CN105859347B - A kind of radix pseudostellariae Special compound microbial fertilizer and the preparation method and application thereof - Google Patents
A kind of radix pseudostellariae Special compound microbial fertilizer and the preparation method and application thereof Download PDFInfo
- Publication number
- CN105859347B CN105859347B CN201610208743.7A CN201610208743A CN105859347B CN 105859347 B CN105859347 B CN 105859347B CN 201610208743 A CN201610208743 A CN 201610208743A CN 105859347 B CN105859347 B CN 105859347B
- Authority
- CN
- China
- Prior art keywords
- radix pseudostellariae
- bacterium
- burkholderia
- bacillus
- special compound
- 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.)
- Active
Links
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
-
- 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
-
- 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
-
- 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
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Soil Sciences (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Environmental Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Botany (AREA)
- Environmental & Geological Engineering (AREA)
- Fertilizers (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Soil Conditioners And Soil-Stabilizing Materials (AREA)
Abstract
The present invention relates to a kind of Special compound microbial fertilizers for preventing and treating radix pseudostellariae continuous cropping obstacle, and by specific complex microbial bacterial agent and organic materials (beans cypress, rice bran, drying pig manure, fish meal, brown sugar weight ratio are 2:1:1:1:0.03), fermentation is made together.The complex micro organism fungicide is mixed by general beneficial bacterium bacterium solution and indigenous beneficial bacterium bacterium solution by the volume ratio of 4:6.Wherein indigenous beneficial bacterium includes separating the burkholderia to radix pseudostellariae rhizosphere soil, pseudomonad, bacillus, is individually mixed and made into indigenous beneficial bacterium composite bacteria liquid by the volume ratio of 4:1:1 after fermentation.Special compound microbial fertilizer is made after specific complex microbial bacterial agent anaerobic fermentation and oxygen consumption ferment two stages in organic materials.The present invention is verified on the spot for a long time, and rationally, stable effect is obvious to prevention and treatment radix pseudostellariae continuous cropping obstacle effect for formula, is a kind of good, pollution-free, safety ecology the microbial manure of biocontrol effect, and development and application prospect is wide.
Description
Technical field
The present invention relates to a kind of radix pseudostellariae Special compound microbial fertilizers and the preparation method and application thereof, 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 technique
Radix pseudostellariae (Pseudostellaria heterophylla) alias child ginseng, caryophyllaceous ginseng, it is Caryophyllaceae perennial grass
This plant is used as medicine with root, is had and is replenished qi to invigorate the spleen, promoting production of body fluid and nourishing the lung and other effects, medication is with a long history, clinical efficacy is definite, city
Field demand is big.But there are serious Soil-sickness Problem in radix pseudostellariae planting process, also referred to as continuous cropping problem or disease is replanted
Evil, i.e., same plot plant radix pseudostellariae for years, radix pseudostellariae root tuber are caused not expand normally, and serious, production occurs for pest and disease damage
Amount quality the problems such as being remarkably decreased (as shown in Figure 1), this seriously inhibits the utilization of radix pseudostellariae medicine resource and its related industries
It is significant to explore a kind of environmentally friendly radix pseudostellariae continuous cropping obstacle decreasing measure for sustainable development.
Early-stage study discovery, radix pseudostellariae continuous cropping cause structure of soil microbial community unbalance, i.e., beneficial bacterium is reduced, pathogen
Mass propagation growth, eventually leads to 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 are considered as solving the main path of radix pseudostellariae continuous cropping obstacle.But micro- life
Object microbial inoculum or biological organic fertilizer do not have broad spectrum activity usually, and the Species of Pathogens or Damage Types that can be prevented and treated are extremely limited.Class
Topic group early period, radix pseudostellariae cultivating disease is prevented and treated by buying commercially available several anti-continuous cropping bacterial manure or microbial bacterial agent, is as a result sent out
Existing control efficiency is extremely limited, this may be with the rhizospere competition of beneficial bacterium in commercially available microbial inoculum and the host specific of pathogen
The correlations such as property.Seminar's early period from morbidity radix pseudostellariae plant and continuous cropping soil separation screening to many strong pathogenic cause of diseases
Bacterium, such as Fusarium oxysporum, fusarium moniliforme, ankle section sickle-like bacteria (as shown in Figure 2), and more pathogen strain bacterium show host spy
The opposite sex only infects radix pseudostellariae without infecting rear stubble other crops such as corn and soybean, rice etc..So separation screening can be short of money
The beneficial bacterium of anti-radix pseudostellariae specialized pathogen bacterium can just effectively prevent radix pseudostellariae cultivating disease.Early-stage study is, it was also found that radix pseudostellariae
Continuous cropping causes soil environment that significant change occurs, and acidification problem such as occurs, and a variety of cometabolisms of radix pseudostellariae sustained release produce
Object and suppression harmful bacteria, these factors all may cause the beneficial bacterium in commercially available microbial inoculum and can not colonize in radix pseudostellariae rhizosphere, therefore adopt
Host specificity and the continuous cropping of radix pseudostellariae soil-borne pathogen need to be comprehensively considered when preventing and treating radix pseudostellariae cultivating disease with microbial bacterial agent
The particularity of soil environment.
