CN105859347A - Special composite microbial fertilizer for radix psuedostellariae, and preparation method and application thereof - Google Patents
Special composite microbial fertilizer for radix psuedostellariae, and preparation method and application thereof Download PDFInfo
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- CN105859347A CN105859347A CN201610208743.7A CN201610208743A CN105859347A CN 105859347 A CN105859347 A CN 105859347A CN 201610208743 A CN201610208743 A CN 201610208743A CN 105859347 A CN105859347 A CN 105859347A
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- radix pseudostellariae
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F1/00—Fertilisers made from animal corpses, or parts thereof
- C05F1/005—Fertilisers made from animal corpses, or parts thereof from meat-wastes or from other wastes of animal origin, e.g. skins, hair, hoofs, feathers, blood
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/50—Treatments combining two or more different biological or biochemical treatments, e.g. anaerobic and aerobic treatment or vermicomposting and aerobic treatment
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
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- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- 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
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The invention relates to a special composite microbial fertilizer for controlling successive cropping obstacles of radix psuedostellariae. The fertilizer is prepared by fermenting a special composite microbial inoculant and organic materials (bean pulp, rice bran, dried pig manure, fish meal and brown sugar in a weight ratio of 2:1:1:1:0.03). The composite microbial inoculant is prepared by mixing a universal beneficial bacterium solution and an aboriginal beneficial bacterium solution in a volume ratio of 4:6. The aboriginal beneficial bacterium comprises an aboriginal beneficial bacterium composite bacterium solution which is formed by independently fermenting Burkholderia, pseudomonad and Bacillus separated to the radix psuedostellariae rhizosphere soil, and mixing in a volume ratio of 4:1:1. The organic materials are subjected to special composite microbial inoculant anaerobic fermentation and oxygen-consuming fermentation to prepare the special composite microbial fertilizer. The long-term on-site verification proves that the microbial fertilizer has the advantages of reasonable formula, stable effects, favorable biocontrol effect and no pollution, has obvious effects on controlling succession cropping obstacles of radix psuedostellariae, is safe and ecological, and thus, has wide development and application prospects.
Description
Technical field
The present invention relates to a kind of Radix Pseudostellariae Special compound microbial fertilizer and preparation method and application, can be used for preventing and treating medicine
With plant Radix Pseudostellariae Soil-sickness Problem, belong to agricultural fertilizer and technical field of biological control.
Background technology
Radix Pseudostellariae (Pseudostellaria heterophylla) another name child's ginseng, Radix Ginseng, for Caryophyllaceae perennial grass
This plant, is used as medicine with root, has the effects such as replenishing QI to invigorate the spleen, promoting the production of body fluid for nourishing the lung, and its medication is with a long history, clinical efficacy definite, city
Field demand is big.But, Radix Pseudostellariae planting process exists serious Soil-sickness Problem, also referred to as continuous cropping problem or replant disease
Harmful, the most same plot is planted Radix Pseudostellariae for years, is caused Radix Pseudostellariae tuber normally to expand, and pest and disease damage occurs serious, produces
Amount quality the problem (as shown in Figure 1) such as is remarkably decreased, and this seriously inhibits Radix Pseudostellariae medicine resource and utilizes and related industry
Sustainable development, explores a kind of environmentally friendly Radix Pseudostellariae continuous cropping obstacle decreasing measure significant.
Early-stage Study finds, Radix Pseudostellariae continuous cropping causes structure of soil microbial community unbalance, i.e. probiotics reduces, pathogen
Amount reproduction grows, and ultimately results in continuous cropping obstacle effect.The continuous cropping of deterioration is improved by microbial bacterial agent or biological organic fertilizer
Soil, the soil ecosystem of reconstruction health is considered as the main path solving Radix Pseudostellariae continuous cropping obstacle.But, micro-life
Thing microbial inoculum or biological organic fertilizer the most do not have broad spectrum activity, it is possible to Species of Pathogens or the Damage Types of preventing and treating are extremely limited.Class
Topic group early stage, prevents and treats Radix Pseudostellariae cultivating disease by buying commercially available several anti-continuous cropping bacterial manure or microbial bacterial agent, and result is sent out
Existing prevention effect is extremely limited, and this may be with the rhizospere competition of probiotics in commercially available microbial inoculum and the host specific of pathogen
Property etc. relevant.Seminar's early stage from morbidity Radix Pseudostellariae plant and continuous cropping soil separation screening to the most pathogenic many cause of disease
Bacterium, as Fusarium oxysporum, fusarium moniliforme, ankle save (as shown in Figure 2) such as Fusarium spp., and many pathogen strains bacterium shows host spy
The opposite sex, the most only infects Radix Pseudostellariae and does not infect rear stubble other crops such as corn and soybean, Oryza sativa L. etc..So, separation screening can be short of money
The probiotics of anti-Radix Pseudostellariae specialized pathogen bacterium could effectively prevent and treat Radix Pseudostellariae cultivating disease.Early-stage Study is it was also found that Radix Pseudostellariae
Continuous cropping causes soil environment generation significant change, and as there is acidifying problem, and the multiple cometabolism of Radix Pseudostellariae sustained release produces
Thing and from poisonous substance matter, these factors all may cause the probiotics in commercially available microbial inoculum surely cannot grow in Radix Pseudostellariae rhizosphere, therefore adopts
Host specificity and the continuous cropping of Radix Pseudostellariae soil-borne pathogen need to be considered during with microbial bacterial agent preventing and treating Radix Pseudostellariae cultivating disease
The particularity of soil environment.
