CN102690755B - Compound microbial bacterial preparation for degrading crop straw and preparation method and application of compound microbial bacterial preparation - Google Patents

Compound microbial bacterial preparation for degrading crop straw and preparation method and application of compound microbial bacterial preparation Download PDF

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CN102690755B
CN102690755B CN2012101789631A CN201210178963A CN102690755B CN 102690755 B CN102690755 B CN 102690755B CN 2012101789631 A CN2012101789631 A CN 2012101789631A CN 201210178963 A CN201210178963 A CN 201210178963A CN 102690755 B CN102690755 B CN 102690755B
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展彬
杨瑞
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Yuanhe Biotechnology Dezhou Co ltd
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Dezhou Yuanhe Agricultural Technology Development Co Ltd
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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
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Abstract

The invention relates to a compound microbial bacterial preparation for degrading crop straw and a preparation method and application of the compound microbial bacterial preparation. The compound microbial bacterial preparation is prepared from bacillus subtilis, bacillus cereus, aspergillus niger, aspergillus flavus, trichoderma reesei, trichoderma longibrachiatum, sporotrichum thermophile and phanerochaete chrysosporium through solid fermentation. The compound microbial bacterial preparation can shorten the decomposition time and improve the decomposition efficiency of the crop straw. Microorganisms generate various effective metabolic products with nitrogen fixation and phosphate solubilizing functions and a high disease and pest resistance effect in the fermentation process, so that the compound microbial bacterial preparation is favorable for improving the soil structure and the yield and the quality of crops.

Description

Complex micro organism fungicide of a kind of crop material of degrading and preparation method thereof and application
Technical field
The present invention relates to a kind of complex micro organism fungicide and preparation method thereof and application of efficient degradation crop material, belong to the microbial fermentation technology field.
Background technology
China is large agricultural country, and agricultural crop straw output is big, distribution is wide, kind is many.Since the reform and opening-up, along with the increasing of central authorities to agriculture support dynamics, China's agricultural develops rapidly, the stalk quantum of output surpasses 700,000,000 tons, but most stalk is arbitrarily abandoned or directly burning, and not only energy utilization rate is low, and serious environment pollution, human health is caused serious harm.Some stalk is directly used in traditional application model of fertilizer, feed without any processing in addition, seriously restricting the further raising of the sharp transformation efficiency of stalk, therefore accelerate to advance the stalk comprehensive utilization, for stablizing the Agro-ecology balance, alleviate resource constraint, alleviating environmental stress and have very important meaning.
The fertilizer that with the stalk is raw material production is at present extensively promoted in the rural area.Traditional direct composting process of organic fertilizer production process using, because microbe population and kind in the material are limited, microorganism active is low, causes the time of becoming thoroughly decomposed long, the fertilizer of generation is of low quality.Utilize microorganism stalk thoroughly can be decomposed, and can obtain a large amount of tropinas by the growth of microorganism self, the multiple effective meta-bolites of microorganisms has good disease and insect resistance effect in the fermenting process, is conducive to the structure of improving the soil, and improves crop yield and quality.Therefore use microbial method to handle stalk, can increase substantially the utilization ratio of stalk, compare with direct composting process and have plurality of advantages.
Chinese patent literature CN101775359A(application number 200910013635.4) special microorganism composite bacterial agent preparation and the application method thereof that a kind of direct decomposing and fermenting crops straws produces biogas disclosed.This special microorganism composite bacterial agent is liquid or solid-state, a strain or a few strain in, the aspergillus niger mould by cured shape gemma bar, subtilis, Bacillus licheniformis, bacillus megaterium, bacillusmusilaginosiengineering, Bacillus circulans, spherical bacillus, clostridium sporogene, acetone clostridium butylicum, Clostridium baratii, Bai Shi clostridium, Clostridium thermocellum, viride, Rui Shi wood, after fermentation culture, make up composite forming.This method is decomposed agricultural crop straw by adopting several microorganisms to make complex micro organism fungicide, has accelerated the speed of fermentation methane production, has improved biogas output.But this composite fungus agent still can't satisfy stalk and produce speed aspect quick decomposition crop material, and seeking the new composite fungus agent that can decompose crop material fast becomes the not high problem of present straw utilization rate that solves.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, a kind of complex micro organism fungicide that can the efficient degradation crop material and preparation method thereof and application are provided.
A kind of complex micro organism fungicide of the crop material of degrading, it is characterized in that, be by subtilis (Bacillus subtilis) CICC10088, bacillus cereus (Bacillus cereus) ACCC03002, aspergillus niger (Aspergillus niger) CICC41130, flavus (Aspergillus flavus) CICC40258, Trichodermareesei (Trichoderma reesei) CICC41491, long handle wood mould (Trichoderma viride) ACCC30150, sporotrichum thermophile (Sporotrichum thermophilic) ACCC30346 and Phanerochaete chrysosporium (Phanerochaete chrysosporium) CICC40719 make through solid fermentation.
