CN105461370A - Microbial growth promoter specially used for ecological planting of wild rice stems and preparation method thereof - Google Patents

Microbial growth promoter specially used for ecological planting of wild rice stems and preparation method thereof Download PDF

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Publication number
CN105461370A
CN105461370A CN201510999966.5A CN201510999966A CN105461370A CN 105461370 A CN105461370 A CN 105461370A CN 201510999966 A CN201510999966 A CN 201510999966A CN 105461370 A CN105461370 A CN 105461370A
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preparation
wild rice
rice stem
bacterial classification
screening
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江瀚
陆瑛
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SHANGHAI CHUANGBO ECOLOGICAL ENGINEERING Co Ltd
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SHANGHAI CHUANGBO ECOLOGICAL ENGINEERING Co Ltd
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    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05FORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F5/00Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
    • C05F5/002Solid waste from mechanical processing of material, e.g. seed coats, olive pits, almond shells, fruit residue, rice hulls
    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05FORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00Other organic fertilisers
    • C05F11/08Organic fertilisers containing added bacterial cultures, mycelia or the like

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  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Mechanical Engineering (AREA)
  • Botany (AREA)
  • Environmental & Geological Engineering (AREA)
  • Biochemistry (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Fertilizers (AREA)

Abstract

The invention provides a preparation method of a microbial growth promoter specially used for ecological planting of wild rice stems. The preparation method comprises the following steps: 1) preliminarily screening pseudomonas, azotobacter chroococcum, bacillus subtilis, methylomonas and streptomycete respectively from wild rice stem planting soil and respectively obtaining strains with high growth rates through further screening; 2) carrying out mixed culture on the strains obtained in the step 1) in a liquid culture medium to obtain mixed bacterium liquid; 3) carrying out solid enlarge culture on the mixed bacterium liquid in a solid enlarge culture medium to obtain an initial fermented product; and 4) drying the initial fermented product, adding matrix carriers and carrying out sieving and subpackaging, thus obtaining the microbial growth promoter. The inventors of the application improve existing wild rice stem planting methods and design the planting methods and fertilizer use according to the law of requirements of wild rice stems for fertilizers in the growth process, thus obtaining higher yield and high-quality fruit and increasing the economic benefits of planting.

Description

A kind ofly be specifically designed to microbial growth promoters of wild rice stem ecologic planting and preparation method thereof
Technical field
The present invention relates to microorganism field, be specifically related to a kind of microbial growth promoters, particularly relate to and be a kind ofly specifically designed to microbial growth promoters of wild rice stem ecologic planting and preparation method thereof.
Background technology
Wild rice stem is a kind of vegetative aquatic perennial root gramineous crop, is commonly called as: hay bamboo shoot, hay melon, its fleshy stem sugary 4%, protein 1.5%, but also is rich in multivitamin, therefore, be well received by consumers.Wild rice stem originates in China and South East Asia, is a kind of comparatively common aquatic vegetable, the loose part of the fusiform that the tender cane for grass wild rice is stimulated by wild rice ustilago and formed.At present, wild rice stem only had China and Vietnam as vegetable growing, wherein again with Chinese cultivated the earliest.Wild rice stem is distributed in various places, China north and south, and all there is plantation on the ground such as Hebei, Jiangsu, Zhejiang, Anhui, Jiangxi, Fujian, Taiwan, Hong Kong, Henan, Hunan, Hubei, Hainan, Guangdong, Guangxi, Sichuan, Yunnan, Heilungkiang.Wherein, Yuyao wild rice stem is the most well-known with He Mudu town, produces wild rice stem 50,000 tons now, and nuisanceless wild rice stem base, ten thousand mu, He Mudu town is successively listed in Ningbo City's secondary Vegetable Base, Shanghai vegetables group wild rice stem support base, Zhejiang Province's high performance and fine agriculture Demonstration Base.
The breeding time of wild rice stem longer, strong adaptability, do not select soil, therefore, general paddy field, bottom land, the shallow water pool, ditch river bank have the place of water all can plant.Although wild rice stem is less demanding to soil property, unsuitable continuous cropping, and preferably with farming layer depth, the clay being rich in organic matter and clayey soil plantation.Further, because wild rice stem plant is tall and big, vegetative period long, fertilizer requirement is large, therefore, also need many bulk applications, gradation is topdressed, and requires sufficient nitrogenous fertilizer and suitable phosphorus potash fertilizer.In addition, in the growth and development process of wild rice stem, the growth and development environment of warm and moist is also prerequisite.
