CN104962489A - Liquid straw decomposition agent and preparation and application methods thereof - Google Patents
Liquid straw decomposition agent and preparation and application methods thereof Download PDFInfo
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- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
The invention provides a liquid straw decomposition agent, and contains main active microbes as follows: Bacillus amyloliquefaciens and Saccharomyces cerevisiae, and the two active microbes are mutually cooperated without antagonistic action, wherein, the effective bacteria number is 2-5 hundred million/ml, the number of effective live bacteria accounts for 50-60% of the number of Bacillus amyloliquefaciens, and the number of effective live bacteria accounts for 40-50% of the number of Saccharomyces cerevisiae. The Bacillus amyloliquefaciens and Saccharomyces cerevisiae are treated by the following steps: (1) test tube bacterial strain slang culture; (2) shake flask and activation culture; (3) first grade seeding tank fermentation; (4) fermentation tank fermentation; the fermentation liquors of Bacillus amyloliquefaciens and Saccharomyces cerevisiae are obtained, and the fermentation liquors are mixed and compounded according to the volume ratio, and the liquid straw decomposition agent is obtained, wherein, the Bacillus amyloliquefaciens fermentation liquor accounts for 50-60% and the Saccharomyces cerevisiae fermentation liquor accounts for 40-50%. The liquid straw decomposition agent provided by the invention is suitable for rapid decomposition of wheat, paddy rice and other crop straw.
Description
Technical field
The present invention relates to a kind of liquid straw decomposing inoculant and preparation and application, be applicable to the quick composting of the agricultural crop straws such as wheat, paddy rice, corn, belong to biological technical field.
Background technology
Straw-returning is an important agricultural measures, and to raising soil organic matter content, preservation of fertility, the Sustainable development realizing agricultural has great importance.In recent years, along with the development of mechanization of agriculture and microbial technique (microbial preparation), straw directly returning to field is generally paid attention to, and large-scale popularization application in China.This correct for the wasting phenomenon of crop straw burning to a certain extent, decreases the destruction to atmospheric environment and ecotope, and soil fertility of simultaneously having fostered and apply fertilizer, effectively improves soil organic matter content.
At present, straw-returning is based on direct returning to farmland, the comprehensive analysis of vegetable fibre is classified, lacked fundamental research to many inner links in aspect such as the residual body decomposition of stalk and soil fertility, plant nutrition, physiological metabolisms, makes it to there is series of problems in straw degradative, dietetic alimentation etc.In addition straw decomposing microbial inoculum is the use complex steps of pulvis, product substantially on the market, and poorly water-soluble, did not mate with the existing agro-farming custom, and common people need to pay more labours and carry out straw-returning.Deficient all the more and cost of labor ever-increasing today rural laborer, this contradiction highlights day by day, and thus powder product can not make common people generally accept and use, and this also counteracts that the development of straw decomposing inoculant industry.
In sum, easy in the urgent need to using method at present and straw decomposing microbial inoculum that is that mate with the existing agro-farming custom, allows common people afford to use, the meeting of learning, visible.
Summary of the invention
The object of the invention is the deficiency overcoming prior art existence, there is provided one mainly for the problem such as powder dose-type product poorly water-soluble, use inconvenience in existing straw directly returning to field process, the cellulase utilizing straw decomposing inoculant to produce, proteolytic enzyme isoreactivity material accelerate the decomposition of stalk, improve soil, be conducive to liquid straw decomposing inoculant and the preparation and application of straw directly returning to field.
The object of the invention is to have come by following technical solution, a kind of liquid straw decomposing inoculant, this liquid straw decomposing inoculant includes chief active microorganism: bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), described two kinds of living microorganisms are mutually collaborative and without antagonistic action, wherein contained effectively bacterium number is 2-5 hundred million/ml, the living bacteria count of wherein said bacillus amyloliquefaciens accounts for 50-60%, and the living bacteria count of yeast saccharomyces cerevisiae accounts for 40-50%.
