CN105218213A - There is biological organic fertilizer of prevention and control soilborne fungal pathogens function and preparation method thereof - Google Patents

There is biological organic fertilizer of prevention and control soilborne fungal pathogens function and preparation method thereof Download PDF

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CN105218213A
CN105218213A CN201510508992.3A CN201510508992A CN105218213A CN 105218213 A CN105218213 A CN 105218213A CN 201510508992 A CN201510508992 A CN 201510508992A CN 105218213 A CN105218213 A CN 105218213A
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prevention
organic fertilizer
biological organic
fungal pathogens
mixed bacteria
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徐永平
王佳宁
续彦龙
龚改林
王丽丽
李晓宇
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Dalian University of Technology
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Dalian University of Technology
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Abstract

There is the preparation method of the biological organic fertilizer of prevention and control soilborne fungal pathogens function, comprise the step of compost fermentation, in described composting material, inoculate the mixed bacteria liquid of Bacillus licheniformis, subtilis, hot-bulb shape urea genus bacillus and bacillus thuringiensis four kinds of Antagonistic Fungis; After compost fermentation terminates, in tunning, add poly-γ L-glutamic acid.The preparation-obtained biological organic fertilizer of the present invention has significant control effect to plant soil-borne fungal disease.The production technique of product of the present invention is simple, and raw material sources are extensive, and production cost is low, and applicable scale operation has the biological organic fertilizer of prevention and control soilborne fungal pathogens function.

Description

There is biological organic fertilizer of prevention and control soilborne fungal pathogens function and preparation method thereof
Technical field
The present invention relates to and a kind of there is biological organic fertilizer of prevention and control soilborne fungal pathogens function and preparation method thereof, belong to agricultural biological technical field.
Background technology
Along with China's aquaculture is separated with plant husbandry, do not have enough farmlands to mass-producing of dissolving, intensive culture industry produce a large amount of Stock raising wastes, ecotope is caused to having a strong impact on.And the chemical fertilizer amount of application cumulative year after year in farmland, the contribution bottleneck of crop yield is highlighted day by day.
China agriculture production fertilising feature be that a large amount of long-term application of chemical fertilizer make crops quality reduce partially to fertilize, causes soil compaction, organic matter decline, soil and crop root Tiny ecosystem destructurized, result in that soilborne fungal pathogens spreads unchecked especially.Common soilborne fungal pathogens has the blight and gray mold etc. of the banded sclerotial blight of wheat and paddy rice, the head smut of corn and soybean blight and melon beans.Current disease prevention and control mainly rely on chemical prevention and control, but long-term application chemical agent also can cause the problems such as the residual and environmental pollution of medicament; In agriculture Routine Management, can there is several years in phytopathogen in soil, and during disease outburst, crop rotation means receive limitation greatly, and the modes such as grafting substantially increase prevention and control cost especially.Therefore, will be the prevention and control trend of following soilborne fungal pathogens in conjunction with the seed selection of disease-resistant variety and the preventing control method such as biological prevention and control of the probiotics that possesses antagonism function.
The use of biological organic fertilizer can improve soil, improve crop quality and by strengthening crop resistance, utilizing microbial diversity to suppress the prevention and control such as pathogenic bacteria to control soilborne fungal pathogens, and nutrition can be provided for Antagonistic Fungi, by Antagonistic Fungi self and meta-bolites thereof, the suppression to pathogenic bacteria reaches good prevention effect.Wherein, many genus bacillus produces subtilyne, polymyxin, nystatin, linear gramicidins isoreactivity material have obvious restraining effect to various pathogens.The comprehensive utilization of biological organic fertilizer and Antagonistic Fungi is the sustainable means of Plant diseases prevention and control.
Summary of the invention
An object of the present invention, is first to provide a kind of preparation method with the biological organic fertilizer of prevention and control soilborne fungal pathogens function.Described method comprises the step of compost fermentation, Bacillus licheniformis (ABTNL-1 is inoculated in composting material, Baclicuslincheniformis), subtilis (ABTNL-2, Bacillussubtilis), hot-bulb shape urea genus bacillus (ABTNL-3, and the mixed bacteria liquid of bacillus thuringiensis (ABTNL-4, Bacillusthuringiensis) four kinds of Antagonistic Fungis Ureibacillusthermosphaericus); After compost fermentation terminates, in compost fermentation product, add poly-γ L-glutamic acid.