Present inventor finds, because the well developed root system of wild rice stem, water requirement are many, therefore appropriate source sufficient, pour water conveniently, deep soft, the fertile soil of soil layer, be rich in the strong glutinous loam of organic matter, preserve moisture and fertility ability or loam.But in the implantation methods that wild rice stem is traditional, because people use chemical fertilizer in a large number, thus cause soil block, the problem such as to harden is especially serious, and then causes the generation of the problems such as soil soil property worsens, Water Bamboo Yield is low, deficiency in economic performance.In fact, the quality of soil aeration directly has influence on the growth of root and the absorption of nutrient.Compare with air, the air feature in soil is that oxygen level is lower, and carbon monoxide content is high, it is reported, the oxygen level of upper soll layer is than 1-2% few in air, and then than 6-30 high in air doubly, this is caused by vegetable root system breathing and the micro-raw biological activity of soil to carbonic acid gas.
Be further advanced by research, present inventor finds, the normal activities of vegetable root system, requires that in soil air, oxygen level is greater than 10%, when oxygen level lower than 10% time, increment just sharply declines, and nutrition absorption is also affected; And, vegetables also have relation with soil oxygen level to the absorption of nitrogen, phosphorus, potassium, when oxygen level in soil air is at 10-20%, the absorption of nitrogen, phosphorus, potassium does not almost have difference, and when soil oxygen level drops to 5%, the absorption of nitrogen, phosphorus, potassium also sharply declines, and its absorbed dose even drops to 2%.
Summary of the invention
Research based on above-mentioned prior art and present inventor finds, the invention provides a kind ofly to be specifically designed to microbial growth promoters of wild rice stem ecologic planting and preparation method thereof.Wherein, with wild rice stem waste for main raw material, total crop return on the spot, be the efficient biologic-organic fertilizer that a kind of nutrient content is high, its special medium carrier can ensure that Soil ventilation is loosened, and is convenient to probiotics growth, and can soil-borne disease be suppressed, other exchanges and forms stable probiotics ecological soil system etc. to improve soil, thus solves all sidedly or a prevention soil continuous cropping difficult problem, is a kind of novel microorganism agrotechnique product.
Technical scheme of the present invention comprises a kind of preparation method being specifically designed to the microbial growth promoters of wild rice stem ecologic planting, it is characterized in that, comprising:
1) from wild rice stem planting soil, preliminary screening goes out pseudomonas, former brown vinelandii, subtilis, Methyldmonas and streptomycete respectively, and is obtained the bacterial classification of fast growth respectively by further screening;
2) by step 1) in carry out mixed culture in the bacterial classification liquid medium within that obtains, obtain mixed bacteria liquid;
3) by step 2) mixed bacteria liquid that obtains carries out solid enlarged culturing in solid enlarged culturing base, obtains the head product that ferments;
4) by step 3) the fermentation head product that obtains carries out drying, adds medium carrier, sieves, packing, obtains described microbial growth promoters.
In one embodiment of the invention, the concrete steps of described preliminary screening are: soak wild rice stem planting soil with sterilized water and obtain leach liquor; Pseudomonas is filtered out with photosynthetic bacteria culture medium respectively from leach liquor, former brown vinelandii are gone out with fixed nitrogen Screening of Media, subtilis, Methyldmonas is filtered out with beef extract-peptone nutrient agar, filter out streptomycete with potato sucrose nutrient agar, thus obtain described pseudomonas, former brown vinelandii, subtilis, Methyldmonas and streptomycete respectively.
More preferably, select well-grown planting soil, and adopt ordinary method to carry out microorganism sterilizing, the sterilized water adding its 10 times of weight parts in soil after sterilizing is soaked 24-26 hour, pseudomonas is filtered out with photosynthetic bacteria culture medium respectively in leach liquor, former brown vinelandii are gone out with fixed nitrogen Screening of Media, subtilis is filtered out with beef extract-peptone nutrient agar, Methyldmonas, streptomycete is filtered out with potato sucrose nutrient agar, thus obtain described pseudomonas respectively, former brown vinelandii, subtilis, Methyldmonas and streptomycete.
In a preferred embodiment of the invention, described further screening is specially: described bacterial classification preliminary screening obtained is diluted to 10 respectively -5-10 -7, be then inverted in nutrient agar plate respectively and cultivate, select the bacterium colony of fast growth, then carry out shake flat experiment respectively, filter out speed of growth bacterial classification faster; Further, then each bacterial classification is transferred respectively and to cultivate into the eggplant bottle that nutrient agar is housed, when band is yellow during eggplant bottle surface lawn is covered with and is white, take out stand-by.