Preferred according to the present invention, the every gram of above-mentioned complex micro organism fungicide contains: subtilis (Bacillus subtilis) CICC10088 viable count 10 8～10 10Individual, bacillus cereus (Bacillus cereus) ACCC03002 viable count 10 8～10 10Individual, aspergillus niger (Aspergillus niger) CICC41130 spore count 10 7～10 9Individual, flavus (Aspergillus niger) CICC40258 spore count 10 7～10 9Individual, Trichodermareesei (Trichoderma reesei) spore count 10 7～10 9Individual, long handle wooden mould (Trichoderma viride) ACCC30150 spore count 10 7～10 9Individual, sporotrichum thermophile (Sporotrichum thermophilic) ACCC30346 spore count 10 7～10 9Individual, Phanerochaete chrysosporium (Phanerochaete chrysosporium) CICC40719 spore count 10 7～10 9Individual.
Described subtilis (Bacillus subtilis) CICC10088, aspergillus niger (Aspergillus niger) CICC41130, flavus (Aspergillus flavus) CICC40258, Trichodermareesei (Trichoderma reesei) CICC41491, Phanerochaete chrysosporium (Phanerochaete chrysosporium) CICC40719 are all available from Chinese industrial microbial strains preservation administrative center;
Described bacillus cereus (Bacillus cereus) ACCC03002, long handle wood mould (Trichoderma viride) ACCC30150, sporotrichum thermophile (Sporotrichum thermophilic) ACCC30346 are all available from Chinese agriculture microbial strains preservation administrative center.
The preparation method of above-mentioned complex micro organism fungicide comprises the steps:
(1) subtilis CICC10088 is inoculated in the LB liquid nutrient medium, liquid culture 18～24h under 30～37 ℃ of conditions, inoculum size by 10 ~ 20wt% is inoculated among the solid medium A then, cultivated 3 ~ 5 days at 30～37 ℃, make and contain subtilis CICC10088 viable count 8～15 * 10 9The solid fungicide A of individual/g;
(2) bacillus cereus ACCC03002 is inoculated in the LB liquid nutrient medium, liquid culture 18～24h under 30～37 ℃ of conditions, inoculum size by 10 ~ 20wt% is inoculated in bacillus cereus among the solid medium B then, cultivated 3 ~ 5 days at 30～37 ℃, make and contain bacillus cereus ACCC03002 viable count 8～15 * 10 9The solid fungicide B of individual/g;
(3) aspergillus niger CICC41130 is inoculated in the PDA solid medium, solid culture is 2～3 days under 25～30 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, the inoculum size by 10 ~ 20wt% is inoculated in aspergillus niger among the solid medium C then, cultivates 5 ~ 7 days at 25～30 ℃, makes and contains aspergillus niger CICC41130 spore count 1～3 * 10 9The solid fungicide C of individual/g;
(4) flavus CICC40258 is inoculated in the PDA solid medium, solid culture is 2～3 days under 25～30 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, the inoculum size by 10 ~ 20wt% is inoculated in flavus among the solid medium D then, cultivates 5 ~ 7 days at 25～30 ℃, makes and contains flavus CICC40258 spore count 1～3 * 10 9The solid fungicide D of individual/g;
(5) Trichodermareesei CICC41491 is inoculated in the PDA solid medium, solid culture is 2～3 days under 25～30 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, the inoculum size by 10 ~ 20wt% is inoculated in Trichodermareesei among the solid medium E then, cultivates 5 ~ 7 days at 25～30 ℃, makes and contains Trichodermareesei CICC41491 spore count 1～3 * 10 9The solid fungicide E of individual/g;
(6) the mould ACCC30150 of long handle wood is inoculated in the PDA solid medium, solid culture is 2～3 days under 25～30 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, then by the inoculum size of 10 ~ 20wt% with mould being inoculated among the solid medium F of long handle wood, cultivated 5 ~ 7 days at 25～30 ℃, make and contain the mould ACCC30150 spore count 1～3 * 10 of long handle wood 9The solid fungicide F of individual/g;
(7) sporotrichum thermophile ACCC30346 is inoculated in the PDA solid medium, solid culture is 2～3 days under 40～45 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, the inoculum size by 10 ~ 20wt% is inoculated in sporotrichum thermophile among the solid medium G then, cultivates 5 ~ 7 days at 40～45 ℃, makes and contains sporotrichum thermophile ACCC30346 spore count 0.5～1.5 * 10 9The solid fungicide G of individual/g;
(8) Phanerochaete chrysosporium CICC40719 is inoculated in the PDA solid medium, solid culture is 2～3 days under 25～30 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, the inoculum size by 10 ~ 20wt% is inoculated in Phanerochaete chrysosporium among the solid medium H then, cultivates 5 ~ 7 days at 25～30 ℃, makes and contains Phanerochaete chrysosporium CICC40719 spore count 0.5～1.5 * 10 9The solid fungicide H of individual/g;
(9) get the solid fungicide H that solid fungicide A that 1～3 weight part step (1) makes, solid fungicide B that 1～3 weight part step (2) makes, solid fungicide C that 1～3 weight part step (3) makes, solid fungicide D that 1～3 weight part step (4) makes, solid fungicide E that 1～3 weight part step (5) makes, solid fungicide F that 1～3 weight part step (6) makes, solid fungicide G that 1～3 weight part step (7) makes and 1～3 weight part step (8) make and mix, make complex micro organism fungicide.