Described bacillus amyloliquefaciens and yeast saccharomyces cerevisiae are respectively through (1) test tube strains slant culture, (2) shaking flask activation culture, (3) first class seed pot fermentation and (4) ferment tank, obtain bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid, after above-mentioned bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid is composite by following volume ratio mixing, obtain liquid straw decomposing inoculant, wherein bacillus amyloliquefaciens fermented liquid 50-60%, fermentation by saccharomyces cerevisiae liquid 40-50%.
As a preparation method for aforesaid liquid straw decomposing inoculant, this preparation method comprises: the composite mixing of the preparation of bacillus amyloliquefaciens fermented liquid, the preparation of fermentation by saccharomyces cerevisiae liquid and bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid, wherein:
A) preparation of bacillus amyloliquefaciens fermented liquid is:
(1) test tube strains slant culture: bacillus amyloliquefaciens is inoculated in LB slant medium 37 DEG C and cultivates 1-2 days;
(2) shaking flask activation culture: activated spawn is inoculated in LB liquid nutrient medium, 37 DEG C, 180rpm/min cultivates living bacteria count 1-2 hundred million/ml in 18h to nutrient solution;
(3) seeding tank fermentation: liquid culture bacterium liquid is inoculated in seed culture medium by 1-2% inoculum size, 37 DEG C, 200rpm/min cultivates living bacteria count 1-2 hundred million/ml in 24h to nutrient solution;
(4) ferment tank: be inoculated in fermention medium by 1-2% inoculum size by seed culture fluid, 37 DEG C, 200rpm/min cultivates living bacteria count 2-4 hundred million/ml in 36h to nutrient solution, obtains bacillus amyloliquefaciens fermented liquid.
B) preparation of fermentation by saccharomyces cerevisiae liquid is:
(1) test tube strains slant culture: yeast saccharomyces cerevisiae is inoculated in PDA slant medium 28 DEG C and cultivates 2-3 days;
(2) shaking flask activation culture: activated spawn is inoculated in PDA liquid nutrient medium, 28 DEG C, 180rpm/min cultivates living bacteria count 1-2 hundred million/ml in 36h to nutrient solution;
(3) seeding tank fermentation: liquid culture bacterium liquid is inoculated in seed culture medium by 5% inoculum size, 28 DEG C, 200rpm/min cultivates living bacteria count 1-2 hundred million/ml in 36h to nutrient solution;
(4) ferment tank: be inoculated in fermention medium by 5% inoculum size by seed culture fluid, 28 DEG C, 200rpm/min cultivates living bacteria count 2-3 hundred million/ml in 48h to nutrient solution, obtains fermentation by saccharomyces cerevisiae liquid.
C) after above-mentioned bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid is composite by following volume ratio mixing, obtain containing the liquid straw decomposing inoculant that effective bacterium number is 2-5 hundred million/ml, wherein bacillus amyloliquefaciens fermented liquid 50-60%, fermentation by saccharomyces cerevisiae liquid 40-50%.
In the preparation of bacillus amyloliquefaciens fermented liquid of the present invention:
LB slant medium described in step (1) is pH value 7.2 solution configured by 10g peptone, 5g yeast extract paste, 10g sodium-chlor, 15g agar, 1000ml water;
Seed culture medium described in LB liquid nutrient medium described in step (2) and step (3) is by pH value 7.2 solution of 10g peptone, 5g yeast extract paste, 10g sodium-chlor, the configuration of 1000ml water;
Fermention medium described in step (4) is by 20g Semen Maydis powder, 30g bean cake powder, 5g glucose, 1g K
2hPO
4, 0.2g manganous sulfate, 0.3g calcium carbonate and 1000ml water configuration form.
In the preparation of fermentation by saccharomyces cerevisiae liquid of the present invention:
The solution that PDA slant medium described in step (1) and the PDA liquid nutrient medium described in step (2) configure by 200g potato, 20g sucrose, 1000ml water;
Seed culture medium described in step (3) is the solution configured by 5g glucose, 5g peptone, 3g yeast extract paste and 1000ml water;
Fermention medium described in step (4) is the solution configured by 20g glucose, 10g yeast extract paste, 5g peptone, 10g calcium carbonate and 1000ml water.
