CN105948844A - Method for producing bacillus licheniformis bacterial fertilizer by using pig raising bedding of microbial fermentation bed - Google Patents
Method for producing bacillus licheniformis bacterial fertilizer by using pig raising bedding of microbial fermentation bed Download PDFInfo
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Abstract
The invention provides a method for producing a bacillus licheniformis bacterial fertilizer by using pig raising bedding of a microbial fermentation bed. The method comprises the following steps: taking bacillus licheniformis FJAT-4 as a strain, and taking an LB liquid culture medium as a culture medium for a liquid strain to prepare and produce the liquid strain; and mixing the pig raising bedding of the microbial fermentation bed with bran or corncob or wheat, then taking the mixture as a solid foundation culture medium for preparing the bacterial fertilizer, packing into tissue culture vessels, sterilizing and then inoculating the liquid culture, culturing and fermenting for 40-52 hours at the constant temperature of 30-35 DEG C, preparing bacillus licheniformis solid fermenting products, and uniformly mixing the bacillus licheniformis solid fermenting products after fermentation is finished to obtain the bacillus licheniformis bacterial fertilizer. A solid fermenting mode is adopted, the pig raising bedding of the microbial fermentation bed is used as one of ingredients of a solid fermenting culture medium, so that required nutrients for fermentation of the strain can be provided, production cost is reduced, and pollution of wastes of agricultural and sideline products to the environment can be reduced.
Description
[technical field]
The present invention relates to a kind of utilize fermentable bed to raise pig bedding and padding to produce Bacillus licheniformis bacterial manure
Method.
[background technology]
The preventing and treating of plant disease at present is mainly by the high anti-kind of selection-breeding, agricultural measures, chemical agent and life
The methods such as thing pesticide are prevented and treated, because chemical pesticide brings the drawback such as ecological environment and human health increasingly
The problems such as seriously, breeding cycle is long, some agricultural measures complex operations, Biological control obtained in recent years
Paying close attention to widely, increasing scientific research personnel starts to dive the biological preparation with fertilizer efficiency and biological and ecological methods to prevent plant disease, pests, and erosion, with
Reduce the use of fertilizer and pesticide, cost-effective, reduce and pollute, protect environment.
Crop droop is to be infected a kind of destruction caused by point born of the same parents Fusarium spp. (Fusarium oxysporum)
Property soil-borne disease, can cause heavy losses, and the preventing and treating to this disease is the most difficult at present.Biological control
One of prevention and controls as this disease, in recent years correlational study get more and more, particularly bacillus because of
It is easily cultivated, survival ability strong, and prevention effect is good, and existing wide research and application, such as hay bud
Born of the same parents bacillus, Brevibacillus brevis, starch liquefacation bacillus etc. prevent and treat have in droop certain should
With.Bacillus licheniformis has that heat-resisting, enzyme system is abundant, yield of enzyme is high and many good characteristics such as safety because of it,
Point born of the same parents Fusarium spp. is had the strongest inhibitory action, prevents in crop droops such as Fructus Musae, Cotton Gossypii and Citrullus vulgariss
Control aspect and have successfully application.
The production of Bacillus licheniformis is fermented frequently with liquid fermentation or liquid-solid two-way fermentation.Lichens bud at present
Spore bacillus mainly uses liquid fermentation to be main, and solid fermentation is less.At Zymolysis Equipment and process aspect, though
Deep fermentation technique is the most ripe, but for the production of agricultural microorganism preparation, exist cost high,
Metabolite can not make full use of, have the drawbacks such as discharging of waste liquid.And solid fermentation tool humidity is low, energy consumption is low,
The superioritys such as culture medium wide material sources, effective production of particularly suitable developing country biological pesticide.At present
Conventional solid fermentation raw material is the agricultural by-products such as wheat bran, Testa oryzae, Semen Glycines muffin, Semen arachidis hypogaeae muffin, exists
The problems such as viscosity increases, sedimentation is inadequate, space is little, subsequent inoculations is difficult after price height, high temperature sterilize,
Therefore it is required for finding a kind of new agricultural by-products as the main medium of solid fermentation.In recent years
The by-product that the microbial fermentation bed pig raising technology risen produces--fermentable bed to raise pig bedding and padding are (following
It is called for short bedding and padding), it is by rice husk by microorganism, and the thing such as sawdust, pig manure, urine mixes through decomposition in situ
Fermentation forms, rich in nutrient, it is contemplated that produce as bacillus licheniformis solid fermentation raw material.
