CN107058151A - A kind of microbial bacteria with fermentation water retaining function and its application - Google Patents
A kind of microbial bacteria with fermentation water retaining function and its application Download PDFInfo
- Publication number
- CN107058151A CN107058151A CN201611074021.3A CN201611074021A CN107058151A CN 107058151 A CN107058151 A CN 107058151A CN 201611074021 A CN201611074021 A CN 201611074021A CN 107058151 A CN107058151 A CN 107058151A
- Authority
- CN
- China
- Prior art keywords
- microbial
- mtz102
- fermentation
- bacterial strain
- mass ratio
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F5/00—Fertilisers 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
- C05F5/002—Solid waste from mechanical processing of material, e.g. seed coats, olive pits, almond shells, fruit residue, rice hulls
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Environmental & Geological Engineering (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Mechanical Engineering (AREA)
- Environmental Sciences (AREA)
- Fertilizers (AREA)
Abstract
The invention discloses a kind of microbial bacteria with fermentation water retaining function and its application, bacterial strain MTZ102 is a kind of bacillus amyloliquefaciens for the functions such as production aliphatic acid, organic acid, capsular polysaccharide of fermenting, above secondary metabolite can be given birth under different fermentation conditions, relative to artificial synthesized water-loss reducer, the secondary metabolite that microbial fermentation is produced can be biodegradable, environmentally friendly, safe, soil environment will not be caused harm and secondary pollution.Bacterial strain MTZ102 can be obviously promoted crowndaisy chrysanthemum, the seeding corn and other crops tolerance low to soil moisture content in the barren Rhizosphere of Crops soil of arid, and for crowndaisy chrysanthemum, seeding corn and other crops, growth provides a kind of excellent liquid bacterial fertilizer in relatively arid soil.
Description
Technical field
The present invention relates to the microbial organic fertilizer field of microbial bacteria, and in particular to a kind of to have the water retaining function that ferments
Microbial bacteria and its application.
Background technology
Arid is to endanger one of disaster the most serious in the world, its occurrence number, duration, coverage, is caused
Loss etc. occupy first of various natural calamities, considerably beyond other meteorological disasters.National crop drought area 3.4 in 2014
Hundred million mu, 1.8 hundred million mu of disaster area, 85,160,000 mu of Disaster Area, total crop failure 22,270,000 mu of area, grain loss 20,060,000 caused by drought
Ton, industrial crops lose 27,600,000,000 yuan, 91,000,000,000 yuan of direct economy total losses.
Over nearly 40 years.With the rise of atmospheric greenhouse gas content, China's weather totally occurs warming the trend of exsiccation
This trend causes great adverse effect to China's agricultural.Research shows, in existing planting system, varieties of plant and life
Under the premise of production is poor.2030-2050, because extreme weather events can make China's grain-production caused by climate change
Potentiality reduction about 10%.Wherein wheat, paddy rice and the big crop of corn 3 are based on the underproduction, and also have certain influence on quality, must
The grain security for having a strong impact on China is surrounded.Therefore, influence and counter-measure of the arid to China's agricultural production are reduced, to development
Sustainable agriculture is particularly important, and with reality and far reaching significance.So, retardance soil erosion and resisting drought saving water turn into China
Agricultural future can sustainable development it is in the urgent need to address the problem of.
Water-loss reducer claims high water absorbent, super-strong moisture absorbing resin (super absorbent polymer, SAP), is that one kind can
Absorb the compound of itself original size and weight several times moisture.The hydrogel formed after water-loss reducer water suction can slowly discharge moisture
For crop utilization, meanwhile, water retention, improved soil structure, influence soil physico-chemical property, crop root growth can be strengthened again
Development etc., and then the migration rule and plant physiology function of moisture and solute in soil are influenceed, improve the utilization rate of chemical fertilizer.Mesh
Water-loss reducer is used for the multiple fields such as industry, agricultural, building, gardening, health by preceding nearly 30 countries, is agriculturally also being obtained
Development, has a extensive future very well.But, begin to occur environmental problem from the use of first generation water-loss reducer.First generation water conservation
Agent is because using acrylonitrile monemer as raw material, raw material residual can produce toxicity, influence Environmental security.By SANYO GS company in
The second generation starch-polyacrylic acid graft copolymer of exploitation in 1975, although cost is low, toxicity is low, but heat endurance is not high, resistance to
Salt is poor, and water absorbing properties can reduce and cause serious soil hardening in high salinity soil.The new guarantor of the mid-80 research and development
Aqua increases in terms of salt tolerance, heat endurance and water-retaining property, and the problem of causing soil hardening has also obtained effectively changing
It is kind.But, water-loss reducer once blindly promotes the use of the environmental problem being likely to result in so that its safety for agricultural use in application process
Evaluating turns into inevitable.Although the Water Retaining Agent on Soil physicochemical properties researched and developed and used at present have not significant impact, with
The progress of modern technologies, it is far from being enough only to carry out evaluation to physiochemical properties of soil (including water retention property).It is special
It is not important component of the edaphon as soil ecosystem, with soil material circulation and the close phase of energy flow
Close.The research of Water Retaining Agent on Soil microbial ecological influence is still within blank (Li Xi etc., 2012).And utilize beneficial microbe
Diaphragm is formed in the capillary gap that the metabolites such as aliphatic acid, organic acid, the capsular polysaccharide that fermentation is produced can be in soil, can
To block the evaporation of interstices of soil moisture, so as to reach the purpose of soil water-retaining drought resisting.