A kind of using method of liquid straw decomposing inoculant described above, this using method is: by direct for stalk chopping and returning after crop harvesting, make stalk uniform spreading at surface layer, 60 kilograms are watered by every mu of ground stalk decomposing agent 2 kilograms, and the urea adding 5-6 kilogram is evenly sprayed on stalk after making it fully dissolve, carry out immediately turning over and irrigating.
In the present invention, described stalk is specially rice straw or wheat, maize straw.
The invention has the beneficial effects as follows: straw decomposing inoculant provided by the present invention, adopt efficient collaborative flora composition, make full use of the cellulase that bacterial classification produces, proteolytic enzyme isoreactivity material, accelerate the maturity of stalk, experiment prove decomposing microbial inoculum of the present invention to stalk degradation rate high, can 89.1% be reached at laboratory condition decomposition effect, also good decomposition effect can be reached under field demonstration condition, within 10 days, can 30% be reached, within 30 days, reach 42%, straw decomposing inoculant degradation rate than direct returning to farmland and use commercial type is higher, liquid dosage form can adopt the mode directly sprayed more to meet agro-farming custom after straw-returning simultaneously, raw work is laborsaving, and preparation cost is cheap, there is higher application prospect.
Embodiment
Below in conjunction with specific embodiment, the present invention will be described in detail: a kind of liquid straw decomposing inoculant of the present invention, it includes chief active microorganism: bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), described two kinds of living microorganisms are mutually collaborative and without antagonistic action, wherein contained effectively bacterium number is 2-5 hundred million/ml, the living bacteria count of wherein said bacillus amyloliquefaciens accounts for 50-60%, and the living bacteria count of yeast saccharomyces cerevisiae accounts for 40-50%.
Bacillus amyloliquefaciens of the present invention and yeast saccharomyces cerevisiae are respectively through (1) test tube strains slant culture, (2) shaking flask activation culture, (3) first class seed pot fermentation and (4) ferment tank, obtain bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid, after above-mentioned bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid is composite by following volume ratio mixing, obtain liquid straw decomposing inoculant, wherein bacillus amyloliquefaciens fermented liquid 50-60%, fermentation by saccharomyces cerevisiae liquid 40-50%.
A preparation method for liquid straw decomposing inoculant described above, this preparation method comprises: the composite mixing of the preparation of bacillus amyloliquefaciens fermented liquid, the preparation of fermentation by saccharomyces cerevisiae liquid and bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid, wherein:
A) preparation of bacillus amyloliquefaciens fermented liquid is:
(1) test tube strains slant culture: bacillus amyloliquefaciens is inoculated in LB slant medium 37 DEG C and cultivates 1-2 days;
(2) shaking flask activation culture: activated spawn is inoculated in LB liquid nutrient medium, 37 DEG C, 180rpm/min cultivates living bacteria count 1-2 hundred million/ml in 18h to nutrient solution;
(3) seeding tank fermentation: liquid culture bacterium liquid is inoculated in seed culture medium by 1-2% inoculum size, 37 DEG C, 200rpm/min cultivates living bacteria count 1-2 hundred million/ml in 24h to nutrient solution;
(4) ferment tank: be inoculated in fermention medium by 1-2% inoculum size by seed culture fluid, 37 DEG C, 200rpm/min cultivates living bacteria count 2-4 hundred million/ml in 36h to nutrient solution, obtains bacillus amyloliquefaciens fermented liquid.
B) preparation of fermentation by saccharomyces cerevisiae liquid is:
(1) test tube strains slant culture: yeast saccharomyces cerevisiae is inoculated in PDA slant medium 28 DEG C and cultivates 2-3 days;
(2) shaking flask activation culture: activated spawn is inoculated in PDA liquid nutrient medium, 28 DEG C, 180rpm/min cultivates living bacteria count 1-2 hundred million/ml in 36h to nutrient solution;
(3) seeding tank fermentation: liquid culture bacterium liquid is inoculated in seed culture medium by 5% inoculum size, 28 DEG C, 200rpm/min cultivates living bacteria count 1-2 hundred million/ml in 36h to nutrient solution;
(4) ferment tank: be inoculated in fermention medium by 5% inoculum size by seed culture fluid, 28 DEG C, 200rpm/min cultivates living bacteria count 2-3 hundred million/ml in 48h to nutrient solution, obtains fermentation by saccharomyces cerevisiae liquid.
