CN104962489B - A kind of liquid straw decomposing inoculant and preparation and application - Google Patents

A kind of liquid straw decomposing inoculant and preparation and application Download PDF

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Publication number
CN104962489B
CN104962489B CN201510178913.7A CN201510178913A CN104962489B CN 104962489 B CN104962489 B CN 104962489B CN 201510178913 A CN201510178913 A CN 201510178913A CN 104962489 B CN104962489 B CN 104962489B
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liquid
bacillus amyloliquefaciens
saccharomyces cerevisiae
fermentation
culture
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CN104962489A (en
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张永江
周波
孟凡国
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Jiaxing Huabin Biotechnology Co ltd
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Jiaxing Huabin Biotechnology Co ltd
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    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02WCLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00Technologies for solid waste management
    • Y02W30/40Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse

Abstract

A kind of liquid straw decomposing inoculant, it, which includes chief active microorganism, is:Bacillus amyloliquefaciens and saccharomyces cerevisiae, described two active microorganisms mutually cooperate with and without antagonisms, effectively bacterium number contained therein is 25 hundred million/ml, wherein the living bacteria count of described bacillus amyloliquefaciens accounts for 50 60%, the living bacteria count of saccharomyces cerevisiae accounts for 40 50%;Described bacillus amyloliquefaciens and saccharomyces cerevisiae pass through respectively:(1)Test tube strains inclined-plane culture,(2)Shaking flask activation culture,(3)First class seed pot ferment and(4)Ferment tank, obtain bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae liquid, after above-mentioned bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae liquid are mixed into compounding by volumes below ratio, obtain liquid straw decomposing inoculant, wherein bacillus amyloliquefaciens zymotic fluid 50 60%, fermentation by saccharomyces cerevisiae liquid 40 50%;It present invention can be suitably applied to the quick composting of the agricultural crop straws such as wheat, rice.

Description

A kind of liquid straw decomposing inoculant and preparation and application
Technical field
The present invention relates to a kind of liquid straw decomposing inoculant and preparation and application, wheat, rice, corn etc. are applicable to The quick composting of agricultural crop straw, belongs to biological technical field.
Background technology
Straw-returning is an important agricultural measures, to improving soil organic matter content, preservation of fertility, realizes agriculture The sustainable development of industry has great importance.In recent years, with mechanization of agriculture and microbial technique (microorganism formulation) Development, straw directly returning to field are commonly available attention, and large-scale popularization application in China.This correct for a certain extent The wasting phenomenon of crop straw burning, reduce the destruction to atmospheric environment and ecological environment, while soil fertility of having fostered and apply fertilizer, effectively increase Soil organic matter content.
At present, straw-returning based on direct returning to farmland, classify, be rotten to stalk residuum by the comprehensive analysis to string Solution lacks basic research with many inner links of soil fertility, plant nutrient, physiological metabolism etc., be allowed to straw degradative, A series of problems be present in nutrient absorption etc..In addition straw decomposing microbial inoculum is essentially pulvis, the use step of product on the market It is cumbersome, poorly water-soluble, it is accustomed to mismatch etc. with existing agro-farming, common people need to pay more labours' progress straw-returnings. Rural laborer is deficient all the more and cost of labor ever-increasing today, and the contradiction increasingly highlights, thus powder product can not make Common people generally receive and used, and this also counteracts that the development of straw decomposing inoculant industry.
In summary, straw decomposing microbial inoculum that is easy there is an urgent need to application method at present and being matched with existing agro-farming custom, Allow common people to afford to use, be the meeting of learning, visible.
The content of the invention
Present invention aims to overcome that the shortcomings of the prior art, there is provided a kind of directly to be gone back primarily directed to existing stalk Powder dose-type product poorly water-soluble during field, in-convenience in use the problems such as, utilize cellulase, egg caused by straw decomposing inoculant White enzyme isoreactivity material accelerates the decomposition of stalk, improves soil, is advantageous to the liquid straw decomposing inoculant and system of straw directly returning to field Standby and application method.