Bacillus amyloliquefaciens are a kind of bacteriums very high with bacillus subtilis affinity, in the growth course of its own
In can produce a series of metabolite, these metabolites bacillus amyloliquefaciens to have widely to suppress true
The activity of bacterium and bacterium is outer, can also produce the biological membrane substance of the formation such as aliphatic acid, organic acid, capsular polysaccharide, be suspended in moisture
Sublist face.Therefore, in recent years a large amount of scholars to the new discoveries of bacillus amyloliquefaciens and new opplication and gene functional research
Substantial amounts of exploration has been done in optimization with fermentation condition.Bacillus amyloliquefaciens can produce a variety of antibacterial materials, wide sterilization spectrum, to disease
Opportunistic pathogen has preventive and therapeutic effect, therefore it prevents and treats crop, vegetables, melon and fruit disease effect highly significant, while to reduce chemical pesticide, protecting
Demonstrate,prove food security and provide an effective way.Our research of what is more important finds bacillus amyloliquefaciens in fermentation one
High-molecular biologic ester film can be produced during a little specific organic matters, suspends and hydrone surface, the evaporation of hydrone can be reduced, reached
To the purpose of water conservation drought resisting.
The content of the invention
In view of the above-mentioned problems, the present invention is intended to provide a kind of fast growth, and resistance of reverse strong to environmental suitability are strong
Microbial bacteria, the high-molecular biologic ester film produced by strain fermentation organic matter, is suspended in the hydrone in soil pore space
Surface, reduces soil water evaporation, improves the water content of soil, so as to significantly improve vegetables and crop yield and quality
Microbial bacteria and its application with fermentation water retaining function.
To realize the technical purpose, the solution of the present invention is：A kind of microbial bacteria with fermentation water retaining function, including bacterium
Strain MTZ102, is bacillus amyloliquefaciens (Bacillus amyloliquefaciens), and preserving number is GDMCC No.
60099, preservation date is on October 31st, 2016；
Bacterial strain MTZ102 form and physio-biochemical characteristics is as follows：
Bacterial strain MTZ102 is gram-positive bacteria, and cell can produce gemma in shaft-like；
Fast growth on LB agar plates, bacterium colony is circular, middle small raised (37 DEG C, grow 36 hours), and milky is not
It is transparent, 2-8 millimeters of colony diameter；Cell size 2.7-4.1x 1.5-1.7 μm, shaft-like, tool end life flagellum；Optimum growth temperature
It is 27-40 DEG C, pH growth scope 4-10, optimum growh pH value 6-8；
Amphimicrobian, can be with D- galactolipins, and L- arabitols, sucrose, dextrin, sorbose, D-Fructose, D-Glucose is fine
Dimension disaccharides, turanose, trehalose, mannitol, D-R alcohol, sorbierite, hexitol, melezitose, galactitol, D-sorbite,
L- fructose, sodium gluconate, sodium lactate, sodium succinate, malic acid, glycerine, soybean oil, D-Glucose aldehydic acid sodium, butanediamine, N-
Acetylglucosamine, Pidolidone, ALANINE, Glu, L-Histidine, CYSTINE, aspartic acid is sole carbon
Grown on the culture medium in source；
PH growth scopes are wider, are resistant to pH4-pH10；NaCl concentration can grow less than 2.0%；It is resistant to 300 μ g/mL
Streptomysin, ampicillin, tetracycline, and 100 μ g/mL gentamicin, erythromycin.Kalamycin higher than 50 μ g/mL,
With regard to its growth can be suppressed under chloramphenicol, the concentration conditions of neomycin；Siderophore is produced, indole reaction is positive, and can secrete growth
Plain IAA.
A kind of utilization bacterial strain MTZ102 microbial organic fertilizer, the bacterial strain MTZ102 contents are 108More than cfu/g's
Bacterium number, total nitrogen content mass ratio is 4~5%, and mass ratio more than 90% is organic nitrogen in full nitrogen, and total npk nutrient mass ratio is
6~10%, content of organic matter mass ratio is not less than 45%.
A kind of production method of bacterial strain MTZ102 microbial organic fertilizer,
The first step, bacterial strain MTZ102 is inoculated into the nutrient solution of modified MRS type and carries out fermenting and producing, the bar of fermenting and producing
Part is：28~35 DEG C of cultivation temperature scope, mixing speed is 180~300rpm, and the fermentation middle and later periods forms gemma, made in zymotic fluid
Containing bacterium or gemma amount >=1 × 109cfu·ml-1；
Second step, carries out solid dung fermentation, often by bacterial strain MTZ102 with the agricultural organic waste mixture after becoming thoroughly decomposed
Added in ton SOLID ORGANIC material per turning in 7-12 days 1 time in zymocyte liquid 100-200L, solid fermentation process, while solid is sent out
Ferment temperature is no more than 80 DEG C, and general fermentation terminates after 25-40 days, fermentation bacterial content is reached 1 × 108cfu·g-1More than, obtain
MTZ102 containing zymophyte and agricultural organic waste mixture microbial organic fertilizer.