More preferably, described bacterial classification preliminary screening obtained is the normal saline dilution to 10 of 0.9% respectively by mass concentration -5-10 -7, the bacteria suspension obtained is inverted cultivation 48 hours respectively in nutrient agar plate under 35 DEG C of conditions, selects the bacterium colony of fast growth, then carries out shake flat experiment, observes growth speed, filters out the bacterial classification of quantity higher than 5,000,000,000/ml; Pseudomonas, former brown vinelandii, subtilis, Methyldmonas, streptomycete are transferred respectively and be equipped with in the eggplant bottle of nutrient agar, 45-50 hour is cultivated under 35-40 DEG C of condition, when during eggplant bottle surface lawn is covered with and is white, band is yellow, can take out, then the refrigerator putting into 2-6 DEG C is preserved, stand-by.
Wherein, screening the described pseudomonas obtained is Pseudomonas fluorescens.
Further, in one embodiment, step 2) in described mixed culture be specially: by step 1) in carry out mixed culture, when total viable count reaches 5,000,000,000/ml in the bacterial classification liquid medium within that obtains, pH value stops fermentation when reaching 7.5-8.0, obtains the head product that ferments.
More preferably, each bacterial classification is taken by following weight proportion: pseudomonas 10-13 part, former brown vinelandii 20-23 part, subtilis 30-33 part, Methyldmonas 10-13 part, streptomycete 18-30 part; Further, described mixed culture be by the mixing lawn of the above bacterial classification taken according to the part by weight of 1:25-30 be inoculated in through 120-130 DEG C, in the liquid nutrient medium of 30-40min high-temperature sterilization, stir culture in fermentor tank, its rotating speed is 220-240r/min, fermentation air flow maintains 1:1, and temperature is 30-32 DEG C; Ferment every 2 hours monitoring fermented liquid pH value and total viable count after 24 hours, when total viable count reaches 5,000,000,000/ml, when pH value reaches 7.5-8.0, stop fermentation.
In another embodiment, step 3) in described solid enlarged culturing for by step 2) in described mixed bacteria liquid access solid enlarged culturing base in carry out sealing and fermenting cultivation, treat that moisture is 30-40%, when pH value is 7-8.5, stop fermentation.
More preferably, by step 2) in the mixed bacteria liquid of mixed culture that obtains in the ratio access of 1:25-30 through 120-130 DEG C, in the solid enlarged culturing base of 30-45min high-temperature sterilization, stir 30-45min to even, pour sealing and fermenting in sterilization Double-layer Plastic envelope into, temperature 25-35 DEG C, time 8-15 days; Ferment every sampling in 6 hours after 7 days, measure moisture and pH value, treat that moisture is 30-40%, when pH value is 7-8.5, stopping is fermented.
Wherein in an embodiment, described drying is dry in incubator, and temperature controls at 35-45 DEG C, time 12-18 hour, and the moisture controlled of the fermentation head product that drying completes is below 20%.
Further, add medium carrier, mix according to the part by weight of dried fermentation head product 45-50 part, medium carrier 50-55 part, wherein, described medium carrier is sterilizing wood chip and sterilizing wild rice stem waste.
Further, carry out sieving and load with in the bucket of inner bag or plastic packaging bag, tightening sack, fasten nameplate, proceed in stockyard and stack in order.
Preferably, described preparation method also comprises sampling and detects, and main detect parameters is: moisture≤11.8%, viable count 25-30 hundred million/g, and physical behavior is brown powder solid.
Further, preferably, described liquid nutrient medium is the combination that sterilized water adds at least three kinds of mixtures in corn steep liquor, barley starch, monoammonium sulfate, molasses, potassiumphosphate, potassium sulfate, trace element solution again, preferably, its component and weight proportion thereof are corn steep liquor 2-3 part, barley starch 1-2 part, monoammonium sulfate 0.1-0.3 part, molasses 10-14 part, potassiumphosphate 1.2-2 part, potassium sulfate 0-0.8 part, trace element solution 0.3-0.5 part, sterilized water 75-85 part.
Preferably, described solid enlarged culturing base is the combination that sterilized water adds at least four kinds of mixtures in chaff, bean cake powder, yeast powder, Semen Maydis powder, potassium primary phosphate, manganous sulfate, garlic juice clearly again, preferably, its component and weight proportion thereof are: clear chaff 40-40 part, bean cake powder 10-12 part, yeast powder 1-1.2 part, Semen Maydis powder 8-10 part, potassium primary phosphate 0.8-1 part, manganous sulfate 0-0.4 part, garlic juice 1-1.2 part, sterilized water 32.3-39.2 part.