Preferred according to the present invention, described solid medium A is identical with the component of solid medium B, all adopts following raw material to mix, make after sterilization, and raw material is weight part:
600～800 parts of bran powders, 100～300 parts on rice husk, 60～100 parts of Semen Maydis powders, 10～30 parts of bean cake powders, (NH 4) 2SO 44～6 parts, MgSO 47H 20.4～0.6 part of O, KH 2PO 40.4～0.6 part, 16～20 parts of lime powders, 1000～1200 parts in water, pH 7.0～8.0.
Preferred according to the present invention, described solid medium C, solid medium D are identical with the component of solid medium H, all adopt following raw material to mix, make after sterilization, and raw material is weight part:
500～700 parts of straw powder, 300～500 parts of bran powders, (NH 4) 2SO 410～30 parts, KH 2PO 40.5～1.5 parts, MgSO 47H 20.4～0.6 part of O, 1500～2500 parts in water, pH 6.0 ~ 6.5.
Preferred according to the present invention, described solid medium E is identical with the component of solid medium F, all adopts following raw material to mix, make after sterilization, and raw material is weight part:
200～400 parts of bran powders, 600～800 parts of Semen Maydis powders, 3～5 parts of calcium superphosphate, 400～700 parts in water, pH 6.0 ~ 6.5.
Preferred according to the present invention, described solid medium G adopts following raw material to mix, make after sterilization, and raw material is weight part:
300～400 parts of bran powders, 500～600 parts of bagasse, (NH 4) 2SO 410～30 parts, KH 2PO 41～3 part, MgSO 47H 20.4～0.6 part of O, 1000～2000 parts in water, pH 6.0 ~ 6.5.
Above-mentioned LB liquid nutrient medium and PDA solid medium are this area substratum commonly used, and LB liquid nutrient medium component is as follows:
Yeast extract paste 5g, peptone 10g, NaCl 10g, adding distil water regulate pH7.0 ~ 7.2 to 1000ml with NaOH and HCl, the packing sterilization;
PDA solid medium component is as follows:
Potato juice 1000ml, glucose 20g, agar 20g dissolves back packing sterilization;
The application of above-mentioned complex micro organism fungicide in the degraded crop material.
Above-mentioned application, step is as follows:
(1) with crop material after pulverizer is pulverized, add 15 ~ 35 kilograms amount according to stalk per ton and add urea, add water and make the stalk water content reach 60～70wt%, obtain handling stalk;
(2) take by weighing complex micro organism fungicide according to 2～3wt% of stalk weight amount, add in the processing stalk that step (1) makes, mix, composting is in heaps, ferments 20 ~ 35 days.
Preferred according to the present invention, described crop material is the stalk of corn, wheat, paddy rice, jowar and/or beans.
The aforesaid operations step if no special instructions, all by this area routine operation.
Beneficial effect
1. the various plants straws such as corn, wheat, paddy rice, jowar, beans that can become thoroughly decomposed, thus solve the straw transportation and deposited difficulty, avoided the topsoil that causes because of crop straw burning, have good benefits in environment.
2. can shorten stalk and become thoroughly decomposed the time, improve the stalk efficient of becoming thoroughly decomposed.The multiple effective meta-bolites of microorganisms has the effect of fixed nitrogen, phosphorus decomposing, phosphorus decomposing in the fermenting process, and good disease and insect resistance effect, is conducive to the structure of improving the soil, and improves crop yield and quality.
3. complex micro organism fungicide is solid form, bacteria containing amount height in the unit raw material, and microorganism exists with the form of gemma and spore under drying regime, but prolonged preservation is beneficial to packing, transportation and storage.
4. preparation technology is simple, and production process does not need main equipment, and production cost is low, and environmental pollution is low.
Specific implementation method
Be further elaborated below in conjunction with the present invention of embodiment, but institute of the present invention protection domain is not limited thereto.
Microbial material
Subtilis described in the embodiment (Bacillus subtilis) CICC10088, aspergillus niger (Aspergillus niger) CICC41130, flavus (Aspergillus flavus) CICC40258, Trichodermareesei (Trichoderma reesei) CICC41491, Phanerochaete chrysosporium (Phanerochaete chrysosporium) CICC40719 are all available from Chinese industrial microbial strains preservation administrative center;
Described bacillus cereus (Bacillus cereus) ACCC03002, long handle wood mould (Trichoderma viride) ACCC30150, sporotrichum thermophile (Sporotrichum thermophilic) ACCC30346 are all available from Chinese agriculture microbial strains preservation administrative center.