The purpose of the present invention is completed by following technical solution, a kind of liquid straw decomposing inoculant, the liquid stalk Decomposing agent includes chief active microorganism:Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and wine brewing ferment Female (Saccharomyces cerevisiae), described two active microorganisms mutually cooperate with and without antagonisms, wherein institute It is hundred million/ml of 2-5 containing effective bacterium number, wherein the living bacteria count of described bacillus amyloliquefaciens accounts for 50-60%, saccharomyces cerevisiae Living bacteria count accounts for 40-50%.
Described bacillus amyloliquefaciens and saccharomyces cerevisiae pass through respectively:(1) test tube strains inclined-plane culture, (2) shaking flask are lived Change culture, the fermentation of (3) first class seed pot and (4) ferment tank, obtain bacillus amyloliquefaciens zymotic fluid and saccharomyces cerevisiae Zymotic fluid, after above-mentioned bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae liquid are mixed into compounding by volumes below ratio, obtain Liquid straw decomposing inoculant, wherein bacillus amyloliquefaciens zymotic fluid 50-60%, fermentation by saccharomyces cerevisiae liquid 40-50%.
A kind of preparation method of such as above-mentioned liquid straw decomposing inoculant, the preparation method include:The system of bacillus amyloliquefaciens zymotic fluid Standby, fermentation by saccharomyces cerevisiae liquid preparation and the mixing of the compounding of bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae liquid, wherein:
A) preparation of bacillus amyloliquefaciens zymotic fluid is:
(1) test tube strains inclined-plane culture:Bacillus amyloliquefaciens are inoculated in into 37 DEG C of LB slant mediums to cultivate 1-2 days;
(2) shaking flask activation culture:Activated spawn is inoculated into LB fluid nutrient mediums, 37 DEG C, 180rpm/min cultures 18h Hundred million/ml of living bacteria count 1-2 into nutrient solution;
(3) seeding tank ferments:Liquid Culture bacterium solution is inoculated into seed culture medium by 1-2% inoculum concentrations, 37 DEG C, 200rpm/min culture 24h hundred million/ml of living bacteria count 1-2 into nutrient solution;
(4) ferment tank:Seed culture fluid is inoculated into fermentation medium by 1-2% inoculum concentrations, 37 DEG C, 200rpm/min cultivates 36h hundred million/ml of living bacteria count 2-4 into nutrient solution, obtains bacillus amyloliquefaciens zymotic fluid.
B) the preparation of fermentation by saccharomyces cerevisiae liquid is:
(1) test tube strains inclined-plane culture:Saccharomyces cerevisiae is inoculated in into 28 DEG C of PDA slant mediums to cultivate 2-3 days;
(2) shaking flask activation culture:Activated spawn is inoculated into PDA liquid medium, 28 DEG C, 180rpm/min cultures 36h hundred million/ml of living bacteria count 1-2 into nutrient solution;
(3) seeding tank ferments:Liquid Culture bacterium solution is inoculated into seed culture medium by 5% inoculum concentration, 28 DEG C, 200rpm/min culture 36h hundred million/ml of living bacteria count 1-2 into nutrient solution;
(4) ferment tank:Seed culture fluid is inoculated into fermentation medium by 5% inoculum concentration, 28 DEG C, 200rpm/ Min cultivates 48h hundred million/ml of living bacteria count 2-3 into nutrient solution, obtains fermentation by saccharomyces cerevisiae liquid.
C after above-mentioned bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae liquid) are mixed into compounding by volumes below ratio, obtain To containing the liquid straw decomposing inoculant that effective bacterium number is hundred million/ml of 2-5, wherein bacillus amyloliquefaciens zymotic fluid 50-60%, ferment of making wine Female zymotic fluid 40-50%.
In the preparation of bacillus amyloliquefaciens zymotic fluid of the present invention:
LB slant mediums described in step (1) be by 10g peptones, 5g yeast extracts, 10g sodium chloride, 15g agar, The solution of pH value 7.2 that the configuration of 1000ml water forms;
Seed culture medium is by 10g peptones, 5g described in LB fluid nutrient mediums and step (3) described in step (2) The solution of pH value 7.2 that yeast extract, 10g sodium chloride, the configuration of 1000ml water form;
Fermentation medium described in step (4) is by 20g corn flour, 30g bean cake powders, 5g glucose, 1g K2HPO4、 0.2g manganese sulfates, 0.3g calcium carbonate and the configuration of 1000ml water form.