C) after above-mentioned bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid is composite by following volume ratio mixing, obtain containing the liquid straw decomposing inoculant that effective bacterium number is 2-5 hundred million/ml, wherein bacillus amyloliquefaciens fermented liquid 50-60%, fermentation by saccharomyces cerevisiae liquid 40-50%.
In the preparation of bacillus amyloliquefaciens fermented liquid of the present invention:
LB slant medium described in step (1) is pH value 7.2 solution configured by 10g peptone, 5g yeast extract paste, 10g sodium-chlor, 15g agar, 1000ml water;
Seed culture medium described in LB liquid nutrient medium described in step (2) and step (3) is by pH value 7.2 solution of 10g peptone, 5g yeast extract paste, 10g sodium-chlor, the configuration of 1000ml water;
Fermention medium described in step (4) is by 20g Semen Maydis powder, 30g bean cake powder, 5g glucose, 1g K
2hPO
4, 0.2g manganous sulfate, 0.3g calcium carbonate and 1000ml water configuration form.
In the preparation of fermentation by saccharomyces cerevisiae liquid of the present invention:
The solution that PDA slant medium described in step (1) and the PDA liquid nutrient medium described in step (2) configure by 200g potato, 20g sucrose, 1000ml water;
Seed culture medium described in step (3) is the solution configured by 5g glucose, 5g peptone, 3g yeast extract paste and 1000ml water;
Fermention medium described in step (4) is the solution configured by 20g glucose, 10g yeast extract paste, 5g peptone, 10g calcium carbonate and 1000ml water.
A kind of using method of liquid straw decomposing inoculant described above, it is characterized in that described using method is: by direct for stalk chopping and returning after crop harvesting, make stalk uniform spreading at surface layer, 60 kilograms are watered by every mu of ground stalk decomposing agent 2 kilograms, and the urea adding 5-6 kilogram is evenly sprayed on stalk after making it fully dissolve, carry out immediately turning over and irrigating.
The specific embodiment of the following stated is to understand the present invention better, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is ordinary method.Test materials used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative test in following examples, all arranges and repeats experiment for three times, results averaged.
The preparation of embodiment 1, liquid straw decomposing inoculant
One, the separation of microbial strains obtains
Gather land for growing field crops stalk to rot plot stalk mud sample, liquid acclimating is carried out in indoor, and the sample high to the straw degradative efficiency of enrichment carries out the separation of microorganism, obtain there is good straw decomposition efficiency and the bacillus amyloliquefaciens jx-10 of mutual not antagonism and yeast saccharomyces cerevisiae jx-15.
Bacterial classification bacillus amyloliquefaciens (Bacillus amyloliquefaciens) jx-10 and yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) jx-15 that prepare decomposing agent needs have been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC on 02 11st, 2015, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City), deposit number is respectively as CGMCC No.10559 and CGMCC No.10560.
Two, the preparation of bacillus amyloliquefaciens fermented liquid
By bacillus amyloliquefaciens through test tube strains slant culture-shaking flask activation culture-seeding tank fermentation-ferment tank, obtain bacillus amyloliquefaciens fermented liquid.
Actication of culture: bacillus amyloliquefaciens is inoculated in LB (peptone 10g, yeast extract paste 5g, sodium-chlor 10g, agar 15g, water 1000ml, pH7.2) slant medium 37 DEG C and cultivates 1-2 days.
Shaking flask activation culture: activated spawn be inoculated in LB (peptone 10g, yeast extract paste 5g, sodium-chlor 10g, water 1000ml, pH7.2) liquid nutrient medium, 37 DEG C, 180rpm/min cultivates living bacteria count 1-2 hundred million/ml in 18h to nutrient solution.
Seeding tank ferments: be inoculated in seed culture medium (peptone 10g, yeast extract paste 5g, sodium-chlor 10g, water 1000ml, pH7.2) by liquid culture bacterium liquid by 1-2% inoculum size, 37 DEG C, 200rpm/min cultivates living bacteria count 1-2 hundred million/ml in 24h to nutrient solution.
