CN106116883B - A kind of preparation method of tree peony pouring root composite microbic bacterial fertilizer - Google Patents
A kind of preparation method of tree peony pouring root composite microbic bacterial fertilizer Download PDFInfo
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Abstract
The present invention relates to a kind of preparation methods of tree peony pouring root composite microbic bacterial fertilizer, comprising the following steps: carries out the obtained fermentation material I that ferments to sweet potato waste and fruit waste by candida tropicalis；Fermentation material II is made in chicken manure, cow dung, biogas scum and wheat stalk mixing spontaneous fermentation；Fermentation material I is mixed with fermentation material II by proper proportion, access geotrichum candidum, bacillus megaterium and bacillusmusilaginosiengineering, which ferment, is made fermentation material III；Be added in fermentation material III it is suitable containing mycelia inside and outside two kinds of mycorrhizal fungal spores of Glomus intraradices and Glomus mosseae, root and infect mixture and fish meal, the zinc sulfate and bacillus subtilis HS5B5 bacterium solution of plant root segment, mixing obtains tree peony pouring root composite microbic bacterial fertilizer.The present invention is according to tree peony growing nutrient and environmental demand, and using agricultural by-products as raw material, the bacterial manure of preparation can provide protection for tree peony root, inhibits pathogenic microorganism, provides full nutrition for tree peony growth, promotes growth, blooming period prolonging.
Description
Technical field
The present invention relates to microbial-bacterial fertilizer preparation method fields, and in particular to a kind of tree peony pouring root composite microbic bacterial fertilizer
Preparation method.
Background technique
Tree peony is a kind of flowers of famous ornamental class in China, and flower is very large, elegant and poised, very popular.
Tree peony is grown in neutral or slightly alkaline soil, not acidproof, intolerant to heavy soil.Tree peony is fertilizer favorite, especially the elements such as N P and K
More complete fertilizer, currently used fertilizer are the organic fertilizers such as inorganic compound fertilizer or feces of livestock and poultry.Tree peony is fleshy root, is applied
It must be applied with inorganic compound fertilizer away from the farther away place of main root, if fertilising is too close away from main root, easily cause the mistake of tree peony root
Water, and then the absorption of mineral element is influenced, long-time service inorganic composite fertilizer be easy to cause the hardened and groundwater resources of soil
Pollution；Application of organic fertilizers has to pass through that retting is decomposed, can ferment without decomposed organic fertilizer and release organic acid, and then cause male
Red root rots.In addition, tree peony is during the growth process, it is easy to be influenced by unsuitable environmental condition and pathogenic microorganism is invaded
Dye, leaf, branch and the rhizome portion of tree peony can become the position of pathogenic microorganism harm, and currently used solution is usually
Apply chemical pesticide or wipe out disease branches and leaves etc., but a large amount of application chemical pesticides easily make pathogenic microorganism generate anti-medicine for a long time
Property, while environmental pollution and the ecological balance being caused to destroy.For tree peony as ornamental flower, tourist can also be threatened by applying chemical pesticide
Health.
With the development of agricultural, by-product and waste are also increasingly more, and environment is made in the fermentation of their bulk depositions
At serious pollution, the rancid smell and harmful substance released causes adverse effect to people's lives, if substrate can be used as,
It is also a kind of good solution for producing bio-feritlizer.
Bacillus subtilis (Bacillus subtilis) HS5B5 in the present invention, is preserved in China on May 9th, 2012
Microbiological Culture Collection administration committee common micro-organisms center (CGMCC), address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number Institute of Microorganism, Academia Sinica, deposit number: CGMCC No.6088 is (be shown in application No. is 201410163640.4 hairs
Bright patent application specification).Bacillus subtilis HS5B5 is mould to tree peony branch spore for the male of disease fungus by Ant agonism
Red brown spot has inhibitory effect, also has certain inhibitory effect to other Common fungi diseases.
Summary of the invention
The purpose of the present invention is to provide a kind of preparation methods of tree peony pouring root composite microbic bacterial fertilizer, pass through this method
The composite microbic bacterial fertilizer of preparation can not only grow for tree peony provides various nutritional ingredients, but also it is micro- to be able to suppress tree peony cause of disease
The growth of biology provides microorganism protection for tree peony root.
In order to solve the above technical problems, the technical solution adopted by the present invention is that: a kind of tree peony pouring root composite microbial bacteria
The preparation method of fertilizer, comprising the following steps:
Step 1: candida tropicalis is taken to be activated, culture to logarithmic growth phase is obtained mixed containing candida tropicalis
Close object A；1:4 in mass ratio takes sweet potato waste and fruit waste, is crushed to 1.5cm hereinafter, total with sweet potato waste and fruit waste
On the basis of quality, 1% urea is added, is uniformly mixing to obtain mixture B；Mixture A is inoculated into mixture B, inoculum concentration
It is the 1% of mixture B mass, fermentation 4-5 days is stirred at room temperature, fermentation material I is made；
Step 2: 1:1:2-3:2-3 in mass ratio takes chicken manure, cow dung, biogas scum and is crushed to 1cm Wheat Straw below
Stalk is uniformly mixed and obtains mixture C, on the basis of the quality of mixture C, 5% quick lime is added, is exposed in air, from
It so places 3-5 days, fermentation material II is made；
Step 3: fermentation material II made from step 2 is added in fermentation material I made from step 1, wherein fermentation material II
It is 3:1 with I mass ratio of fermentation material, obtains mixture D after mixing；In mixture D inoculation in logarithmic growth phase geotrichum candidum,
Bacillus megaterium and bacillusmusilaginosiengineering, and the inoculum concentration of each bacterial strain is the 1% of mixture D quality, in 30 DEG C of conditions
Lower stirring is fermented 4-6 days, and fermentation material III is made；
Step 4: using clover as host plant, expand numerous Glomus intraradices in expanding numerous matrix, inoculum concentration 3%-5%, when
When Glomus intraradices spore is reached in every gram of soil containing 30 or more, smashes clover root and expand numerous matrix, obtain containing sacculus in root
Mycelia and the mixture E 1 of root segment is infected inside and outside mycorrhiza fungal spore, root；Using clover as host plant, expand in expanding numerous matrix
Numerous Glomus mosseae, inoculum concentration 3%-5% smash lucerne when Glomus mosseae spore reaches in every gram of soil containing 30 or more
Mu root and the numerous matrix of expansion, obtain containing mycelia inside and outside Glomus mosseae mycorrhizal fungal spore, root and infect the mixture E 2 of root segment；
Mixture E 1 and the 1:1 in mass ratio of mixture E 2 are uniformly mixed and obtain mixture E；
Step 5: 100:90:2:1 in mass ratio takes fermentation material III, mixture E, fish meal and zinc sulfate, by mixture E, fish
Powder and zinc sulfate are added in fermentation material III, and the mixture F of semi-solid is made after mixing；
Step 6: taking bacillus subtilis HS5B5, activate in seed culture medium, is transferred in fermentation medium after activation
Fermentation, when bacillus subtilis HS5B5 grows to stationary phase, obtains the mixture G containing bacillus subtilis HS5B5；
Step 7: mixture F made from step 5 is mixed with mixture G 3-5:1 in mass ratio made from step 6, stir
It mixes and uniformly obtains tree peony pouring root composite microbic bacterial fertilizer.