In consideration of it, Special compound microbial fertilizer of the present invention be by radix pseudostellariae specific complex microbial bacterial agent with have
Machine material ferments together to be made.The radix pseudostellariae specific complex microbial bacterial agent is by general beneficial bacterium (can directly buy) and soil
It writes beneficial bacteria (separation to radix pseudostellariae rhizosphere soil) to mix by proportion optimizing, it is therefore an objective to guarantee beneficial bacterium in radix pseudostellariae root
The success on border colonizes, can activating soil nutrient, the suppression harmful bacteria of degradation radix pseudostellariae release, and radix pseudostellariae specialized form disease can be prevented and treated
Opportunistic pathogen (as shown in Figure 3).
Summary of the invention
The object of the present invention is to provide a kind of radix pseudostellariae Special compound microbial fertilizers and the preparation method and application thereof, are used for
The Soil-sickness Problem of biological control medicinal plant radix pseudostellariae plays the dual purpose for promoting root tuber growth and controlling disease.
Technical scheme is as follows:
A kind of Special compound microbial fertilizer is fermented together by complex micro organism fungicide and organic materials and is made.Described
Complex micro organism fungicide is made of general beneficial bacterium (EM) and indigenous beneficial bacterium (IM), and viable count is greater than 1.0 × 109 CFU/
mL.Wherein, general beneficial bacterium includes lactic acid bacteria, photosynthetic bacteria, saccharomycete, actinomyces, can use commercially available EM stoste routinely side
Method expands culture and obtains EM composite bacteria liquid；The indigenous beneficial bacterium includes separating to the Burkholder of radix pseudostellariae rhizosphere soil
Bacterium, pseudomonad, bacillus, respectively individually fermentation.The multiple beneficial bacterium of the same category mixes system after respectively fermenting in equal volume
At belonging to composite bacteria liquid.EM composite bacteria liquid and each indigenous beneficial bacterium (are finally followed successively by burkholderia, pseudomonad, bud
Spore bacillus) composite bacteria liquid be by volume that the ratio of 4:4:1:1 mixes.
Lactic acid bacteria in the specific complex microbial bacterial agent can be lactobacillus plantarum, streptococcus lactis, cheese cream
Bacillus；Photosynthetic bacteria can be Rhodopseudomonas palustris, hydrogenlike silicon ion；Saccharomycete can be saccharomyces cerevisiae, Candida utilis ferment
It is female；Actinomyces can be streptomyces griseus, streptomyces albus.Burkholderia can beBurkholderia 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.), new primary gram of Hall of onion
One of moral bacterium is several；Pseudomonad 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.), South Korea pseudomonad
(Pseudomonas koreensisOne or more of)；Bacillus can be bacillus amyloliquefaciens, bacillus subtilis
One or more of bacterium, bacillus cereus, bacillus pumilus, bacillus megaterium, bacillus mycoides.
The organic materials include dregs of beans, rice bran, drying pig manure, fish meal, brown sugar, by weight 2:1:1:1:0.03 into
Row mixing is mixed thoroughly as fermentation bottom material.
The preparation method of Special compound microbial fertilizer:
A kind of Special compound microbial fertilizer is fermented together by specific complex microbial bacterial agent and organic materials and is made.Its
In dedicated complex micro organism fungicide by general beneficial bacterium bacterium solution (EM composite bacteria liquid) and indigenous beneficial bacterium bacterium solution (IM composite bacteria liquid)
It is mixed by volume for the ratio of 4:6.
(1) EM composite bacteria liquid: being by volume the ratio of 1:1:1:1 including lactic acid bacteria, photosynthetic bacteria, saccharomycete, actinomyces
Example joint expands culture, or, expansion culture activated with commercially available EM stoste is made.
