as the raw material of industry or agricultural fuel in addition, remain quite a few and burned, this processing mode is not only wasted resource but also serious environment pollution.Although various government decrees forbid burning straw, remain incessant after repeated prohibition because of suffering from no suitable outer reason mode.On the other hand; because crops are caused the underproduction even total crop failure through the invasion that regular meeting is subjected to various damage by disease and insect; more present method is to use chemical pesticide, insecticide to kill or suppresses damage by disease and insect; a large amount of uses along with agricultural chemicals; various pest resistance to insecticide increase; pesticide dosage is also continuing to increase, and its result forms vicious circle, causes damage by disease and insect to be difficult to control more and the agricultural product persticide residue is increased.In recent years, discover the trichoderma as biological preparation as the edaphon microbial inoculum, be used to prevent and treat the fungoid soil-borne diseases of crops and economic crops such as gray mold, damping off, fusarium wilt, rotten mildew etc., good control efficiency arranged, and can not produce the side effect of chemicals.But the problem that exists is at present: prepare wooden mould bactericide and need use the more expensive raw materials of price such as corn flour, molasses, wheat bran, make the production cost of wooden mould bactericide increase, be difficult to extensively promote the use of.
To sum up, if can utilize cheap raw material such as straw to prepare wooden mould bactericide, both can turn waste into wealth solves straw and utilizes problem, can use the wooden mould bactericidal agent for preventing and treating insect pest of environmental protection at a low price again.
Summary of the invention
The present invention is used to solve the defective of above-mentioned prior art and a kind of preparation method who uses cheap corn stalk powder to be used to produce meter stalk trichoderma as biological antiseptic as raw material is provided.
The alleged problem of the present invention is dissatisfied by the following technical programs determines:
A kind of preparation method of corn stalk trichoderma as biological antiseptic, it is raw material with the corn straw powder and prepares according to following steps:
A. raw material processing: selecting does not have the maize straw that goes mouldy, and it is standby to be ground into 20 order powderies;
B. liquid fermentation: medium component is: wheat bran 0.5～0.8 gram, (NH
4)
2SO
402.～0.4 gram, KH
2O
40.1～0.3 gram, CaCO
30.1～0.3 gram, glucose 1～2 gram is with 100 milliliters of above-mentioned substance obtaining liq fermentation mediums, adjusting pH value is 5.5～6, autoclaving 30～40 minutes is treated material cooling back access antagonism Trichoderma kind, stops to cultivate standby after 36～48 hours in 26～30 ℃ of shaken cultivation;
C. solid fermentation: get corn stalk powder 8～10 grams that crush, (NH
4)
2SO
40.12～0.14 gram, MgSO
40.002～0.004 gram, KH
2PO
40.004～0.005 gram, above-mentioned mineral salt are added the dissolving of 200 ml waters, put into maize straw again, stir, autoclaving 30～40 minutes treats that material cooling back inserts 1～1.5 milliliter of the culture fluid of step b, stirs, be put in and carry out dark illumination in 12 hours in the constant incubator and alternately cultivated 90～96 hours, cultured solid fermentation material is air-dry standby in being lower than 50 ℃ environment;
D. fungicide preparation: get that drying solid microbial inoculum 1 gram adds 99 ml sterile waters and an amount of bead among the step c, vibrated on the shaking table 30～40 minutes, measure the spore number, being diluted to spore content with the corn stalk powder that crushes again is 10
8Individual/gram spore is the trichoderma as biological antiseptic of score packing.
The preparation method of above-mentioned corn stalk trichoderma as biological antiseptic, described antagonism Trichoderma kind is a Trichoderma viride, and through the UV-irradiation mutagenic and breeding.
The preparation method of above-mentioned corn stalk trichoderma as biological antiseptic, the constant temperature culture temperature is 28～30 ℃ in the described solid fermentation step, humidity 65～80%.