Further, fruit waste described in step 1 be apple shedding, apple corruption fruit, the fresh slag of apple, in pineapple bran
Any one or any mixture.
Further, fruit waste described in step 1 is the mixture of apple fresh slag and apple shedding, and apple is fresh
Slag and the mass ratio of apple shedding are 1:3.
Further, sweet potato waste described in step 1 and fruit waste are crushed to 0.8cm or less.
Further, the mass ratio of chicken manure, cow dung described in step 2, biogas scum and wheat stalk is 1:1:2:3.
Further, step 4 expands the numerous matrix of expansion used in numerous Glomus intraradices and Glomus mosseae by river sand and soil by matter
Amount is made than sterilizing after 1:1 mixing.
Further, seed culture medium described in step 6 is nutrient broth medium.
Further, the activation temperature of bacillus subtilis HS5B5 described in step 6 is 30 DEG C, and a length of 11 is small when activation
When.
Further, fermentation medium described in step 6 is using every addition 1000ml water as reference, component and its addition
Amount is respectively as follows: corn flour 15.0g, glucose 3.0g, soy meal 15.0g, peptone 3.0g, MgSO4·7H2O 0.5g,
KH2PO4·2H2O 1.0g, water 1000ml adjust pH to 7.0.
Further, the condition that bacillus subtilis HS5B5 ferments in the fermentation medium described in step 6 are as follows: 30 DEG C
Aerobic fementation 25 hours.
The utility model has the advantages that
Various raw materials and microorganism in the present invention make prepared tree peony pouring root with again under the processing of related process
The various composition closed in microbial-bacterial fertilizer mutually cooperates with, and is finally reached and promotes tree peony growth, blooming period prolonging, prevents and treats Peony disease
Purpose；
The tree peony pouring root composite microbic bacterial fertilizer prepared through the invention enters soil through pouring root, various micro- in bacterial manure
Form delicate dynamic equilibrium between biology and edaphon, make in bacterial manure various organic matters convert between each microorganism with
It utilizes, finally releases the nutriment that tree peony can directly be absorbed and utilized and be absorbed and utilized into soil, and then by tree peony.In root
Sacculus is mould and Glomus mosseae is in soil-grown reproductive process, and most of mycelia spreads around tree peony rhizosphere and and tree peony root
It is symbiosis, increases tree peony root system to the absorption area of nutriment, be conducive to absorption of the tree peony to mineral elements such as phosphorus, simultaneously
Glomus intraradices and Glomus mosseae are mutually cooperateed with bacillus subtilis HS5B5, resist pathogenic microorganism jointly to tree peony
Invasion；The zinc sulfate of addition can significantly improve itself resistance of tree peony and the resistivity of multiple diseases, accelerate chlorophyll
Formation, it is flourishing, promote the nutrient growth of tree peony；
The fresh slag of the raw materials used in the present invention sweet potato waste, apple, apple shedding, chicken manure, cow dung, biogas scum, wheat stalk are equal
Can not only it alleviate after method provided by the invention processing for agricultural by-products cheap and easy to get, waste or house refuse
The pressure of environmental pollution, and bacterial manure obtained can satisfy demand of the tree peony to nutritional ingredient, while penetrating to soil is improved
Property, enhancing soil fertility have good effect, tree peony root growth can also be promoted, blooming period prolonging, inhibit the micro- life of tree peony cause of disease
The growth of object provides microorganism protection for tree peony root.
Specific embodiment
The present invention will be further described in detail below with reference to specific embodiments.