Another object of the present invention, is to provide the biological organic fertilizer with prevention and control soilborne fungal pathogens function prepared by the method for the invention described above.By its living bacteria count >5 × 10 of the fertilizer prepared by aforesaid method of the present invention 8cFUg -1, organic content >45%, water ratio is 15 ~ 30%.
Again on the one hand, the present invention aims to provide a kind of method of prevention and control soilborne fungal pathogens, and described method uses the biological organic fertilizer with prevention and control soilborne fungal pathogens function of the present invention mentioned above.
The preparation method of biological organic fertilizer of the present invention matches four kinds of Antagonistic Fungis and participates in composting fermentation process, these bacterium can have the effect of competition nutritional autagonism to soil-borne plant pathogen, and can produce during the fermentation multiplely has obvious inhibiting meta-bolites to soil-borne plant pathogen, prepared biological organic fertilizer is to wheat hypochnus, muskmelon gray mold, maize head smut, soybean blight, the multiple soil-borne plant pathogen such as cucumber fusarium axysporum has obvious restraining effect, can be used for prevention and control Plant diseases, reduce the applications of pesticide, promote the sustainable use of biological raw material.Meanwhile, the biological organic fertilizer prepared by profit of the present invention can be increase soil fertility, and is conducive to Zinc fractions crumb structure thus improvement soil property, promote plant seed germination and growth, strengthen plant stress-resistance ability, improve product quality, make product reach green or organic standard.
Embodiment
If no special instructions, involved in the present invention biological material specimens comprises:
ABTNL-1: Bacillus licheniformis (Bacilluslicheniformis), CGMCCNO.10037.
ABTNL-2: subtilis (Bacillussubtilis), CGMCCNO.10038.
ABTNL-3: hot-bulb shape urea genus bacillus (Ureibacillusthermosphaericus), CGMCCNO.10039.
ABTNL-4: bacillus thuringiensis (Bacillusthuringiensis), CGMCCNO.10036.
First the present invention provides a kind of preparation method with the biological organic fertilizer of prevention and control soilborne fungal pathogens function.Described method comprises the step of compost fermentation, inoculates the mixed bacteria liquid of four kinds of Antagonistic Fungis in composting material; After compost fermentation terminates, in compost fermentation product, add poly-γ L-glutamic acid.
Four kinds of described Antagonistic Fungis are respectively: Bacillus licheniformis (ABTNL-1, Baclicuslincheniformis), subtilis (ABTNL-2, Bacillussubtilis), hot-bulb shape urea genus bacillus (ABTNL-3, and bacillus thuringiensis (ABTNL-4, Bacillusthuringiensis) Ureibacillusthermosphaericus).
In embodiment, described mixed bacteria liquid is prepared by the following method: with pre-culture medium activation culture four kinds of Antagonistic Fungis respectively, obtain cell concentration and be 10 8~ 10 9cFUml -1pre-culture; Then in the pre-culture of four kinds of Antagonistic Fungis of equal-volume ratio mixing gained; Described pre-culture medium is prepared with distilled water, sodium chloride-containing 10.0gL -1, Tryptones 10.0gL -1with yeast powder 5.0gL -1, through 121 DEG C of sterilizings, 20 minutes gained.
In the preparation method of above-mentioned biological organic fertilizer, in described composting material, the inoculative proportion of mixed bacteria liquid is 0.01 ~ 0.05Lkg -1(volume of the mixed bacteria liquid inoculated in per kilogram composting material).Wherein, especially preferred 0.03Lkg -1.
In preferred embodiment, also comprise in compost fermentation product according to 0.08 ~ 0.12Lkg in the preparation method of described biological organic fertilizer -1ratio add the step of mixed bacteria liquid.
In the preparation method of above-mentioned biological organic fertilizer, described poly-γ L-glutamic acid is preferably prewired for 200mgL with distilled water -1poly-γ glutamic acid solution, addition is 0.03 ~ 0.05Lkg -1(volume of the poly-γ L-glutamic acid-distilled water solution added in per kilogram tunning).
In another embodiment, the preferred composting material of the present invention is the mixture of animal excrement and plant residue 3 ~ 4.5:1 ~ 1.5 in mass ratio, and water ratio is 55 ~ 65%.Particularly preferably, described animal excrement are selected from pig manure, chicken manure, cow dung or its mixture, and water ratio is 70 ~ 80% (quality ratios); Plant residue is selected from withered grass, branch, stalk or its mixture, and water ratio is 5 ~ 10% (quality ratios).