Culture medium raw material described in the embodiment is this area raw material commonly used, but all market is buied.
Embodiment 1
The preparation method of the complex micro organism fungicide of degraded crop material comprises the steps:
(1) subtilis CICC10088 is inoculated in the LB liquid nutrient medium, liquid culture 24h under 37 ℃ of conditions, inoculum size by 10wt% is inoculated in subtilis among the solid medium A then, cultivates 3 days at 37 ℃, makes and contains subtilis CICC10088 viable count 10 10The solid fungicide A of individual/g;
(2) bacillus cereus ACCC03002 is inoculated in the LB liquid nutrient medium, liquid culture 24h under 30 ℃ of conditions, inoculum size by 10wt% is inoculated in bacillus cereus among the solid medium B then, cultivates 3 days at 30 ℃, makes and contains bacillus cereus ACCC03002 viable count 8 * 10 9The solid fungicide B of individual/g;
(3) aspergillus niger CICC41130 is inoculated in the PDA solid medium, under 28 ℃ of conditions, cultivates 60h, add sterilized water and adjust spore concentration to 10 8Individual/ml, the inoculum size by 10wt% is inoculated in aspergillus niger among the solid medium C then, cultivates 5 days at 28 ℃, makes and contains aspergillus niger CICC41130 spore count 2 * 10 9The solid fungicide C of individual/g;
(4) flavus CICC40258 is inoculated in the PDA solid medium, under 28 ℃ of conditions, cultivates 60h, add sterilized water and adjust spore concentration to 10 8Individual/ml, the inoculum size by 10wt% is inoculated in flavus among the solid medium D then, cultivates 5 days at 28 ℃, makes and contains flavus CICC40258 spore count 2 * 10 9The solid fungicide D of individual/g;
(5) Trichodermareesei CICC41491 is inoculated in the PDA solid medium, under 28 ℃ of conditions, cultivates 72h, add sterilized water and adjust spore concentration to 10 8Individual/ml, the inoculum size by 15wt% is inoculated in Trichodermareesei among the solid medium E then, cultivates 7 days at 28 ℃, makes and contains Trichodermareesei CICC41491 spore count 1.5 * 10 9The solid fungicide E of individual/g;
(6) the mould ACCC30150 of long handle wood is inoculated in the PDA solid medium, under 28 ℃ of conditions, cultivates 72h, add sterilized water and adjust spore concentration to 10 8Individual/ml, then by the inoculum size of 15wt% with mould being inoculated among the solid medium F of long handle wood, cultivated 7 days at 28 ℃, make and contain the mould ACCC30150 spore count 1.5 * 10 of long handle wood 9The solid fungicide F of individual/g;
(7) sporotrichum thermophile ACCC30346 is inoculated in the PDA solid medium, under 45 ℃ of conditions, cultivates 60h, add sterilized water and adjust spore concentration to 10 8Individual/ml, the inoculum size by 10wt% is inoculated in sporotrichum thermophile among the solid medium G then, cultivates 7 days at 45 ℃, makes and contains sporotrichum thermophile ACCC30346 spore count 10 9The solid fungicide G of individual/g;
(8) Phanerochaete chrysosporium CICC40719 is inoculated in the PDA solid medium, under 30 ℃ of conditions, cultivates 60h, add sterilized water and adjust spore concentration to 10 8Individual/ml, the inoculum size by 10wt% is inoculated in Phanerochaete chrysosporium among the solid medium H then, cultivates 7 days at 30 ℃, makes and contains Phanerochaete chrysosporium CICC40719 spore count 10 9The solid fungicide H of individual/g;
(9) get the solid fungicide H that solid fungicide A that equal weight step (1) makes, solid fungicide B that step (2) makes, solid fungicide C that step (3) makes, solid fungicide D that step (4) makes, solid fungicide E that step (5) makes, solid fungicide F that step (6) makes, solid fungicide G that step (7) makes and step (8) make and mix, make complex micro organism fungicide.
Described solid medium A is identical with the component of solid medium B, all adopts following raw material to mix, make after sterilization:
Bran powder 80kg, rice husk 10kg, Semen Maydis powder 8kg, bean cake powder 2kg, (NH 4) 2SO 40.5kg, MgSO 47H 2O 0.05kg, KH 2PO 40.05kg, lime powder 1.8kg, water 100L, pH 7.4.
Described solid medium C, solid medium D are identical with the component of solid medium H, all adopt following raw material to mix, make after sterilization:
Straw powder 60kg, bran powder 40kg, (NH 4) 2SO 42kg, KH 2PO 40.1kg, MgSO 47H 2O 0.05kg, water 200L, pH 6.0.
Described solid medium E is identical with the component of solid medium F, all adopts following raw material to mix, make after sterilization, and raw material is weight part:
Bran powder 30kg, Semen Maydis powder 70kg, calcium superphosphate 0.4kg, water 50L, pH 6.0.