A kind of preparation method of tree peony pouring root composite microbic bacterial fertilizer, comprising the following steps:
Step 1: candida tropicalis is taken (malt extract powder 130g to be added in every 1000ml water, chlorine is mould in malt extract medium
Plain 0.1g；PH is adjusted to being activated in 6.0), is cultivated to logarithmic growth phase, is obtained mixture A, contain malt in mixture A
Juice culture medium and the rapid Candida tropicalis body of growth and breeding and its metabolite；Growth is in the false silk in the torrid zone that logarithm increases
Yeast accesses in new culture medium, and candida tropicalis can be made to adapt to new growing environment faster, shortens period of delay, to contract
Short fermentation time；
1:4 in mass ratio takes sweet potato waste and fruit waste, is crushed to 1.5cm hereinafter, with sweet potato waste and fruit waste
On the basis of gross mass, 1% urea is added, is uniformly mixing to obtain mixture B；Mixture B is substantially with sweet potato waste and fruit
Waste is a kind of culture medium that can be used to breed Candida tropicalis body of raw material preparation, not only solves agriculture waste
The problem of object (i.e. sweet potato waste and fruit waste) accumulation corruption, pollution environment, alleviate environmental pressure, and sweet potato waste and water
Fruit waste also for candida tropicalis growth and breeding provide the various nutrition such as available carbon source, nitrogen source, inorganic salts, water at
Point, 1% urea, which is added, makes carbon-nitrogen ratio reach optimum state, is conducive to Candida tropicalis body growth and breeding；Sweet potato waste and
Fruit waste is crushed to 1.5cm hereinafter, when stirring fermentation, and candida tropicalis can be allowed to come into full contact with mixture B, had
Conducive to candida tropicalis growth；
Mixture A is inoculated into mixture B, inoculum concentration is the 1% of mixture B mass, fermentation 4-5 days is stirred at room temperature,
Fermentation material I is made；Using the inoculum concentration after stirring fermentation under room temperature, fermentation 4-5 days, candida tropicalis is by sweet potato
Most of macro-nutrients in slag and fruit waste are decomposed into small molecule compound easy to use, in growth course
In can synthesize a large amount of mycoproteins, after candida tropicalis growth cycle, mycoprotein again can by subsequent geotrichum candidum,
Bacillus megaterium, bacillusmusilaginosiengineering and edaphon utilize；Candida tropicalis meeting during growth and breeding
The fermentation material I for generating a large amount of organic acid substance not being decomposed in a large amount of water and carbon dioxide and pomace, therefore obtaining
It is a kind of mixture of the semisolid of slant acidity；
Step 2: 1:1:2-3:2-3 in mass ratio takes chicken manure, cow dung, biogas scum and is crushed to 1cm Wheat Straw below
Stalk is uniformly mixed and obtains mixture C, and chicken manure, cow dung and biogas scum can be employed as the organic fertilizer of plant, itself contain largely
Organic matter, if three uses individually, it will cause a large number of nutrients losses, and utilization rate is low, and contain a large amount of diseases
Opportunistic pathogen spore and parasitic ovum, peculiar smell is big, and improper use is also easy to cause burn seedlings phenomenon；In chicken manure, cow dung and biogas scum
Containing a large amount of different types of microorganisms, including aerobic microbiological, anaerobe and amphimicrobian type microorganism, chicken manure,
Cow dung and biogas scum three mixing enrich microbe species and metabolite in mixture C more, are conducive to nutriment
Conversion between various microorganisms, using and be discharged into soil, be conducive to absorption and benefit of the tree peony to various nutritional ingredients
With；Proper proportion is added in mixture C and is crushed to 1cm wheat stalk below, mixture C can be made to keep certain ventilation
Property, loosing soil can be made ventilative, meet the growth needs of tree peony fleshy root, while meeting aerobic microbiological and amphimicrobian type
Demand of the microorganism to oxygen；On the basis of the quality of mixture C, 5% quick lime is added, is exposed in air, puts naturally
It sets 3-5 days, fermentation material II is made, quick lime encounters the water in chicken manure, cow dung and biogas scum, the white lime of alkalinity is obtained, and
High temperature, the alkaline most of pathogen that can not only be killed in mixture C of high temperature and white lime are formed with a large amount of heat
And parasitic ovum, moreover it is possible to so that wheat stalk further decomposed, isolated and degrades, it is big in mixture C after spontaneous fermentation 3-5 days
Fraction of pathogens bacterium and parasitic ovum are killed, and the mixing between various raw materials is also more uniform, and the macromolecular substances in raw material exist
Under the action of microorganism, it is converted into the nutriment that more can be directly absorbed and utilized by tree peony；
Step 3: fermentation material II made from step 2 is added in fermentation material I made from step 1, the alkali of fermentation material II
Property can be neutralized with the acidity of fermentation material I, wherein fermentation material II and I mass ratio of fermentation material are 3:1, make the mixing obtained after mixing
The pH value of object D is suitble to the requirement of subsequent microbial fermentation, and finally obtained tree peony pouring root is not damaged male with composite microbic bacterial fertilizer
Red fleshy root；Inoculation is in geotrichum candidum, bacillus megaterium and the gel-shaped gemma bar of logarithmic growth phase in mixture D
Bacterium, and the inoculum concentration of each bacterial strain is the 1% of mixture D quality, 30 DEG C of stirrings are fermented 4-6 days, and fermentation material III is made；By step
After the effect of quick lime in rapid two, wheat stalk initial isolation and degradation, the macromolecular substances such as cellulose and lignin therein
It is further decomposed by geotrichum candidum and bacillus megaterium, generates dextrin, oligosaccharides and monosaccharide, it can be micro- for fermentation strain and soil
Biological utilisation；Bacillusmusilaginosiengineering can release potassium element in slightly solubility sylvite and other during growth and breeding
Middle microelement is absorbed and utilized these elements conducive to tree peony, has positive effect, gel-shaped gemma bar to peony forescence is extended
The substances such as the organic acid in the metabolite of bacterium can be directly absorbed and utilized by tree peony, promote the growth of tree peony；
Step 4: using clover as host plant, expand numerous Glomus intraradices in expanding numerous matrix, inoculum concentration 3%-5%, when
When Glomus intraradices spore is reached in every gram of soil containing 30 or more, mycorrhizal fungi largely infects root system of plant at this time, beats