Again in a concrete embodiment, the condition of described compost fermentation controls to comprise:
In compost container, composting material is placed in layering, and fully mixes laggard aerobe fermentation of acting charitably after mixed bacteria liquid of evenly splashing; Carry out with fermentation, heap temperature rises, and enters the pliotherm period when heap temperature reaches 45 DEG C, and turning in every five days once and keep the skin wet; Heap temperature reaches more than 60 DEG C and maintains 5-7 days; Temperature starts slow decline subsequently, terminates fermentation when windrow temperature near room temperature.
More specifically in embodiment, the preparation method of biological organic fertilizer of the present invention comprises the steps:
(1) prepare composting material: after animal excrement and plant residue being pulverized, the mixing of 3 ~ 4.5:1 ~ 1.5 in mass ratio, controlling mixture water ratio is 55 ~ 65%, obtains composting material;
Wherein the preferred water ratio of animal excrement be 70 ~ 80% pig manure, chicken manure, cow dung or its mixture; The preferred water ratio of plant residue be 5 ~ 10% withered grass, branch, stalk or its mixture; Preferred mistake 1 order mesh screen after two class raw material pulverizing.
(2) mixed bacteria liquid is prepared: with pre-culture medium to Bacillus licheniformis (ABTNL-1, Baclicuslincheniformis), subtilis (ABTNL-2, Bacillussubtilis), hot-bulb shape urea genus bacillus (ABTNL-3, and bacillus thuringiensis (ABTNL-4 Ureibacillusthermosphaericus), Bacillusthuringiensis) carry out activation culture, obtain cell concentration respectively and be 10 8~ 10 9cFUml - 1pre-culture; Then in the pre-culture of four kinds of Antagonistic Fungis of equal-volume ratio mixing gained;
Described pre-culture medium is prepared with distilled water, sodium chloride-containing 10.0gL -1, Tryptones 10.0gL -1with yeast powder 5.0gL -1, through 121 DEG C of sterilizings, 20 minutes gained;
(3) compost fermentation: composting material prepared by step (1) is placed in layering in compost container, and mixed bacteria liquid prepared by step (2) of evenly splashing, the fully laggard aerobe fermentation of acting charitably of mixing; Fermentation starts, and every day surveys a temperature sooner or later, and heap temperature is passed in time and rises, and enters the pliotherm period when heap temperature reaches 45 DEG C, and heap body turns over for every five days once to be piled and keeps the skin wet.Temperature reaches more than 60 DEG C and maintains 5-7 days, and temperature starts slow decline subsequently.Stop turning when windrow temperature near room temperature, obtain the compost fermentation product become thoroughly decomposed;
(4) in the compost fermentation product of step (3) gained according to 0.03 ~ 0.05Lkg -1ratio to add concentration be 200mgL -1poly-γ glutamic acid aqueous solution, fully dry in the shade to water content after mixing and be 15 ~ 30%, the biological organic fertilizer of prevention and control soilborne fungal pathogens function must be had.
The present invention provides the method for prevention and control soilborne fungal pathogens on the other hand, and the method uses the biological organic fertilizer with prevention and control soilborne fungal pathogens function prepared by above-mentioned either method.
In the preferred implementation of the method for above-mentioned prevention and control soilborne fungal pathogens, described biological organic fertilizer amount of application is 8 ~ 12thm -2.
On the other hand, as preferably, the disease of above-mentioned soilborne fungal pathogens especially caused by fungal pathogen bacterium.More preferably for the disease caused by the fungal pathogen bacterium such as wheat hypochnus fungi, Sporisorium reilianum, soybean blight bacterium, cucumber fusarium axysporum.
The present invention further provides following embodiment, bio-organic fertilizer of the present invention, its preparation method and application are described further.
Embodiment 1
Preparation has the biological organic fertilizer I of prevention and control soilborne fungal pathogens function, comprises the steps:
1, the preparation of windrow
After being pulverized by the plant residues such as the animal excrement such as pig manure, chicken manure or cow dung and withered grass, branch or stalk, cross 1 order mesh screen, then by animal excrement: plant residue mass ratio 4:1 mixes, adjustment composting material water ratio is 60%, saves backup.Selected animal excrement and plant residue water content are 75% and 5% (percent mass hundred) respectively.