In consideration of it, the Special compound microbial fertilizer that the present invention relates to be by Radix Pseudostellariae specific complex microbial bacterial agent with have
Machine material ferments together and makes.Described Radix Pseudostellariae specific complex microbial bacterial agent is by general probiotics (can directly buy) and soil
Write probiotics (separating to Radix Pseudostellariae rhizosphere soil) to be mixed by proportion optimizing, it is therefore an objective to ensure that probiotics is at Radix Pseudostellariae root
The success on border is grown surely, can activating soil nutrient, degraded Radix Pseudostellariae release from poisonous substance matter, Radix Pseudostellariae specialized form can be prevented and treated again sick
Former bacterium (as shown in Figure 3).
Summary of the invention
It is an object of the invention to provide a kind of Radix Pseudostellariae Special compound microbial fertilizer and preparation method and application, be used for
The Soil-sickness Problem of Biological control medicinal plants Radix Pseudostellariae, plays and promotes tuber growth and the dual purpose of controlling disease.
Technical scheme is as follows:
A kind of Special compound microbial fertilizer, is fermented together with organic materials by complex micro organism fungicide and makes.Described is compound
Microbial bacterial agent, is made up of with original inhabitants' probiotics (IM) general probiotics (EM), and viable count is more than 1.0 × 109 CFU/mL.Its
In, general probiotics includes lactic acid bacteria, photosynthetic bacteria, yeast, actinomycetes, and available commercially available EM stock solution expands according to a conventional method
Cultivate and obtain EM composite bacteria liquid；Described indigenous probiotics includes burkholderia, the vacation separated to Radix Pseudostellariae rhizosphere soil
Zymomonas mobilis, bacillus cereus, the most individually fermentation.After the multiple beneficial bacterium of same genus is each fermented, equal-volume is mixed and belongs to together
Composite bacteria liquid.EM composite bacteria liquid and each original inhabitants' probiotics (being followed successively by burkholderia, pseudomonas, bacillus cereus) the most at last
Composite bacteria liquid mix for the ratio of 4:4:1:1 by volume.
The described lactic acid bacteria in specific complex microbial bacterial agent can be Lactobacillus plantarum, streptococcus acidi lactici, cheese milk
Bacillus；Photosynthetic bacteria can be Rhodopseudomonas palustris, hydrogenlike silicon ion；Yeast can be saccharomyces cerevisiae, Candida utilis ferment
Female；Actinomycetes can be streptomyces griseus, streptomyces albus.Burkholderia can be Burkholderia ambifaria
(Burkholderia ambifaria sp. nov., a novel member of the Burkholderia cepacia
Complex including biocontrol and cystic brosis-related isolates.Tom Coenye,
Eshwar Mahenthiralingam、Deborah Henry, et. International Journal of
Systematic and Evolutionary Microbiology, 2001 (51): 1481-1490.), primary gram of Hall of new Bulbus Allii Cepae
One or several in moral bacterium；Pseudomonas can be pseudomonas fluorescens, pseudomonas putida, Pseudomonas
Protegens(Pseudomonas protegens sp. nov., widespread plant-protecting
bacteria producing the biocontrol compounds 2,4-diacetylphloroglucinol and
pyoluteorin.Alban Ramette, Michele Frapolli, Marion Fischer-Le Saux.
Et.Systematic and Applied Microbiology, 2011 (34): 180-188.), Korea S pseudomonas
One or more in (Pseudomonas koreensis)；Bacillus cereus can be bacillus amyloliquefaciens, bacillus subtilis
One or more in bacterium, bacillus cereus, Bacillus pumilus, bacillus megaterium, bacillus mycoides.