In the preparation of fermentation by saccharomyces cerevisiae liquid of the present invention:
The PDA liquid medium described in PDA slant mediums and step (2) described in step (1) is by 200g horses The solution that bell potato, 20g sucrose, the configuration of 1000ml water form;
Seed culture medium described in step (3) is matched somebody with somebody by 5g glucose, 5g peptones, 3g yeast extracts and 1000ml water The solution put;
Fermentation medium described in step (4) be by 20g glucose, 10g yeast extracts, 5g peptones, 10g calcium carbonate with And the solution that the configuration of 1000ml water forms.
A kind of application method of liquid straw decomposing inoculant as described above, the application method are:By stalk after crop harvesting Direct chopping and returning, makes stalk uniformly be layered on surface layer, is watered 60 kilograms by every mu of 2 kilograms of ground stalk decomposing agent, and add 5-6 kilograms of urea makes it be uniformly sprayed at after fully dissolving on stalk, is turned over and is irrigated immediately.
In the present invention, the stalk is specially rice straw or wheat, maize straw.
The beneficial effects of the invention are as follows:Straw decomposing inoculant provided by the present invention, using the flora composition efficiently cooperateed with, fill Divide and utilize cellulase, protease isoreactivity material caused by strain, accelerate the maturity of stalk, experiment proves the present invention's Decomposing microbial inoculum is high to the degradation rate of stalk, in laboratory condition decomposition effect up to 89.1%, under the conditions of field demonstration Preferable decomposition effect can be reached, 30% can be reached within 10 days, reach 42% within 30 days, sold on the market than direct returning to farmland and use The straw decomposing inoculant degradation rate sold is higher, while liquid dosage form can more meet agriculture after straw-returning by the way of directly spraying Custom is ploughed, raw work is laborsaving, and preparation cost is cheap, has higher application prospect.
Embodiment
Below in conjunction with specific embodiment, the present invention will be described in detail:A kind of liquid straw decomposing of the present invention Agent, it, which includes chief active microorganism, is:Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) and wine brewing ferment Female (Saccharomyces cerevisiae), described two active microorganisms mutually cooperate with and without antagonisms, wherein institute It is hundred million/ml of 2-5 containing effective bacterium number, wherein the living bacteria count of described bacillus amyloliquefaciens accounts for 50-60%, saccharomyces cerevisiae Living bacteria count accounts for 40-50%.
Bacillus amyloliquefaciens and saccharomyces cerevisiae of the present invention pass through respectively:(1) test tube strains inclined-plane culture, (2) Shaking flask activation culture, the fermentation of (3) first class seed pot and (4) ferment tank, obtain bacillus amyloliquefaciens zymotic fluid and wine Brewer yeast zymotic fluid, above-mentioned bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae liquid are mixed into compounding by volumes below ratio Afterwards, liquid straw decomposing inoculant, wherein bacillus amyloliquefaciens zymotic fluid 50-60%, fermentation by saccharomyces cerevisiae liquid 40-50% are obtained.
A kind of preparation method of liquid straw decomposing inoculant as described above, the preparation method include:Bacillus amyloliquefaciens are sent out The preparation of zymotic fluid, the preparation of fermentation by saccharomyces cerevisiae liquid and the compounding of bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae liquid Mixing, wherein:
A) preparation of bacillus amyloliquefaciens zymotic fluid is:
(1) test tube strains inclined-plane culture:Bacillus amyloliquefaciens are inoculated in into 37 DEG C of LB slant mediums to cultivate 1-2 days;
(2) shaking flask activation culture:Activated spawn is inoculated into LB fluid nutrient mediums, 37 DEG C, 180rpm/min cultures 18h Hundred million/ml of living bacteria count 1-2 into nutrient solution;
(3) seeding tank ferments:Liquid Culture bacterium solution is inoculated into seed culture medium by 1-2% inoculum concentrations, 37 DEG C, 200rpm/min culture 24h hundred million/ml of living bacteria count 1-2 into nutrient solution;
(4) ferment tank:Seed culture fluid is inoculated into fermentation medium by 1-2% inoculum concentrations, 37 DEG C, 200rpm/min cultivates 36h hundred million/ml of living bacteria count 2-4 into nutrient solution, obtains bacillus amyloliquefaciens zymotic fluid.