Described solid medium G adopts following raw material to mix, make after sterilization, and raw material is weight part:
Bran powder 35kg, bagasse 65kg, (NH 4) 2SO 42kg, KH 2PO 40.2kg, MgSO 47H 2O 0.05kg, water 150L, pH 6.0.
After testing, in the complex micro organism fungicide of the degraded crop material that makes, every gram complex micro organism fungicide contains: subtilis (Bacillus subtilis) viable count 1.25 * 10 9Individual, bacillus cereus (Bacillus cereus) viable count 10 9Individual, aspergillus niger (Aspergillus niger) spore count 2.5 * 10 8Individual, flavus (Aspergillus niger) spore count 2.5 * 10 8Individual, Trichodermareesei (Trichoderma reesei) spore count 1.87 * 10 8Individual, long handle wooden mould (Trichoderma viride) spore count 1.87 * 10 8Individual, sporotrichum thermophile (Sporotrichum thermophilic) spore count 1.25 * 10 8Individual, Phanerochaete chrysosporium (Phanerochaete chrysosporium) spore count 1.25 * 10 8Individual.
Embodiment 2
Adopt as embodiment 1 described method, difference is that the mixed weight ratio of step (9) solid fungicide A, solid fungicide B, solid fungicide C, solid fungicide D, solid fungicide E, solid fungicide F, solid fungicide G and solid fungicide H is 3: 1: 3: 1: 3: 1: 3: 1.
In the complex micro organism fungicide of the degraded crop material that makes, every gram complex micro organism fungicide contains: subtilis (Bacillus subtilis) viable count 1.87 * 10 9Individual, bacillus cereus (Bacillus cereus) viable count 5 * 10 8Individual, aspergillus niger (Aspergillus niger) spore count 3.75 * 10 8Individual, flavus (Aspergillus niger) spore count 1.25 * 10 8Individual, Trichodermareesei (Trichoderma reesei) spore count 2.81 * 10 8Individual, long handle wooden mould (Trichoderma viride) spore count 9.37 * 10 7Individual, sporotrichum thermophile (Sporotrichum thermophilic) spore count 1.87 * 10 8Individual, Phanerochaete chrysosporium (Phanerochaete chrysosporium) spore count 6.25 * 10 7Individual.
Embodiment 3
Adopt as embodiment 1 described method, difference is that the mixed weight ratio of step (9) solid fungicide A, solid fungicide B, solid fungicide C, solid fungicide D, solid fungicide E, solid fungicide F, solid fungicide G and solid fungicide H is 2: 1: 1: 1: 2: 1: 1: 1.
In the complex micro organism fungicide of the degraded crop material that makes, every gram complex micro organism fungicide contains: subtilis (Bacillus subtilis) viable count 2 * 10 9Individual, bacillus cereus (Bacillus cereus) viable count 8 * 10 8Individual, aspergillus niger (Aspergillus niger) spore count 2 * 10 8Individual, flavus (Aspergillus niger) spore count 2 * 10 8Individual, Trichodermareesei (Trichoderma reesei) spore count 3 * 10 8Individual, long handle wooden mould (Trichoderma viride) spore count 1.5 * 10 8Individual, sporotrichum thermophile (Sporotrichum thermophilic) spore count 10 8Individual, Phanerochaete chrysosporium (Phanerochaete chrysosporium) spore count 10 8Individual.
Embodiment 4
The application of above-mentioned complex micro organism fungicide in the degraded crop material, step is as follows:
(1) with the 500kg maize straw after pulverizer is pulverized, mix with 10kg urea, add water and make the stalk water content reach 65wt%, obtain handling stalk;
(2) take by weighing the 15kg complex micro organism fungicide, add in the processing stalk that step (1) makes, mix, wide 2 meters by heap, to pile highly 1.6 meters, composting is in heaps to ferment, and ferments about 20 days to make.
The complex micro organism fungicide that respectively embodiment 1, embodiment 2 and embodiment 3 is made is tested as stated above, is respectively and handles 1, handles 2, handles 3.
Stalk is in degradation process, temperature and change in color have shown the action effect of microorganism to stalk, during the fermentation, record temperature and the color of stalk at set intervals, wherein color is divided into five grades, be respectively: pale yellow, deep yellow, yellowish-brown, brown, dark brown, the explanation stalk becomes thoroughly decomposed fully when the stalk color becomes chocolate.The result is as shown in table 1.
The quality of process degradation effect is the evaluation of handling Mierocrystalline cellulose, hemicellulose, xylogen (being called for short three elements) content in the stalk of back by measuring.
The present invention after fermentative processing, is dried maize straw to constant weight through 60 ℃, measures " three elements " content, is contrast with " three elements " content of untreated maize straw.
According to Wang Jin master's detection method in " mensuration of Mierocrystalline cellulose, hemicellulose and xylogen in the maize straw " (" Shandong food fermentation ", 03 phase in 2010) maize straw is carried out the detection of palliating degradation degree, palliating degradation degree is as shown in table 3.