2, the preparation of mixed bacteria liquid and poly-γ glutamic acid solution:
Cultivate Antagonistic Fungi Bacillus licheniformis ABTNL-1, subtilis ABTNL-2, hot-bulb shape urea genus bacillus ABTNL-3 and bacillus thuringiensis ABTNL-4 respectively with liquid nutrient medium, be cultured to cell concentration in often kind of Antagonistic Fungi liquid and be 10 8~ 10 9cFUml -1, four kinds of culture equal-volume ratio mixing, obtained mixed bacteria liquid; Wherein liquid culture basigamy is sodium-chlor 10.0g, Tryptones 10.0g, yeast powder 5.0g be dissolved in 1L distilled water, through 121 DEG C, uses after sterilizing in 20 minutes.
Another is that solvent prepares concentration for 200mgL with distilled water -1poly-γ glutamic acid solution.
3, the foundation of compost and condition control
In compost container, material is placed in layering, by the mixed bacteria liquid of preparation in step 2 with 0.03Lkg -1ratio to add in composting material and fully mixing, then carry out microorganism aerobic fermentation.Every day surveys a temperature sooner or later, and heap temperature is passed in time and rises, and enters the pliotherm period when heap temperature reaches 45 DEG C, and heap body turns over for every five days once to be piled and keeps the skin wet.Temperature reaches more than 60 DEG C and maintains 5-7 days, and temperature starts slow decline subsequently.Turning is stopped when windrow temperature near room temperature.The compost free from extraneous odour now obtained, do not attract mosquito.When its rate of emergence >80%, worm's ovum content are 0%, compost matter is saved backup.
4, there is the preparation of the biological organic fertilizer of prevention and control soilborne fungal pathogens function
According to 0.04Lkg in step 3 gained compost fermentation thing -1ratio add poly-γ glutamic acid solution prepared by step 2, fully dry in the shade after mixing to water content be 30%, and living bacteria count >5 × 10 8cFUg -1, organic content >45%.By product grinding and sieving, make powder, can need to carry out granulation according to business, the obtained biological organic fertilizer I with prevention and control soilborne fungal pathogens function.
Embodiment 2
Preparation has the biological organic fertilizer II of prevention and control soilborne fungal pathogens function, comprises the steps:
1, the preparation of windrow
With embodiment 1 step 1.
2, the preparation of mixed bacteria liquid and poly-γ glutamic acid solution:
Cultivate Antagonistic Fungi Bacillus licheniformis ABTNL-1, subtilis ABTNL-2, hot-bulb shape urea genus bacillus ABTNL-3 and bacillus thuringiensis ABTNL-4 respectively with liquid nutrient medium, be cultured to cell concentration in often kind of Antagonistic Fungi liquid and be 10 8~ 10 9cFUml -1, four kinds of culture equal-volume ratio mixing, obtained mixed bacteria liquid; Wherein liquid culture basigamy is sodium-chlor 10.0g, Tryptones 10.0g, yeast powder 5.0g be dissolved in 1L distilled water, through 121 DEG C, uses after sterilizing in 20 minutes.
Another is that solvent prepares concentration for 200mgL with distilled water -1poly-γ glutamic acid solution.
3, the foundation of compost and condition control
In compost container, material is placed in layering, by the mixed bacteria liquid of preparation in step 2 with 0.03Lkg -1ratio to add in composting material and fully mixing, then carry out microorganism aerobic fermentation.Every day surveys a temperature sooner or later, and heap temperature is passed in time and rises, and enters the pliotherm period when heap temperature reaches 45 DEG C, and heap body turns over for every five days once to be piled and keeps the skin wet.Temperature reaches more than 60 DEG C and maintains 5-7 days, and temperature starts slow decline subsequently.Turning is stopped when windrow temperature near room temperature.The compost free from extraneous odour now obtained, do not attract mosquito.When its rate of emergence >80%, worm's ovum content are 0%, compost fermentation product is saved backup.
4, there is the preparation of the biological organic fertilizer of prevention and control soilborne fungal pathogens function
According to 0.04Lkg in step 3 gained compost fermentation product -1ratio add poly-γ glutamic acid solution prepared by step 2, and according to 0.1Lkg -1ratio add mixed bacteria liquid prepared by step 2, fully dry in the shade after mixing to water content be 30%, and living bacteria count >5 × 10 8cFUg -1, organic content >45%.By product grinding and sieving, make powder, can need to carry out granulation according to business, the obtained biological organic fertilizer II with prevention and control soilborne fungal pathogens function.