Described organic materials includes bean cake, Testa oryzae, drying pig manure, fish flour, brown sugar, enters by weight 2:1:1:1:0.03
Row mixing is mixed thoroughly as fermentation bed material.
The preparation method of Special compound microbial fertilizer:
A kind of Special compound microbial fertilizer, is fermented together with organic materials by specific complex microbial bacterial agent and makes.Its special secondary school
Body is pressed by general probiotics bacterium solution (EM composite bacteria liquid) and original inhabitants' probiotics bacterium solution (IM composite bacteria liquid) with complex micro organism fungicide
The ratio that long-pending ratio is 4:6 is mixed.
(1) EM composite bacteria liquid: include lactic acid bacteria, photosynthetic bacteria, yeast, actinomycetes, is the ratio of 1:1:1:1 by volume
Example associating amplification culture, or activated by commercially available EM stock solution, amplification culture prepared.
(2) IM composite bacteria liquid: by burkholderia composite bacteria liquid, pseudomonas composite bacteria liquid, bacillus cereus compound bacteria
Liquid is that the ratio of 4:1:1 mixes by volume.
A. burkholderia composite bacteria liquid includes following bacterium: Burkholderia ambifaria, new Bulbus Allii Cepae primary gram
Hall moral bacterium, after each bacterium is individually fermented, equal-volume mixes.
B. pseudomonas composite bacteria liquid includes following bacterium: pseudomonas fluorescens, pseudomonas putida, Pseudomonas
Protegens, Korea S pseudomonas, after each bacterium is individually fermented, equal-volume mixes.
C. bacillus cereus composite bacteria liquid includes following bacterium: bacillus amyloliquefaciens, bacillus subtilis, waxy spore
Bacillus, Bacillus pumilus, bacillus megaterium, bacillus mycoides, after each bacterium is individually fermented, equal-volume mixes.
(3) ratio that EM composite bacteria liquid and IM composite bacteria liquid are 4:6 by volume is mixed and made into specific complex microbial bacterial agent.
(4) organic materials bean cypress, Testa oryzae, dry pig manure, fish flour, brown sugar are carried out by weight the ratio of 2:1:1:1:0.03
Mixing is mixed thoroughly.
(5) anaerobic fermentation: by volume mass ratio 1:10 adds specific complex microbial bacterial agent in the organic materials mixed thoroughly,
Adding water and fully mix thoroughly, water content control is 30%, and subpackage carries out closing anaerobic fermentation, 37 DEG C of bottom fermentation 48h, then room temperature place 2～
Within 4 weeks, carry out after-ripening.
(6) oxygen consumption fermentation: access special microorganism liquid according still further to volume mass than 1:30 in above-mentioned anaerobic fermentation product
Body microbial inoculum, airtight and ventilate equipment in 37 DEG C fermentation 48h, prepare Special compound microbial fertilizer, total viable count 1.0～
5.0 hundred million/g.
Special compound microbial fertilizer of the present invention has several advantages that
(1) Special compound microbial fertilizer of the present invention is fermented system by specific complex microbial bacterial agent together with organic materials
?.Wherein specific complex microbial bacterial agent contains Radix Pseudostellariae rhizosphere original inhabitants' Antagonistic Fungi, including burkholderia, pseudomonas and
Bacillus cereus, is all to screen to obtain from Radix Pseudostellariae rhizosphere soil, and during Field information, its rhizospere competition is good and safety can
Lean on, and to other crops of rear stubble (such as corn and soybean, Oryza sativa L., vegetable etc.) all without the infection ability that causes a disease.
(2) original inhabitants' Antagonistic Fungi of the present invention is to be obtained by substantial amounts of screening operation, is thin from 3000 strain soil
In bacterium, screening obtains, and has Radix Pseudostellariae specialized pathogen bacterium (such as Fusarium oxysporum, fusarium moniliforme, ankle joint mycete etc.) very
Strong antagonistic effect (as shown in Figure 3), the wherein strong new Burkholderia cepacia of Antagonistic Fungi, Burkholderia
Ambifaria, Pseudomonas protegens, Korea S pseudomonas are preserved in China Microbiological on January 28th, 2016
Culture presevation administration committee common micro-organisms center, deposit number is respectively as follows: CGMCC NO. 12119, CGMCC NO.
12120、CGMCC NO. 12121、CGMCC NO. 12122。
(3) during original inhabitants' Antagonistic Fungi screening of the present invention toward culture medium adds higher concentration from poisonous substance matter, make
For selection pressure, the major part original inhabitants' probiotics being sieved to has Degradation and Transformation function to what Radix Pseudostellariae root system was secreted from poisonous substance matter.