Preferably, in second step used microbial organic fertilizer after becoming thoroughly decomposed, wherein quality of organic matter than content >=
35%, organic nitrogen quality is 1.2-2%, water content mass ratio more than 25% than content.
Preferably, nutrient solution proportioning is in the first step：0.15~0.25g of dusty yeast, molasses 6~14g, KH2PO4
0.01~0.02g, K2HPO40.01~0.02g, 3~9g of dregs of beans, 0.5~1.5g of wheat bran, add water to 1L, PH6-8；Nutrient solution is matched somebody with somebody
After the completion of putting, 121 DEG C of sterilizing 20min.
Preferably, total nitrogen content mass ratio is matter in 4~5%, full nitrogen in microbial organic fertilizer in the second step
Amount is organic nitrogen than more than 90%, and total npk nutrient mass ratio is that 6~10%, content of organic matter mass ratio is not less than 45%.
Preferably, agricultural organic waste is stalk or cavings or dregs of beans or wheat bran in the second step
In one or more combinations.
With application of the bacterial strain MTZ102 microbial organic fertilizer in crowndaisy chrysanthemum and maize culture.
Beneficial effects of the present invention, 1. bacterial strain MTZ102 are the functions such as a kind of fermentation production aliphatic acid, organic acid, capsular polysaccharide
Bacillus amyloliquefaciens, above secondary metabolite can be given birth under different fermentation conditions, relative to artificial synthesized guarantor
Aqua, the secondary metabolite that microbial fermentation is produced can be biodegradable, environmentally friendly, safe, and soil environment will not be caused
Harm and secondary pollution.
2. bacterial strain MTZ102 is in the barren Rhizosphere of Crops soil of arid, the work such as crowndaisy chrysanthemum, corn can be obviously promoted
The thing tolerance low to soil moisture content, for crowndaisy chrysanthemum, seeding corn and other crops in relatively arid soil growth to provide one kind excellent
Liquid bacterial fertilizer.
3. microbial organic fertilizer prepared by bacterial strain MTZ102 contains the organic molecule film of retardance moisture evaporation, soil is kept
Water content, can significantly improve crowndaisy chrysanthemum, seeding corn and other crops to mineral matter and organotrophic absorption, make crop leaf tissue
Chlorophyll content, blade Vc contents and yield, with increase quality and increase yield good result.
Embodiment
The present invention is described in further details with reference to specific embodiment.
The microbial bacteria with functions such as fermentation production aliphatic acid, organic acid, capsular polysaccharides of the present invention is by using 80
Non- bacillus is killed in DEG C processing, with modified MRS culture (1L)：Peptone 10g, beef extract 10g, yeast extract 5g, hydrogen citrate
Two ammonium 2.0g, glucose 20g, dipotassium hydrogen phosphate 2g, ammonium sulfate 1.63g, sodium acetate 5g, magnesium sulfate 0.58g, manganese sulfate 0.25g,
Tween 80 mL, calcium carbonate 1g, agar 18g, plus distilled water is to 1000mL, the scholars 0.2 of pH 6.4, respectively under the conditions of anaerobic and aerobic
Culture.Separated from the corn plant grown on the dead soil of Guangzhou, purify the bacillus for obtaining 5 plants of production capsular polysaccharides.
The abilities such as fermentation production aliphatic acid, organic acid, capsular polysaccharide to 5 plants of bacterium are further screened, and obtain fast growth, production pod membrane is more
Sugared ability is strong, and can produce under different culture media formula capsular polysaccharide, and the stable bacterial strain MTZ102 of production capsular polysaccharide；Should
The zymotic fluid of bacterial strain is as Inoculant, using the soil of drought and water shortage, can promote crowndaisy chrysanthemum, the growth of seeding corn and other crops.
First, the microbial bacteria with fermentation water retaining function, including bacterial strain MTZ102,
Bacterial strain MTZ102 is bacillus amyloliquefaciens (Bacillus amyloliquefaciens), and preserving number is GDMCC
No.60099, preservation date is on October 31st, 2016；Depositary institution：Guangdong Province's Culture Collection, address：Guangzhou
5 building, the building of compound the 59th of city martyr Road 100.