Broken clover root and the numerous matrix of expansion, obtain containing mycelia inside and outside Glomus intraradices mycorrhizal fungal spore, root and infect the mixing of root segment
Object E1；Using clover as host plant, expand numerous Glomus mosseae in expanding numerous matrix, inoculum concentration 3%-5% works as Glomus mosseae
When spore is reached in every gram of soil containing 30 or more, mycorrhizal fungi largely infects root system of plant at this time, smash clover root and
Expand numerous matrix, obtains containing mycelia inside and outside Glomus mosseae mycorrhizal fungal spore, root and infect the mixture E 2 of root segment；It will mixing
The object E1 and 1:1 in mass ratio of mixture E 2, which is uniformly mixed, obtains mixture E；Glomus intraradices and Glomus mosseae are in soil-grown
In reproductive process, most of mycelia spread around tree peony rhizosphere and with tree peony Root Symbiont, increase tree peony root system to nutrition
The absorption area of substance, is conducive to absorption of the tree peony to mineral elements such as phosphorus, at the same Glomus intraradices and Glomus mosseae with it is withered
Careless bacillus HS5B5 is mutually cooperateed with, and resists invasion of the pathogenic microorganism to tree peony in soil jointly；
Step 5: 100:90:2:1 in mass ratio takes fermentation material III, mixture E, fish meal and zinc sulfate, by mixture E, fish
Powder and zinc sulfate are added in fermentation material III, and the mixture F of semi-solid is made after mixing；Protein content compares in fish meal
Height, generates a large amount of small peptide and amino acid after degradation, tree peony can be enhanced the metabolic function of tree peony, improve photosynthetic work after absorbing
With blooming period prolonging promotes the growth of tree peony root system, improves soil microenvironment；Zinc sulfate can significantly improve oneself of tree peony
Body resistance and the ability for resisting multiple diseases, accelerate the formation of chlorophyll, flourishing, promote the nutrient growth of tree peony；
Step 6: taking bacillus subtilis HS5B5, activate in seed culture medium, is transferred in fermentation medium after activation
Fermentation, when bacillus subtilis HS5B5 grows to stationary phase, obtains the mixture G containing bacillus subtilis HS5B5, this
When fermentation medium in nutrient consumption totally, a large amount of thallus (﹥ 10 is mainly contained in mixture G8CFU/ml), gemma and secondary
Raw metabolite；Bacillus subtilis HS5B5 has significant antagonism bacteriostasis to the pathogenic microorganism of tree peony multiple diseases, can
Mutually to cooperate with enhancing tree peony root system disease resistance with Glomus intraradices and Glomus mosseae；Meanwhile using finally obtained
During composite microbic bacterial fertilizer, bacillus subtilis HS5B5 microbial inoculum also is sprayed to the aerial part of tree peony, after handling in this way
To tree peony root, stem, leaf at omnibearing protection, than using the disease resisting effect of bacterial manure pouring root more obvious merely；
Step 7: mixture F made from step 5 is mixed with mixture G 3-5:1 in mass ratio made from step 6, stir
It mixes and uniformly obtains tree peony pouring root composite microbic bacterial fertilizer.
Further, fruit waste described in step 1 be apple shedding, apple corruption fruit, the fresh slag of apple, in pineapple bran
Any one or any mixture, containing a large amount of acid organic in apple shedding, apple corruption fruit, the fresh slag of apple and pineapple bran
Substance, acid and beneficial microbe colony can inhibit the breeding of tree peony pathogenic microorganism, improve saline Land degree, show
Work is built up fertility.
Further, fruit waste described in step 1 is the mixture of apple fresh slag and apple shedding, the fresh slag of apple
Mass ratio with apple shedding is 1:3；The fresh slag of sweet potato waste, apple and pectin, protein and cellulose in apple shedding are through bacterium
The substances such as the available amino acid of microbial degradation, polypeptide and fatty acid in fertilizer are conducive to the growth of tree peony absorption enhancement tree peony,
Improve tree peony resistance.
Further, sweet potato waste described in step 1 and fruit waste are crushed to 0.8cm hereinafter, when being conducive to shorten fermentation
Between.
Further, the mass ratio of chicken manure, cow dung described in step 2, biogas scum and wheat stalk is 1:1:2:3.
Further, step 4 expands the numerous matrix of expansion used in numerous Glomus intraradices and Glomus mosseae by river sand and soil by matter
Amount is made than sterilizing after 1:1 mixing.
Further, seed culture medium described in step 6 is nutrient broth medium；Fermented and cultured described in step 6
For base using every addition 1000ml water as reference, component and its additive amount are respectively as follows: corn flour 15.0g, glucose 3.0g, soy meal
15.0g, peptone 3.0g, MgSO4·7H2O 0.5g, KH2PO4·2H2O 1.0g, water 1000ml adjust pH to 7.0；Described
The activation temperature of bacillus subtilis HS5B5 is 30 DEG C, and when activation is 11 hours a length of；Bacillus subtilis described in step 6
The condition that HS5B5 ferments in the fermentation medium are as follows: 30 DEG C aerobic fementation 25 hours.
There is provided through the invention method preparation tree peony pouring root composite microbic bacterial fertilizer while to tree peony pouring root,
Tree peony cauline leaf is sprayed with bacillus subtilis HS5B5 microbial inoculum, peony plant can be made healthy and strong, enhance disease resistance, improve peony
Piece quality, blooming period prolonging increase appreciation effect.
Embodiment:
Candida tropicalis used in following embodiment, geotrichum candidum, bacillus megaterium, bacillusmusilaginosiengineering, in root
Sacculus is mould and Glomus mosseae is purchased from Shandong abyssopelagic organism Science and Technology Co., Ltd., each culture presevation address and deposit number point
Not are as follows: candida tropicalis (Candida tropicalis) is preserved in China General Microbiological culture presevation administrative center, protects
Hiding number is CGMCC 2.2735.Geotrichum candidum (Geotrichum candidum) is preserved in China General Microbiological culture presevation pipe
Reason center, preservation bacterium number are CGMCC 2.1132.Bacillus megaterium (Bacillus megaterium) is preserved in Chinese common
Microbiological Culture Collection administrative center, preservation bacterium number are CGMCC 1.2416.Gel-shaped series bacillus (Bacillus
Mucilaginosus it) is preserved in Chinese agriculture Microbiological Culture Collection administrative center, deposit number is ACCC 10012.Ball in root
Capsule mould (Glomus intraradices) and Glomus mosseae (Glomus mosseae) are that European Culture Collection Center registration is protected
Hiding, deposit number is respectively BEG 141 and BEG 167.