Embodiment 5
The application of above-mentioned complex micro organism fungicide in the degraded crop material, step is as follows:
(1) with the 500kg wheat stalk after pulverizer is pulverized, mix with 15kg urea, add water and make the stalk water content reach 65wt%, obtain handling stalk;
(2) take by weighing the 15kg complex micro organism fungicide, add in the processing stalk that step (1) makes, mix, wide 2 meters by heap, to pile highly 1.6 meters, composting is in heaps to ferment, and ferments about 20 days to make.
The complex micro organism fungicide that respectively embodiment 1, embodiment 2 and embodiment 3 is made is tested as stated above, is respectively and handles 4, handles 5, handles 6.
Detection method is identical with embodiment 4, and temperature and change in color result are as shown in table 2, and palliating degradation degree is as shown in table 4.
Reference examples 1
(1) with the 500kg maize straw after pulverizer is pulverized, mix with 10kg urea, add water and make the stalk water content reach 65wt%, obtain handling stalk, wide 2 meters by heap, pile highly 1.6 meters, composting is in heaps to ferment.The result is shown in table 1, table 3.
Reference examples 2
With reference to the technical scheme preparation contrast microbial inoculum that embodiment 1 among the Chinese patent literature CN101775359A puts down in writing, use above-mentioned contrast microbial inoculum according to the method for the embodiment of the present application 4.
Wherein bacterial classification adopts the agent of becoming thoroughly decomposed bacterial classification commonly used: subtilis (Bacillus subtilis) CICC10088 is available from Chinese industrial microbial strains preservation administrative center.Bacillus licheniformis (Bacillus licheniformis) SDMCC10292 is available from DSMZ of Shandong Agricultural University.Cured shape genus bacillus (Bacillus cereus) ACCC10606 is available from Chinese agriculture microbial strains preservation administrative center.Bacillus megaterium (Bacillus megaterium) CICC20611 is available from Chinese industrial microbial strains preservation center.Bacillusmusilaginosiengineering (Bacillus mucilaginosus) CGMCC1.232 is available from Chinese common micro-organisms DSMZ.Bacillus circulans (Bacillus circulans) CICC20133 is available from Chinese industrial microbial strains preservation administrative center.Spherical bacillus (Bacillus sphaericus) CICC20687 is available from Chinese industrial microbial strains preservation center.Clostridium sporogene (Clostridium sporogenes) CGMCC1.2157 is available from Chinese common micro-organisms DSMZ.Acetone clostridium butylicum (Clostridium acetobutylicum) CGMCC1.70 is available from Chinese common micro-organisms DSMZ.Clostridium baratii (Clostridium pasterrianum) ACCC00100 is available from Chinese agriculture microbial strains preservation administrative center.Bai Shi clostridium (Clostridium beijerincki) ACCC00174 is available from Chinese agriculture microbial strains preservation administrative center.Clostridium thermocellum (Clostridium thermocellum) ACCC00168 is available from Chinese agriculture microbial strains preservation administrative center.Viride (Trichoderma viride) ACCC30793 is available from Chinese agriculture microbial strains preservation administrative center.Rui Shi wood mould (Trichoderma reesei) ATCC13631 U.S. representative microbial DSMZ, aspergillus niger (Aspergillus niger) ACCC30786 are available from Chinese agriculture microbial strains preservation administrative center.
Detection method is identical with embodiment 4, and the result is shown in table 1, table 3.
Reference examples 3
(1) with the 500kg wheat stalk after pulverizer is pulverized, mix with 15kg urea, add water and make the stalk water content reach 65wt%, obtain handling stalk, wide 2 meters by heap, pile highly 1.6 meters, composting is in heaps to ferment.
Detection method is identical with embodiment 4, and palliating degradation degree is shown in table 2, table 4.
Reference examples 4
With reference to the technical scheme preparation contrast microbial inoculum that embodiment 1 among the Chinese patent literature CN101775359A puts down in writing, use above-mentioned contrast microbial inoculum according to the method for the embodiment of the present application 5.
Wherein bacterial classification is shown in reference examples 2.
Detection method is identical with embodiment 4, and the result is shown in table 2, table 4.
Detected result
Table 1 different treatment is to the influence of maize straw temperature and color
Figure BDA00001718114600081
Interpretation of result
As can be seen from Table 1, use complex micro organism fungicide of the present invention to handle maize straw and play heat soon than control group, all rise rapidly in the 4th day temperature of compost, and reached the highest temperature at the 13rd day, all become chocolate in the 20th day stalk color, illustrate that stalk becomes thoroughly decomposed fully.Utilize 2 heat of contrast of microbial inoculum described in the CN 101775359A slower, reached the highest temperature on the 16th day at compost, became thoroughly decomposed fully at the 25th day, a hot time and the time of becoming thoroughly decomposed fully all are slower than microbial inoculum of the present invention.Explanation thus, complex micro organism fungicide of the present invention plays thermal velocity faster than microbial inoculum described in the patent CN 101775359A.