(1), bacterial strain MTZ102 form and physio-biochemical characteristics are as follows：
Bacterial strain MTZ102 is gram-positive bacteria, and cell can produce gemma in shaft-like；
Fast growth on LB agar plates, bacterium colony is circular, middle small raised (37 DEG C, grow 36 hours), and milky is not
It is transparent, 2-8 millimeters of colony diameter；2.7-4.1x1.5-1.7 μm of cell size, shaft-like, tool end life flagellum；Optimum growth temperature is
27-40 DEG C, pH growth scope 4-10, optimum growh pH value 6-8；
Amphimicrobian, can be with D- galactolipins, and L- arabitols, sucrose, dextrin, sorbose, D-Fructose, D-Glucose is fine
Dimension disaccharides, turanose, trehalose, mannitol, D-R alcohol, sorbierite, hexitol, melezitose, galactitol, D-sorbite,
L- fructose, sodium gluconate, sodium lactate, sodium succinate, malic acid, glycerine, soybean oil, D-Glucose aldehydic acid sodium, butanediamine, N-
Acetylglucosamine, Pidolidone, ALANINE, Glu, L-Histidine, CYSTINE, aspartic acid is sole carbon
Grown on the culture medium in source；
PH growth scopes are wider, are resistant to pH4-pH10；NaCl concentration can grow less than 2.0%；It is resistant to 300 μ g/mL
Streptomysin, ampicillin, tetracycline, and 100 μ g/mL gentamicin, erythromycin.Kalamycin higher than 50 μ g/mL,
With regard to its growth can be suppressed under chloramphenicol, the concentration conditions of neomycin；Siderophore is produced, indole reaction is positive, and can secrete growth
Plain IAA.
(2), the determination of bacillus amyloliquefaciens MTZ102 of the present invention molecular classification status
Performing PCR amplification is entered using bacterial 16 S rDNA gene universal primers 25f and 1492r, PCR primer is directly carried out just
Opposite direction sequencing, the DNA sequence dna input GenBank of acquisition is compared, primarily determines that the bacillus of the present invention exists
Kind, category position in taxology.As a result find that the bacillus of the present invention belongs to bacillus (Bacillus sp.), with solution
Bacillus amyloliquefacienses (Bacillus amyloliquefacien) type strain LMG 12234 has 98.3% similitude.Physiology
Biochemical test feature and 16S rRNA gene sequencings show that bacillus amyloliquefaciens MTZ102 of the invention is with solving starch bud
Spore bacillus type strain LMG12234 has difference.With reference to above-mentioned physio-biochemical characteristics, 16S rDNA the sequencing results,
The bacillus amyloliquefaciens of the present invention should belong to bacillus (Bacillus sp.), be a new strains of the category, tool
There are fermentation production aliphatic acid, organic acid, capsular polysaccharide ability.It judges to refer to following document：Borriss,R.,et al.,
Relationship of Bacillus amyloliquefaciens clades associated with strains
DSM7T and FZB42T:a proposal for Bacillus amyloliquefaciens
subsp.amyloliquefaciens subsp.nov.and Bacillus amyloliquefaciens
subsp.plantarum subsp.nov.based on complete genome sequence comparisons.Int J
Syst Evol Microbiol,2011.61(Pt 8):p.1786-801；Fan,B.,et al.,Transcriptomic
profiling of Bacillus amyloliquefaciens FZB42in response to maize root
exudates.BMC Microbiol,2012.12:p.116；Yuan,J., et al.,Plant growth-promoting
rhizobacteria strain Bacillus amyloliquefaciens NJN-6-enriched bio-organic
fertilizer suppressed Fusarium wilt and promoted the growth of banana
plants.J Agric Food Chem,2013.61(16):p.3774-80.
(3), the isolation and purification method of strain of the present invention
3.1 enrichment：The Rhizosphere of Crops soil 30g of lean soil growth is weighed in the triangular flask of the sterilized water containing 50mL, is filled
Divide vibration to break up, suspension is made, 1 is pressed with enrichment nutrient solution：In 2 ratios dress people's triangular flask, it is statically placed in incubator, 30 ± 2
DEG C, cultivate 3 days, the careful supernatant that takes out is heated to 80 DEG C and kills non-Bacillus in the beaker of another sterilizing.
Take the μ L to 250mL of heating bacteria suspension 500 in this first generation blake bottle to be enriched with nutrient solution progress second to be commissioned to train
Support, cultural method is identical with the first generation.
3.2 isolate and purify：Draw second generation bacterial culture fluid 1ml and be made 10 in 9ml sterilized water test tubes-1Bacterium solution is diluted again,
This concentration bacteria suspension such as method is taken to be diluted to 10-2,10-3 times again, until being diluted to 10-6Times.10 are drawn respectively-4、10-5、10-6Three
The μ L of bacteria suspension 100 of individual gradient, use spread plate method and pour into and cultivated at methods,anaerobic, 30 ± 2 DEG C successively, observation
The growing state of different cultural method hypothalluses, picking growing way is preferably and the bacterium colony with different characteristic is entered with plate streak
Capable Secondary Culture repeatedly, until shape, color, quality, the transparency of bacterium colony are consistent.Finally by simple dyeing and microscopy, enter
One step observes its form, so that length is homogeneous, width is consistent, staining conditions are unified for bacterial strain purification standard.
3.3 preserve：Bacterial strain after isolating and purifying is cultivated after 2~4d on isolation medium flat board and inclined-plane, collects flat
Thalline on plate.Part is stored in 15% sterile glycerol (- 20 DEG C) and (- 80 DEG C).
Wherein enrichment culture liquid (1L)：KH2PO40.5g, K2HPO40.6g, MgSO47H2O0.2g, NaCl 0.2g, ferment
Female cream 0.5g, peptone 5.0g, calcium carbonate 0.5g, plus deionized water is to 1L), pH is naturally, about 7.0 ± 0.2.