Embodiment 1
A kind of preparation method of tree peony pouring root composite microbic bacterial fertilizer, comprising the following steps:
Step 1: candida tropicalis is taken (malt extract powder 130g to be added in every 1000ml water, chlorine is mould in malt extract medium
Plain 0.1g；PH is adjusted to being activated in 6.0), cultivates to logarithmic growth phase, obtains the mixture A containing candida tropicalis；It presses
Mass ratio 1:1:3 takes the fresh slag of sweet potato waste, apple and apple shedding, is crushed to 0.8cm hereinafter, with sweet potato waste, the fresh slag of apple and apple
On the basis of fruit shedding gross mass, 1% urea is added, is uniformly mixing to obtain mixture B；Mixture A is inoculated into mixture B
In, inoculum concentration is the 1% of mixture B mass, and fermentation 5 days is stirred at room temperature, and fermentation material I is made；
Step 2: 1:1:2:3 in mass ratio takes chicken manure, cow dung, biogas scum and is crushed to 1cm wheat stalk below,
It is uniformly mixed and obtains mixture C, on the basis of the quality of mixture C, 5% quick lime is added, is exposed in air and puts naturally
It sets 5 days, fermentation material II is made；
Step 3: fermentation material II made from step 2 is added in fermentation material I made from step 1, wherein fermentation material II
It is 3:1 with I mass ratio of fermentation material, obtains mixture D after mixing；In mixture D inoculation in logarithmic growth phase geotrichum candidum,
Bacillus megaterium and bacillusmusilaginosiengineering, and the inoculum concentration of each bacterial strain is the 1% of mixture D quality, in 30 DEG C of conditions
Lower stirring is fermented 6 days, and fermentation material III is made；
Step 4: expanding numerous Glomus intraradices, inoculum concentration 5%, when in root in expanding numerous matrix using clover as host plant
When the mould spore of sacculus is reached in every gram of soil containing 30 or more, smashes clover root and expand numerous matrix, obtain and contain Glomus intraradices bacterium
Mycelia and the mixture E 1 of root segment is infected inside and outside mycorrhiza fungi spore, root；Using clover as host plant, expand numerous rub in expanding numerous matrix
Western sacculus is mould, inoculum concentration 5%, when Glomus mosseae spore reaches in every gram of soil containing 30 or more, smashes clover root and expansion
Numerous matrix obtains containing mycelia inside and outside Glomus mosseae mycorrhizal fungal spore, root and infects the mixture E 2 of root segment；By mixture
The E1 and 1:1 in mass ratio of mixture E 2, which is uniformly mixed, obtains mixture E；Expand expansion used in numerous Glomus intraradices and Glomus mosseae
Numerous matrix is to be made by sterilizing after river sand and soil 1:1 in mass ratio mixing；
Step 5: 100:90:2:1 in mass ratio takes fermentation material III, mixture E, fish meal and zinc sulfate, by mixture E, fish
Powder and zinc sulfate are added in fermentation material III, and the mixture F of semi-solid is made after mixing；
Step 6: taking bacillus subtilis HS5B5, is activated 11 hours for 30 DEG C in seed culture medium, be then transferred to fermentation
30 DEG C aerobic fementation 25 hours, make bacillus subtilis HS5B5 grow to stationary phase in culture medium, are made and contain bacillus subtilis
The mixture G of bacterium HS5B5；
The seed culture medium is nutrient broth medium, using every addition 1000ml water as reference, component and its is added
Dosage is respectively as follows: beef extract 5.0g, peptone 10.0g, NaC1 5.0g, water 1000ml, adjusts pH to 7.0；
For the fermentation medium using every addition 1000ml water as reference, component and its additive amount are respectively as follows: corn flour
15.0g, glucose 3.0g, soy meal 15.0g, peptone 3.0g, MgSO4·7H2O 0.5g, KH2PO4·2H2O 1.0g, water
1000ml adjusts pH to 7.0；
Step 7: mixture F made from step 5 is mixed with mixture G 5:1 in mass ratio made from step 6, stir
Uniformly obtain tree peony pouring root composite microbic bacterial fertilizer.