The present invention uses cheap corn stalk powder to be raw material, substitutes the class raw materials such as starch that use at present, adopts liquid-solid two-phase zymotechnique, the preparation biological bactericide.This bactericide does not produce harm and pollution to people and animals and ecotope, the fungoid soil-borne diseases of crops such as gray mold, damping off, fusarium wilt, rotten mildew etc. there is good control efficiency, also the fiber of maize straw is have good decomposability simultaneously, make it to convert to useful nutriment, this to the generation of conidium of Trichoderma and the plant growing after using highly beneficial.Can obviously promote crop growth, increase crop yield 10-20%, have ill diseases prevention, the effect of anosis volume increase.This is that the conidium quantity that produces is big, by illumination in 12 hours and dark hand over to dive cultivate, can make the trichoderma conidium number of the every gram dry product of fermentation product reach 200～50,000,000,000 (being generally 2～500,000,000 every grams).In addition, present used high sporopollen of trichoderma all is direct application, or with using after the dilution of inertia fill materials such as soil, because the mycostasis of soil can make the mould control efficiency to plant disease of wood reduce and instability.The present invention makes dilution, nutritional agents with corn stalk powder, makes the Trichoderma that applies can grow more, long mycelia.What can extend to the plant rhizosphere goes to infect germ in a big way, suppresses disease and takes place, and this not only can eliminate or reduce the soil mycostasis, and Trichoderma agent control efficiency is improved, stable lasting, and has improved the wooden mould disease effect of preventing and treating; With the corn straw powder is raw material, can utilize discarded maize straw in a large number, reduces cost several times～tens times, turns waste into wealth, and reduces to burn and pollutes, and has fine social benefit and the economic benefit of getting, and is that development is efficient, green, the desirable biopesticide of the ecological agriculture.
Embodiment
The present invention selects the antagonism Trichoderma-Trichoderma viride of the fermentation growth in corn stalk powder that suits for use through repeated screening and UV-irradiation mutagenic and breeding.This bacterial strain not only has the good restraining effect to plurality of plant diseases.And also the fiber to maize straw have fine decomposability, and product spore amount is big.Usually the microbial inoculum spore number behind undercoating liquid fermentation and the solid fermentation can reach 10
10/ gram spore, and the spore content of finished product trichoderma as biological antiseptic is 10
8Individual/gram, obviously, need a large amount of corn stalk powders as nutritional agents and thinner, therefore can the discarded in a large number maize straws of digestibility and utilization, reduce annual because of the crop straw burning pollution on the environment, for the comprehensive utilization of stalk provides a new approach.
Several embodiment below are provided:
Embodiment 1:
A. raw material processing: selecting does not have the maize straw that goes mouldy, and it is standby to be ground into 20 order powderies behind the dryness in the sun;
B. liquid fermentation: medium component: wheat bran 0.5 gram, (NH
4)
2SO
40.2 gram, KH
2PO
40.1 gram, CaCO
30.1 gram, glucose 1.5 grams.100 milliliters of above-mentioned substance obtaining liq fermentation mediums, the pH value of adjusting liquid is 5.5, puts into 300 milliliters of triangular flask mesohigh sterilizations 30 minutes.The good antagonism Trichoderma kind of screening is inserted in material cooling back, in 28 ℃ of shaken cultivation 40 hours, stops to cultivate standby.
C. solid fermentation: get corn stalk powder 9 grams that crush, (NH
4)
2SO
40.12 gram, MgSO
40.002 gram, KH
2PO
40.004 gram.Above-mentioned mineral salt are put into 300 milliliters of triangular flasks add the dissolving of 200 ml waters, put into corn stalk powder again, stir, build bottle stopper.Autoclaving 30 minutes is treated the 1 milliliters of liquid culture fluid that material cooling back access step b makes.Stir, be put in 28 ℃ the constant incubator, keep humidity 65% to carry out solid culture 90 hours.Cultured solid fermentation material temperature in less than 50 ℃ of environment air-dry back standby.
D. fungicide preparation: get the dry good solid fungicide of a gram and put into 300 milliliters of triangular flasks, add 99 ml sterile waters and 10～20 pisiformis beades, the effect of bead is to improve the shake power that stirs.Be placed on 180-200 rev/min the shaking table vibration 30 minutes.Measure spore number 10 with blood counting chamber
10/ gram spore, being diluted to spore content with the corn stalk powder that crushes is 10
8/ gram spore promptly gets trichoderma as biological antiseptic.