(4) original inhabitants' Antagonistic Fungi of the present invention also has dissolving P capacity in addition to can suppressing pathogen, it is possible to effectively
Activating soil nutrient (as shown in Figure 4), improves soil physico-chemical property, promotes that plant nutrient absorption aspect has important function.
(5) present invention is through verifying on the spot for a long time, and formula is reasonable, stable effect, bright to preventing and treating Radix Pseudostellariae continuous cropping obstacle effect
Aobvious, improve the yield and quality (as shown in Figure 5) of continuous cropping Radix Pseudostellariae.
Accompanying drawing explanation
Fig. 1 is continuous cropping and first planting Radix Pseudostellariae underground part growing way compares (A) and incidence (B).
Fig. 2 is the colonial morphology of the pathogen that Radix Pseudostellariae plant diseases root is separated to. A: Fusarium oxysporum；B: beading
Fusarium spp.；C: ankle joint mycete.
Fig. 3 is the opposite culture photo of original inhabitants' Antagonistic Fungi and Radix Pseudostellariae specialized pathogen fungus (fusarium moniliforme). A: only
Inoculation pathogen fusarium moniliforme；B: original inhabitants' probiotics and fusarium moniliforme opposite culture.
Fig. 4 is that original inhabitants' probiotics dissolving P capacity measures.Arrow instruction has the indigenous probiotics of dissolving P capacity.
Fig. 5 is that continuous cropping, the first planting and special microorganism microbial inoculum process lower Radix Pseudostellariae tuber growing state comparison in difference.
Detailed description of the invention
Below in conjunction with specific embodiment, technical scheme is described in detail.Following example are used only for
The description and interpretation present invention, and do not constitute the restriction to technical solution of the present invention.
The screening of embodiment 1 original inhabitants' Antagonistic Fungi and qualification thereof
It is respectively adopted pseudomonas selective medium, bacillus cereus selective medium to Radix Pseudostellariae rhizosphere pseudomonas, bud
Bacillus separates spore, uses inorganic medium (Radix Pseudostellariae containing higher concentration is from poisonous substance matter) to carry out from poisonous substance matter degradation bacteria
Separate.
The method that the single bacterium colony of random choose on above-mentioned different culture media uses PDA plate face-off is evaluated them respectively
Antagonistic effect to Radix Pseudostellariae specialized pathogen bacterium (such as Fusarium oxysporum, fusarium moniliforme, ankle joint mycete etc.).To have the most short of money
The antibacterial of anti-ability uses 16s rRNA gene or 16s-23s rRNA interval PCR qualification to carry out Molecular Identification.16s rRNA base
Because PCR amplification upstream and downstream primer is respectively as follows: 27f(5'-AGAGTTTGATCCTGGCTCAG-3') and 1522r(5'-
AAGGAGGTGAT CCAGCCGCA-3')；16s-23s rRNA gene PCR amplification upstream and downstream primer is respectively as follows: 1407f(5'-
TTGTACACACCG CCCGTC-3') and 456r(5'-CCTTTCCCTCAC GGTACTG-3').
Described culture medium prescription is:
(1) pseudomonas selective medium: weigh 20g Soybean-casein digest agar culture medium, adds the double steaming of 495.5mL
Water heated and stirred, after fully dissolving, adds 1mL 0.1%(wt/vol) crystal violet storing solution, 121 ° of C autoclaving 15min,
When being cooled to about 50 ° of C, in culture medium, add 3.5mL 5%(wt/vol again) Nitrofurantoin storing solution (use N, N-dimethyl
Methanamide makees solvent), a kind of rhyme scheme in Chinese operas serving as the prelude to a complete score for voices cools down, standby.
(2) bacillus cereus selective medium: every liter of peptone Han 5.0g, 1.5g yeast extract, 1.5g beef
Cream, 5.0g NaCl, 20g agar, pH 7.2.
(3) from poisonous substance matter degradation bacteria culture medium: every liter contains NH4NO31.0g, NaCl 0.5g, K2HPO4·3H2O 1.5g,
MgSO47H2O 0.5g, (NH4)2SO40.5g, pH 7.2.Weigh each component to be dissolved in 1000mL distilled water, 121 ° of C height
Pressure sterilizing 15min, when being cooled to 45～50 ° of C, identify in addition Radix Pseudostellariae continuous cropping soil is various from poisonous substance matter (Galla Turcica (Galla Helepensis)
Acid, coumaric acid, PCA, P-hydroxybenzoic acid, vanillic acid, syringic acid, vanillin, ferulic acid, benzoic acid),
Total phenols concentration 1000mg/L, after inoculating for 10% Radix Pseudostellariae rhizosphere soil suspending liquid in enrichment cultivation number generation, screening has from poison mass degradation
Ability and the beneficial bacteria of antagonism Radix Pseudostellariae pathogen.