Embodiment 2
A kind of preparation method of tree peony pouring root composite microbic bacterial fertilizer, comprising the following steps:
Step 1: candida tropicalis is taken (malt extract powder 130g to be added in every 1000ml water, chlorine is mould in malt extract medium
Plain 0.1g；PH is adjusted to being activated in 6.0), cultivates to logarithmic growth phase, obtains the mixture A containing candida tropicalis；It presses
Mass ratio 1:1:3 takes the fresh slag of sweet potato waste, apple and apple shedding, is crushed to 0.8cm hereinafter, with sweet potato waste, the fresh slag of apple and apple
On the basis of fruit shedding gross mass, 1% urea is added, is uniformly mixing to obtain mixture B；Mixture A is inoculated into mixture B
In, inoculum concentration is the 1% of mixture B mass, and fermentation 4 days is stirred at room temperature, and fermentation material I is made；
Step 2: 1:1:2:3 in mass ratio takes chicken manure, cow dung, biogas scum and is crushed to 1cm wheat stalk below,
It is uniformly mixed and obtains mixture C, on the basis of the quality of mixture C, 5% quick lime is added, is exposed in air and puts naturally
It sets 3 days, fermentation material II is made；
Step 3: fermentation material II made from step 2 is added in fermentation material I made from step 1, wherein fermentation material II
It is 3:1 with I mass ratio of fermentation material, obtains mixture D after mixing；In mixture D inoculation in logarithmic growth phase geotrichum candidum,
Bacillus megaterium and bacillusmusilaginosiengineering, and the inoculum concentration of each bacterial strain is the 1% of mixture D quality, in 30 DEG C of conditions
Lower stirring is fermented 4 days, and fermentation material III is made；
Step 4: expanding numerous Glomus intraradices, inoculum concentration 3%, when in root in expanding numerous matrix using clover as host plant
When the mould spore of sacculus is reached in every gram of soil containing 30 or more, smashes clover root and expand numerous matrix, obtain and contain Glomus intraradices bacterium
Mycelia and the mixture E 1 of root segment is infected inside and outside mycorrhiza fungi spore, root；Using clover as host plant, expand numerous rub in expanding numerous matrix
Western sacculus is mould, inoculum concentration 3%, when Glomus mosseae spore reaches in every gram of soil containing 30 or more, smashes clover root and expansion
Numerous matrix obtains containing mycelia inside and outside Glomus mosseae mycorrhizal fungal spore, root and infects the mixture E 2 of root segment；By mixture
The E1 and 1:1 in mass ratio of mixture E 2, which is uniformly mixed, obtains mixture E；Expand expansion used in numerous Glomus intraradices and Glomus mosseae
Numerous matrix is to be made by sterilizing after river sand and soil 1:1 in mass ratio mixing；
Step 5: 100:90:2:1 in mass ratio takes fermentation material III, mixture E, fish meal and zinc sulfate, by mixture E, fish
Powder and zinc sulfate are added in fermentation material III, and the mixture F of semi-solid is made after mixing；
Step 6: taking bacillus subtilis HS5B5, is activated 11 hours for 30 DEG C in seed culture medium, be then transferred to fermentation
In culture medium: 30 DEG C aerobic fementation 25 hours, grow to bacillus subtilis HS5B5 to stationary phase, be made and contain withered grass bud
The mixture G of spore bacillus HS5B5；
The seed culture medium is nutrient broth medium, using every addition 1000ml water as reference, component and its is added
Dosage is respectively as follows: beef extract 5.0g, peptone 10.0g, NaC1 5.0g, water 1000ml, adjusts pH to 7.0；
For the fermentation medium using every addition 1000ml water as reference, component and its additive amount are respectively as follows: corn flour
15.0g, glucose 3.0g, soy meal 15.0g, peptone 3.0g, MgSO4·7H2O 0.5g, KH2PO4·2H2O 1.0g, water
1000ml adjusts pH to 7.0；
Step 7: mixture F made from step 5 is mixed with mixture G 3:1 in mass ratio made from step 6, stir
Uniformly obtain tree peony pouring root composite microbic bacterial fertilizer.
Test:
With the tree peony pouring root composite microbic bacterial fertilizer of the method preparation provided by embodiment 1, (hereinafter referred to as tree peony is special
With bacterial manure) and intermediate product obtained is tested during the preparation process.
It tests one, in the peony forescence previous moon, chooses the identical tree peony of Luoyang world peony garden Peony growing head of district's gesture
400 plants, 1., 2., 3., 4., 5., 6., 7. and 8. eight groups are randomly divided into, every group 50 plants, each component other places reason is as follows:
1. spraying tree peony branches and leaves with the dedicated bacterial manure pouring root of tree peony, while with bacillus subtilis HS5B5 microbial inoculum；
2. spraying tree peony branches and leaves with the dedicated bacterial manure pouring root of tree peony, while with water；
3. spraying tree peony branches and leaves with III pouring root of fermentation material, while with water；
4. spraying tree peony branches and leaves with mixture D pouring root, while with water；
5. spraying tree peony branches and leaves with water pouring root, while with bacillus subtilis HS5B5 microbial inoculum；
6. spraying tree peony branches and leaves with water pouring root, while with water；
7. spraying tree peony branches and leaves with common micro-organisms bacterial manure pouring root, while with bacillus subtilis HS5B5 microbial inoculum；
8. spraying tree peony branches and leaves with common micro-organisms bacterial manure pouring root, while with water；
In above-mentioned processing the dedicated bacterial manure of the tree peony of pouring root, fermentation material III, mixture D and common micro-organisms bacterial manure by
80g/ plants of uses are diluted with water to 1000ml and carry out pouring root；6. in 5., every plant of tree peony applies 1000ml water pouring root；
It is in 1 preparation process of embodiment that the bacillus subtilis HS5B5 microbial inoculum of tree peony branches and leaves is sprayed in above-mentioned processing
Mixture G is used by 10g/ plants, is diluted with water to 500ml and uniformly sprays branches and leaves；2., 3., 4., 6. and 8. in, every plant of tree peony is applied
Branches and leaves are sprayed with 500ml water.
Reprocessing in every 15 days is primary, and florescence, shoot length, abloom rate, Hua Jing, single flower quality are observed and recorded after 30 days
(fungal disease refers mainly to the common tree peony fungal disease such as tree peony brown spot, as described below with infection fungal disease strain number
Fungal disease is identical with this), the results are shown in Table 1:
Table 1 is various to handle the influence bloomed to tree peony
In table 1, tree peony is ornamental class flowers, and the whole strain peony forescence time is longer, then bring income is higher；Shoot long generation
Table current year blooms the growing way of branch, and branch is longer, and flower is upright, then is improved the case where safron in leaf, and sight flower can be increased
Effect；Abloom rate reflects bud nutritional development situation；Hua Jing and single flower quality reflect flower size, and flower is bigger, then watches
Effect is preferable；A, b, c and d indicate the conspicuousness of difference between different data, and difference is not significant between same letter, between different letters
Significant difference.