Comparative example 3 (based on embodiment 2, lacking Bacillus licheniformis ABTNL-1)
Preparation has the biological organic fertilizer III of prevention and control soilborne fungal pathogens function, comprises the steps:
1, the preparation of windrow
With embodiment 1 step 1.
2, the preparation of mixed bacteria liquid and poly-γ glutamic acid solution:
Cultivate Antagonistic Fungi subtilis ABTNL-2, hot-bulb shape urea genus bacillus ABTNL-3 and bacillus thuringiensis ABTNL-4 respectively with liquid nutrient medium, be cultured to cell concentration in often kind of Antagonistic Fungi liquid and be 10 8~ 10 9cFUml -1, three kinds of culture equal-volume ratio mixing, obtained mixed bacteria liquid; Wherein liquid culture basigamy is sodium-chlor 10.0g, Tryptones 10.0g, yeast powder 5.0g be dissolved in 1L distilled water, through 121 DEG C, uses after sterilizing in 20 minutes.
Another is that solvent prepares concentration for 200mgL with distilled water -1poly-γ glutamic acid solution.
3, the foundation of compost and condition control
In compost container, material is placed in layering, by the mixed bacteria liquid of preparation in step 2 with 0.03Lkg -1ratio to add in composting material and fully mixing, then carry out microorganism aerobic fermentation.Every day surveys a temperature sooner or later, and heap temperature is passed in time and rises, and enters the pliotherm period when heap temperature reaches 45 DEG C, and heap body turns over for every five days once to be piled and keeps the skin wet.Temperature reaches more than 60 DEG C and maintains 5-7 days, and temperature starts slow decline subsequently.Turning is stopped when windrow temperature near room temperature.The compost free from extraneous odour now obtained, do not attract mosquito.When its rate of emergence >80%, worm's ovum content are 0%, compost fermentation product is saved backup.
4, there is the preparation of the biological organic fertilizer of prevention and control soilborne fungal pathogens function
According to 0.04Lkg in step 3 gained compost fermentation product -1ratio add poly-γ glutamic acid solution prepared by step 2, and according to 0.1Lkg -1ratio add mixed bacteria liquid prepared by step 2, fully dry in the shade after mixing to water content be 30%, and living bacteria count >5 × 10 8cFUg -1, organic content >45%.By product grinding and sieving, make powder, can need to carry out granulation according to business, the obtained biological organic fertilizer III with prevention and control soilborne fungal pathogens function.
Comparative example 4 (based on embodiment 2, lacking subtilis ABTNL-2)
Preparation has the biological organic fertilizer IV of prevention and control soilborne fungal pathogens function, comprises the steps:
1, the preparation of windrow
With embodiment 1 step 1.
2, the preparation of mixed bacteria liquid and poly-γ glutamic acid solution:
Cultivate Antagonistic Fungi Bacillus licheniformis ABTNL-1, hot-bulb shape urea genus bacillus ABTNL-3 and bacillus thuringiensis ABTNL-4 respectively with liquid nutrient medium, be cultured to cell concentration in often kind of Antagonistic Fungi liquid and be 10 8~ 10 9cFUml -1, three kinds of culture equal-volume ratio mixing, obtained mixed bacteria liquid; Wherein liquid culture basigamy is sodium-chlor 10.0g, Tryptones 10.0g, yeast powder 5.0g be dissolved in 1L distilled water, through 121 DEG C, uses after sterilizing in 20 minutes.
Another is that solvent prepares concentration for 200mgL with distilled water -1poly-γ glutamic acid solution.
3, the foundation of compost and condition control
In compost container, material is placed in layering, by the mixed bacteria liquid of preparation in step 2 with 0.03Lkg -1ratio to add in composting material and fully mixing, carry out microorganism aerobic fermentation.Every day surveys a temperature sooner or later, and heap temperature is passed in time and rises, and enters the pliotherm period when heap temperature reaches 45 DEG C, and heap body turns over for every five days once to be piled and keeps the skin wet.Temperature reaches more than 60 DEG C and maintains 5-7 days, and temperature starts slow decline subsequently.Turning is stopped when windrow temperature near room temperature.The compost free from extraneous odour now obtained, do not attract mosquito.When its rate of emergence >80%, worm's ovum content are 0%, compost fermentation product is saved backup.