Wherein isolation medium (1L)：Yeast extract 2g, peptone 4g, calcium carbonate 0.5g, agar powder 17g, pH 7.0 ±
0.2。
2nd, the microbial organic fertilizer component ratio of microbial bacteria fermentation：
A kind of utilization bacterial strain MTZ102 microbial organic fertilizer, the bacterial strain MTZ102 contents are 108More than cfu/g's
Bacterium number, total nitrogen content mass ratio is 4~5%, and mass ratio more than 90% is organic nitrogen in full nitrogen, and total npk nutrient mass ratio is
6~10%, content of organic matter mass ratio is not less than 45%.
3rd, prepared by the microbial organic fertilizer of microbial bacteria fermentation：
A kind of production method of bacterial strain MTZ102 microbial organic fertilizer,
The first step, bacterial strain MTZ102 is inoculated into the nutrient solution of modified MRS type and carries out fermenting and producing, the bar of fermenting and producing
Part is：28~35 DEG C of cultivation temperature scope, mixing speed is 180~300rpm, and the fermentation middle and later periods forms gemma, made in zymotic fluid
Containing bacterium or gemma amount >=1 × 109cfu·ml-1；
Second step, carries out solid dung fermentation, often by bacterial strain MTZ102 with the agricultural organic waste mixture after becoming thoroughly decomposed
Added in ton SOLID ORGANIC material per turning in 7-12 days 1 time in zymocyte liquid 100-200L, solid fermentation process, while solid is sent out
Ferment temperature is no more than 80 DEG C, and general fermentation terminates after 25-40 days, fermentation bacterial content is reached 1 × 108cfu·g-1More than, obtain
MTZ102 containing zymophyte and agricultural organic waste mixture microbial organic fertilizer.
In order to which the proportioning for the N P and K for ensureing organic fertilizer meets microbial organic fertilizer in the growth of plant, second step used
Material is after becoming thoroughly decomposed, and wherein quality of organic matter is than content >=35%, and organic nitrogen quality is 1.2-2%, water content mass ratio than content
More than 25%.
In order to ensure that bacterial strain MTZ102 fast and stable grows, nutrient solution proportioning is in the first step：Dusty yeast 0.15
~0.25g, molasses 6~14g, KH2PO40.01~0.02g, K2HPO40.01~0.02g, 3~9g of dregs of beans, wheat bran 0.5~
1.5g, adds water to 1L, PH6-8；After the completion of nutrient solution configuration, 121 DEG C of sterilizing 20min
In order to which the proportioning for the N P and K for ensureing organic fertilizer meets microbial organic fertilizer in the growth of plant, the second step
Total nitrogen content mass ratio is 4~5% in material, and mass ratio more than 90% is organic nitrogen in full nitrogen, and total npk nutrient mass ratio is 6
~10%, content of organic matter mass ratio is not less than 45%.
For more efficient utilization agricultural organic waste, in the second step agricultural organic waste be stalk or
One or more combinations in cavings or dregs of beans or wheat bran.
Specific fermentation medium is prepared and condition is following (1L)：Dusty yeast 0.2g, molasses 10g, KH2PO40.01g,
K2HPO40.01g, dregs of beans 5g, wheat bran 1g, add water to 1L, pH natures.Fermentation seed liquid is first prepared with LB cultures, is cultivated 2-3 days,
When bacterial strain concentration reaches 108 cells/more than mL, with 1：10 ratio, which is inoculated into fermentation culture medium, expands culture, in 28-
Under the conditions of 30 DEG C, fermented and cultured 20 days, at this moment zymotic fluid surface be observed that the formation of biomembrane.Constituent analysis to the film
The predominantly mixture of aliphatic acid, organic acid, pod membrane and polysaccharose substance.Another bacillus amyloliquefaciens for not producing film are located at
Fermented under same culture conditions.The liquid surface evaporation speed of fermentation production film bacterial strain is not significantly lower than film bacterial strain is produced, so as to reach water conservation
Effect.
The microbial organic fertilizer produced with bacterial strain MTZ102, the bacterial strain MTZ102 contents are × more than 108cfu/g
Bacterial strain, total nitrogen content mass ratio is 4~5%, and mass ratio more than 90% is organic nitrogen in full nitrogen, and total npk nutrient mass ratio is
6~10%, content of organic matter mass ratio is not less than 45%.
4th, potted plant experiment：
(1) crowndaisy chrysanthemum potted plant experiment
1.1 use bacterial strain MTZ102 water retaining function bacteria fermentation culture medium YAW (1L)：Dusty yeast 0.2g, molasses 10g, KH2PO4
0.01g, K2HPO40.01g, dregs of beans 5g, wheat bran 1g, add water to 1L, pH natures.
1.2 using：Soil is well mixed consistent, and the original initial pH value of soil is 6.5, and field capacity is 60%.Will activation
Obtained single bacterium is fallen within 15ml fresh LB seed fluid nutrient mediums of saccharomycete, 37 DEG C, 180rpm shaken cultivation 24h, then will be cultivated
Liquid seeds nutrient solution be transferred to by 5% inoculum concentration in fermentation medium YAW, 30 DEG C, cultivate 7d, it is standby.Pot experiment does 4
Individual processing, 3 repeat.CK1：Control 1, it is nonvaccinated that the YAW nutrient solution 1mL for stating preparation are added per kg soil, normally cultivate 20 days,
Period does not water；CK3：Control 3, nonvaccinated every kg soil adds the YAW nutrient solution 1mL for stating preparation, normally cultivation 20 days, often
Suitable quantity of water was sprayed every 5 days；Processing 1-1-1 and 1-1-2 is that the zymotic fluid 1mL for stating preparation is added per kg soil, normal cultivation 20 days, phase
Between do not water.It is consistent that the above four handles other daily managements.