(2) IM composite bacteria liquid: by burkholderia composite bacteria liquid, pseudomonad composite bacteria liquid, bacillus 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 primary gram of onion
Hall moral bacterium, each bacterium mix in equal volume after individually fermenting.
B. pseudomonad composite bacteria liquid includes following bacterium: Pseudomonas fluorescens, pseudomonas putida,Pseudomonas protegens, South Korea pseudomonad, each bacterium mixes in equal volume after individually fermenting.
C. bacillus composite bacteria liquid includes following bacterium: bacillus amyloliquefaciens, bacillus subtilis, waxy gemma
Bacillus, bacillus pumilus, bacillus megaterium, bacillus mycoides, each bacterium mix in equal volume after individually fermenting.
(3) ratio that EM composite bacteria liquid and IM composite bacteria liquid are 4:6 by volume is mixed and made into specific complex microorganism
Microbial inoculum.
(4) organic materials beans cypress, rice bran, dry pig manure, fish meal, brown sugar are carried out by weight the ratio of 2:1:1:1:0.03
Mixing is mixed thoroughly.
(5) anaerobic fermentation: being added specific complex microbial bacterial agent by volume mass ratio 1:10 into the organic materials mixed thoroughly,
Water is added to mix well, 30%, packing carries out closed anaerobic fermentation for water content control, and ferment 48h at 37 DEG C, then it is placed at room temperature for 2~
Progress after-ripening in 4 weeks.
(6) oxygen consumption ferments: accessing special microorganism liquid according still further to volume mass ratio 1:30 in above-mentioned anaerobic fermentation product
Body microbial inoculum, 37 DEG C of fermentation 48h in closed and ventilation equipment, obtained 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 by one starting of specific complex microbial bacterial agent and organic materials
Ferment is made.Wherein specific complex microbial bacterial agent contains radix pseudostellariae rhizosphere original inhabitants' Antagonistic Fungi, including burkholderia, false unit cell
Bacterium and bacillus are to screen to obtain from radix pseudostellariae rhizosphere soil, its rhizospere competition is good when Field information and pacifies
It is complete reliable, and to rear stubble, other crops (such as corn and soybean, rice, vegetables) are equal without the infection ability that causes a disease.
(2) indigenous Antagonistic Fungi of the present invention is obtained by a large amount of screening operation, is thin from 3000 plants of soil
Screening obtains in bacterium, has very to radix pseudostellariae specialized pathogen bacterium (such as Fusarium oxysporum, fusarium moniliforme, ankle section mould)
Strong antagonistic effect (as shown in Figure 3), wherein the new Burkholderia cepacia of strong Antagonistic Fungi,Burkholderia ambifaria、Pseudomonas protegens, South Korea pseudomonad be preserved in China Microbiological on January 28th, 2016
Culture presevation administration committee common micro-organisms center, deposit number are respectively as follows: CGMCC NO. 12119, CGMCC NO.
12120、CGMCC NO. 12121、CGMCC NO. 12122。
(3) suppression harmful bacteria that joined higher concentration when indigenous Antagonistic Fungi screening of the present invention into culture medium, makees
To select pressure, the suppression harmful bacteria that the most of indigenous beneficial bacterium being sieved to secretes radix pseudostellariae root system has the function of Degradation and Transformation.
(4) indigenous Antagonistic Fungi of the present invention also has dissolving P capacity other than being able to suppress pathogen, can be effective
Activating soil nutrient (as shown in Figure 4) improves soil physico-chemical property, and plant nutrient absorption aspects is promoted to play a significant role.
(5) present invention is verified on the spot for a long time, and rationally, stable effect is bright to prevention and treatment radix pseudostellariae continuous cropping obstacle effect for formula
It is aobvious, improve the yield and quality (as shown in Figure 5) of continuous cropping radix pseudostellariae.
Detailed description of the invention
Fig. 1 is that continuous cropping and first planting radix pseudostellariae underground part growing way compare (A) and incidence (B).
Fig. 2 is the colonial morphology A: Fusarium oxysporum for the pathogen that radix pseudostellariae plant diseases root is separated to；B a: beading
Sickle-like bacteria；C: ankle section mould.
Fig. 3 is the opposite culture photo A of indigenous Antagonistic Fungi and radix pseudostellariae specialized pathogen fungi (fusarium moniliforme): only
It is inoculated with pathogen fusarium moniliforme；B: indigenous beneficial bacterium and fusarium moniliforme opposite culture.