The shortcoming of prior art: at Zymolysis Equipment and process aspect, though deep fermentation technique is the most more
Maturation, but for the production of agricultural microorganism preparation, have that cost is high, metabolite can not make full use of,
There are the drawbacks such as discharging of waste liquid.The most conventional solid fermentation raw material is wheat bran, Testa oryzae, Semen Glycines muffin, Semen arachidis hypogaeae
The agricultural by-products such as muffin, after there is price height, high temperature sterilize, viscosity increases, sedimentation is inadequate, space
The problems such as little, subsequent inoculations is difficult, therefore find a kind of new agricultural by-products master as solid fermentation
It is required for wanting culture medium.
[summary of the invention]
The technical problem to be solved in the present invention, is to provide one to utilize fermentable bed to raise pig bedding and padding raw
The method producing Bacillus licheniformis bacterial manure, uses solid fermentation mode, utilizes fermentable bed to raise pig pad
Material, as one of solid-state fermentation culture medium composition, can provide required nutrient for strain fermentation, reduce and produce
Cost, can reduce again the pollution to environment of the agricultural byproducts garbage.
The present invention is achieved in that
Utilizing the method that fermentable bed to raise pig bedding and padding produce Bacillus licheniformis bacterial manure, its feature exists
In: described method step is as follows:
Step (1) produces with the preparation of liquid spawn: with Bacillus licheniformis FJAT-4 (Bacillus
Licheniformis) being strain, with LB fluid medium for liquid spawn culture medium, preparation produces to be used
Liquid spawn；
The system of step (2) Bacillus licheniformis FJAT-4 (Bacillus licheniformis) solid bacterial manure
Standby: raw as preparing bacterial manure after mixing using fermentable bed to raise pig bedding and padding and wheat bran or corn cob or wheat grain
The total amount of binder culture medium produced, is sub-packed in tissue culture bottle, after sterilizing, prepared by inoculation step (1)
Liquid spawn, 30-35 DEG C of constant temperature culture fermentation 40-52h, prepare Bacillus licheniformis solid fermentation product,
Mixed thoroughly after fermentation ends, obtained Bacillus licheniformis fertile；
Described Bacillus licheniformis FJAT-4 (Bacillus licheniformis), November 1 in 2010
Day is preserved in China typical culture collection center, and address is Luo Jia Shan, Wuhan City Wuhan University, and preservation is compiled
Number it is CCTCC NO:M 2010285.
Further, in described step (1), production is specific as follows with liquid spawn making step:
The activation of step A. lichem bacillus strain FJAT-4: with inoculating loop by Bacillus licheniformis
Strain FJAT-4 lines in NA culture medium, and cultivates 48h in constant incubator, and cultivation temperature sets
It is set to 30 DEG C；
The preparation of step B. fermentation liquid: the lichem bacillus strain after the activation that step A is obtained
Single colony inoculation of FJAT-4 is in LB fluid medium, and is placed on constant-temperature table shaken cultivation,
Rotating speed 200rpm, temperature is set as 30 DEG C, i.e. obtains production and use liquid spawn after cultivating 24h.
Further, in described step A, the component of NA culture medium is: beef extract 0.1%, albumen
Peptone 0.5%, yeast extract 0.5%, sodium chloride 0.5%, agar 1.8%, prepare with water, pH 7.0-7.2；
In described step B, LB fluid medium component is: beef extract 0.1%, peptone 0.5%, yeast
Cream 0.5%, sodium chloride 0.5%, prepare with water, pH 7.0-7.2；And the percentage ratio in each nutrient media components
All it is weight percentage.
Further, in described step (2), the preparation process of Bacillus licheniformis solid bacterial manure is:
The total amount of binder culture medium preparation with fermentable bed to raise pig bedding and padding as Main Ingredients and Appearance of step C.:
Described fermentable bed to raise pig bedding and padding are prepared by weight for 0:10～7:3 with wheat bran, described microorganism
Fermentation bed to raise pig bedding and padding form total amount of binder cultivation with corn cob or wheat grain by weight preparing for 3:7～7:3
Base；
The interpolation of other compositions in step D. basal medium: in total amount of binder culture medium in mass ratio
Add K2HPO40.05%, MgSO40.025%, add water after stirring, adjusting aqueous volume ratio is
40%～45%, proceed in tissue culture bottle, in tissue culture bottle, charge controls in 5%-25% volume range,
Sterilizing；
The cultivation of step E. Bacillus licheniformis solid bacterial manure and preparation: connect in the culture medium of step D
Plant the liquid spawn of preparation in step (1), inoculate by inoculum concentration 3% (v/g) volume, 30～35 DEG C
Constant temperature bottom fermentation 16-52 hour, i.e. Bacillus licheniformis solid fermentation product, by it after fermentation ends
Mix thoroughly after aeration-drying, obtain Bacillus licheniformis fertile.