1.3 result：
Specific effect see the table below：
1.3.1 gemma fermentation water conservation bacterium handles the reduced parameter of crowndaisy chrysanthemum
| Sequence number | Plant average height cm | Chlorophyll content mg/g | Vc contents mg/g | Yield g/ |
| CK1 | 10.5±0.5 | 1.2±0.02 | 0.18±0.02 | 40.2±0.08 |
| CK3 | 15.8±0.6 | 1.7±0.02 | 0.25±0.01 | 61.3±0.07 |
| 1-1-1 | 14.6±0.4 | 1.5±0.01 | 0.25±0.01 | 60.5±0.06 |
| 1-1-2 | 15.5±0.5 | 1.7±0.03 | 0.24±0.02 | 59.1±0.09 |
1.3.2 gemma fermentation water conservation bacterium processing crowndaisy chrysanthemum 1-1-1 and 1-1-2 and CK1 contrasts lifting capacity (%)
| 1-1-1 | 1-1-2 | CK3 | |
| Plant height increases | 39.05 | 47.62 | 50.48 |
| Chlorophyll content is improved | 25.00 | 41.67 | 41.67 |
| Vc contents are improved | 38.89 | 33.33 | 38.89 |
| Output increased | 50.50 | 47.01 | 52.49 |
It is seen from the above data that the Tong planted with the bacterial strain MTZ102 water retaining function bacteria fermentation culture medium YAW kinds produced
Wormwood artemisia (1-1-1 and 1-1-2) yield, the average content for planting high, Vc and chlorophyll are substantially far above the CK1 not watered；With normally pouring
The CK3 of water remains basically stable compared to parameters such as yield, the average contents for planting high, Vc and chlorophyll, illustrates in the feelings do not watered
The water retaining function bacteria fermentation culture medium YAW that bacterial strain MTZ102 is produced under condition, which increases crowndaisy chrysanthemum quality and increase yield, good effect
Really.
Two) corn potted plant experiment
2.1 use bacterial strain MTZ102 water retaining function bacteria fermentation culture medium YAW (1L)：Dusty yeast 0.2g, molasses 10g, KH2PO4
0.01g, K2HPO40.01g, dregs of beans 5g, wheat bran 1g, add water to 1L, pH natures.
2.2 soil are well mixed consistent, and the original initial pH value of soil is 6.5, and field capacity is 60%.It will activate what is obtained
Single bacterium is fallen within 15ml fresh LB seed fluid nutrient mediums of saccharomycete, 37 DEG C, 180rpm shaken cultivation 24h, then by cultured liquid
Seed culture fluid is transferred in fermentation medium YAW by 5% inoculum concentration, 30 DEG C, cultivates 7d, standby.Pot experiment is done at 4
Reason, 3 repeat.CK1：Control 1, nonvaccinated every kg soil adds the YAW nutrient solution 1mL for stating preparation, normally cultivation 20 days, during which
Do not water；CK2：Control 2, it is nonvaccinated that the YAW nutrient solution 1mL for stating preparation, normal cultivation 20 days, every 5 are added per kg soil
Its spray suitable quantity of water；Processing A1 and A2 is that the zymotic fluid 1mL for stating preparation is added per kg soil, and during which normal cultivation 20 days does not water.
It is consistent that the above four handles other daily managements.
2.3 result：
2.3.1 gemma fermentation water conservation bacterium handles corn reduced parameter
2.3.2 gemma fermentation water conservation bacterium processing corn A1 and A2 and CK1 contrasts lifting capacity (%)
| A1 | A2 | CK2 | |
| Plant height increases | 43.63 | 46.75 | 50.12 |
| Chlorophyll-a Content is improved | 45.10 | 43.14 | 47.06 |
| Content of chlorophyll b is improved | 50.00 | 50.00 | 56.25 |
| Ground dry weight is improved | 44.44 | 42.22 | 42.22 |
It is seen from the above data that the jade planted with the bacterial strain MTZ102 water retaining function bacteria fermentation culture medium YAW kinds produced
Rice (A1 and A2) ground dry weight, the average content for planting high, chlorophyll a and chlorophyll b are substantially far above the CK1 not watered；With just
The CK2 often watered remains basically stable compared to parameters such as ground dry weight, the average contents for planting high, chlorophyll a and chlorophyll b, says
The water retaining function bacteria fermentation culture medium YAW of the bright productions of the bacterial strain MTZ102 in the case where not watering increases quality and increase to corn
Yield has good result.
Described above, only presently preferred embodiments of the present invention is not intended to limit the invention, every skill according to the present invention
Any trickle amendment, equivalent substitution and improvement that art is substantially made to above example, should be included in technical solution of the present invention
Protection domain within.