Table 2 different treatment is to the influence of wheat stalk temperature and color
Figure BDA00001718114600082
Interpretation of result
As can be seen from Table 2, use complex micro organism fungicide of the present invention to handle wheat stalk and contrasted heat soon, all rise rapidly in the 4th day temperature of compost, and reached the highest temperature at the 13rd day, all become chocolate in the 20th day stalk color, illustrate that stalk becomes thoroughly decomposed fully.Utilize 4 heat of contrast of microbial inoculum described in the CN 101775359A slower, arrived the highest temperature on the 16th day at compost, become thoroughly decomposed fully at the 25th day stalk, a hot time and the time of becoming thoroughly decomposed fully all are slower than microbial inoculum of the present invention.Explanation thus, complex micro organism fungicide of the present invention plays thermal velocity faster than microbial inoculum described in the patent CN 101775359A.
Table 3 is the maize straw degradation rate (%) of different treatment group
Group Mierocrystalline cellulose Hemicellulose Xylogen
Handle 1 35.16% 41.36% 43.04%
Handle 2 35.12% 40.24% 42.20%
Handle 3 36.56% 43.42% 40.24%
Contrast 1 17.12% 25.22% 18.18%
Contrast 2 31.42% 36.46% 35.26%
Interpretation of result
The degradation rate of the maize straw " three elements " of complex micro organism fungicide degraded of the present invention all is higher than control group as can be seen from Table 3, illustrates that microbial inoculum action effect of the present invention is better than microbial inoculum described in the CN 101775359A.
Table 4 is the wheat stalk degradation rate (%) of different treatment group
Group Mierocrystalline cellulose Hemicellulose Xylogen
Handle 4 34.12% 40.16% 41.24%
Handle 5 34.14% 42.24% 42.14%
Handle 6 33.26% 41.12% 39.24%
Contrast 3 15.12% 21.22% 15.14%
Contrast 4 30.42% 34.46% 33.28%
Interpretation of result
The degradation rate of the wheat stalk " three elements " of complex micro organism fungicide degraded of the present invention all is higher than control group as can be seen from Table 4, illustrates that microbial inoculum action effect of the present invention is better than microbial inoculum described in the CN 101775359A.

Claims (2)

1. the complex micro organism fungicide of the crop material of degrading is characterized in that, be by subtilis ( Bacillus subtilis) CICC10088, bacillus cereus ( Bacillus cereus) ACCC03002, aspergillus niger ( Aspergillus niger) CICC41130, flavus ( Aspergillus flavus) CICC40258, Trichodermareesei ( Trichoderma reesei) CICC41491, long handle wood mould ( Trichoderma viride) ACCC30150, sporotrichum thermophile ( Sporotrichum thermophilic) ACCC30346 and Phanerochaete chrysosporium ( Phanerochaete chrysosporium) CICC40719 makes through solid fermentation.
2. complex micro organism fungicide as claimed in claim 1 is characterized in that, every gram complex micro organism fungicide contains: subtilis ( Bacillus subtilis) CICC10088 viable count 10 8～10 10Individual, bacillus cereus ( Bacillus cereus) ACCC03002 viable count 10 8～10 10Individual, aspergillus niger ( Aspergillus niger)CICC41130 spore count 10 7～10 9Individual, flavus ( Aspergillus niger) CICC40258 spore count 10 7～10 9Individual, Trichodermareesei ( Trichoderma reesei) CICC41491 spore count 10 7～10 9Individual, long handle wood mould ( Trichoderma viride) ACCC30150 spore count 10 7～10 9Individual, sporotrichum thermophile ( Sporotrichum thermophilic) ACCC30346 spore count 10 7～10 9Individual, Phanerochaete chrysosporium ( Phanerochaete chrysosporium) CICC40719 spore count 10 7～10 9Individual.