Further, in described step C, described fermentable bed to raise pig bedding and padding and wheat bran or Semen Maydis
Core or wheat grain form total amount of binder culture medium by weight for 7:3 preparation；
In described step D, adjusting aqueous volume ratio is 45%, proceeds in tissue culture bottle, fills in tissue culture bottle
Doses controls at 25% volume range；
In described step E, after having inoculated, at 35 DEG C of constant temperature bottom fermentations 40 hours, i.e. lichens spore
Bacillus solid fermentation product, mixes thoroughly after its aeration-drying after fermentation ends, obtains Bacillus licheniformis
Fertile.
Present invention have the advantage that
One aspect of the present invention provides a new utilization ways for fermentable bed padding garbage, solves again
The environmental pollution that garbage of having determined causes, it is achieved resource reutilization；On the other hand it is again Bacillus licheniformis
Solid fermentation screened cheap fermentation raw material, for technique produce cost-effective.This fermentation is produced
The existing good organic fertilizer of thing, contains again abundant Bacillus licheniformis biocontrol microorganisms, and bacterial content can reach
2.63×109Cfu/g, can not only be used for bacterial manure and uses, again can be as Fertilizer application, effectively by bacterium and fertilizer
Organically combine.Therefore the present invention has good economic benefits and environmental effect.
In a word, the present invention uses solid fermentation mode, utilizes fermentable bed to raise pig bedding and padding as solid-state
One of fermentation medium composition, can provide required nutrient for strain fermentation, reduce production cost, again may be used
Reduce the pollution to environment of the agricultural byproducts garbage.
[detailed description of the invention]
Below in conjunction with specific embodiment, the present invention is further illustrated.
Embodiment
One, the acquisition of bacterial strain
Bacillus licheniformis FJAT-4 of the present invention (Bacillus licheniformis) is from Fujian Province
The Rhizosphere Soil of the crops such as the Fructus Lycopersici esculenti of 6 counties (city) such as Yongtai, Fuzhou City, Fuqing, Minhou, Citrullus vulgaris, Semen vignae sinensis
Earth separates and screens the bacterial strain obtaining having fine Biological control effect.
1. the separation of bacterial strain:
(1) take rhizosphere soil sample 10g in 90mL sterilized water, fully draw 1mL after vibration
Soil supension carries out gradient dilution；
(2) the soil dilution liquid that step (1) obtains is coated on NA culture medium flat plate, then will
NA culture medium flat plate cultivates 72h in being placed in 30 DEG C of constant incubators；
(3) single bacterium colony that first picking step (2) cultivates acquisition carries out smear, mirror after basic fuchsin dyeing
Inspection, chooses and can produce the bacterial strain of spore and proceed to slant medium and preserve, as test strains, then will
Test strains is inoculated in BPY culture medium, and this BPY culture medium is placed in rotating speed 200rpm, 30 DEG C
Lower shaken cultivation 72h, collects culture fluid；The wherein component of BPY culture medium: peptone 1.0%, cattle
Meat extractum 0.5%, yeast extract 0.3%, glucose 0.5%, NaCl 0.5%, joins with distilled water
System, pH 7.2；It addition, the percentage ratio in this BPY nutrient media components is weight ratio.
(4) with Citrullus vulgaris Fusarium oxysporum as indicator bacteria, the cultivation that step (3) is collected by cup-plate method is used
Test strains in liquid screens, concrete operations: with PDA plate cultivate Citrullus vulgaris Fusarium oxysporum, 7
Collect spore with sterilized water after d, make spore suspension；The spore suspension prepared is joined temperature fall
To about 50 DEG C of molten state PDA culture medium, pour in culture dish after mixing rapidly；Treat in culture dish
After liquid solidification, the flat board of band pathogen spore is placed pipe dish, takes collected by step (3)
Culture fluid 200 μm, in pipe dish, is observed after cultivating 48h, and compare with sterilized water at 30 DEG C,
Measure antibacterial circle diameter by diameter interior extrapolation method, thus filter out pathogen spore inhibited
Bacterial strain.