B) the preparation of fermentation by saccharomyces cerevisiae liquid is:
(1) test tube strains inclined-plane culture:Saccharomyces cerevisiae is inoculated in into 28 DEG C of PDA slant mediums to cultivate 2-3 days;
(2) shaking flask activation culture:Activated spawn is inoculated into PDA liquid medium, 28 DEG C, 180rpm/min cultures 36h hundred million/ml of living bacteria count 1-2 into nutrient solution;
(3) seeding tank ferments:Liquid Culture bacterium solution is inoculated into seed culture medium by 5% inoculum concentration, 28 DEG C, 200rpm/min culture 36h hundred million/ml of living bacteria count 1-2 into nutrient solution;
(4) ferment tank:Seed culture fluid is inoculated into fermentation medium by 5% inoculum concentration, 28 DEG C, 200rpm/ Min cultivates 48h hundred million/ml of living bacteria count 2-3 into nutrient solution, obtains fermentation by saccharomyces cerevisiae liquid.
C after above-mentioned bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae liquid) are mixed into compounding by volumes below ratio, obtain To containing the liquid straw decomposing inoculant that effective bacterium number is hundred million/ml of 2-5, wherein bacillus amyloliquefaciens zymotic fluid 50-60%, ferment of making wine Female zymotic fluid 40-50%.
In the preparation of bacillus amyloliquefaciens zymotic fluid of the present invention:
LB slant mediums described in step (1) be by 10g peptones, 5g yeast extracts, 10g sodium chloride, 15g agar, The solution of pH value 7.2 that the configuration of 1000ml water forms;
Seed culture medium is by 10g peptones, 5g described in LB fluid nutrient mediums and step (3) described in step (2) The solution of pH value 7.2 that yeast extract, 10g sodium chloride, the configuration of 1000ml water form;
Fermentation medium described in step (4) is by 20g corn flour, 30g bean cake powders, 5g glucose, 1g K2HPO4、 0.2g manganese sulfates, 0.3g calcium carbonate and the configuration of 1000ml water form.
In the preparation of fermentation by saccharomyces cerevisiae liquid of the present invention:
The PDA liquid medium described in PDA slant mediums and step (2) described in step (1) is by 200g horses The solution that bell potato, 20g sucrose, the configuration of 1000ml water form;
Seed culture medium described in step (3) is matched somebody with somebody by 5g glucose, 5g peptones, 3g yeast extracts and 1000ml water The solution put;
Fermentation medium described in step (4) be by 20g glucose, 10g yeast extracts, 5g peptones, 10g calcium carbonate with And the solution that the configuration of 1000ml water forms.
A kind of application method of liquid straw decomposing inoculant as described above, it is characterised in that described application method is:Crop By the direct chopping and returning of stalk after harvest, stalk is uniformly layered on surface layer, be watered by every mu of 2 kilograms of ground stalk decomposing agent 60 kilograms, and the urea for adding 5-6 kilograms makes it be uniformly sprayed at after fully dissolving on stalk, is turned over and is irrigated immediately.
Specific embodiments described below be for a better understanding of the present invention, but limit the present invention.Following implementations Experimental method in example, is conventional method unless otherwise specified.Test material used in following embodiments, such as without special Illustrate, be to be commercially available from routine biochemistry reagent shop.Quantitative test in following examples, is respectively provided with and repeats reality three times Test, results averaged.
The preparation of embodiment 1, liquid straw decomposing inoculant
First, the separation of microorganism fungus kind obtains
The rotten plot stalk mud sample product of crop field stalk, indoor carry out liquid acclimating are gathered, and the stalk of enrichment is dropped The sample for solving efficiency high carries out the separation of microorganism, obtains the solution starch bud with preferable straw decomposition efficiency and mutual not antagonism Spore bacillus jx-10 and saccharomyces cerevisiae jx-15.
Prepare decomposing agent needs strain bacillus amyloliquefaciens (Bacillus amyloliquefaciens) jx-10 and Saccharomyces cerevisiae (Saccharomyces cerevisiae) jx-15 was preserved in Chinese microorganism strain on 02 11st, 2015 (abbreviation CGMCC, address are preservation administration committee common micro-organisms center:Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), Deposit number is CGMCC No.10559 and CGMCC No.10560.