3 .Preparation method according to claim 1 or 2 described complex micro organism fungicides is characterized in that, comprises the steps:
(1) subtilis CICC10088 is inoculated in the LB liquid nutrient medium, liquid culture 18～24h under 30～37 ℃ of conditions, inoculum size by 10 ~ 20wt% is inoculated among the solid medium A then, cultivated 3 ~ 5 days at 30～37 ℃, make and contain subtilis CICC10088 viable count 8～15 * 10 9The solid fungicide A of individual/g;
(2) bacillus cereus ACCC03002 is inoculated in the LB liquid nutrient medium, liquid culture 18～24h under 30～37 ℃ of conditions, inoculum size by 10 ~ 20wt% is inoculated in bacillus cereus among the solid medium B then, cultivated 3 ~ 5 days at 30～37 ℃, make and contain bacillus cereus ACCC03002 viable count 8～15 * 10 9The solid fungicide B of individual/g;
(3) aspergillus niger CICC41130 is inoculated in the PDA solid medium, solid culture is 2～3 days under 25～30 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, the inoculum size by 10 ~ 20wt% is inoculated in aspergillus niger among the solid medium C then, cultivates 5 ~ 7 days at 25～30 ℃, makes and contains aspergillus niger CICC41130 spore count 1～3 * 10 9The solid fungicide C of individual/g;
(4) flavus CICC40258 is inoculated in the PDA solid medium, solid culture is 2～3 days under 25～30 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, the inoculum size by 10 ~ 20wt% is inoculated in flavus among the solid medium D then, cultivates 5 ~ 7 days at 25～30 ℃, makes and contains flavus CICC40258 spore count 1～3 * 10 9The solid fungicide D of individual/g;
(5) Trichodermareesei CICC41491 is inoculated in the PDA solid medium, solid culture is 2～3 days under 25～30 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, the inoculum size by 10 ~ 20wt% is inoculated in Trichodermareesei among the solid medium E then, cultivates 5 ~ 7 days at 25～30 ℃, makes and contains Trichodermareesei CICC41491 spore count 1～3 * 10 9The solid fungicide E of individual/g;
(6) the mould ACCC30150 of long handle wood is inoculated in the PDA solid medium, solid culture is 2～3 days under 25～30 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, then by the inoculum size of 10 ~ 20wt% with mould being inoculated among the solid medium F of long handle wood, cultivated 5 ~ 7 days at 25～30 ℃, make and contain the mould ACCC30150 spore count 1～3 * 10 of long handle wood 9The solid fungicide F of individual/g;
(7) sporotrichum thermophile ACCC30346 is inoculated in the PDA solid medium, solid culture is 2～3 days under 40～45 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, the inoculum size by 10 ~ 20wt% is inoculated in sporotrichum thermophile among the solid medium G then, cultivates 5 ~ 7 days at 40～45 ℃, makes and contains sporotrichum thermophile ACCC30346 spore count 0.5～1.5 * 10 9The solid fungicide G of individual/g;
(8) Phanerochaete chrysosporium CICC40719 is inoculated in the PDA solid medium, solid culture is 2～3 days under 25～30 ℃ of conditions, adds sterilized water and adjusts spore concentration to 10 8Individual/ml, the inoculum size by 10 ~ 20wt% is inoculated in Phanerochaete chrysosporium among the solid medium H then, cultivates 5 ~ 7 days at 25～30 ℃, makes and contains Phanerochaete chrysosporium CICC40719 spore count 0.5～1.5 * 10 9The solid fungicide H of individual/g;
(9) get the solid fungicide H that solid fungicide A that 1～3 weight part step (1) makes, solid fungicide B that 1～3 weight part step (2) makes, solid fungicide C that 1～3 weight part step (3) makes, solid fungicide D that 1～3 weight part step (4) makes, solid fungicide E that 1～3 weight part step (5) makes, solid fungicide F that 1～3 weight part step (6) makes, solid fungicide G that 1～3 weight part step (7) makes and 1～3 weight part step (8) make and mix, make complex micro organism fungicide;
Described solid medium A is identical with the component of solid medium B, all adopts following raw material to mix, make after sterilization, and raw material is weight part:
600～800 parts of bran powders, 100～300 parts on rice husk, 60～100 parts of Semen Maydis powders, 10～30 parts of bean cake powders, (NH 4) 2SO 44～6 parts, MgSO 47H 20.4～0.6 part of O, KH 2PO 40.4～0.6 part, 16～20 parts of lime powders, 1000～1200 parts in water, pH 7.0～8.0;
Described solid medium C, solid medium D are identical with the component of solid medium H, all adopt following raw material to mix, make after sterilization, and raw material is weight part:
500～700 parts of straw powder, 300～500 parts of bran powders, (NH 4) 2SO 410～30 parts, KH 2PO 40.5～1.5 parts, MgSO 47H 20.4～0.6 part of O, 1500～2500 parts in water, pH nature;
Described solid medium E is identical with the component of solid medium F, all adopts following raw material to mix, make after sterilization, and raw material is weight part:
200～400 parts of bran powders, 600～800 parts of Semen Maydis powders, 3～5 parts of calcium superphosphate, 400～700 parts in water, pH 6.0.
Described solid medium G adopts following raw material to mix, make after sterilization, and raw material is weight part:
300～400 parts of bran powders, 500～600 parts of bagasse, (NH 4) 2SO 410～30 parts, KH 2PO 41～3 part, MgSO 47H 20.4～0.6 part of O, 1000～2000 parts in water, pH 6.0.
4 .The application of the described complex micro organism fungicide of claim 1 in the degraded crop material.
5 .Application as claimed in claim 4 is characterized in that, step is as follows:
(1) with crop material after pulverizer is pulverized, add 15 ~ 35 kilograms amount according to stalk per ton and add urea, add water and make the stalk water content reach 60～70wt%, obtain handling stalk;
(2) take by weighing complex micro organism fungicide according to 2～3wt% of stalk weight amount, add in the processing stalk that step (1) makes, mix, composting is in heaps, ferments 20 ~ 35 days.
6 .Application as claimed in claim 5 is characterized in that, described crop material is the stalk of corn, wheat, paddy rice, jowar and/or beans.
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