2. the qualification of bacterial strain:
The bacterial strain to pathogen spore with high inhibition effect obtained is carried out physics and chemistry and biology
The mensuration of characteristic, the Main Morphology and the biological property that obtain this bacterial strain are as follows: bacteria colony white, shriveling,
Opaque, Gram’s staining is positive, and the thin spore of trophosome is shaft-like；Having spore, spore is the most oval, in
Raw or secondary end life, does not expands.The following is its concrete Physiology and biochemistry identification of indicator:
Note :+be shown with effect or respond；-show without effect or reactionless
Each index and Bacillus licheniformis (Bacillus is found after above-mentioned each index is analyzed comparison
Licheniformis) completely the same, then judge that this bacterial strain is as Bacillus licheniformis (Bacillus
licheniformis)。
Two, fermentable bed to raise pig bedding and padding are utilized to produce the preparation of Bacillus licheniformis solid bacterial manure
A kind of method utilizing fermentable bed to raise pig bedding and padding to produce Bacillus licheniformis bacterial manure, specifically walks
Rapid as follows:
The activation of step A. lichem bacillus strain FJAT-4: with inoculating loop by Bacillus licheniformis
Strain FJAT-4 lines in NA culture medium, and cultivates 48h in constant incubator, and cultivation temperature sets
It is set to 30 DEG C；
The preparation of step B. fermentation liquid: the lichem bacillus strain after the activation that step A is obtained
Single colony inoculation of FJAT-4 is in LB fluid medium, and is placed on constant-temperature table shaken cultivation,
Rotating speed 200rpm, temperature is set as 30 DEG C, i.e. obtains production and use liquid spawn after cultivating 24h.
In described step A, the component of NA culture medium is: beef extract 0.1%, peptone 0.5%, ferment
Female cream 0.5%, sodium chloride 0.5%, agar 1.8%, prepare with water, pH 7.0-7.2；Described step B
Middle LB fluid medium component is: beef extract 0.1%, peptone 0.5%, yeast extract 0.5%, chlorine
Change sodium 0.5%, prepare with water, pH 7.0-7.2；And the percentage ratio in each nutrient media components is weight hundred
Proportion by subtraction.
The total amount of binder culture medium preparation with fermentable bed to raise pig bedding and padding as Main Ingredients and Appearance of step C.:
Described fermentable bed to raise pig bedding and padding form total amount of binder culture medium with wheat bran 7:3 (mass ratio) preparation；
The interpolation of other compositions in step D. basal medium: in total amount of binder culture medium in mass ratio
Add K2HPO40.05%, MgSO40.025%, add water after stirring, adjusting aqueous volume ratio is
45%, proceed in tissue culture bottle, in tissue culture bottle, charge controls in 25% volume range, sterilizing；
The cultivation of step E. Bacillus licheniformis solid bacterial manure and preparation: in the culture medium of step D
The liquid spawn of preparation in inoculation step B, is inoculated by inoculum concentration 3% (v/g) volume, 35 DEG C of constant temperature
Bottom fermentation 40 hours, i.e. Bacillus licheniformis solid fermentation product, by its aeration-drying after fermentation ends
After mix thoroughly, obtain Bacillus licheniformis fertile.
Mentioned microorganism fermentation bed to raise pig bedding and padding, are a kind of organic matters become thoroughly decomposed, and this technology is with fermentation
Bed is carrier, and microorganism fungus kind, rice husk, sawdust, wood flour etc. are mixed, by a certain percentage as pig house
Bedding and padding, are layed in pig house.By microbial action, the organic substance in Pig dung and urine is fully divided
Solve, convert, ferment, finally remove abnormal flavour and reach innoxious, be a kind of no pollution, fermentable
The New Environment Protection Technology of bed.