Further, described matrix is made up of sterilizing wood chip, sterilizing wild rice stem waste, and its weight proportion is: wood chip 55-60, wild rice stem waste 40-45.
Technical scheme of the present invention also comprises the microbial growth promoters that a kind of preparation method described above prepares.
Application contriver improves existing wild rice stem implantation methods, according in wild rice stem process of growth to the demand rule of fertilizer, design implantation methods and the use of fertilizer, thus obtain higher output and the fruit of high-quality, improve the economic return of plantation.Particularly, adopt with wild rice stem waste for main raw material, be equipped with the composition such as feces of livestock and poultry, clear chaff, fermentation maturity forms.Wherein, the bacterial classification of fermentation uses from soil and can improve the beneficial microbe colony of soil, finds that it for a long time constantly for crop provides growth nutrient, can improve the output of crop through research.Simultaneously, because medium carrier is by the preparation of exclusive solid, liquid synchronous fermentation process, thus ensure that the ventilative loose of soil, be convenient to probiotics growth, also inhibits soil-borne disease, other exchanges and defines stable probiotics ecological soil system to improve soil.
Embodiment
The invention provides a kind of preparation method being specifically designed to the microbial growth promoters of wild rice stem ecologic planting, it is characterized in that, comprising:
1) from wild rice stem planting soil, preliminary screening goes out pseudomonas, former brown vinelandii, subtilis, Methyldmonas and streptomycete respectively, and is obtained the bacterial classification of fast growth respectively by further screening;
2) by step 1) in carry out mixed culture in the bacterial classification liquid medium within that obtains, obtain mixed bacteria liquid;
3) by step 2) mixed bacteria liquid that obtains carries out solid enlarged culturing in solid enlarged culturing base, obtains the head product that ferments;
4) by step 3) the fermentation head product that obtains carries out drying, adds medium carrier, sieves, packing, obtains described microbial growth promoters.
In one embodiment of the invention, the concrete steps of described preliminary screening are: soak wild rice stem planting soil with sterilized water and obtain leach liquor; Pseudomonas is filtered out with photosynthetic bacteria culture medium respectively from leach liquor, former brown vinelandii are gone out with fixed nitrogen Screening of Media, subtilis, Methyldmonas is filtered out with beef extract-peptone nutrient agar, filter out streptomycete with potato sucrose nutrient agar, thus obtain described pseudomonas, former brown vinelandii, subtilis, Methyldmonas and streptomycete respectively.
More preferably, select well-grown planting soil, and adopt ordinary method to carry out microorganism sterilizing, the sterilized water adding its 10 times of weight parts in soil after sterilizing is soaked 24-26 hour, pseudomonas is filtered out with photosynthetic bacteria culture medium respectively in leach liquor, former brown vinelandii are gone out with fixed nitrogen Screening of Media, subtilis is filtered out with beef extract-peptone nutrient agar, Methyldmonas, streptomycete is filtered out with potato sucrose nutrient agar, thus obtain described pseudomonas respectively, former brown vinelandii, subtilis, Methyldmonas and streptomycete.
In a preferred embodiment of the invention, described further screening is specially: described bacterial classification preliminary screening obtained is diluted to 10 respectively -5-10 -7, be then inverted in nutrient agar plate respectively and cultivate, select the bacterium colony of fast growth, then carry out shake flat experiment respectively, filter out speed of growth bacterial classification faster; Further, then each bacterial classification is transferred respectively and to cultivate into the eggplant bottle that nutrient agar is housed, when band is yellow during eggplant bottle surface lawn is covered with and is white, take out stand-by.
More preferably, described bacterial classification preliminary screening obtained is the normal saline dilution to 10 of 0.9% respectively by mass concentration -5-10 -7, the bacteria suspension obtained is inverted cultivation 48 hours respectively in nutrient agar plate under 35 DEG C of conditions, selects the bacterium colony of fast growth, then carries out shake flat experiment, observes growth speed, filters out the bacterial classification of quantity higher than 5,000,000,000/ml; Pseudomonas, former brown vinelandii, subtilis, Methyldmonas, streptomycete are transferred respectively and be equipped with in the eggplant bottle of nutrient agar, 45-50 hour is cultivated under 35-40 DEG C of condition, when during eggplant bottle surface lawn is covered with and is white, band is yellow, can take out, then the refrigerator putting into 2-6 DEG C is preserved, stand-by.