Fig. 4 is indigenous beneficial bacterium dissolving P capacity measurement.Arrow instruction has the indigenous beneficial bacterium of dissolving P capacity.
Fig. 5 is that continuous cropping, the first planting and special microorganism microbial inoculum handle lower radix pseudostellariae root tuber growing state comparison in difference.
Specific embodiment
Technical solution of the present invention is described in detail below in conjunction with specific embodiment.Following embodiment is used only for
The description and interpretation present invention, without constituting the limitation to technical solution of the present invention.
The screening and its identification of the indigenous Antagonistic Fungi of embodiment 1
Pseudomonad selective medium, bacillus selective medium is respectively adopted to radix pseudostellariae rhizosphere vacation unit cell
Bacterium, bacillus are separated, and are degraded using inorganic medium (the radix pseudostellariae suppression harmful bacteria containing higher concentration) to suppression harmful bacteria
Bacterium is separated.
The single bacterium colony selected at random in above-mentioned different culture medium is evaluated them using the method for PDA plate face-off respectively
To the antagonistic effect of radix pseudostellariae specialized pathogen bacterium (such as Fusarium oxysporum, fusarium moniliforme, ankle section mould).To have strong short of money
The bacterium of anti-ability carries out Molecular Identification using 16s rRNA gene or 16s-23s rRNA section PCR 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').
The culture medium prescription are as follows:
(1) pseudomonad selective medium: 20g Soybean-casein digest agar medium is weighed, 495.5mL is added
Distilled water heating stirring adds 1mL 0.1%(wt/vol after completely dissolution) crystal violet stock solution, 121 °C of high pressure sterilizations
15min adds 3.5mL 5%(wt/vol into culture medium when being cooled to about 50 °C again) Nitrofurantoin stock solution (use N, N-
Dimethylformamide makees solvent), a kind of rhyme scheme in Chinese operas serving as the prelude to a complete score for voices is cooling, spare.
(2) bacillus selective medium: every liter of peptone containing 5.0g, 1.5g yeast extract, 1.5g beef
Cream, 5.0g NaCl, 20g agar, pH 7.2.
(3) suppression harmful bacteria degradation bacterium culture medium: every liter contains NH4NO31.0g, NaCl 0.5g, K2HPO4·3H2O 1.5g,
MgSO47H2O 0.5g, (NH4)2SO40.5g, pH 7.2.It weighs each component to be dissolved in 1000mL distilled water, 121 °C of height
Pressure sterilizing 15min, when being cooled to 45~50 °C, is added the various suppression harmful bacteria (galla turcicas identified in radix pseudostellariae continuous cropping soil
Acid, coumaric acid, 3,4-Dihydroxybenzoic acid, P-hydroxybenzoic acid, vanillic acid, syringic acid, vanillic aldehyde, ferulic acid, benzoic acid),
Total phenol concentration 1000mg/L, after 10% radix pseudostellariae rhizosphere soil suspending liquid in enrichment culture number generation of inoculation, screening has suppression harmful bacteria degradation
The beneficial bacteria of ability and antagonism radix pseudostellariae pathogen.
It obtains 12 plants of radix pseudostellariae Rhizosphere Soils with strong antagonistic ability altogether through screening and identification and writes beneficial bacteria (as shown in Figure 3),
Including 2 plants of burkholderias (Burkholderia ambifaria, new Burkholderia cepacia)；4 pseudomonas are (glimmering
Light pseudomonad, pseudomonas putida,Pseudomonas protegens, South Korea pseudomonad)；6 bacillus (Xie Dian
Afnyloliquefaciens, bacillus subtilis, bacillus cereus, bacillus pumilus, bacillus megaterium, bacillus mycoides).
Wherein South Korea pseudomonad be South Korea pseudomonad (Pseudomonas koreensis) CJ-361, in 2016
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on January 28, in, culture presevation number is CGMCC
NO. 12122, address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
New Burkholderia cepacia be 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
Number is CGMCC NO. 12119, and address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
Pseudomonad be pseudomonad (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 number is CGMCC NO. 12121, ground
Location is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
Burkholderia be burkholderia (Burkholderia ambifaria) LK-P4, in January, 2016
It is preserved within 28th China Committee for Culture Collection of Microorganisms's common micro-organisms center, culture presevation number is CGMCC NO.
12120, address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City.