Three, Bacillus licheniformis FJAT-4 solid fermentation bacterial manure application in tomato planting is cultivated
1. Bacillus licheniformis FJAT-4 solid fermentation bacterial manure application in tomato seedling
1.1 method
Add 5% Bacillus licheniformis FJAT-4 solid fermentation bacterial manure of above-mentioned preparation in seedling medium,
With pure matrix treatments for comparison.Carry out tomato seeds accelerating germination in advance, after seed shows money or valuables one carries unintentionally, be sowed at respectively and contain
In the seedling medium hole tray of Bacillus licheniformis FJAT-4 solid fermentation bacterial manure and the hole tray of pure seedling medium
In, to cultivate 20 days in 30 DEG C, the statistics root length of tomato seedling, plant height, stem be thick and survival rate.
1.2 result and analysis
The application result of tomato seedling is as shown in table 1, and result shows, uses Bacillus licheniformis FJAT-4
Solid fermentation bacterial manure processes its root length of tomato seedling educated and plant height is respectively 6.58cm and 11.37cm,
More slightly higher than 4.92cm and 9.72cm of pure Matrix controls group；Bacillus licheniformis FJAT-4 bacterial manure processes
Kind shoot survival percent up to 98.52%, be better than pure matrix treatments comparison 96.20%.Two kinds process to kind
The impact that eggplant Seedling stem is thick is the most notable.
Table 1 Bacillus licheniformis FJAT-4 solid fermentation bacterial manure
On the impact of plant strain growth character in tomato seedling
In table, data are average ± standard error.With letter representations different after column data through DuncanShi duncan's new multiple range method method
Check notable at P < 0.05 level difference.
The fertilizer efficiency of Fructus Lycopersici esculenti Potted orchard is tested by 2 Bacillus licheniformis FJAT-4 solid fermentation bacterial manure
2.1 method
Bacillus licheniformis FJAT-4 solid fermentation bacterial manure is mixed the most according to a certain volume with substrate soil
After, it is used for planting tomato seedling, if following 6 process groups:
Containing 5% Bacillus licheniformis FJAT-4 solid fermentation bacterial manure in process group 1 substrate soil
Containing 10% Bacillus licheniformis FJAT-4 solid fermentation bacterial manure in process group 2 substrate soil
Containing 15% Bacillus licheniformis FJAT-4 solid fermentation bacterial manure in process group 3 substrate soil
Containing 20% Bacillus licheniformis FJAT-4 solid fermentation bacterial manure in process group 4 substrate soil
The pure substrate soil of process group 5
More than process tomato seedling and often organize 30 strains, routine observation tomato plant strain growth situation, screen potted plant
The Bacillus licheniformis FJAT-4 solid fermentation bacterial manure addition of the suitableeest Growth of Tomato Seedling of process.
Select optimum content two process, compare Bacillus licheniformis FJAT-4 solid fermentation product with
The impact on plant strain growth situation respectively of culture before fermentation.
2.2 results and analysis
Bacillus licheniformis FJAT-4 solid fermentation bacterial manure is mixed the most according to a certain volume with substrate soil
After, it being used for planting tomato seedling, result of the test is shown in Table 2, adds the lichens of different content in seedling medium
Bacillus cereus FJAT-4 solid fermentation bacterial manure, transplants after cultivating 50d, the lichens spore bar of different content
Bacterium FJAT-4 solid fermentation bacterial manure has impact in various degree to Fructus Lycopersici esculenti Potted orchard.In terms of plant height, base
In matter, Bacillus licheniformis FJAT-4 solid fermentation bacterial manure addition is 5% (process group 1) and 10%
(process group 2) growing way is preferable, and plant height is respectively 21.50cm and 22.64cm, other process group plant heights
Between 12.55-20.92cm, substantially less than process group 1 and process group 2；In the thick aspect of stem, process group
The stem of 2 is slightly 2.89mm, and the stem of other process groups is slightly between 2.18-2.70mm, and process group 2 is notable
Higher than other process groups；In terms of the number of blade, process group 1 and process group 2 growing way are preferable, mean leaf
Number is respectively 7.77 and 7.87, and other process group blades are between 5.66-6.35 sheet.From plant height, stem
Thick and three aspects of the number of blade are comprehensively analyzed, and process group 2 and other process groups all exist significant difference
(p < 0.05), illustrates that in substrate, Bacillus licheniformis FJAT-4 solid fermentation bacterial manure addition is 10%
Maximum to tomato growth facilitation, secondly it is 5%.
The growth indexes of Fructus Lycopersici esculenti Potted orchard between table 2 different disposal
In table, data are average ± standard error.With letter representations different after column data through DuncanShi duncan's new multiple range method method
Check notable at P < 0.05 level difference.