Claims (8)
1. a kind of microbial bacteria with fermentation water retaining function, it is characterised in that：Including bacterial strain MTZ102, for solution starch gemma bar
Bacterium (Bacillus amyloliquefaciens), preserving number is GDMCC No. 60099, and preservation date is October 31 in 2016
Day；
Bacterial strain MTZ102 form and physio-biochemical characteristics is as follows：
Bacterial strain MTZ102 is gram-positive bacteria, and cell can produce gemma in shaft-like；
Fast growth on LB agar plates, bacterium colony is circular, middle small raised (37 DEG C, grow 36 hours), and milky is opaque,
2-8 millimeters of colony diameter；2.7-4.1x1.5-1.7 μm of cell size, shaft-like, tool end life flagellum；Optimum growth temperature is 27-40
DEG C, pH growth scope 4-10, optimum growh pH value 6-8；
Amphimicrobian, can be with D- galactolipins, L- arabitols, sucrose, dextrin, sorbose, D-Fructose, D-Glucose, fiber two
Sugar, turanose, trehalose, mannitol, D-R alcohol, sorbierite, hexitol, melezitose, galactitol, D-sorbite, L- fruits
Sugar, sodium gluconate, sodium lactate, sodium succinate, malic acid, glycerine, soybean oil, D-Glucose aldehydic acid sodium, butanediamine, N- acetyl
Gucosamine, Pidolidone, ALANINE, Glu, L-Histidine, CYSTINE, aspartic acid is sole carbon source
Grown on culture medium；
PH growth scopes are wider, are resistant to pH4-pH10；NaCl concentration can grow less than 2.0%；It is resistant to 300 μ g/mL chain
Mycin, ampicillin, tetracycline, and 100 μ g/mL gentamicin, erythromycin.Kalamycin, chlorine higher than 50 μ g/mL is mould
With regard to its growth can be suppressed under element, the concentration conditions of neomycin；Siderophore is produced, indole reaction is positive, and can secrete auxin
IAA。
2. the microbial organic fertilizer of microbial bacteria described in a kind of utilization claim 1, it is characterised in that：The bacterial strain MTZ102
Content is 108More than cfu/g bacterium number, total nitrogen content mass ratio is 4~5%, and mass ratio more than 90% is organic nitrogen in full nitrogen,
Total npk nutrient mass ratio is 6~10%, and content of organic matter mass ratio is not less than 45%.
3. the production method of the microbial organic fertilizer of microbial bacteria described in a kind of claim 2, it is characterised in that：
The first step, bacterial strain MTZ102 is inoculated into the nutrient solution of modified MRS type and carries out fermenting and producing, the condition of fermenting and producing
For：28~35 DEG C of cultivation temperature scope, mixing speed is 180~300rpm, and the fermentation middle and later periods forms gemma, makes to contain in zymotic fluid
Bacterium or gemma amount >=1 × 109cfu·ml-1；
Second step, solid dung fermentation is carried out by bacterial strain MTZ102 with the agricultural organic waste mixture after becoming thoroughly decomposed, per ton solid
Added in body organic materials per turning in 7-12 days 1 time in zymocyte liquid 100-200L, solid fermentation process, while solid fermentation temperature
Degree is no more than 80 DEG C, and general fermentation terminates after 25-40 days, fermentation bacterial content is reached 1 × 108cfu·g-1More than, obtain containing hair
Yeast-like fungi MTZ102 and agricultural organic waste mixture microbial organic fertilizer.
4. the production method of the microbial organic fertilizer of microbial bacteria according to claim 3, it is characterised in that：Used
Microbial organic fertilizer is after becoming thoroughly decomposed in two steps, and wherein quality of organic matter is than content >=35%, and organic nitrogen quality is than content
1.2-2%, water content mass ratio more than 25%.
5. the production method of the microbial organic fertilizer of microbial bacteria according to claim 3, it is characterised in that：Described
Nutrient solution proportioning is in one step：0.15~0.25g of dusty yeast, molasses 6~14g, KH2PO40.01~0.02g, K2HPO4 0.01
~0.02g, 3~9g of dregs of beans, 0.5~1.5g of wheat bran, add water to 1L, PH6-8；After the completion of nutrient solution configuration, 121 DEG C of sterilizings
20min。
6. the production method of the microbial organic fertilizer of microbial bacteria according to claim 3, it is characterised in that：Described
Total nitrogen content mass ratio is 4~5% in microbial organic fertilizer in two steps, and mass ratio more than 90% is organic nitrogen in full nitrogen, always
Npk nutrient mass ratio is that 6~10%, content of organic matter mass ratio is not less than 45%.
7. the production method of the microbial organic fertilizer of microbial bacteria according to claim 3, it is characterised in that：Described
Agricultural organic waste is one or more combinations in stalk or cavings or dregs of beans or wheat bran in two steps.