Wherein, screening the described pseudomonas obtained is Pseudomonas fluorescens.
Further, in one embodiment, step 2) in described mixed culture be specially: by step 1) in carry out mixed culture, when total viable count reaches 5,000,000,000/ml in the bacterial classification liquid medium within that obtains, pH value stops fermentation when reaching 7.5-8.0, obtains the head product that ferments.
More preferably, each bacterial classification is taken by following weight proportion: pseudomonas 10-13 part, former brown vinelandii 20-23 part, subtilis 30-33 part, Methyldmonas 10-13 part, streptomycete 18-30 part; Further, described mixed culture be by the mixing lawn of the above bacterial classification taken according to the part by weight of 1:25-30 be inoculated in through 120-130 DEG C, in the liquid nutrient medium of 30-40min high-temperature sterilization, stir culture in fermentor tank, its rotating speed is 220-240r/min, fermentation air flow maintains 1:1, and temperature is 30-32 DEG C; Ferment every 2 hours monitoring fermented liquid pH value and total viable count after 24 hours, when total viable count reaches 5,000,000,000/ml, when pH value reaches 7.5-8.0, stop fermentation.
In another embodiment, step 3) in described solid enlarged culturing for by step 2) in described mixed bacteria liquid access solid enlarged culturing base in carry out sealing and fermenting cultivation, treat that moisture is 30-40%, when pH value is 7-8.5, stop fermentation.
More preferably, by step 2) in the mixed bacteria liquid of mixed culture that obtains in the ratio access of 1:25-30 through 120-130 DEG C, in the solid enlarged culturing base of 30-45min high-temperature sterilization, stir 30-45min to even, pour sealing and fermenting in sterilization Double-layer Plastic envelope into, temperature 25-35 DEG C, time 8-15 days; Ferment every sampling in 6 hours after 7 days, measure moisture and pH value, treat that moisture is 30-40%, when pH value is 7-8.5, stopping is fermented.
Wherein in an embodiment, described drying is dry in incubator, and temperature controls at 35-45 DEG C, time 12-18 hour, and the moisture controlled of the fermentation head product that drying completes is below 20%.
Further, add medium carrier, mix according to the part by weight of dried fermentation head product 45-50 part, medium carrier 50-55 part, wherein, described medium carrier is sterilizing wood chip and sterilizing wild rice stem waste.
Further, carry out sieving and load with in the bucket of inner bag or plastic packaging bag, tightening sack, fasten nameplate, proceed in stockyard and stack in order.
Preferably, described preparation method also comprises sampling and detects, and main detect parameters is: moisture≤11.8%, viable count 25-30 hundred million/g, and physical behavior is brown powder solid.
Further, preferably, described liquid nutrient medium is the combination that sterilized water adds at least three kinds of mixtures in corn steep liquor, barley starch, monoammonium sulfate, molasses, potassiumphosphate, potassium sulfate, trace element solution again, preferably, its component and weight proportion thereof are corn steep liquor 2-3 part, barley starch 1-2 part, monoammonium sulfate 0.1-0.3 part, molasses 10-14 part, potassiumphosphate 1.2-2 part, potassium sulfate 0-0.8 part, trace element solution 0.3-0.5 part, sterilized water 75-85 part.
Preferably, described solid enlarged culturing base is the combination that sterilized water adds at least four kinds of mixtures in chaff, bean cake powder, yeast powder, Semen Maydis powder, potassium primary phosphate, manganous sulfate, garlic juice clearly again, preferably, its component and weight proportion thereof are: clear chaff 40-40 part, bean cake powder 10-12 part, yeast powder 1-1.2 part, Semen Maydis powder 8-10 part, potassium primary phosphate 0.8-1 part, manganous sulfate 0-0.4 part, garlic juice 1-1.2 part, sterilized water 32.3-39.2 part.
Further, described matrix is made up of sterilizing wood chip, sterilizing wild rice stem waste, and its weight proportion is: wood chip 55-60, wild rice stem waste 40-45.
Present invention also offers the microbial growth promoters that a kind of preparation method described above prepares.Wherein, it should be noted that, the proportioning related in the present invention and content, when there is no specified otherwise, referring to weight proportion or weight content.
According to technique scheme of the present invention, existing by following specific embodiment content of the present invention explained further and illustrate.