Select to add at two of 5% and 10% Bacillus licheniformis FJAT-4 solid fermentation bacterial manure content
Reason group, before being respectively compared its tunning and fermentation, the impact of plant strain growth situation is shown by culture, knot
Fruit as shown in table 3, the plant that tunning processes on plant height apparently higher than medium treatment before fermentation
, stem is slightly the most notable with the impact of number of blade aspect.
Before table 3 different content tunning and fermentation, culture processes the growing state of Fructus Lycopersici esculenti Potted orchard
In table, data are average ± standard error.Check through DuncanShi duncan's new multiple range method method with letter representations different after column data
Notable at P < 0.05 level difference.
3. Bacillus licheniformis FJAT-4 solid fermentation bacterial manure is to tomato wilt Potted orchard sickness rate
Impact
3.1 method
Tomato wilt former bacterium FJAT-282 uses shake flask fermentation, and fermentation liquid is diluted to 106
Cfu/mL, standby.Test and set following 5 process groups:
Process group 1 tomato seedling is planted by pure substrate soil, hinders root pouring clear water, as the moon after 7d
Property comparison；
Process group 2 tomato seedling is with containing 10% Bacillus licheniformis fertilizer ground substance soil plantation, after 7d
Hinder root pouring clear water；
Process group 3 tomato seedling is with containing 10% Bacillus licheniformis fertilizer ground substance soil plantation, after 7d
Hinder root pouring inoculation withered pathogenic bacteria；
Process group 4 tomato seedling, with containing 10% Bacillus licheniformis fertilizer ground substance soil plantation, waters simultaneously
Inoculation withered pathogenic bacteria；
Process group 5 tomato seedling is planted by pure substrate soil, hinders the root withered cause of disease of pouring inoculation after 7d
Bacterium, as positive control；
More than processing and often organize 30 strain tomato seedling, the cultivation management mode that often group processes is all consistent, connects bacterium
Rear observation tomato plant strain growth and incidence, investigate its sickness rate when the 50th day, calculates prevention effect.
Prevention effect (%)=(positive controls sickness rate-process group sickness rate)/positive controls morbidity
Rate × 100%
3.2 results and analysis
Result of the test is as shown in table 4, and result shows, process group 5 20d morbidity is 35%, during 40d
Sickness rate be 100%.Process group 3 relatively process group 4 postpones morbidity, the sickness rate of the two of 50d
It is respectively 30.00% and 66.67%.Other process 1 and morbidity strain do not occur with processing 2.Real as can be seen here
Test and indoor carry out that tomato seedling is potted plant has preventing and treating to droop by adding Bacillus licheniformis solid bacterial manure
Effect, the highest preventive effect is up to 70%.
The sickness rate of the Potted orchard of tomato wilt between table 4 different disposal
4. the field plot of tomato wilt is prevented and treated by Bacillus licheniformis FJAT-4 solid fermentation bacterial manure
Effect
4.1 method
Test and carry out in time booth proving ground, Fujian, Fuzhou City, Fujian Province, toward annual morbidity 20%～30%,
Before transplanting, experimental field soil has tedded and appropriate quick lime immersion treatment, and within 26th, moves in JIUYUE in 2014
Plant.Test and set 3 process:
Process 1 Bacillus licheniformis solid bacterial manure and mix soil by 10%, process of spreading manuer in holes；
Process 2 50% carbendazol wettable powders to process by 800 times of liquid pourings；
Process 3 clear water comparisons.
Often group processes plantation 200 strain Fructus Lycopersici esculenti seedlings, if 3 repetitions.In time transplanting and after transplanting
15, respectively use during 30d 1 time, process 60d " Invest, Then Investigate " sickness rate, within every 30 days, investigate 1 time, Yu Yi
After cultivation, 180d investigation terminates, and calculates prevention effect.Correcting controling effect=(comparison sickness rate-process
Sickness rate)/comparison sickness rate × 100%.
4.2 results and analysis
Incidence investigation to tomato seedling shows as shown in table 5, processes 1 and (uses Bacillus licheniformis
FJAT-4 solid fermentation bacterial manure processes) and process the sickness rate of 2 (using carbendazim to process) respectively
7.17% and 9.50%, it is below processing the 28.83% of 3, correcting controling effect is respectively 75.13% He
67.05%, the prevention effect that Bacillus licheniformis processes is close with what carbendazim processed, and lichens bud is described
Spore bacillus solid fermentation bacterial manure is better than chemical agent carbendazim and processes anti-the prevention effect of tomato wilt
Control effect.