Embodiment 2:
A. raw material processing: selecting does not have the maize straw that goes mouldy, and it is standby to be ground into 20 order powderies behind the dryness in the sun;
B. liquid fermentation: medium component: wheat bran 0.8 gram, (NH
4)
2SO
40.35 gram, KH
2PO
40.25 gram, CaCO
30.2 gram, glucose 1 gram.100 milliliters of above-mentioned substance obtaining liq fermentation mediums, the pH value of adjusting liquid is 6, puts into 300 milliliters of triangular flask mesohigh sterilizations 40 minutes.The good Hangzhoupro property Trichoderma kind short of money of screening is inserted in material cooling back, in 29 ℃ of shaken cultivation 42 hours, stops to cultivate standby.
C. solid fermentation: get corn stalk powder 10 grams that crush, (NH
4)
2SO
40.14 gram, MgSO
40.004 gram, KH
2PO
40.045 gram.Above-mentioned mineral salt are put into 300 milliliters of triangular flasks add the dissolving of 200 ml waters, put into corn stalk powder again, stir, build bottle stopper.Autoclaving 35 minutes is treated the 1.5 milliliters of liquid culture fluids that material cooling back access step b makes.Stir, be put in 30 ℃ the constant incubator, keep humidity 80% to carry out dark illumination in 12 hours and alternately cultivated solid culture 96 hours.Cultured solid fermentation material temperature in less than 50 ℃ of environment air-dry back standby.
D. fungicide preparation: with embodiment 1.
Embodiment 3:
A. raw material processing: selecting does not have the beautiful stalk stalk that goes mouldy, and it is standby to be ground into 20 order powderies behind the dryness in the sun;
B. liquid fermentation: medium component: wheat bran 0.7 gram, (NH
4)
2SO
40.4 gram, KH
2PO
40.3 gram, CaCO
30.3 gram, glucose 2 grams.100 milliliters of above-mentioned substance obtaining liq fermentation mediums, the pH value of adjusting liquid is 5.5, puts into 300 milliliters of triangular flask mesohigh sterilizations 35 minutes.The good antagonism Trichoderma kind of screening is inserted in material cooling back, in 30 ℃ of shaken cultivation 48 hours, stops to cultivate standby.
C. solid fermentation: get corn stalk powder 8 grams that crush, (NH
4)
2SO
40.13 gram, MgSO
40.003 gram, KH
2PO
40.005 gram.Above-mentioned mineral salt are put into 300 milliliters of triangular flasks add the dissolving of 200 ml waters, put into corn stalk powder again, stir, build bottle stopper.Autoclaving 35 minutes is treated the 1.3 milliliters of liquid culture fluids that material cooling back access step b makes.Stir, be put in 28 ℃ the constant incubator, keep humidity 70% to carry out dark illumination in 12 hours and alternately cultivated solid culture 93 hours.Cultured solid fermentation material temperature in less than 50 ℃ of environment air-dry back standby.
D. fungicide preparation: with embodiment 1.
Used antagonism Trichoderma kind is a Trichoderma viride among the present invention, and through the UV-irradiation mutagenic and breeding.So-called UV-irradiation mutagenic and breeding is meant the seed selection of being undertaken by following technology: (potato 200 restrains preparation PDA, glucose 20 grams earlier, agar 18 grams, water 1000ml) culture medium flat plate is seeded in the antagonism Trichoderma on the flat board, produces the Trichoderma viride conidium in 5 days 25 ℃ of constant temperature culture.Add 10 milliliters of wash-out spores of aqua sterilisa, making concentration is 10
6The suspension of/ml spore.Get the PDA flat board, every plate is drawn 0.1 milliliter of suspension, is evenly coated in planar surface, is placed in the constant incubator 25 ℃ of constant temperature culture one day, is covering under the optical condition then, and 30 watts of uviol lamps shone 80 minutes apart from fluorescent tube 30cm place.Good with black paper bag then, be placed in 4 ℃ of incubators low temperature and cultivated 12 hours.Put into 25 ℃ of constant temperature culture dark again.Choose the bacterial strain fast of growing after 3 days, insert PDA medium purifying and cultivate, as the bacterial classification of next ultraviolet mutagenesis.Up to selecting better resistance short of money, can fine cellulolytic trichoderma strain.
The present invention's use of can dressing seed also can be stained with root when transplanting, can also when put in order ground, execute on the ridge, every mu the 10-20 kilogram.