The preparation of 2 Special compound microbial fertilizer of embodiment
A kind of Special compound microbial fertilizer of the present invention, together by specific complex microbial bacterial agent and organic materials
Fermentation is made.Wherein specific complex microbial bacterial agent is by general beneficial bacterium bacterium solution (EM composite bacteria liquid) and indigenous beneficial bacterium bacterium solution
(IM composite bacteria liquid) is mixed by volume for the ratio of 4:6.It is specific the preparation method is as follows:
The preparation of 2.1 EM composite bacteria liquids
300g brown sugar and 50mL MS mother liquor are added into the well water (tap water uses after need at least placing 2 days) of 18L, then
The activated commercially available EM liquid spawn of 1L is added, is placed in 37 °C of indoor sealed fermenting 3 weeks, every one since fermentation second day
Its openable closed container mouth deflates decompression once, can smell sour-sweet taste when opening vessel port, and be more and more obvious, this is just
Often occur as, that is, indicate that fermentation spreads cultivation success, it is spare.Successful finished product fermentation liquid has sour-sweet taste, and pH 3.0~4.0, viable count is big
In 10,000,000,000/mL.
2.2 indigenous beneficial bacterium activation and expansion culture
1) actication of culture
By Pseudomonas fluorescens, pseudomonas putida,Pseudomonas protegens, South Korea pseudomonad connects respectively
Kind is placed in 30 °C of incubators in pseudomonad Selective solid culture medium and activates 16h.
By bacillus amyloliquefaciens, bacillus subtilis, bacillus cereus, bacillus pumilus, bacillus megaterium,
Bacillus mycoides are inoculated in respectively in bacillus selective medium, are placed in 37 °C of incubators and are activated 12h.
It willBurkholderia ambifaria, new Burkholderia cepacia be inoculated in LB solid culture respectively, set
14h is activated in 37 °C of incubators.
2) expand culture
Activated various indigenous beneficial bacteriums are inoculated in LB Liquid Culture and carry out fermented and cultured, pseudomonad is placed in
Expand culture under 30 °C, other indigenous beneficial bacteriums, which are placed under 37 °C, expands culture, when viable count is higher than 1.0 × 109 When CFU/mL
Stop expanding culture.The multiple beneficial bacterium of the same category is mixed in equal volume and belongs to composite bacteria liquid, such as burkholderia
Composite bacteria liquid, pseudomonad composite bacteria liquid, bacillus composite bacteria liquid, it is spare.
The preparation of 2.3 complex micro organism fungicides
It is by volume 4:1:1's by indigenous beneficial bacterium, that is, burkholderia, pseudomonad, bacillus composite bacteria liquid
Ratio is mixed and made into indigenous beneficial bacterium composite bacteria liquid (IM composite bacteria liquid).
Further, that the ratio that EM composite bacteria liquid and IM composite bacteria liquid are 4:6 by volume is mixed and made into radix pseudostellariae is dedicated
Complex micro organism fungicide.
The preparation of 2.4 composite microbiological fertilizers
Organic materials beans cypress, rice bran, drying pig manure, fish meal, brown sugar are carried out mixing by weight 2:1:1:1:0.03 to mix
It is even, specific complex microbial liquid microbial inoculum (1L/10kg) is added, is added water and stirred according to the degree of drying of material, water content control
System 30%(with hand pinch easily it is agglomerating but be not discharged be advisable), after mixing well pack or barrelling carry out closed anaerobic fermentation, 37 °C,
48h, fermentation finish, and are placed at room temperature for progress after-ripening in 3 weeks.Special microorganism liquid bacterial agent is accessed again in above-mentioned anaerobic fermentation product
(1L/30kg), 37 °C of oxygen consumptions fermentation 48h, are made Special compound microbial fertilizer, total viable count in closed and ventilation equipment
1.0~5.0 hundred million/g.
3 Special compound microbial fertilizer of embodiment prevents and treats the evaluation of radix pseudostellariae continuous cropping obstacle field efficacy
Radix pseudostellariae continuous cropping soil is improved in advance using the Special compound microbial fertilizer of preparation as base fertilizer first, it
Site preparation plays furrow afterwards, plants radix pseudostellariae, and timely and appropriate discovery pouring root is imposed for the dedicated of fermentation fertilizer since radix pseudostellariae seedling stage
Complex microorganism liquid bacterial agent carries out volume variance comparison in harvest time.Specific control measure is as follows:
The organic base fertilizer preparation of 3.1 microorganisms
Organic materials beans cypress, rice bran, drying pig manure, fish meal, brown sugar are carried out mixing by weight 2:1:1:1:0.03 to mix
It is even, specific complex microbial liquid microbial inoculum (1L/10kg) is added, water content control is 30%, and packing carries out close after mixing well
Anaerobic fermentation, 37 DEG C, 48h are closed, fermentation finishes, and is placed at room temperature for 3 weeks with after-ripening.Later, it is connect again in above-mentioned anaerobic fermentation product
Enter special microorganism liquid bacterial agent (1L/30kg), 37 DEG C of oxygen consumptions fermentation 48h, are made dedicated multiple in closed and ventilation equipment
Close microbial manure, total 1.0~5.0 hundred million/g of viable count.