2nd, the preparation of bacillus amyloliquefaciens zymotic fluid
Bacillus amyloliquefaciens are passed through into test tube strains inclined-plane culture-shaking flask activation culture-seeding tank fermentation-fermentation Tank ferments, and obtains bacillus amyloliquefaciens zymotic fluid.
Actication of culture:Bacillus amyloliquefaciens are inoculated in LB (peptone 10g, yeast extract 5g, sodium chloride 10g, agar 15g, water 1000ml, pH7.2) culture 1-2 days of 37 DEG C of slant medium.
Shaking flask activation culture:Activated spawn is inoculated into LB (peptone 10g, yeast extract 5g, sodium chloride 10g, water 1000ml, pH7.2) in fluid nutrient medium, 37 DEG C, 180rpm/min culture 18h hundred million/ml of living bacteria count 1-2 into nutrient solution.
Seeding tank ferments:Liquid Culture bacterium solution is inoculated into seed culture medium (peptone 10g, yeast by 1-2% inoculum concentrations Cream 5g, sodium chloride 10g, water 1000ml, pH7.2) in, 37 DEG C, 200rpm/min culture 24h living bacteria count 1- into nutrient solution 200000000/ml.
Ferment tank:Seed culture fluid is inoculated into fermentation medium (corn flour 20g, bean cake powder by 1-2% inoculum concentrations 30g, glucose 5g, K2HPO41g, manganese sulfate 0.2g, calcium carbonate 0.3g, water 1000ml) in, 37 DEG C, 200rpm/min cultures 36h Hundred million/ml of living bacteria count 2-4 into nutrient solution, obtain bacillus amyloliquefaciens zymotic fluid.
3rd, the preparation of fermentation by saccharomyces cerevisiae liquid
Saccharomyces cerevisiae is passed through into test tube strains inclined-plane culture-shaking flask activation culture-seeding tank fermentation-ferment tank, Obtain bacillus amyloliquefaciens zymotic fluid.
Actication of culture:Saccharomyces cerevisiae is inoculated in PDA (potato 200g, sucrose 20g, water 1000ml, pH are natural) inclined-plane 28 DEG C of culture medium is cultivated 2-3 days.
Shaking flask activation culture:Activated spawn is inoculated into PDA (potato 200g, sucrose 20g, water 1000ml, pH are natural)) In fluid nutrient medium, 28 DEG C, 180rpm/min culture 36h hundred million/ml of living bacteria count 1-2 into nutrient solution.
Seeding tank ferments:Liquid Culture bacterium solution is inoculated into seed culture medium (glucose 5g, peptone by 5% inoculum concentration 5g, yeast extract 3g, water 1000ml) in, 28 DEG C, 200rpm/min culture 36h hundred million/ml of living bacteria count 1-2 into nutrient solution.
Ferment tank:Seed culture fluid is inoculated into fermentation medium (glucose 20g, yeast extract by 5% inoculum concentration 10g, peptone 5g, calcium carbonate 10g, water 1000ml) in, 28 DEG C, 200rpm/min culture 48h living bacteria counts into nutrient solution Hundred million/ml of 2-3, obtain fermentation by saccharomyces cerevisiae liquid.
4th, the compounding of liquid decomposing agent
Bacillus amyloliquefaciens zymotic fluid obtained above and fermentation by saccharomyces cerevisiae liquid are mixed in proportion, obtain mixed liquor Body decomposing agent;Bacillus amyloliquefaciens zymotic fluid accounts for 50-60% (volume ratio), fermentation by saccharomyces cerevisiae liquid wherein in mixed bacteria liquid Account for 40-50% (volume ratio).
Embodiment 2 prepares decomposing agent decomposition effect in straw-returning
Test place:Jiaxing City academy of agricultural sciences
Liquid straw decomposing inoculant provided by the invention and certain commercially available brand decomposing agent are crushed in wheat stalk Field decomposition contrast test is carried out after returning to the field, experimental design is as follows.