Table 5 Bacillus licheniformis prevents and treats sickness rate and the prevention effect that tomato wilt respectively processes
In summary, one aspect of the present invention provides a new utilization for fermentable bed padding garbage
Approach, solves again the environmental pollution that garbage causes, it is achieved resource reutilization；On the other hand it is ground again
The solid fermentation of clothing bacillus cereus has screened cheap fermentation raw material, produces for technique and saves into
This.The existing good organic fertilizer of this tunning, contains again abundant Bacillus licheniformis biocontrol microorganisms,
Bacterial content can reach 2.63 × 109Cfu/g, can not only be used for bacterial manure and uses, can have as Fertilizer application again
Effect bacterium and fertilizer are organically combined.Therefore the present invention has good economic benefits and environmental effect.
In a word, the present invention uses solid fermentation mode, utilizes fermentable bed to raise pig bedding and padding as solid-state
One of fermentation medium composition, can provide required nutrient for strain fermentation, reduce production cost, again may be used
Reduce the pollution to environment of the agricultural byproducts garbage.
Although the foregoing describing the detailed description of the invention of the present invention, but it is familiar with the technology people of the art
Member should be appreciated that our described specific embodiment is merely exemplary rather than for this
The restriction of bright scope, those of ordinary skill in the art are in the equivalence made according to the spirit of the present invention
Modify and change, all should contain in the scope of the claimed protection of the present invention.
Claims (5)
1. utilize the method that fermentable bed to raise pig bedding and padding produce Bacillus licheniformis bacterial manure, its feature
It is: described method step is as follows:
Step (1) produces with the preparation of liquid spawn: with Bacillus licheniformis FJAT-4 (Bacillus
Licheniformis) being strain, with LB fluid medium for liquid spawn culture medium, preparation produces to be used
Liquid spawn；
The system of step (2) Bacillus licheniformis FJAT-4 (Bacillus licheniformis) solid bacterial manure
Standby: raw as preparing bacterial manure after mixing using fermentable bed to raise pig bedding and padding and wheat bran or corn cob or wheat grain
The total amount of binder culture medium produced, is sub-packed in tissue culture bottle, after sterilizing, prepared by inoculation step (1)
Liquid spawn, 30-35 DEG C of constant temperature culture fermentation 40-52h, prepare Bacillus licheniformis solid fermentation product,
Mixed thoroughly after fermentation ends, obtained Bacillus licheniformis fertile；
Described Bacillus licheniformis FJAT-4 (Bacillus licheniformis), November 1 in 2010
Day is preserved in China typical culture collection center, and address is Luo Jia Shan, Wuhan City Wuhan University, and preservation is compiled
Number it is CCTCC NO:M 2010285.
The most according to claim 1 utilize fermentable bed to raise pig bedding and padding produce lichens spore bar
The method of bacterium bacterial manure, it is characterised in that: described step (1) produces and has with liquid spawn making step
Body is as follows:
The activation of step A. lichem bacillus strain FJAT-4: with inoculating loop by Bacillus licheniformis
Strain FJAT-4 lines in NA culture medium, and cultivates 48h in constant incubator, and cultivation temperature sets
It is set to 30 DEG C；
The preparation of step B. fermentation liquid: the lichem bacillus strain after the activation that step A is obtained
Single colony inoculation of FJAT-4 is in LB fluid medium, and is placed on constant-temperature table shaken cultivation,
Rotating speed 200rpm, temperature is set as 30 DEG C, i.e. obtains production and use liquid spawn after cultivating 24h.
The most according to claim 2 utilize fermentable bed to raise pig bedding and padding produce lichens spore bar
The method of bacterium bacterial manure, it is characterised in that: in described step A, the component of NA culture medium is: beef soaks
Cream 0.1%, peptone 0.5%, yeast extract 0.5%, sodium chloride 0.5%, agar 1.8%, prepare with water,
pH 7.0-7.2；In described step B, LB fluid medium component is: beef extract 0.1%, peptone
0.5%, yeast extract 0.5%, sodium chloride 0.5%, prepare with water, pH 7.0-7.2；And each nutrient media components
In percentage ratio be all weight percentage.