Embodiment 1
Be specifically designed to a preparation method for the microbial growth promoters of wild rice stem ecologic planting, particularly:
The first step: fermentation strain seed selection
A. bacterial screening: select well-grown planting soil, sterilized water soil being added 10 times of weight parts soaks 24 hours, rhodopseudomonas is filtered out with photosynthetic bacteria culture medium in leach liquor, with the former brown vinelandii of fixed nitrogen Screening of Media, with beef extract-peptone nutrient agar screening subtilis, Methyldmonas, with potato sucrose nutrient agar screening streptomycete;
B. shaking flask counting screening: be the normal saline dilution to 10 of 0.9% by the above bacterial classification concentration screened -5after bacteria suspension in nutrient agar plate 35 DEG C be inverted and cultivate 48H, the bacterium colony selecting fast growth carries out shake flat experiment, observes growth speed, and screening quantity is higher than the bacterial classification of 5,000,000,000/ml;
C. the making of bacterial classification is produced: Pseudomonas fluorescens, former brown vinelandii, subtilis, Methyldmonas, streptomycete are transferred respectively and be equipped with in the eggplant bottle of nutrient agar, cultivate 45 hours at 35 DEG C of temperature, treat that eggplant bottle surface lawn is covered with, and can take out during white middle band Huang, the refrigerator then putting into 2 DEG C is preserved;
Second step: the liquid culture of useful bacterial strain
A. each bacterial classification is taken by following weight proportion: Pseudomonas fluorescens 10 parts, former 20 parts, brown vinelandii, subtilis 30 parts, Methyldmonas 10 parts, streptomycete 18 parts;
B. the liquid culture of bacterial classification: by the mixing lawn taking above bacterial classification according to the ratio of 1:25 be inoculated in through 120 DEG C, in the liquid nutrient medium of 30min high-temperature sterilization, stir culture in fermentor tank, its rotating speed is 220r/min, and fermentation air flow maintains 1:1, temperature 30 DEG C;
C. every 2 hours monitoring fermented liquid pH value and total viable count after fermenting 24 hours, when total viable count reaches 5,000,000,000/ml, when pH value reaches 7.5, fermentation is stopped;
3rd step: the solid enlarged culturing of bacterial classification
A. the ratio in 1:25 after the mixing of the mixed bacteria liquid of step 2 mixed culture is accessed through 120 DEG C, in the solid enlarged culturing base of 30min high-temperature sterilization, stir 30min to evenly, pour sealing and fermenting in sterilization Double-layer Plastic envelope into, temperature 25 DEG C, 8 days time;
B. every sampling in 6 hours after fermenting 7 days, measure moisture and pH value, measuring moisture is 30%, stops fermentation when pH value is 7;
4th step: pulverize and add medium carrier
A. dry: the head product completely that step 3 fermented in chronological sequence puts into incubator drying, and temperature controls at 35 DEG C, and 12 hours time, the material moisture that drying completes controls below 20%;
B. add medium carrier: by dried fermentation head product according to: fermentation head product 45 parts, medium carrier 50-55 part ratio mixing add, medium carrier is sterilizing wood chip and sterilizing wild rice stem waste;
C. load after sieving with in the bucket of inner bag or plastic packaging bag, tighten sack, fasten nameplate, proceed to stockyard and stack in order; Sampling detects, significant parameter: moisture≤11.8%, the ripe 2,500,000,000/g of viable bacteria, and physical behavior is brown powder solid;
Wherein, the weight proportion of the component of the liquid nutrient medium in described step 2 is corn steep liquor 2 parts, barley starch 1 part, monoammonium sulfate 0.1 part, 10 parts, molasses, potassiumphosphate 1.2 parts, potassium sulfate 0 part, trace element solution 0.3 part, sterilized water 75 parts;
The weight proportion of the component of the solid enlarged culturing base in described step 3 is: clear chaff 40 parts, bean cake powder 10 parts, yeast powder 1 part, Semen Maydis powder 8 parts, potassium primary phosphate 0.8 part, manganous sulfate 0 part, garlic juice 1 part, sterilized water 32.3 parts;
Described matrix is made up of aseptic wood chip, aseptic wild rice stem waste, and its weight proportion is: wood chip 55, wild rice stem waste are 40.