Control numbering Returning total stalks into fields Decomposing microbial inoculum Synchronous fertilising
T1 It is It is no It is
T2 It is Certain brand It is
T3 It is The present invention It is
Straw harvesting shreds after harvesting wheat, direct chopping and returning, stalk is uniformly layered on surface layer, by every mu of ground stalk With decomposing agent, 2 kilograms are watered 60 kilograms, and the urea for adding 5-6 kilograms makes it be uniformly sprayed at after fully dissolving on stalk, with Turned over and irrigated.
It is accustomed to rice cultivation of pouring water according to normal agro-farming after being handled according to aforesaid operations method.Respectively at the 10 of processing My god, the weight-loss ratio of rear measurement stalk after 20 days, 30 days, 60 days, 100 days.
As a result it is as follows:
Stalk buries a bag weight-loss ratio (percentage)
Processing 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, weight-loss ratio 41.72% after being handled 10 days using decomposing agent of the present invention, weight-loss ratio reaches within 20 days 49.17%, 30 days weight-loss ratios reach 58.11%, and degradation effect shows this hair apparently higher than using other decomposing microbial inoculums on the market The straw decomposing inoculant of bright offer can fast and effectively decomposition stalk, and liquid dosage form is convenient, saves labor.
Decomposition effect of the decomposing agent of embodiment 3 in field demonstration
Demonstration place:Anji County emperor Hu Zhen Xiao Yun villages
Liquid straw decomposing inoculant provided by the invention is applied after wheat stalk chopping and implements straw-returning, control design It is as follows:
Control numbering Returning total stalks into fields Decomposing microbial inoculum Synchronous fertilising
Ck It is no It is no It is no
T1 It is It is no It is
T2 It is Certain brand It is
T3 It is The present invention It is
Field is presented below by burying bag operation acquisition decomposition effect:
Stalk buries a bag weight-loss ratio (percentage)
Field demonstration result is shown:Using stalk weight-loss ratio is reachable after 10 days after the decomposing agent soil conditioning of the present invention Reach 46.71% after 39.19%, 20 days, 52.73% is reached after 30 days, and compare using other microbial inoculums on the market after 30 days For 43.68%, difference analysis reaches notable.The straw decomposing inoculant provided is provided, can be fast compared to existing commercially available decomposing agent Cellulose, lignin component in speed destruction stalk, decomposition effect are good.

Claims (5)

1. a kind of liquid straw decomposing inoculant, it is characterised in that chief active microorganism is:Deposit number is CGMCC No.10559 Bacillus amyloliquefaciens(Bacillus amyloliquefaciens)With the wine brewing that deposit number is CGMCC No.10560 Yeast(Saccharomyces cerevisiae), two kinds of active microorganisms mutually cooperate with and without antagonisms, contained therein Effective bacterium number is hundred million/ml of 2-5, wherein the living bacteria count of described bacillus amyloliquefaciens accounts for 50-60%, saccharomyces cerevisiae has Effect viable count accounts for 40-50%;
Described bacillus amyloliquefaciens and saccharomyces cerevisiae pass through respectively:(1)Test tube strains inclined-plane culture,(2)Shaking flask activation training Support,(3)First class seed pot ferment and(4)Ferment tank, obtain bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae Liquid, after above-mentioned bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae liquid are mixed into compounding by volumes below ratio, obtain liquid Straw decomposing inoculant, wherein bacillus amyloliquefaciens zymotic fluid 50-60%, fermentation by saccharomyces cerevisiae liquid 40-50%.