Obtain 12 strains altogether through Screening and Identification and there is Radix Pseudostellariae Rhizosphere Soil work probiotics (as shown in Figure 3) of strong antagonistic ability,
Including 2 strain burkholderias (Burkholderia ambifaria, new Burkholderia cepacia)；4 pseudomonas are (glimmering
Light pseudomonas, pseudomonas putida, Pseudomonas protegens, Korea S pseudomonas)；6 bacillus (Xie Dian
Afnyloliquefaciens, bacillus subtilis, bacillus cereus, Bacillus pumilus, bacillus megaterium, bacillus mycoides).
Wherein Korea S pseudomonas is Korea S pseudomonas (Pseudomonas koreensis) CJ-361, in 2016
On January 28, in is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, the numbered CGMCC of culture presevation
NO. 12122, address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
New Burkholderia cepacia is new Burkholderia cepacia (Burkholderia cenocepacia) CJ-
P8, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, culture presevation on January 28th, 2016
Numbered CGMCC NO. 12119, address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
Pseudomonas is pseudomonas (Pseudomonas protegens) LK-313, in preservation on January 28 in 2016
In China Committee for Culture Collection of Microorganisms's common micro-organisms center, culture presevation numbered CGMCC NO. 12121, ground
Location is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
Burkholderia is burkholderia (Burkholderia ambifaria) LK-P4, in January, 2016
Within 28th, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, culture presevation numbered CGMCC NO.
12120, address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
The preparation of embodiment 2 Special compound microbial fertilizer
A kind of Special compound microbial fertilizer of the present invention, is fermented together with organic materials by specific complex microbial bacterial agent
Make.Wherein (IM is multiple by general probiotics bacterium solution (EM composite bacteria liquid) and original inhabitants' probiotics bacterium solution for specific complex microbial bacterial agent
Close bacterium solution) it is mixed for the ratio of 4:6 by volume.Concrete preparation method is as follows:
Prepared by 2.1 EM composite bacteria liquids
In the well water (tap water uses after need at least placing 2 days) of 18L, add 300g brown sugar and 50mL MS mother solution, add
The commercially available EM liquid spawn that 1L activates, is placed in the indoor sealed fermenting 3 weeks of 37 ° of C, within second day, starts every other day may be used from fermentation
Open the venting decompression of hermetic container mouth once, sour-sweet taste when opening vessel port, can be smelt, and more and more obvious, and this is normal existing
As, i.e. represent that fermentation spreads cultivation successfully, standby.Successfully finished product fermentation liquid has sour-sweet taste, pH 3.0～4.0, and viable count is more than 100
Hundred million/mL.
2.2 original inhabitants' probiotics activation and amplification culture
1) actication of culture
Pseudomonas fluorescens, pseudomonas putida, Pseudomonas protegens, Korea S pseudomonas are inoculated in respectively
In pseudomonas Selective solid culture medium, it is placed in 30 ° of C incubators activation 16h.
By bacillus amyloliquefaciens, bacillus subtilis, bacillus cereus, Bacillus pumilus, bacillus megaterium,
Bacillus mycoides is inoculated in bacillus cereus selective medium respectively, is placed in 37 ° of C incubators activation 12h.
Burkholderia ambifaria, new Burkholderia cepacia are inoculated in LB solid culture respectively, put
14h is activated in 37 ° of C incubators.
2) amplification culture
The various original inhabitants probiotics activated being inoculated in LB liquid culture and carry out fermentation culture, pseudomonas is placed under 30 ° of C
Amplification culture, other original inhabitants' probioticss are placed in amplification culture under 37 ° of C, when viable count is higher than 1.0 × 109 Stop during CFU/mL expanding
Big cultivation.The multiple beneficial bacterium equal-volume of same genus is mixed and belongs to composite bacteria liquid together, such as burkholderia compound bacteria
Liquid, pseudomonas composite bacteria liquid, bacillus cereus composite bacteria liquid, standby.
Prepared by 2.3 complex micro organism fungicides
It is the ratio of 4:1:1 by volume by original inhabitants probiotics i.e. burkholderia, pseudomonas, bacillus cereus composite bacteria liquid
It is mixed and made into original inhabitants' probiotics composite bacteria liquid (IM composite bacteria liquid).
Further, by EM composite bacteria liquid and IM composite bacteria liquid by volume ratio for 4:6 to be mixed and made into Radix Pseudostellariae special
Complex micro organism fungicide.
Prepared by 2.4 composite microbiological fertilizers
Organic materials bean cypress, Testa oryzae, drying pig manure, fish flour, brown sugar by weight 2:1:1:1:0.03, are carried out mixing and mixes thoroughly, then
Adding specific complex microbial liquid microbial inoculum (1L/10kg), add water stirring according to the degree of drying of material, water content control exists
30%(pinches the most agglomerating but not water outlet with hands and is advisable), after fully mixing thoroughly, pack or barrelling carry out closing anaerobic fermentation, 37 ° of C, 48h,
Fermenting complete, room temperature is placed 3 weeks and is carried out after-ripening.Special microorganism liquid bacterial agent (1L/ is accessed again in above-mentioned anaerobic fermentation product
30kg), in equipment that is airtight and that ventilate, 37 ° of C oxygen consumption fermentation 48h, prepare Special compound microbial fertilizer, total viable count 1.0
～5.0 hundred million/g.
Embodiment 3 Special compound microbial fertilizer preventing and treating Radix Pseudostellariae continuous cropping obstacle field efficacy is evaluated
First Radix Pseudostellariae continuous cropping soil is improved in advance by the Special compound microbial fertilizer of preparation as base fertilizer, the most whole
Ground plays furrow, plants Radix Pseudostellariae, and starts timely and appropriate discovery pouring root seedling stage from Radix Pseudostellariae and impose the specific complex for fermentation fertilizer
Microbial liquid microbial inoculum, carries out volume variance harvest time and compares.Concrete prophylactico-therapeutic measures is as follows:
Prepared by the 3.1 organic base fertilizer of microorganism
Organic materials bean cypress, Testa oryzae, drying pig manure, fish flour, brown sugar by weight 2:1:1:1:0.03, are carried out mixing and mixes thoroughly, then
Adding specific complex microbial liquid microbial inoculum (1L/10kg), water content control, 30%, is fully mixed rear subpackage thoroughly and is carried out airtight detesting
Aerobe fermentation, 37 DEG C, 48h, to ferment complete, room temperature places 3 weeks with after-ripening.Afterwards, access again specially in above-mentioned anaerobic fermentation product
With microbial liquid microbial inoculum (1L/30kg), in equipment that is airtight and that ventilate, 37 DEG C of oxygen consumption fermentation 48h, prepare specific complex micro-
Bio-feritlizer, total viable count 1.0～5.0 hundred million/g.
3.2 soil improvement
Microbial manure process group: after Radix Pseudostellariae results, carry out plot the most wholely, coordinating applies fertilizer to the subsoil every mu applies in advance
Fermenting the Special compound microbial fertilizer 1200kg of (starting fermentation at least in advance for 1 month), deep ploughing is ploughed under in soil, harrows even
Furrow, cover black mulch film, and furrow irrigation is spread and sunk in, and water is micro-wet (but not flooding to furrow face) to furrow face i.e. cuts off the water, and overlay film ferments at least 1 month, with
The Continuous Cropping Soil Environment deteriorated is repaired in improvement, plants Radix Pseudostellariae the most again.During test, other field management measures are right with the positive
Keep consistent according to group, negative control group.
Positive controls: never planting the plot top time plantation Radix Pseudostellariae of Radix Pseudostellariae, period does not applies the present invention
Described complex micro organism fungicide or composite microbiological fertilizer, as first planting Radix Pseudostellariae.
Negative control group: plant Radix Pseudostellariae on a plot for years planting Radix Pseudostellariae again, and do not apply this
Complex micro organism fungicide described in invention or composite microbiological fertilizer, as continuous cropping Radix Pseudostellariae.
3.3 Radix Pseudostellariae plantations
Plant the last week, carry out second time the most wholely, whole good furrow face is opened by line-spacing 15 centimetres the craspedodrome bar ditch of deep 9 centimetres.
Before plantation, compound fertilizer 20kg, phosphate fertilizer 25kg, potash fertilizer 10kg is used to make seed manure in applying plantation ditch after mixing for every mu, a little blindings a little,
Then tiltedly arranging in furrow bank by spacing in the rows 6 centimetres by seed ginseng, head-up, bud is away from Horizon about 5cm, light earthing for bud.In late November extremely
December is planted, and every mu with seed ginseng 25～35 kilograms.
3.4 impose bacterium solution
In mid or late Febuary, after coming up, pouring root applies the specific complex microbial inoculums 1 time of 100 times of dilutions, mid-March and
Mid-April, then pouring root applies 50 times of specific complex microbial inoculums diluted each 1 time respectively, to resist soil-borne disease generation.Root
According to Radix Pseudostellariae growing way and incidence, adjusting pouring root number of times, if there being disease symptom, can suitably increase pouring root number of times.Positive, negative
Matched group pouring root applies the water of equivalent.
3.5 water and fertilizer management
The first tenday period of a month in March, in conjunction with in weeding carry out topdressing for the first time, use compoiste fertilizer 25kg and carbamide for every mu
2.5kg.Carrying out second time in mid or late April to topdress, every mu sprays 5kg potassium dihydrogen phosphate, needs first to be made into 0.5% before spraying
Solution, sprayed every 8 days 1 time, sprays 3 times.
Radix Pseudostellariae Seedling Stage requires ground moistening, to water in time during weather arid, and the middle and late growth stage meets high temperature and drought season
Joint still needs to carry out as required sooner or later irrigating, and keeps furrow face to moisten, beneficially root of hair and plant strain growth.But Radix Pseudostellariae is afraid of flood, and chance is many
Rainy day gas, will dredge gutter in time, note draining, in order to avoid rotten.
3.6 results and volume variance compare
Wither the time of falling Seedling in late June Radix Pseudostellariae plant, select fine day to excavate Radix Pseudostellariae.More than 15 centimetres are deep-cut with spud,
Radix Pseudostellariae tuber is completely dug out, it is to avoid damage tuber, sort out, clean, dry, claim fresh weight.
Result of the test shows for many years, and compared with continuous cropping Radix Pseudostellariae, using invented Special compound microbial fertilizer can show
Write suppression incidence, improve Radix Pseudostellariae per mu yield, increase production about 30～65%, yield and first planting Radix Pseudostellariae quite (as it is shown in figure 5,
Table 1).Visible, Special compound microbial fertilizer of the present invention is notable to preventing and treating Radix Pseudostellariae continuous cropping obstacle field efficacy, effect
Stable, market application foreground is huge.
The impact on Radix Pseudostellariae yield of table 1 different disposal
The foregoing is only presently preferred embodiments of the present invention, all equalizations done according to scope of the present invention patent change and repair
Decorations, all should belong to the covering scope of the present invention.
SEQUENCE LISTING
<110>University Of Agriculture and Forestry In Fujian
<120>a kind of Radix Pseudostellariae Special compound microbial fertilizer and preparation method and application
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<170> PatentIn version 3.3
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Claims (8)
1. a Radix Pseudostellariae Special compound microbial fertilizer, it is characterised in that: organic materials is added specific complex microbial bacteria
Agent is made after anaerobic fermentation and oxygen consumption are fermented.
A kind of Radix Pseudostellariae Special compound microbial fertilizer the most according to claim 1, it is characterised in that: the micro-life of specific complex
Thing microbial inoculum is mixed by the ratio that general probiotics bacterium solution and original inhabitants' probiotics bacterium solution are 4:6 by volume.
A kind of Radix Pseudostellariae Special compound microbial fertilizer the most according to claim 2, it is characterised in that: described general have
Benefit bacterium bacterium solution includes lactic acid bacteria, photosynthetic bacteria, yeast, actinomycetes, and the ratio for 1:1:1:1 combines expansion training by volume
Support, or prepare through amplification culture by commercially available EM stock solution.
A kind of Radix Pseudostellariae Special compound microbial fertilizer the most according to claim 2, it is characterised in that: original inhabitants probiotics bacterium
Liquid is by the ratio that burkholderia composite bacteria liquid, pseudomonas composite bacteria liquid, bacillus cereus composite bacteria liquid are 4:1:1 by volume
Example is mixed；Described burkholderia composite bacteria liquid includes Burkholderia ambifaria, new Bulbus Allii Cepae Bai Kehuo
That moral bacterium (Burkholderia cenocepacia), each bacterium after individually fermenting, equal-volume is mixed；Described false unit cell
Bacterium composite bacteria liquid includes pseudomonas fluorescens, pseudomonas putida, Pseudomonas protegens, Korea S pseudomonas
(Pseudomonas koreensis), each bacterium after individually fermenting, equal-volume is mixed；Described bacillus cereus composite bacteria liquid
Including bacillus amyloliquefaciens, bacillus subtilis, bacillus cereus, Bacillus pumilus, bacillus megaterium, gill fungus shape bud
Spore bacillus, each bacterium after individually fermenting, equal-volume is mixed.
A kind of Radix Pseudostellariae Special compound microbial fertilizer the most according to claim 1, it is characterised in that: organic materials is by bean
The ratio mixing of the dregs of rice, Testa oryzae, dry pig manure, fish flour, brown sugar 2:1:1:1:0.03 by weight is mixed thoroughly and is made.
A kind of Radix Pseudostellariae Special compound microbial fertilizer the most according to claim 1, it is characterised in that anaerobic fermentation is concrete
Step is: in the organic materials mixed thoroughly, by volume mass ratio 1:10 adds specific complex microbial bacterial agent, adds water and fully mixes
Even, water content control is 30%, and subpackage carries out closing anaerobic fermentation, 37 DEG C of bottom fermentation 48h, then room temperature place after within 2～4 weeks, carrying out
Ripe, prepare anaerobic fermentation product.
A kind of Radix Pseudostellariae Special compound microbial fertilizer the most according to claim 1, it is characterised in that oxygen consumption fermentation is concrete
Step is: access specific complex microorganism according still further to volume mass than 1:30 in the anaerobic fermentation product described in claim 6
Microbial inoculum, in equipment that is airtight and that ventilate, 37 DEG C of fermentation 48h, prepare Special compound microbial fertilizer.
8. the Radix Pseudostellariae Special compound microbial fertilizer as claimed in claim 1 application on preventing and treating Radix Pseudostellariae continuous cropping obstacle.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106433676A (en) * | 2016-09-23 | 2017-02-22 | 河海大学 | Soil conditioner and application thereof in organic agriculture fertilizing for newly cultivated coastal beach saline-alkali soil |
| CN107556093A (en) * | 2017-08-03 | 2018-01-09 | 甘肃农业大学 | A kind of special hybrid bacterial strain microbial manure of solanaceous vegetables and preparation method thereof |
| CN107721539A (en) * | 2017-09-12 | 2018-02-23 | 甘肃农业大学 | A kind of samphire is special to mix bacterial manure and preparation method |
| CN108530147A (en) * | 2018-07-20 | 2018-09-14 | 贵州珍安药业有限公司 | A kind of radix pseudostellariae special fertilizer and preparation method thereof |
| CN110981591A (en) * | 2019-12-20 | 2020-04-10 | 海南科大林业有限公司 | Special growth-promoting fertilizer for oil tea, and preparation and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103667158A (en) * | 2013-12-27 | 2014-03-26 | 中国热带农业科学院香料饮料研究所 | Bacillus cereus and application of bacillus cereus |
| CN104761308A (en) * | 2014-12-04 | 2015-07-08 | 安徽汇丰园生物科技有限公司 | Preparation and application of soil ecosystem nursing microorganism bacterial fertilizer |
| CN105199995A (en) * | 2015-10-27 | 2015-12-30 | 山西省农业科学院生物技术研究中心 | Micro-ecological composite inoculant special for overcoming watermelon continuous cropping obstacle and application method |
| CN105199963A (en) * | 2015-08-13 | 2015-12-30 | 山西省农业科学院生物技术研究中心 | Micro-ecological compound microbial agent and method for processing solid crop culture medium through micro-ecological compound microbial agent |
-
2016
- 2016-04-06 CN CN201610208743.7A patent/CN105859347B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103667158A (en) * | 2013-12-27 | 2014-03-26 | 中国热带农业科学院香料饮料研究所 | Bacillus cereus and application of bacillus cereus |
| CN104761308A (en) * | 2014-12-04 | 2015-07-08 | 安徽汇丰园生物科技有限公司 | Preparation and application of soil ecosystem nursing microorganism bacterial fertilizer |
| CN105199963A (en) * | 2015-08-13 | 2015-12-30 | 山西省农业科学院生物技术研究中心 | Micro-ecological compound microbial agent and method for processing solid crop culture medium through micro-ecological compound microbial agent |
| CN105199995A (en) * | 2015-10-27 | 2015-12-30 | 山西省农业科学院生物技术研究中心 | Micro-ecological composite inoculant special for overcoming watermelon continuous cropping obstacle and application method |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106433676A (en) * | 2016-09-23 | 2017-02-22 | 河海大学 | Soil conditioner and application thereof in organic agriculture fertilizing for newly cultivated coastal beach saline-alkali soil |
| CN107556093A (en) * | 2017-08-03 | 2018-01-09 | 甘肃农业大学 | A kind of special hybrid bacterial strain microbial manure of solanaceous vegetables and preparation method thereof |
| CN107721539A (en) * | 2017-09-12 | 2018-02-23 | 甘肃农业大学 | A kind of samphire is special to mix bacterial manure and preparation method |
| CN108530147A (en) * | 2018-07-20 | 2018-09-14 | 贵州珍安药业有限公司 | A kind of radix pseudostellariae special fertilizer and preparation method thereof |
| CN110981591A (en) * | 2019-12-20 | 2020-04-10 | 海南科大林业有限公司 | Special growth-promoting fertilizer for oil tea, and preparation and application thereof |
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