Embodiment 2
Be specifically designed to a preparation method for the microbial growth promoters of wild rice stem ecologic planting, particularly:
The first step: fermentation strain seed selection
A. bacterial screening: select well-grown planting soil, sterilized water soil being added 10 times of weight parts soaks 26 hours, rhodopseudomonas is filtered out with photosynthetic bacteria culture medium in leach liquor, with the former brown vinelandii of fixed nitrogen Screening of Media, with beef extract-peptone nutrient agar screening subtilis, Methyldmonas, with potato sucrose nutrient agar screening streptomycete;
B. shaking flask counting screening: be bacteria suspension 35 DEG C of inversion cultivation 48H in nutrient agar plate of normal saline dilution to 10-7 of 0.9% by the above bacterial classification concentration screened, the bacterium colony selecting fast growth carries out shake flat experiment, observe growth speed, screening quantity is higher than the bacterial classification of 5,000,000,000/ml;
C. the making of bacterial classification is produced: Pseudomonas fluorescens, former brown vinelandii, subtilis, Methyldmonas, streptomycete are transferred respectively and be equipped with in the eggplant bottle of nutrient agar, cultivate 50 hours at 40 DEG C of temperature, treat that eggplant bottle surface lawn is covered with, and can take out during white middle band Huang, the refrigerator then putting into 6 DEG C is preserved;
Second step: the liquid culture of useful bacterial strain
A. each bacterial classification is taken by following weight proportion: Pseudomonas fluorescens 13 parts, former 23 parts, brown vinelandii, subtilis 33 parts, Methyldmonas 13 parts, streptomycete 30 parts;
B. the liquid culture of bacterial classification: by the mixing lawn taking above bacterial classification according to the ratio of 1:30 be inoculated in through 130 DEG C, in the liquid nutrient medium of 40min high-temperature sterilization, stir culture in fermentor tank, its rotating speed 240r/min, fermentation air flow maintains 1:1, temperature 32 DEG C;
C. every 2 hours monitoring fermented liquid pH value and total viable count after fermenting 24 hours, when total viable count reaches 5,000,000,000/ml, when pH value reaches 7.5-8.0, fermentation is stopped;
3rd step: the solid enlarged culturing of bacterial classification
A. the ratio in 1:30 after the mixing of the mixed bacteria liquid of step 2 mixed culture is accessed through 130 DEG C, in the solid enlarged culturing base of 45min high-temperature sterilization, stir 45min to evenly, pour sealing and fermenting in sterilization Double-layer Plastic envelope into, temperature 35 DEG C, 15 days time;
B. every sampling in 6 hours after fermenting 7 days, measure moisture and pH value, measuring moisture is 40%, stops fermentation when pH value is 8.5;
4th step: pulverize and add medium carrier
A. dry: it is dry that the head product completely that step 3 fermented in chronological sequence puts into incubator, and temperature controls at 45 DEG C, and 18 hours time, the material moisture that drying completes controls below 20%;
B. add medium carrier: by dried fermentation head product according to: fermentation head product 50 parts, medium carrier 55 parts ratio mixing add, medium carrier is sterilizing wood chip and sterilizing wild rice stem waste;
C. load after sieving with in the bucket of inner bag or plastic packaging bag, tighten sack, fasten nameplate, proceed to stockyard and stack in order; Sampling detects, significant parameter: moisture≤11.8%, the ripe 3,000,000,000/g of viable bacteria, and physical behavior is brown powder solid;
Wherein, the weight proportion of the component of the liquid nutrient medium in described step 2 is corn steep liquor 3 parts, barley starch 2 parts, monoammonium sulfate 0.3 part, 14 parts, molasses, potassiumphosphate 2 parts, potassium sulfate 0.8 part, trace element solution 0.5 part, sterilized water 85 parts;
The weight proportion of the component of the solid enlarged culturing base in described step 3 is: clear chaff 40 parts, bean cake powder 12 parts, yeast powder 1.2 parts, Semen Maydis powder 8-10 part, potassium primary phosphate 1 part, manganous sulfate 0.4 part, garlic juice 1.2 parts, sterilized water 39.2 parts;
Described matrix is made up of aseptic wood chip, aseptic wild rice stem waste, and its weight proportion is: wood chip 60, wild rice stem waste are 45.
Be described in detail specific embodiments of the invention above, but it is just as example, the present invention is not restricted to specific embodiment described above.To those skilled in the art, any equivalent modifications that the present invention is carried out and substituting also all among category of the present invention.Therefore, equalization conversion done without departing from the spirit and scope of the invention and amendment, all should contain within the scope of the invention.