Claims (3)
1. the preparation method of a corn stalk trichoderma as biological antiseptic, it is characterized in that: described bactericide is that raw material prepares according to following steps with the corn straw powder:
A. raw material processing: selecting does not have the maize straw that goes mouldy, and it is standby to be ground into 20 order powderies;
B. liquid fermentation: medium component is: wheat bran 0.5～0.8 gram, (NH
4)
2SO
402.～0.4 gram, KH
2PO
40.1～0.3 gram, CaCO
30.1～0.3 gram, glucose 1～2 gram is with 100 milliliters of above-mentioned substance obtaining liq fermentation mediums, adjusting pH value is 5.5～6, autoclaving 30～40 minutes is treated material cooling back access antagonism Trichoderma kind, stops to cultivate standby after 36～48 hours in 26～30 ℃ of shaken cultivation;
C. solid is sent out alcohol: get corn stalk powder 8～10 grams that crush, (NH
4)
2SO
40.12～0.14 gram, MgSO
40.002～0.004 gram, KH
2PO
40.004～0.005 gram, above-mentioned mineral salt are added the dissolving of 200 ml waters, put into corn stalk powder again, stir, autoclaving 30～40 minutes treats that material cooling back inserts 1～1.5 milliliter of the culture fluid of step b, stirs, be put in and carry out dark illumination in 12 hours in the constant incubator and alternately cultivated 90～96 hours, cultured solid fermentation material is air-dry standby in being lower than 50 ℃ environment;
D. fungicide preparation: get that drying solid microbial inoculum 1 gram adds 99 ml sterile waters and bead among the step c, vibrated on the shaking table 30～40 minutes, measure the spore number, being diluted to spore content with the corn stalk powder that crushes again is 10
8/ gram spore promptly gets trichoderma as biological antiseptic.
2. according to the preparation method of the described corn stalk trichoderma as biological antiseptic of claim 1, it is characterized in that: described antagonism Trichoderma kind is a Trichoderma viride, and through the UV-irradiation mutagenic and breeding.
3. according to the preparation method of the described corn stalk trichoderma as biological antiseptic of claim 2, it is characterized in that: the constant temperature culture temperature is 28～30 ℃ in the described solid fermentation step, humidity 65～80%.
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|---|---|---|---|
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|---|---|---|---|
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| CN1294816C true CN1294816C (en) | 2007-01-17 |
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101438766B (en) * | 2007-11-23 | 2011-07-20 | 西安新天地草业有限公司 | High-efficient straw stored material fermentation agent |
| CN101525577B (en) * | 2009-03-02 | 2011-07-20 | 河北省生物研究所 | Solid culture method of metarrhizium anisopliae |
| CN105505799A (en) * | 2016-01-26 | 2016-04-20 | 李晓白 | Fermentation method and application of trichoderma viride mutant strains |
| CN106135294A (en) * | 2016-07-22 | 2016-11-23 | 吉林省农业科学院 | A kind of Novel ball beauveria bassiana granule |
| CN109022291A (en) * | 2018-08-15 | 2018-12-18 | 山东泰诺药业有限公司 | A kind of efficient biocontrol agent of Trichoderma |
| CN110591921B (en) * | 2018-12-19 | 2023-12-05 | 扬州工业职业技术学院 | Trichoderma liquid fermentation chlamydospore and preparation method thereof |
| CN113519569B (en) * | 2021-09-03 | 2022-07-05 | 重庆文理学院 | Preparation method of medicament for early-stage defoliation of kiwi fruits |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1593156A (en) * | 2004-07-05 | 2005-03-16 | 郝锁振 | Preparation method of composite humic acid |
| CN1632101A (en) * | 2004-11-26 | 2005-06-29 | 山东大学 | Method for preparing medicine-fertilizer double purpose soil microbial community trichoderm-virida preparation |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1593156A (en) * | 2004-07-05 | 2005-03-16 | 郝锁振 | Preparation method of composite humic acid |
| CN1632101A (en) * | 2004-11-26 | 2005-06-29 | 山东大学 | Method for preparing medicine-fertilizer double purpose soil microbial community trichoderm-virida preparation |
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Granted publication date: 20070117 Termination date: 20101014 |