Ferment tank: seed culture fluid is inoculated into fermention medium (Semen Maydis powder 20g, bean cake powder 30g, glucose 5g, K by 1-2% inoculum size
2hPO
41g, manganous sulfate 0.2g, calcium carbonate 0.3g, water 1000ml) in, 37 DEG C, 200rpm/min cultivates living bacteria count 2-4 hundred million/ml in 36h to nutrient solution, obtains bacillus amyloliquefaciens fermented liquid.
Three, the preparation of fermentation by saccharomyces cerevisiae liquid
By yeast saccharomyces cerevisiae through test tube strains slant culture-shaking flask activation culture-seeding tank fermentation-ferment tank, obtain bacillus amyloliquefaciens fermented liquid.
Actication of culture: yeast saccharomyces cerevisiae is inoculated in PDA (potato 200g, sucrose 20g, water 1000ml, pH nature) slant medium 28 DEG C and cultivates 2-3 days.
Shaking flask activation culture: activated spawn is inoculated into PDA (potato 200g, sucrose 20g, water 1000ml, pH nature)) in liquid nutrient medium, 28 DEG C, 180rpm/min cultivates living bacteria count 1-2 hundred million/ml in 36h to nutrient solution.
Seeding tank ferments: be inoculated in seed culture medium (glucose 5g, peptone 5g, yeast extract paste 3g, water 1000ml) by liquid culture bacterium liquid by 5% inoculum size, 28 DEG C, and 200rpm/min cultivates living bacteria count 1-2 hundred million/ml in 36h to nutrient solution.
Ferment tank: seed culture fluid is inoculated in fermention medium (glucose 20g, yeast extract paste 10g, peptone 5g, calcium carbonate 10g, water 1000ml) by 5% inoculum size, 28 DEG C, 200rpm/min cultivates living bacteria count 2-3 hundred million/ml in 48h to nutrient solution, obtains fermentation by saccharomyces cerevisiae liquid.
Four, liquid decomposing agent is composite
Bacillus amyloliquefaciens fermented liquid obtained above and fermentation by saccharomyces cerevisiae liquid are mixed in proportion, obtain mixing liquid decomposing agent; Wherein in mixed bacteria liquid, bacillus amyloliquefaciens fermented liquid accounts for 50-60% (volume ratio), fermentation by saccharomyces cerevisiae liquid accounts for 40-50% (volume ratio).
Embodiment 2 prepares decomposing agent decomposition effect in straw-returning
Experiment place: Jiaxing City academy of agricultural sciences
Certain brand decomposing agent that liquid straw decomposing inoculant provided by the invention and market are buied is carried out field decomposition simultaneous test after wheat stalk chopping and returning, and experimental design is as follows.
| Contrast numbering | Returning total stalks into fields | Decomposing microbial inoculum | Synchronous fertilising |
| T1 | Be | No | Be |
| T2 | Be | Certain brand | Be |
| T3 | Be | The present invention | Be |
Straw harvesting chopping after harvesting wheat, direct chopping and returning, makes stalk uniform spreading at surface layer, is watered 60 kilograms by every mu of ground stalk decomposing agent 2 kilograms, and the urea adding 5-6 kilogram is evenly sprayed on stalk after making it fully dissolve, carry out immediately turning over and irrigating.
To pour water rice cultivation according to normal agro-farming custom according to after the process of aforesaid operations method.Respectively at 10 days of process, 20 days, 30 days, 60 days, within 100 days, measure the rate of weight loss of stalk afterwards.
Result is as follows:
Stalk buries a bag rate of weight loss (per-cent)
| Process | 10 days | 20 days | 30 days | 60 days | 100 days |
| T1 | 35.65％ | 42.19％ | 48.35％ | 60.72％ | 64.52％ |
| T2 | 36.43％ | 43.24％ | 50.20％ | 61.55％ | 68.58％ |
| T3 | 41.72％ | 49.17％ | 58.11％ | 73.57％ | 77.79％ |
As seen from the above table, adopt decomposing agent process of the present invention rate of weight loss 41.72% after 10 days, within 20 days, rate of weight loss reaches 49.17%, within 30 days, rate of weight loss reaches 58.11%, degradation effect is apparently higher than use other decomposing microbial inoculums on the market, show that straw decomposing inoculant provided by the invention can decomposition stalk fast and effectively, and liquid dosage form is convenient, saves labor.