A kind of 2. preparation method of liquid straw decomposing inoculant as claimed in claim 1, it is characterised in that described preparation method bag Include:The preparation of bacillus amyloliquefaciens zymotic fluid, the preparation of fermentation by saccharomyces cerevisiae liquid and bacillus amyloliquefaciens zymotic fluid and The compounding mixing of fermentation by saccharomyces cerevisiae liquid, wherein:
A)The preparation method of bacillus amyloliquefaciens zymotic fluid is:
(1)Test tube strains inclined-plane culture:Bacillus amyloliquefaciens described in claim 1 are inoculated in 37 DEG C of LB slant mediums Culture 1-2 days;
(2)Shaking flask activation culture:Activated spawn is inoculated into LB fluid nutrient mediums, 37 DEG C, 180rpm/min culture 18h to training Hundred million/ml of living bacteria count 1-2 in nutrient solution;
(3)Seeding tank ferments:Liquid Culture bacterium solution is inoculated into seed culture medium by 1-2% inoculum concentrations, 37 DEG C, 200rpm/ Min culture 24h hundred million/ml of living bacteria count 1-2 into nutrient solution;
(4)Ferment tank:Seed culture fluid is inoculated into fermentation medium by 1-2% inoculum concentrations, 37 DEG C, 200rpm/min 36h hundred million/ml of living bacteria count 2-4 into nutrient solution are cultivated, obtain bacillus amyloliquefaciens zymotic fluid;
B)The preparation method of fermentation by saccharomyces cerevisiae liquid is:
(1)Test tube strains inclined-plane culture:Saccharomyces cerevisiae described in claim 1 is inoculated in the 28 DEG C of cultures of PDA slant mediums 2-3 days;
(2)Shaking flask activation culture:Activated spawn is inoculated into PDA liquid medium, 28 DEG C, 180rpm/min cultures 36h is extremely Hundred million/ml of living bacteria count 1-2 in nutrient solution;
(3)Seeding tank ferments:Liquid Culture bacterium solution is inoculated into seed culture medium by 5% inoculum concentration, 28 DEG C, 200rpm/min Cultivate 36h hundred million/ml of living bacteria count 1-2 into nutrient solution;
(4)Ferment tank:Seed culture fluid is inoculated into fermentation medium by 5% inoculum concentration, 28 DEG C, 200rpm/min trainings 48h hundred million/ml of living bacteria count 2-3 into nutrient solution are supported, obtain fermentation by saccharomyces cerevisiae liquid;
C)After above-mentioned bacillus amyloliquefaciens zymotic fluid and fermentation by saccharomyces cerevisiae liquid are mixed into compounding by volumes below ratio, contained The liquid straw decomposing inoculant that effective bacterium number is hundred million/ml of 2-5, wherein bacillus amyloliquefaciens zymotic fluid 50-60%, saccharomyces cerevisiae hair Zymotic fluid 40-50%.
3. the preparation method of liquid straw decomposing inoculant according to claim 2, it is characterised in that the bacillus amyloliquefaciens
In the preparation method of zymotic fluid:
Step(1)Described in LB slant mediums be by 10g peptones, 5g yeast extracts, 10g sodium chloride, 15g agar, The solution of pH value 7.2 that the configuration of 1000ml water forms;
Step(2)Described in LB fluid nutrient mediums and step(3)Described in seed culture medium by 10g peptones, 5g yeast The solution of pH values 7.2 that cream, 10g sodium chloride, the configuration of 1000ml water form;
Step(4)Described in fermentation medium be by 20g corn flour, 30g bean cake powders, 5g glucose, 1gK2 HPO4,0.2g Manganese sulfate, 0.3g calcium carbonate and the configuration of 1000ml water form.
4. the preparation method of liquid straw decomposing inoculant according to claim 2, it is characterised in that the fermentation by saccharomyces cerevisiae liquid Preparation method in:
Step(1)Described in PDA slant mediums and step(2)Described in PDA liquid medium be by 200g Ma Ling The solution that potato, 20g sucrose, the configuration of 1000ml water form;
Step(3)Described in seed culture medium be by 5g glucose, 5g peptones, 3g yeast extracts and 1000ml water configure The solution formed;
Step(4)Described in fermentation medium be by 20g glucose, 10g yeast extracts, 5g peptones, 10g calcium carbonate and The solution that the configuration of 1000ml water forms.
5. a kind of application method of liquid straw decomposing inoculant as claimed in claim 1, it is characterised in that described application method is: By the direct chopping and returning of stalk after crop harvesting, stalk is set uniformly to be layered on surface layer, by 2 kilograms of decomposing agent of every mu of ground stalk 60 kilograms are watered, and the urea for adding 5-6 kilograms makes it be uniformly sprayed at after fully dissolving on stalk, is turned over and is filled immediately Irrigate.
CN201510178913.7A 2015-04-15 2015-04-15 A kind of liquid straw decomposing inoculant and preparation and application Expired - Fee Related CN104962489B (en)

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