8. application of the microbial organic fertilizer of the microbial bacteria described in claim 2 in crowndaisy chrysanthemum and maize culture.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611074021.3A CN107058151B (en) | 2016-11-29 | 2016-11-29 | Microbial bacterium with fermentation water retention function and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611074021.3A CN107058151B (en) | 2016-11-29 | 2016-11-29 | Microbial bacterium with fermentation water retention function and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107058151A true CN107058151A (en) | 2017-08-18 |
| CN107058151B CN107058151B (en) | 2021-08-20 |
Family
ID=59618495
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611074021.3A Active CN107058151B (en) | 2016-11-29 | 2016-11-29 | Microbial bacterium with fermentation water retention function and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107058151B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108485976A (en) * | 2018-03-16 | 2018-09-04 | 华中科技大学 | A kind of screening technique of heavy metal resistance microorganism |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101875910A (en) * | 2010-04-20 | 2010-11-03 | 山东省食品发酵工业研究设计院 | Bacillus amyloliquefaciens for producing gamma-polyglutamic acid |
| CN103805204A (en) * | 2013-07-22 | 2014-05-21 | 河海大学 | Farmland soil microorganism water-holding product and preparation method thereof |
| CN104744182A (en) * | 2015-01-21 | 2015-07-01 | 河海大学 | Tobacco water retention biological organic fertilizer and preparation method thereof |
-
2016
- 2016-11-29 CN CN201611074021.3A patent/CN107058151B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101875910A (en) * | 2010-04-20 | 2010-11-03 | 山东省食品发酵工业研究设计院 | Bacillus amyloliquefaciens for producing gamma-polyglutamic acid |
| CN103805204A (en) * | 2013-07-22 | 2014-05-21 | 河海大学 | Farmland soil microorganism water-holding product and preparation method thereof |
| CN104744182A (en) * | 2015-01-21 | 2015-07-01 | 河海大学 | Tobacco water retention biological organic fertilizer and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| JIAHUI SHAO: "Analysis and Cloning of the Synthetic Pathway of the Phytohormone indole-3-acetic Acid in the Plant-Beneficial Bacillus Amyloliquefaciens SQR9", 《MICROB CELL FACT》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108485976A (en) * | 2018-03-16 | 2018-09-04 | 华中科技大学 | A kind of screening technique of heavy metal resistance microorganism |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107058151B (en) | 2021-08-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103184162B (en) | Trichoderma asperellum and applications thereof | |
| CN103484396B (en) | New strain of streptomyces thermocarboxydus and application thereof | |
| CN104164394A (en) | Antagonistic phytopathogen strain and application thereof | |
| CN103045515A (en) | Biological agent of bacillus methylotrophicus as well as preparation method and application thereof | |
| CN104232532B (en) | A kind of plant endogenesis bacillus pumilus and its microbial inoculum, preparation method and application | |
| CN101864376A (en) | Pseudomonas fluorescens strain, microbial inoculum and use thereof as seedling culture medium for controlling tomato bacterial wilt | |
| CN113980836B (en) | Bacillus belgii and application thereof in prevention and treatment of strawberry root rot | |
| CN113699075B (en) | Bacillus atrophaeus capable of producing protease and decomposing potassium and application thereof | |
| CN107164261A (en) | Rhizobium and its application that one plant of promotion villose vetch increases | |
| CN104593297B (en) | It is a kind of that there is the microbial bacteria for producing acid function and its organic fertilizer of application | |
| CN105462881A (en) | Paenibacillus polymyxa for preventing and treating vertieillium wilt in crops and application of paenibacillus polymyxa | |
| CN104651267B (en) | A kind of organic fertilizer with the microbial bacteria and its application of fermentation production alkali | |
| CN109957535B (en) | Bacillus simplex, microbial agent prepared by using bacillus simplex, biological fertilizer and application | |
| CN107628894A (en) | Composite bacteria agent increase soil fertility and its preparation method and application | |
| CN107502565A (en) | One plant of fritillary bulb rhizosphere Paenibacillus polymyxa and its application | |
| CN105018393B (en) | One plant of bacillus megaterium and its application | |
| CN107467075B (en) | Application of bacillus pumilus as rice growth promoter | |
| CN104789494B (en) | The method for improving turf salt-resistance using garbage compost microbial bacterial agent is strengthened | |
| CN101525584B (en) | Degradation bacteria for removing fomesafen pesticide residue and microbial inoculum prepared by same | |
| CN101560482B (en) | Buprofezin pesticide residual degrading bacteria and produced microbial inoculum thereof | |
| CN109207393A (en) | One bacillus amyloliquefaciens and its application in prevention and treatment celery root rot | |
| CN107058151A (en) | A kind of microbial bacteria with fermentation water retaining function and its application | |
| CN101486985B (en) | Nicosulfuron pesticide residue degrading bacterium and inocula produced therefrom | |
| CN103243056B (en) | Paracoccus MXX-04 for bromoxynil degradation and application thereof | |
| CN104195082B (en) | A kind of travelling germ and its microbial inoculum and preparation and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: A kind of microbial bacteria with the function of fermentation and water retention and its application Effective date of registration: 20220815 Granted publication date: 20210820 Pledgee: Bank of China Limited by Share Ltd. Guangzhou Liwan branch Pledgor: GUANGDONG SENDU ECOLOGICAL AGRICULTURE TECHNOLOGY CO.,LTD. Registration number: Y2022440000203 |
|
| PE01 | Entry into force of the registration of the contract for pledge of patent right |