The decomposition effect of embodiment 3 decomposing agent in field demonstration
Demonstration place: Anji County emperor Hu Zhen Xiao Yun village
Liquid straw decomposing inoculant provided by the invention is used enforcement straw-returning after wheat stalk chopping, and control design is as follows:
| Contrast numbering | Returning total stalks into fields | Decomposing microbial inoculum | Synchronous fertilising |
| Ck | No | No | No |
| T1 | Be | No | Be |
| T2 | Be | Certain brand | Be |
| T3 | Be | The present invention | Be |
Field is shown as follows by burying bag operation acquisition decomposition effect:
Stalk buries a bag rate of weight loss (per-cent)
Field demonstration result shows: after using decomposing agent soil conditioning of the present invention, after 10 days, stalk rate of weight loss can reach 39.19%, 46.71% is reached after 20 days, reach 52.73% after 30 days, and contrast uses other microbial inoculum to be on the market 43.68% after 30 days, difference analysis reaches remarkable.The straw decomposing inoculant provided is described, compared to existing commercially available decomposing agent, can Mierocrystalline cellulose, lignin component in rapid damage stalk, decomposition effect is good.
Claims (6)
1. a liquid straw decomposing inoculant, it is characterized in that he includes chief active microorganism and is: bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), described two kinds of living microorganisms are mutually collaborative and without antagonistic action, wherein contained effectively bacterium number is 2-5 hundred million/ml, the living bacteria count of wherein said bacillus amyloliquefaciens accounts for 50-60%, and the living bacteria count of yeast saccharomyces cerevisiae accounts for 40-50%.
2. liquid straw decomposing inoculant according to claim 1, it is characterized in that described bacillus amyloliquefaciens and yeast saccharomyces cerevisiae are respectively through (1) test tube strains slant culture, (2) shaking flask activation culture, (3) first class seed pot fermentation and (4) ferment tank, obtain bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid, after above-mentioned bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid is composite by following volume ratio mixing, obtain liquid straw decomposing inoculant, wherein bacillus amyloliquefaciens fermented liquid 50-60%, fermentation by saccharomyces cerevisiae liquid 40-50%.
3. the preparation method of a liquid straw decomposing inoculant as claimed in claim 1 or 2, it is characterized in that described preparation method comprises: the composite mixing of the preparation of bacillus amyloliquefaciens fermented liquid, the preparation of fermentation by saccharomyces cerevisiae liquid and bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid, wherein:
A) preparation of bacillus amyloliquefaciens fermented liquid is:
(1) test tube strains slant culture: bacillus amyloliquefaciens is inoculated in LB slant medium 37 DEG C and cultivates 1-2 days;
(2) shaking flask activation culture: activated spawn is inoculated in LB liquid nutrient medium, 37 DEG C, 180rpm/min cultivates living bacteria count 1-2 hundred million/ml in 18h to nutrient solution;
(3) seeding tank fermentation: liquid culture bacterium liquid is inoculated in seed culture medium by 1-2% inoculum size, 37 DEG C, 200rpm/min cultivates living bacteria count 1-2 hundred million/ml in 24h to nutrient solution;
(4) ferment tank: seed culture fluid is inoculated in fermention medium by 1-2% inoculum size, 37 DEG C, 200rpm/min cultivates living bacteria count 2-4 hundred million/ml in 36h to nutrient solution, obtains bacillus amyloliquefaciens fermented liquid;
B) preparation of fermentation by saccharomyces cerevisiae liquid is:
(1) test tube strains slant culture: yeast saccharomyces cerevisiae is inoculated in PDA slant medium 28 DEG C and cultivates 2-3 days;
(2) shaking flask activation culture: activated spawn is inoculated in PDA liquid nutrient medium, 28 DEG C, 180rpm/min cultivates living bacteria count 1-2 hundred million/ml in 36h to nutrient solution;
(3) seeding tank fermentation: liquid culture bacterium liquid is inoculated in seed culture medium by 5% inoculum size, 28 DEG C, 200rpm/min cultivates living bacteria count 1-2 hundred million/ml in 36h to nutrient solution;
(4) ferment tank: seed culture fluid is inoculated in fermention medium by 5% inoculum size, 28 DEG C, 200rpm/min cultivates living bacteria count 2-3 hundred million/ml in 48h to nutrient solution, obtains fermentation by saccharomyces cerevisiae liquid;
C) after above-mentioned bacillus amyloliquefaciens fermented liquid and fermentation by saccharomyces cerevisiae liquid is composite by following volume ratio mixing, obtain containing the liquid straw decomposing inoculant that effective bacterium number is 2-5 hundred million/ml, wherein bacillus amyloliquefaciens fermented liquid 50-60%, fermentation by saccharomyces cerevisiae liquid 40-50%.