4, there is the preparation of the biological organic fertilizer of prevention and control soilborne fungal pathogens function
According to 0.04Lkg in step 3 gained compost fermentation product -1ratio add poly-γ glutamic acid solution prepared by step 2, and according to 0.1Lkg -1ratio add mixed bacteria liquid prepared by step 2, fully dry in the shade after mixing to water content be 30%, and living bacteria count >5 × 10 8cFUg -1, organic content >45%.By product grinding and sieving, make powder, can need to carry out granulation according to business, the obtained biological organic fertilizer IV with prevention and control soilborne fungal pathogens function.
Comparative example 5 (based on embodiment 2, lacking hot-bulb shape urea genus bacillus ABTNL-3)
Preparation has the biological organic fertilizer V of prevention and control soilborne fungal pathogens function, comprises the steps:
1, the preparation of windrow
With embodiment 1 step 1.
2, the preparation of mixed bacteria liquid and poly-γ glutamic acid solution:
Cultivate Antagonistic Fungi Bacillus licheniformis ABTNL-1, subtilis ABTNL-2 and bacillus thuringiensis ABTNL-4 respectively with liquid nutrient medium, be cultured to cell concentration in often kind of Antagonistic Fungi liquid and be 10 8~ 10 9cFUml -1, three kinds of culture equal-volume ratio mixing, obtained mixed bacteria liquid; Wherein liquid culture basigamy is sodium-chlor 10.0g, Tryptones 10.0g, yeast powder 5.0g be dissolved in 1L distilled water, through 121 DEG C, uses after sterilizing in 20 minutes.
Another is that solvent prepares concentration for 200mgL with distilled water -1poly-γ glutamic acid solution.
3, the foundation of compost and condition control
In compost container, material is placed in layering, by the mixed bacteria liquid of preparation in step 2 with 0.03Lkg -1ratio to add in composting material and fully mixing, carry out microorganism aerobic fermentation.Every day surveys a temperature sooner or later, and heap temperature is passed in time and rises, and enters the pliotherm period when heap temperature reaches 45 DEG C, and heap body turns over for every five days once to be piled and keeps the skin wet.Temperature reaches more than 60 DEG C and maintains 5-7 days, and temperature starts slow decline subsequently.Turning is stopped when windrow temperature near room temperature.The compost free from extraneous odour now obtained, do not attract mosquito.When its rate of emergence >80%, worm's ovum content are 0%, compost fermentation product is saved backup.
4, there is the preparation of the biological organic fertilizer of prevention and control soilborne fungal pathogens function
According to 0.04Lkg in step 3 gained compost fermentation product -1ratio add poly-γ glutamic acid solution prepared by step 2, and according to 0.1Lkg -1ratio add mixed bacteria liquid prepared by step 2, fully dry in the shade after mixing to water content be 20%, and living bacteria count >5 × 10 8cFUg -1, organic content >45%.By product grinding and sieving, make powder, can need to carry out granulation according to business, the obtained biological organic fertilizer V with prevention and control soilborne fungal pathogens function.
Comparative example 6 (based on embodiment 2, lacking bacillus thuringiensis ABTNL-4)
Preparation has the biological organic fertilizer VI of prevention and control soilborne fungal pathogens function, comprises the steps:
1, the preparation of windrow
With embodiment 1 step 1.
2, the preparation of mixed bacteria liquid and poly-γ glutamic acid solution:
Cultivate Antagonistic Fungi Bacillus licheniformis ABTNL-1, subtilis ABTNL-2 and hot-bulb shape urea genus bacillus ABTNL-3 respectively with liquid nutrient medium, be cultured to cell concentration in often kind of Antagonistic Fungi liquid and be 10 8~ 10 9cFUml -1, three kinds of culture equal-volume ratio mixing, obtained mixed bacteria liquid; Wherein liquid culture basigamy is sodium-chlor 10.0g, Tryptones 10.0g, yeast powder 5.0g be dissolved in 1L distilled water, through 121 DEG C, uses after sterilizing in 20 minutes.
Another is that solvent prepares concentration for 200mgL with distilled water -1poly-γ glutamic acid solution.