The result shows that: 1. organizing the average entire florescence is 8.21 days, and shoot longest, corolla is very large, and abloom rate 100% indicates to plant
All shoots can illustrate that nutrition status of the plant is preferable at flower on strain top, no bud abortion situation.Only 1 plant of tree peony sense simultaneously
Contaminate fungal disease；2. group and 1. group difference are unobvious, but there are 8 plants of tree peony infection fungal diseases, illustrate with the dedicated bacterial manure of tree peony
Pouring root can form tree peony root and protect, and preventive and therapeutic effect be played to tree peony fungal disease to a certain extent, if simultaneously
Tree peony branches and leaves are sprayed with bacillus subtilis HS5B5 microbial inoculum, then can form omnibearing protection to tree peony branch, leaf and root, stem,
Fungal pathogens can preferably be inhibited, it is also more preferable to the control efficiency of tree peony fungal disease；3. group is compared with 2. group, the florescence shortens
Nearly 1 day, all than 1. organizing significant decrease, introduced disease plant number was also more for abloom rate, Hua Jing and single flower quality, illustrated mixture
Arbuscular mycorrhiza mixture and fish meal, zinc sulfate in E can increase and the absorption of abundant nutrition, promotes photosynthesis, extends
At the florescence, the resistance of tree peony is improved, and reduces the generation of disease；4. group application mixture D, 3. indices are than organizing summary
Difference, analysis shows, in mixture D containing a certain amount of nitrogen for being not easy to be absorbed and utilized by tree peony, carbohydrate, protein and
A small amount of biochemical fulvic acid；5. branches and leaves are sprayed in group application bacillus subtilis HS5B5 microbial inoculum (i.e. mixture G), can certain journey
Preventing the generation of tree peony fungal disease on degree, but only carry out pouring root with water simultaneously, is compared with 1. group, indices are substantially reduced,
Tree peony has lacked a large number of nutrients, reduces the resistance of plant.1. 7. or 2. with compared with 8., average longer blooming period
20% or more, abloom rate and the quality spent also are significantly increased, and illustrate the dedicated bacterial manure of tree peony to raising tree peony quality, extension peony
Phase is more preferable than common micro-organisms bacterial manure effect.In summary it analyzes, after the dedicated bacterial manure application of tree peony prepared by embodiment 1,
It can be improved the resistance of tree peony and the quality of flower, blooming period prolonging promotes tree peony to bloom, and increases quantity of blooming.Mixture G is
After bacillus subtilis HS5B5 microbial inoculum sprays branches and leaves, it can be also substantially reduced the probability of tree peony infection fungal disease, because it is liquid
The characteristic of body state can be sprayed and pouring root compounding application in practical application.
It tests two, in tree peony vegetative growth phase, chooses the identical tree peony of Luoyang world peony garden Peony growing head of district's gesture
400 plants, eight groups are randomly divided into, specific grouping and each group processing are identical as test one.
Reprocessing in every 15 days is primary, tree peony nutrient growth situation is recorded after 45 days, the results are shown in Table 2:
Influence of the various processing of table 2 to tree peony nutrient growth
In table 2, SPAD indicates Ye Lvdu, reflects chlorophyll content in leaf blades height, plant is embodied together with Net Photosynthetic Rate
The photosynthesis assimilation ability of strain；Stomatal conductance increases, and can mitigate stomata limitation, promote photosynthetic increase, but can not also keep away
That exempts from increases rising dehydration；Water use efficiency increases, and illustrates that then transpiration enhances for stomatal conductance increase, but promote photosynthetic
The effect of effect becomes apparent；A, b, c and d indicate the conspicuousness of difference between different data, and difference is not significant between same letter,
Significant difference between different letters.
As the result is shown: 1. group tree peony growing way is best, and also only 2 plants of tree peonies infect fungal diseases；2. group and 1. group difference are not
Obviously, but there are 7 plants of tree peony infection fungal diseases, while further illustrating bacterial manure pouring root dedicated with tree peony, with withered grass gemma
Bacillus HS5B5 microbial inoculum spray tree peony branches and leaves can better Antagonistic Fungus pathogen, to the control efficiency of tree peony fungal disease
More preferably；3. group is compared with 2. group, photosynthetic rate is substantially reduced, and is illustrated to have lacked arbuscular mycorrhiza and fish meal, zinc sulfate, not only be made
Rhizosphere absorption area is reduced, and lack amino acid and zinc element, causes photosynthesis and plant resistance to reduce, tree peony nutrition is raw
Long indices have conspicuousness reduction；4. the indices of group application mixture D, nutrient growth are poorer than 3. organizing, into one
Step illustrates that the nutriment in mixture D is easy to be absorbed and utilized by tree peony not as good as 3. group；5. group application mixture G sprays branches and leaves,
It can prevent the generation of tree peony fungal disease to a certain extent, but nutriment is insufficient, reduces the resistance of plant.1. with 7.
Or 2. compared with 8., chlorophyll content is high, and photosynthesis enhancing illustrates that plant is more healthy and strong, disease resistance enhancing.In summary
Analysis, the dedicated bacterial manure of tree peony are capable of providing nutriment abundant, promote photosynthesis and the nutrient growth of tree peony, improve plant
Resistance, effectively prevent tree peony fungal disease generation.
In conclusion 2., 3., 4. with 6. four groups compare, 2. organize the tree peony chlorophyll content highest of processing, photosynthetic rate is most
By force, average florescence longest, susceptible strain number is also minimum, and it is male to illustrate that the bacterial manure of the method provided by embodiment 1 preparation can promote
Red root growth, blooming period prolonging also have certain control efficiency to tree peony fungal disease；1., 2. compare with 5. three groups, 1. organize male
Red growing way is best, and average florescence longest, shoot longest, corolla is very large, and abloom rate 100% (indicates that all shoots in plant top are equal
Can be at flower), only one two plants of tree peony introduced diseases illustrate that bacterial manure cooperation bacillus subtilis HS5B5 microbial inoculum use is anti-to tree peony
Only the effect of fungal disease is more preferable；7. or 2. 1. with compared with 8., illustrating the dedicated bacterial manure of tree peony prepared by embodiment 1 than general
Logical microbial-bacterial fertilizer has more specific aim, and the nutrient growth and blooming period prolonging to tree peony have good effect.