3.2 soil improvement
Microbial manure processing group: after radix pseudostellariae harvest, first time site preparation is carried out to plot, cooperation applies fertilizer to the subsoil to be applied per acre
The Special compound microbial fertilizer 1200kg for fermenting in advance and (at least shifting to an earlier date 1 month and start to ferment), deep ploughing are ploughed under in soil, are harrowed even
Furrow are played, cover black mulch film, furrow irrigation water to furrow face micro- wet (but not flooding to furrow face) of spreading and sinking in is cut off the water, and overlay film ferments at least one moon,
To improve the Continuous Cropping Soil Environment repaired and deteriorated, radix pseudostellariae is planted again later.During test, other field management measures and the positive
Control group, negative control group are consistent.
Positive controls: in the plot top for never planting radix pseudostellariae time plantation radix pseudostellariae, the present invention is not during which applied
The complex micro organism fungicide or composite microbiological fertilizer, as first planting radix pseudostellariae.
Negative control group: planting radix pseudostellariae on the plot that one is planted radix pseudostellariae for years again, and does not apply this
The invention complex micro organism fungicide or composite microbiological fertilizer, as continuous cropping radix pseudostellariae.
The plantation of 3.3 radix pseudostellariaes
The last week is planted, second of site preparation is carried out, opens deep 9 centimetres of straight trip item by 15 centimetres of line-spacing in whole good furrow face
Ditch.Before plantation, is applied in kind of planting ditch after using compound fertilizer 20kg, phosphate fertilizer 25kg, potash fertilizer 10kg to mix per acre and make seed manure, slightly a little lids
Then soil is tiltedly arranged seed ginseng by 6 centimetres of spacing in the rows in furrow bank, upward, bud is away from Horizon 5cm or so, light earthing for bud head.In late November
It is planted to December, uses 25~35 kilograms of seed ginseng per acre.
3.4 impose bacterium solution
In mid or late Febuary, after coming up, pouring root applies 100 times in diluted specific complex microbial inoculum 1 time, 3 months
Ten days and mid-April, then respectively pouring root apply 50 times diluted specific complex microbial inoculum each 1 time, with resist soil-borne disease hair
It is raw.According to radix pseudostellariae growing way and incidence, pouring root number can suitably be increased by adjusting pouring root number if there is disease symptom.Sun
Property, negative control group pouring root apply equivalent water.
3.5 water and fertilizer management
The first tenday period of a month in March carry out first time topdressing in conjunction with tillage and weeding, apply organic compound fertilizer 25kg and urea per acre
2.5kg.Second of top dressing is carried out in mid or late April, 5kg potassium dihydrogen phosphate is sprayed per acre, needs first to be made into 0.5% before spraying
Solution sprayed 1 time every 8 days, sprayed 3 times.
Radix pseudostellariae Seedling Stage requires ground moistening, and when weather arid will water in time, and the middle and late growth stage meets high temperature and drought season
Section still needs to carry out morning and evening irrigation as needed, keeps furrow face wet, is conducive to root of hair and plant strain growth.But radix pseudostellariae is afraid of flood, meets more
Rainy day gas will dredge gutter in time, pay attention to draining, in order to avoid rotten.
3.6 harvests and volume variance compare
When late June radix pseudostellariae plant withers sprout tumble, fine day is selected to excavate radix pseudostellariae.With spud deep-cut 15 centimetres with
On, radix pseudostellariae root tuber is completely dug out, root tuber is avoided damage to, sorts out, clean, dry, claim fresh weight.
Many years test result shows that compared with continuous cropping radix pseudostellariae, applying invented Special compound microbial fertilizer can show
Write and inhibit incidence, improve radix pseudostellariae per mu yield, volume increase about 30~65%, yield and first planting radix pseudostellariae quite (as shown in figure 5,
Table 1).As it can be seen that Special compound microbial fertilizer of the present invention is significant to prevention and treatment radix pseudostellariae continuous cropping obstacle field efficacy, effect
Stablize, market application prospect is huge.