3, the foundation of compost and condition control
In compost container, material is placed in layering, by the mixed bacteria liquid of preparation in step 2 with 0.03Lkg -1ratio to add in composting material and fully mixing, carry out microorganism aerobic fermentation.Every day surveys a temperature sooner or later, and heap temperature is passed in time and rises, and enters the pliotherm period when heap temperature reaches 45 DEG C, and heap body turns over for every five days once to be piled and keeps the skin wet.Temperature reaches more than 55 DEG C and maintains 5-7 days, and temperature starts slow decline subsequently.Turning is stopped when windrow temperature near room temperature.The compost free from extraneous odour now obtained, do not attract mosquito.When its rate of emergence >80%, worm's ovum content are 0%, compost fermentation product is saved backup.
4, there is the preparation of the biological organic fertilizer of prevention and control soilborne fungal pathogens function
According to 0.04Lkg in step 3 gained compost fermentation product -1ratio add poly-γ glutamic acid solution prepared by step 2, and according to 0.1Lkg -1ratio add mixed bacteria liquid prepared by step 2, fully dry in the shade after mixing to water content be 20%, and living bacteria count >5 × 10 8cFUg -1, organic content >45%.By product grinding and sieving, make powder, can need to carry out granulation according to business, the obtained biological organic fertilizer VI with prevention and control soilborne fungal pathogens function.
Embodiment 7. effect of fertilizer is tested:
Test point A, B, C, D, E, F and G totally 7 groups carry out the planting experiment of wheat, muskmelon, corn, soybean and cucumber respectively, often organize 100 strain crops, and arrange three repetitions.A group uses biological organic fertilizer I prepared by embodiment 1; B group uses biological organic fertilizer II prepared by embodiment 2; Biological organic fertilizer III, IV, V, VI that C, D, E and F group uses comparative example's example 3,4,5 and 6 to prepare respectively; G group is control group, and soil does not apply fertilizer (using equivalent soil).
The preparation of soil and seed:
Save backup after wherein affecting the various microorganisms of seed growth through fine and closely woven soil 121 DEG C of sterilizing 20min of 8 order stainless steel mesh screenings with removing.
Choose full seed, wheat that shape is complete, corn, muskmelon, soybean and each 300 of cucumber seeds, save backup after repeatedly rinsing well with water after potassium permanganate solution (0.05%) sterilization 30min, and facilitate 24h under 37 DEG C of conditions.
Planting experiment: (biological organic fertilizer is fertile in 10thm to add the soil being evenly mixed into biological organic fertilizer in planting groove -2dosage use) sow afterwards, and bury seed with a certain amount of soil.Cultivate in constant incubator after more than having operated, carry out conventional management of watering every day, when 15 ~ 20 days, crop is in Seedling Stage.Respectively enough specific soil is inoculated to the planting soil of wheat, corn, soybean and cucumber seedling and pass pathogenic bacterium, wherein wheat is banded sclerotial blight fungi, corn is head smut bacterium, soybean is parasitica, cucumber is wilt.Four kinds of diseases are above crop main fungal venereal disease evil.After inoculation germ, basic management basic in 5 ~ 10 days is constant, and the ill strain number of observed and recorded also calculates ratio.
Add up the disease state of each group of plant, with G group for contrast (diseases prevention rate is for being designated as 0) calculates each group of morbidity and diseases prevention rate, result is as shown in table 1.
From the result in table 1, through using biological organic fertilizer A group and B group prepared by embodiment 1 and embodiment 2 compared to the control group do not applied fertilizer, wheat, soybean, corn and cucumber ill strain number is obviously on the low side, more than 85% is reached to the prevention and control rate of wheat hypochnus, soybean blight, maize head smut and cucumber fusarium axysporum, and lack wherein any one Antagonistic Fungi time, it reduces greatly to the control effect of soilborne fungal pathogens.This biological organic fertilizer showing prepared by the present invention passes plant fungal disease to soil and has effective control effect, and this is owing to wherein there is a large amount of distinctive subtilis, Bacillus licheniformis, bacillus amyloliquefaciens and Bacillus flexus and meta-bolites thereof to the Inhibiting and protective function of soil borne plant pathogen.In sum, using of biological organic fertilizer prepared by the present invention has significant control effect to common soil biography plant fungal disease, can be applicable in agricultural production practice.
Table 1 biological organic fertilizer of the present invention passes the control effect test-results of plant disease to soil
The above; be only the present invention's preferably embodiment; but protection scope of the present invention is not limited thereto; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses; be equal to according to technical scheme of the present invention and inventive concept thereof and replace or change, all should be encompassed within protection scope of the present invention.

Claims (9)

1. there is the preparation method of the biological organic fertilizer of prevention and control soilborne fungal pathogens function, comprise the step of compost fermentation, it is characterized in that, in composting material, inoculate the mixed bacteria liquid of Bacillus licheniformis, subtilis, hot-bulb shape urea genus bacillus and bacillus thuringiensis four kinds of Antagonistic Fungis; After compost fermentation terminates, in compost fermentation product, add poly-γ L-glutamic acid.
2. method according to claim 1, is characterized in that, described mixed bacteria liquid is prepared by the following method: with pre-culture medium activation culture four kinds of Antagonistic Fungis respectively, obtain cell concentration and be 10 8~ 10 9cFUml -1pre-culture; Then in the pre-culture of four kinds of Antagonistic Fungis of equal-volume ratio mixing gained;
Described pre-culture medium is prepared with distilled water, sodium chloride-containing 10.0gL -1, Tryptones 10.0gL -1with yeast powder 5.0gL -1, through 121 DEG C of sterilizings, 20 minutes gained.
3. method according to claim 2, is characterized in that, in described composting material, the inoculative proportion of mixed bacteria liquid is 0.01 ~ 0.05Lkg -1.
4. method according to claim 2, is characterized in that, also comprises in compost fermentation product according to 0.08 ~ 0.12Lkg -1ratio add the step of mixed bacteria liquid.
5. method according to claim 1, is characterized in that, described poly-γ L-glutamic acid is prewired is 200mgL -1poly-γ glutamic acid solution, addition is 0.03 ~ 0.05Lkg -1.
6. method according to claim 1, is characterized in that, described composting material is the mixture of animal excrement and plant residue 3 ~ 4.5:1 ~ 1.5 in mass ratio, and water ratio is 55 ~ 65%.
7. method according to claim 1, comprises the steps:
(1) prepare composting material: after animal excrement and plant residue being pulverized, the mixing of 3 ~ 4.5:1 ~ 1.5 in mass ratio, controlling mixture water ratio is 55 ~ 65%, obtains composting material;
(2) prepare mixed bacteria liquid: with pre-culture medium, activation culture is carried out to Bacillus licheniformis, subtilis, hot-bulb shape urea genus bacillus and bacillus thuringiensis, obtain cell concentration respectively and be 10 8~ 10 9cFUml -1pre-culture; Then in the pre-culture of four kinds of Antagonistic Fungis of equal-volume ratio mixing gained;
Described pre-culture medium is prepared with distilled water, sodium chloride-containing 10.0gL -1, Tryptones 10.0gL -1with yeast powder 5.0gL -1, through 121 DEG C of sterilizings, 20 minutes gained;
(3) compost fermentation: composting material prepared by step (1) is placed in layering in compost container, and mixed bacteria liquid prepared by step (2) of evenly splashing, the fully laggard aerobe fermentation of acting charitably of mixing; Carry out with fermentation, heap temperature rises, and enters the pliotherm period when heap temperature reaches 45 DEG C, and turning in every five days once and keep the skin wet; Heap temperature reaches more than 60 DEG C and maintains 5-7 days; Temperature starts slow decline subsequently, terminates fermentation when windrow temperature near room temperature;
(4) in the compost fermentation product of step (3) gained according to 0.03 ~ 0.05Lkg -1ratio to add concentration be 200mgL -1poly-γ glutamic acid aqueous solution, and according to 0.1Lkg -1ratio add mixed bacteria liquid prepared by step (2), fully dry in the shade to water content after mixing and be 15 ~ 30%, the biological organic fertilizer of prevention and control soilborne fungal pathogens function must be had.
8. the biological organic fertilizer with prevention and control soilborne fungal pathogens function prepared by the method described in the arbitrary claim of claim 1 ~ 7.
9. the method for prevention and control soilborne fungal pathogens, it is characterized in that using the biological organic fertilizer with prevention and control soilborne fungal pathogens function described in claim 8.
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CN110934054B (en) * 2019-11-11 2021-08-31 济宁香平生物科技有限公司 Special tobacco seedling culture substrate and preparation method thereof
CN111004741A (en) * 2019-12-10 2020-04-14 枣庄市杰诺生物酶有限公司 Deep-sea-derived campylobacter R29-2, microecological preparation and application thereof

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