Claims (5)
1. a kind of preparation method of tree peony pouring root composite microbic bacterial fertilizer, which comprises the following steps:
Step 1: candida tropicalis is taken to be activated, cultivates to logarithmic growth phase, obtain the mixture containing candida tropicalis
A；1:1:3 in mass ratio takes the fresh slag of sweet potato waste, apple and apple shedding, is crushed to 0.8cm hereinafter, with sweet potato waste, the fresh slag of apple
On the basis of apple shedding gross mass, 1% urea is added, is uniformly mixing to obtain mixture B；Mixture A is inoculated into mixture
In B, inoculum concentration is the 1% of mixture B mass, and fermentation 4-5 days is stirred at room temperature, and fermentation material I is made；
Step 2: 1:1:2:3 in mass ratio takes chicken manure, cow dung, biogas scum and is crushed to 1cm wheat stalk below, mixing
Mixture C uniformly is obtained, on the basis of the quality of mixture C, 5% quick lime is added, is exposed in air, places 3-5 naturally
It, is made fermentation material II；
Step 3: fermentation material II made from step 2 is added in fermentation material I made from step 1, wherein fermentation material II and hair
I mass ratio of ferment object is 3:1, obtains mixture D after mixing；Geotrichum candidum, huge of the inoculation in logarithmic growth phase in mixture D
Bacillus and bacillusmusilaginosiengineering, and the inoculum concentration of each bacterial strain is the 1% of mixture D quality, is stirred under the conditions of 30 DEG C
Fermentation 4-6 days is mixed, fermentation material III is made；
Step 4: expanding numerous Glomus intraradices in expanding numerous matrix using clover as host plant, expand numerous matrix by river sand and soil by matter
Amount is made than sterilizing after 1:1 mixing, and Glomus intraradices inoculum concentration is 3%-5%, is contained when Glomus intraradices spore reaches in every gram of soil
At 30 or more, smashes clover root and expand numerous matrix, obtain containing mycelia inside and outside Glomus intraradices mycorrhizal fungal spore, root and invade
Contaminate the mixture E 1 of root segment；Using clover as host plant, expand numerous Glomus mosseae in expanding numerous matrix, expand numerous matrix by river sand and
It sterilizes and is made after soil 1:1 in mass ratio mixing, Glomus mosseae inoculum concentration is 3%-5%, when Glomus mosseae spore reaches every gram
When containing 30 or more in soil, smashes clover root and expand numerous matrix, obtain containing bacterium inside and outside Glomus mosseae mycorrhizal fungal spore, root
Silk and the mixture E 2 for infecting root segment；Mixture E 1 and the 1:1 in mass ratio of mixture E 2 are uniformly mixed and obtain mixture E；
Step 5: 100:90:2:1 in mass ratio takes fermentation material III, mixture E, fish meal and zinc sulfate, by mixture E, fish meal and
Zinc sulfate is added in fermentation material III, and the mixture F of semi-solid is made after mixing；
Step 6: taking bacillus subtilis HS5B5, activated in seed culture medium, be transferred in fermentation medium and ferment after activation,
When bacillus subtilis HS5B5 grows to stationary phase, the mixture G containing bacillus subtilis HS5B5 is obtained；
Step 7: mixture F made from step 5 is mixed with mixture G 3-5:1 in mass ratio made from step 6, stirring is equal
It is even to obtain tree peony pouring root composite microbic bacterial fertilizer.
2. a kind of preparation method of tree peony pouring root composite microbic bacterial fertilizer according to claim 1, it is characterised in that: step
Seed culture medium described in rapid six is nutrient broth medium.
3. a kind of preparation method of tree peony pouring root composite microbic bacterial fertilizer according to claim 2, it is characterised in that: step
The activation temperature of bacillus subtilis HS5B5 described in rapid six is 30 DEG C, and when activation is 11 hours a length of.
4. a kind of preparation method of tree peony pouring root composite microbic bacterial fertilizer according to claim 1, it is characterised in that: step
For fermentation medium described in rapid six using every addition 1000ml water as reference, component and its additive amount are respectively as follows: corn flour
15.0g, glucose 3.0g, soy meal 15.0g, peptone 3.0g, MgSO4·7H2O 0.5g, KH2PO4·2H2O 1.0g, water
1000ml adjusts pH to 7.0.
5. a kind of preparation method of tree peony pouring root composite microbic bacterial fertilizer according to claim 4, it is characterised in that: step
The condition that bacillus subtilis HS5B5 ferments in the fermentation medium described in rapid six are as follows: 30 DEG C aerobic fementation 25 hours.
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| CN101948751A (en) * | 2010-08-06 | 2011-01-19 | 孙晗笑 | Microorganism bacteria liquid beneficial for growth of plants, preparation method and applications thereof |
| CN104630102A (en) * | 2015-01-27 | 2015-05-20 | 山东苏柯汉生物工程股份有限公司 | Complex bacterium preparation for fixing nitrogen, solubilizing phosphorus and potassium and rooting and preparation method thereof |
| CN105036997A (en) * | 2015-08-27 | 2015-11-11 | 合肥科友生物科技有限公司 | Culture soil for peony flowers and preparation method thereof |
| CN105053015A (en) * | 2015-07-15 | 2015-11-18 | 河南科技大学 | Biopesticide for controlling cercospora varrireolorWinter, and preparation method for biopesticide |
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| CN101948751A (en) * | 2010-08-06 | 2011-01-19 | 孙晗笑 | Microorganism bacteria liquid beneficial for growth of plants, preparation method and applications thereof |
| CN104630102A (en) * | 2015-01-27 | 2015-05-20 | 山东苏柯汉生物工程股份有限公司 | Complex bacterium preparation for fixing nitrogen, solubilizing phosphorus and potassium and rooting and preparation method thereof |
| CN105053015A (en) * | 2015-07-15 | 2015-11-18 | 河南科技大学 | Biopesticide for controlling cercospora varrireolorWinter, and preparation method for biopesticide |
| CN105036997A (en) * | 2015-08-27 | 2015-11-11 | 合肥科友生物科技有限公司 | Culture soil for peony flowers and preparation method thereof |
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