4. the preparation method of liquid straw decomposing inoculant according to claim 3, is characterized in that in the preparation of described bacillus amyloliquefaciens fermented liquid:
LB slant medium described in step (1) is pH value 7.2 solution configured by 10g peptone, 5g yeast extract paste, 10g sodium-chlor, 15g agar, 1000ml water;
Seed culture medium described in LB liquid nutrient medium described in step (2) and step (3) is by pH value 7.2 solution of 10g peptone, 5g yeast extract paste, 10g sodium-chlor, the configuration of 1000ml water;
Fermention medium described in step (4) is by 20g Semen Maydis powder, 30g bean cake powder, 5g glucose, 1g K
2hPO
4, 0.2g manganous sulfate, 0.3g calcium carbonate and 1000ml water configuration form.
5. the preparation method of liquid straw decomposing inoculant according to claim 3, is characterized in that in the preparation of described fermentation by saccharomyces cerevisiae liquid:
The solution that PDA slant medium described in step (1) and the PDA liquid nutrient medium described in step (2) configure by 200g potato, 20g sucrose, 1000ml water;
Seed culture medium described in step (3) is the solution configured by 5g glucose, 5g peptone, 3g yeast extract paste and 1000ml water;
Fermention medium described in step (4) is the solution configured by 20g glucose, 10g yeast extract paste, 5g peptone, 10g calcium carbonate and 1000ml water.
6. the using method of a liquid straw decomposing inoculant as claimed in claim 1, it is characterized in that described using method is: by direct for stalk chopping and returning after crop harvesting, make stalk uniform spreading at surface layer, 60 kilograms are watered by every mu of ground stalk decomposing agent 2 kilograms, and the urea adding 5-6 kilogram is evenly sprayed on stalk after making it fully dissolve, carry out immediately turning over and irrigating.
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| CN108841743A (en) * | 2018-06-10 | 2018-11-20 | 东北农业大学 | Cold ground straw decomposing bacterium bacterial strain and its preparation method and application |
| CN109749971A (en) * | 2019-03-13 | 2019-05-14 | 深圳市励泽农业高科技发展有限公司 | It is a kind of to degenerate the composite bacteria agent decomposed for stalk and its apply method |
| CN110283870A (en) * | 2019-06-21 | 2019-09-27 | 盐城工学院 | A kind of method of double bacterial strains mixed solid fermentation corn stover |
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| CN110283870A (en) * | 2019-06-21 | 2019-09-27 | 盐城工学院 | A kind of method of double bacterial strains mixed solid fermentation corn stover |
| CN110628675A (en) * | 2019-09-29 | 2019-12-31 | 哈尔滨谷润生态科技发展有限公司 | Straw field-returning decomposition agent and preparation method thereof |
| CN112299922A (en) * | 2020-11-18 | 2021-02-02 | 燕淑海 | Wheat and corn crop rotation field soil microbial regulator and preparation method thereof |
| CN113860968A (en) * | 2021-11-17 | 2021-12-31 | 林猷宪 | Method for promoting softening and decomposition of straw fibers by microbial fermentation of organic acid |
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