Influence of 1 different disposal of table to radix pseudostellariae yield
The foregoing is merely presently preferred embodiments of the present invention, all equivalent changes done according to scope of the present invention patent with
Modification, is all covered by the present invention.
SEQUENCE LISTING
<110>University Of Agriculture and Forestry In Fujian
<120>a kind of radix pseudostellariae Special compound microbial fertilizer and the preparation method and application thereof
<130> 4
<160> 4
<170> PatentIn version 3.3
<210> 1
<211> 20
<212> DNA
<213>artificial sequence
<400> 1
agagtttgat cctggctcag 20
<210> 2
<211> 20
<212> DNA
<213>artificial sequence
<400> 2
aaggaggtga tccagccgca 20
<210> 3
<211> 18
<212> DNA
<213>artificial sequence
<400> 3
ttgtacacac cgcccgtc 18
<210> 4
<211> 19
<212> DNA
<213>artificial sequence
<400> 4
cctttccctc acggtactg 19
Claims (6)
1. a kind of radix pseudostellariae Special compound microbial fertilizer, it is characterised in that: specific complex microbial bacteria is added in organic materials
Agent is made after anaerobic fermentation and oxygen consumption fermentation；
Specific complex microbial bacterial agent is mixed by the ratio that general beneficial bacterium bacterium solution and indigenous beneficial bacterium bacterium solution are 4:6 by volume
It is made；
Indigenous beneficial bacterium bacterium solution is pressed by burkholderia composite bacteria liquid, pseudomonad composite bacteria liquid, bacillus composite bacteria liquid
Volume ratio is that the ratio of 4:1:1 is mixed；The burkholderia composite bacteria liquid includesBurkholderia ambifaria、New Burkholderia cepacia (Burkholderia cenocepacia), it is isometric after individually being fermented by each bacterium
It is mixed；The pseudomonad composite bacteria liquid include Pseudomonas fluorescens, pseudomonas putida,Pseudomonas protegens, South Korea pseudomonad (Pseudomonas koreensis), it is mixed in equal volume after individually being fermented by each bacterium；
The bacillus composite bacteria liquid includes bacillus amyloliquefaciens, bacillus subtilis, bacillus cereus, short and small gemma bar
Bacterium, bacillus megaterium, bacillus mycoides are mixed in equal volume after individually being fermented by each bacterium；
Wherein South Korea pseudomonad be South Korea pseudomonad (Pseudomonas koreensis) CJ-361, in January, 2016
It is preserved within 28th China Committee for Culture Collection of Microorganisms's common micro-organisms center, culture presevation number is CGMCC NO.
12122；
New Burkholderia cepacia be new Burkholderia cepacia (Burkholderia cenocepacia) CJ-P8,
China Committee for Culture Collection of Microorganisms's common micro-organisms center, culture presevation number are preserved on January 28th, 2016
For CGMCC NO. 12119；
Pseudomonad be pseudomonad (Pseudomonas protegens) LK-313, it has been preserved on January 28th, 2016
State's Microbiological Culture Collection administration committee common micro-organisms center, culture presevation number is CGMCC NO. 12121；
Burkholderia be burkholderia (Burkholderia ambifaria) LK-P4, on January 28th, 2016
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, culture presevation number is CGMCC NO.
12120。
2. a kind of radix pseudostellariae Special compound microbial fertilizer according to claim 1, it is characterised in that: described general have
Beneficial bacteria bacterium solution includes lactic acid bacteria, photosynthetic bacteria, saccharomycete, actinomyces, expands training by volume for the ratio joint of 1:1:1:1
It supports, or is made with commercially available EM stoste through expanding culture.
3. a kind of radix pseudostellariae Special compound microbial fertilizer according to claim 1, it is characterised in that: organic materials is by beans
The dregs of rice, rice bran, dry pig manure, fish meal, brown sugar are mixed thoroughly by weight the ratio mixing of 2:1:1:1:0.03 and are made.
4. a kind of radix pseudostellariae Special compound microbial fertilizer according to claim 1, which is characterized in that anaerobic fermentation is specific
Step are as follows: specific complex microbial bacterial agent is added by volume mass ratio 1:10 into the organic materials mixed thoroughly, adds water and sufficiently mixes
Even, 30%, packing carries out closed anaerobic fermentation for water content control, and ferment 48h at 37 DEG C, then after being placed at room temperature for progress in 2~4 weeks
It is ripe, anaerobic fermentation product is made.