The most according to claim 1 utilize fermentable bed to raise pig bedding and padding produce lichens spore bar
The method of bacterium bacterial manure, it is characterised in that: the system of Bacillus licheniformis solid bacterial manure in described step (2)
Standby step is:
The total amount of binder culture medium preparation with fermentable bed to raise pig bedding and padding as Main Ingredients and Appearance of step C.:
Described fermentable bed to raise pig bedding and padding are prepared by weight for 0:10～7:3 with wheat bran, described microorganism
Fermentation bed to raise pig bedding and padding form total amount of binder cultivation with corn cob or wheat grain by weight preparing for 3:7～7:3
Base；
The interpolation of other compositions in step D. basal medium: in total amount of binder culture medium in mass ratio
Add K2HPO40.05%, MgSO40.025%, add water after stirring, adjusting aqueous volume ratio is
40%～45%, proceed in tissue culture bottle, in tissue culture bottle, charge controls in 5%-25% volume range,
Sterilizing；
The cultivation of step E. Bacillus licheniformis solid bacterial manure and preparation: connect in the culture medium of step D
Plant the liquid spawn of preparation in step (1), inoculate by inoculum concentration 3% (v/g) volume, 30～35 DEG C
Constant temperature bottom fermentation 16-52 hour, i.e. Bacillus licheniformis solid fermentation product, by it after fermentation ends
Mix thoroughly after aeration-drying, obtain Bacillus licheniformis fertile.
The most according to claim 4 utilize fermentable bed to raise pig bedding and padding produce lichens spore bar
The method of bacterium bacterial manure, it is characterised in that:
In described step C, described fermentable bed to raise pig bedding and padding and wheat bran or corn cob or wheat grain are pressed
Weight ratio is that 7:3 preparation forms total amount of binder culture medium；
In described step D, adjusting aqueous volume ratio is 45%, proceeds in tissue culture bottle, fills in tissue culture bottle
Doses controls at 25% volume range；
In described step E, after having inoculated, at 35 DEG C of constant temperature bottom fermentations 40 hours, i.e. lichens spore
Bacillus solid fermentation product, mixes thoroughly after its aeration-drying after fermentation ends, obtains Bacillus licheniformis
Fertile.
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| CN108179126A (en) * | 2018-01-30 | 2018-06-19 | 广西驰胜农业科技有限公司 | A kind of method using waste edible fungus bacteria stick production bacillus licheniformis bacterial manure |
| CN108504600A (en) * | 2018-04-10 | 2018-09-07 | 福建省农业科学院农业生物资源研究所 | Bacterial wilt plant vaccine produces the fermentation process of bacterium FJAT-1458 |
| CN108841767A (en) * | 2018-09-21 | 2018-11-20 | 云南星耀生物制品有限公司 | A kind of complex micro organism fungicide and microbial-bacterial fertilizer |
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| CN101544960A (en) * | 2009-05-06 | 2009-09-30 | 俞嵘 | A leaven dedicated for fermenting bed of microorganism engineering and fermenting bed pad manufacture method |
| CN103910545A (en) * | 2014-03-18 | 2014-07-09 | 福建省农业科学院农业生物资源研究所 | Method for using pig manure waste to produce trichoderma harzianum fertilizer |
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| CN101298604A (en) * | 2008-06-25 | 2008-11-05 | 天津科技大学 | High-temperature acid-resistant alpha-amylase mutant strain and construction method thereof |
| CN101544960A (en) * | 2009-05-06 | 2009-09-30 | 俞嵘 | A leaven dedicated for fermenting bed of microorganism engineering and fermenting bed pad manufacture method |
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| CN108179126A (en) * | 2018-01-30 | 2018-06-19 | 广西驰胜农业科技有限公司 | A kind of method using waste edible fungus bacteria stick production bacillus licheniformis bacterial manure |
| CN108504600A (en) * | 2018-04-10 | 2018-09-07 | 福建省农业科学院农业生物资源研究所 | Bacterial wilt plant vaccine produces the fermentation process of bacterium FJAT-1458 |
| CN108841767A (en) * | 2018-09-21 | 2018-11-20 | 云南星耀生物制品有限公司 | A kind of complex micro organism fungicide and microbial-bacterial fertilizer |
| CN108841767B (en) * | 2018-09-21 | 2019-06-14 | 云南星耀生物制品有限公司 | A kind of complex micro organism fungicide and microbial-bacterial fertilizer |
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