Claims (10)

1. be specifically designed to a preparation method for the microbial growth promoters of wild rice stem ecologic planting, it is characterized in that, comprise the following steps:
1) from wild rice stem planting soil, preliminary screening goes out pseudomonas, former brown vinelandii, subtilis, Methyldmonas and streptomycete respectively, and is obtained the bacterial classification of fast growth respectively by further screening;
2) by step 1) in carry out mixed culture in the bacterial classification liquid medium within that obtains, obtain mixed bacteria liquid;
3) by step 2) mixed bacteria liquid that obtains carries out solid enlarged culturing in solid enlarged culturing base, obtains the head product that ferments;
4) by step 3) the fermentation head product that obtains carries out drying, adds medium carrier, sieves, packing, obtains described microbial growth promoters.
2. preparation method according to claim 1, is characterized in that, the concrete steps of described preliminary screening are: soak wild rice stem planting soil with sterilized water and obtain leach liquor; Pseudomonas is filtered out with photosynthetic bacteria culture medium respectively from leach liquor, former brown vinelandii are gone out with fixed nitrogen Screening of Media, filter out subtilis, Methyldmonas with beef extract-peptone nutrient agar, filter out streptomycete with potato sucrose nutrient agar.
3. preparation method according to claim 1, is characterized in that, described further screening concrete steps are: described bacterial classification preliminary screening obtained is diluted to 10 respectively -5-10 -7, be then inverted in nutrient agar plate and cultivate, select the bacterium colony of fast growth, then carry out shake flat experiment respectively, filter out speed of growth bacterial classification faster; Then each bacterial classification is transferred respectively and to cultivate into the eggplant bottle that nutrient agar is housed, when band is yellow during eggplant bottle surface lawn is covered with and is white, take out stand-by.
4. preparation method according to claim 1, it is characterized in that, step 2) in described mixed culture be specially: by step 1) in carry out mixed culture in the bacterial classification liquid medium within that obtains, when total viable count reaches 5,000,000,000/ml, when pH value reaches 7.5-8.0, stop fermentation.
5. preparation method according to claim 1, it is characterized in that, step 3) in described solid enlarged culturing for by step 2) in described mixed bacteria liquid access solid enlarged culturing base in carry out sealing and fermenting cultivation, treat that moisture is 30-40%, stop fermentation when pH value is 7-8.5, obtain the head product that ferments.
6. preparation method according to claim 1, is characterized in that, step 4) in, described drying is dry in incubator, and temperature controls at 35-45 DEG C, time 12-18 hour, and the moisture controlled of the fermentation head product that drying completes is below 20%; Described interpolation medium carrier mixes according to the part by weight of dried fermentation head product 45-50 part, medium carrier 50-55 part, wherein, described medium carrier is sterilizing wood chip and sterilizing wild rice stem waste, and its weight proportion is wood chip 55-60, wild rice stem waste 40-45; Further, carry out sieving and load with in the bucket of inner bag or plastic packaging bag, tightening sack, fasten nameplate, proceed in stockyard and stack in order.
7. preparation method according to claim 1, is characterized in that, described preparation method also comprises sampling and detects, and main detect parameters is: moisture≤11.8%, viable count 25-30 hundred million/g, and physical behavior is brown powder solid.
8. preparation method according to claim 1, it is characterized in that, step 2) in, described liquid nutrient medium is the combination that sterilized water adds corn steep liquor, barley starch, monoammonium sulfate, molasses, potassiumphosphate, potassium sulfate, trace element solution again, and its weight proportion is corn steep liquor 2-3 part, barley starch 1-2 part, monoammonium sulfate 0.1-0.3 part, molasses 10-14 part, potassiumphosphate 1.2-2 part, potassium sulfate 0-0.8 part, trace element solution 0.3-0.5 part, sterilized water 75-85 part.
9. preparation method according to claim 1, it is characterized in that, step 3) in, described solid enlarged culturing base is the combination that sterilized water adds chaff, bean cake powder, yeast powder, Semen Maydis powder, potassium primary phosphate, sulphur, sour manganese, garlic juice clearly again, and its weight proportion is: clear chaff 40-40 part, bean cake powder 10-12 part, yeast powder 1-1.2 part, Semen Maydis powder 8-10 part, potassium primary phosphate 0.8-1 part, manganous sulfate 0-0.4 part, garlic juice 1-1.2 part, sterilized water 32.3-39.2 part.
10. the microbial growth promoters prepared as the preparation method in claim 1-9 as described in any one.
CN201510999966.5A 2015-12-28 2015-12-28 Microbial growth promoter specially used for ecological planting of wild rice stems and preparation method thereof Pending CN105461370A (en)

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CN112544162A (en) * 2020-12-02 2021-03-26 无锡市三阳生态农业发展有限公司 Growth-promoting rhizobacteria seedling culture method for improving growth rate of bud seedlings

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