5. a kind of radix pseudostellariae Special compound microbial fertilizer according to claim 1, which is characterized in that oxygen consumption fermentation is specific
Step are as follows: access specific complex microorganism according still further to volume mass ratio 1:30 into anaerobic fermentation product as claimed in claim 4
Microbial inoculum, 37 DEG C of fermentation 48h in closed and ventilation equipment, is made Special compound microbial fertilizer.
6. application of the radix pseudostellariae Special compound microbial fertilizer as described in claim 1 on prevention and treatment radix pseudostellariae continuous cropping obstacle.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610208743.7A CN105859347B (en) | 2016-04-06 | 2016-04-06 | A kind of radix pseudostellariae Special compound microbial fertilizer and the preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610208743.7A CN105859347B (en) | 2016-04-06 | 2016-04-06 | A kind of radix pseudostellariae Special compound microbial fertilizer and the preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105859347A CN105859347A (en) | 2016-08-17 |
| CN105859347B true CN105859347B (en) | 2019-05-03 |
Family
ID=56627153
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610208743.7A Active CN105859347B (en) | 2016-04-06 | 2016-04-06 | A kind of radix pseudostellariae Special compound microbial fertilizer and the preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105859347B (en) |
Families Citing this family (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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105859347A (en) | 2016-08-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105779305B (en) | Special compound microbial agent for eliminating successive cropping obstacles of radix pseudostellariae and preparation method thereof | |
| CN105859347B (en) | A kind of radix pseudostellariae Special compound microbial fertilizer and the preparation method and application thereof | |
| CN101659932B (en) | Antagonistic bacteria preventing and removing continuous cropping tobacco bacterial wilt and microbial organic fertilizer thereof | |
| CN101659934B (en) | Antagonistic bacteria preventing and removing continuous cropping banana Panama wilt disease and microbial organic fertilizer thereof | |
| CN101864376B (en) | Pseudomonas fluorescens strain, microbial inoculum and use thereof as seedling culture medium for controlling tomato bacterial wilt | |
| CN101659933A (en) | Antagonistic bacteria preventing and removing continuous cropping tomato bacterial wilt and microbial organic fertilizer thereof | |
| CN101696395B (en) | Antagonistic bacterium for preventing and killing off continuous cropping tobacco black shank and microbial organic fertilizer thereof | |
| CN106591193A (en) | Bacillus amyloliquefaciens with broad spectrum growth-promoting and stress-resisting effects | |
| CN102876576A (en) | Acidic soil conditioner, and preparation method and application thereof | |
| CN104531533A (en) | Composite microbial inoculant for controlling plant continuous-cropping diseases and preparation method and application thereof | |
| CN109971680A (en) | A kind of preparation and application of the complex micro organism fungicide with disease-resistant growth-promoting function | |
| CN106497831A (en) | A kind of preventing and treating Phytophthora nicotianae disease composite bacteria agent capable and its preparation method and application | |
| CN109852565A (en) | A kind of salt-soda soil composite modifying agent and its method of administration | |
| CN104016745B (en) | microbial organic fertilizer and preparation method thereof | |
| CN105199995B (en) | One kind overcoming the special micro-ecological complex bacterial agent of watermelon continuous cropping and application process | |
| CN101948780B (en) | Antagonist bacterium for preventing and treating continuous cropping hot pepper epidemic disease and microbial organic fertilizer thereof | |
| CN109810924A (en) | A kind of heavy salinized ground modification method | |
| CN101886055B (en) | Antagonistic bacteria NJL-14 for preventing and controlling continuous-cropping tobacco bacterial wilt | |
| CN108552235A (en) | A kind of bio-bacterial manure and its preparation and application for preventing flue-cured tobacco root rot and improveing soil | |
| CN110577916A (en) | Compound microbial inoculum for preventing and treating crop root-knot nematode and preparation method thereof | |
| CN107325821A (en) | A kind of biomass carbon base microbe microbial inoculum for preventing and treating watermelon blight and preparation method and application | |
| CN105439725A (en) | Paenibacillus polymyxa pesticide-fertilizer for farm onsite fermentation and applications thereof | |
| CN106007824B (en) | Composite bacterial fertilizer and preparation method and application thereof | |
| CN108676748A (en) | One plant of pear tree rhizosphere growth-promoting bacillus mycoides Lzh-Z7 and its application | |
| CN108118000A (en) | A kind of high concentration disease-resistant compound microbial bacterial manure and production method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |