CN108432805A - A kind of complex microbial inoculum and preparation method thereof for preventing vegetables root knot - Google Patents
A kind of complex microbial inoculum and preparation method thereof for preventing vegetables root knot Download PDFInfo
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- CN108432805A CN108432805A CN201810300313.7A CN201810300313A CN108432805A CN 108432805 A CN108432805 A CN 108432805A CN 201810300313 A CN201810300313 A CN 201810300313A CN 108432805 A CN108432805 A CN 108432805A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
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Abstract
The invention discloses a kind of complex microbial inoculum and preparation method thereof for preventing vegetables root knot, which includes the following steps：Prepare soil supension；Soil supension is diluted and is cultivated, microbial composite bacteria is prepared；The microbial composite bacteria being prepared is mixed and cultivated with water, culture medium, cow dung is added and carries out mixed fermentation；The complex microbial inoculum for preventing vegetables root knot is prepared.The present invention will apply after multifunctional microbial compound bacteria and cow dung mixture and fermentation, pass through the interaction of multifunctional microbial compound bacteria and cow dung, soil and root nodule germ, vegetables root nodule germ can be killed, improve soil texture, nutrient needed for plant growth is improved, the yield and quality of agricultural product is improved.
Description
Technical field
The invention belongs to phytopathogen fungicide technical fields, specifically, being related to a kind of for preventing vegetables root nodule
The complex microbial inoculum and preparation method thereof of disease.
Background technology
Root knot, also referred to as root knot are the diseases caused by a kind of bacterium of Agrobacterium.The disease occurs mainly in root
On neck, lateral root and at graft union.China's most area is distributed.It falls ill serious field, plant can be caused to wilt in flakes
And death, it is very big to yield effect.Originally more than 100 kind cultivation of evil of being critically ill and wild crucifer.Chinese cabbage group seedling arrives
Adult plant can be aggrieved, only endangers root, and tumour is formed after being killed using root as main feature.Diseased plant shows short Huang, and leaf color becomes
Light, slow-growing, fine day noon diseased plant tune withers, and rear whole strain is dead.Dig out the visible master of diseased plant, formed in lateral root it is not of uniform size swollen
Tumor, tumour is big and measure few on main root, and tumour is small and measure more in lateral root.Early stage tumour is smooth, spheroidal or subsphaeroidal, the later stage
Become coarse, cracking；Aerial part symptom unobvious at the initial stage of a disease, later stage show as the water shortages fertilizer deficiency such as growth retardation, dwarfing
Symptom, diseased plant are wilted since radical leaves, and initial daytime wilts, and can restore night or rainy days, then grave illness plot
Diseased plant cannot restore, gradual yellow fading, wilting, death.Sick portion easily by bacterial invasions such as soft rot, cause tissue rot to send out it is smelly
Taste, it is dead in flakes.Growth of the root knot for crops, especially vegetable cultivation are a great harm, study root knot
Control method be of great significance with measure.
Modern microbiological theory is thought：One microbial population, which can generate another population, to be inhibited or poisons to make
With i.e. antagonism.We isolated from forest soil with antibacterial, decomposition of cellulose, decompose carbohydrate, reduce fat etc. it is more
The microbiologic population of kind function cooperatively forms complex microbial inoculum, and the excrement mixture for the milk cow fed with organic feed is simultaneously
Fermentation a period of time, mixture contains supports needed for the various plants such as fungicide and humic acid, amino acid, nucleotide, trace element
Point, vegetables root knot is not only killed, but also improve soil texture, promote plant growth, there is good promotion to agricultural production
Effect.
Invention content
In view of this, the present invention provides a kind of complex microbial inoculum and its preparation sides for preventing vegetables root knot
Method, the present invention are using the microbiologic population for being isolated from forest, with the cow dung with improvement soil texture with the various nutrients of offer
Mixture simultaneously ferment a period of time, to provide one kind can have both kill vegetables root knot and provide needed for each plant growth support
The complex microbial inoculum divided reaches sterilization, improvement soil, promotes plant growth and improve the multipurpose of quality.
In order to solve the above-mentioned technical problem, the invention discloses a kind of microbial composite bacterias for preventing vegetables root knot
The preparation method of agent, includes the following steps：
Step 1, selection fertile soil, scalp topsoil, fetch earth earth, spare, and the soil for scalping topsoil is put into nothing
In bacterium water and triangular flask with bead, setting shaking table oscillation 3-8min, so that soil sample is dispersed in dilution outstanding as soil
Liquid；
Step 2 is drawn in the test tube that soil supension injection fills sterile water, and pressure-vaccum 3 times vibrates mixing, it is dilute that soil is made
Release liquid；Soil dilution liquid is put on meat soup peptone culture medium tablet, with sterile glass apply stick media surface lightly
Coating is uniform；Ware bottom certain angle is rotated, continues to be coated with, until uniformly；Tablet is inverted in incubator and is cultivated；It will cultivate
To microorganism handled and be sealed with desivac, microbial composite bacteria is prepared；
The microbial composite bacteria being prepared is mixed and is cultivated with water, culture medium by step 3, add cow dung into
Row mixed fermentation；The complex microbial inoculum for preventing vegetables root knot is prepared.
Optionally, the mass fraction of the soil supension is 0.5%-1.5%.
Optionally, the volumetric concentration of the soil dilution liquid is 10-1-10-5。
Optionally, the cultivation temperature in the step 2 is 37 DEG C, incubation time 45-50h.
Optionally, the mass ratio of the microbial composite bacteria in the step 3 and water, culture medium is 1:1000:100-3:
1000:100, incubation time is 8-12 days, and cultivation temperature is 8-12 days.
Optionally, the mass ratio (kg/g) of the cow dung in the step 3 and microbial composite bacteria is 1000:5-1000:15,
The water content of cow dung is 25%-35%.
Optionally, in the step 3 during the fermentation, the water content of fermentate is maintained at 60%-70%；It is fermenting
Urea is added in object and supplements nitrogen source, it is 25: 1 to keep carbon-nitrogen ratio；And suitable lime or calacareous soil is added, keep it is neutral or
Alkalescence environment.
The invention also discloses the microorganisms for preventing vegetables root knot that a kind of above-mentioned preparation method is prepared
Composite bacteria agent.
The invention also discloses a kind of applications of the above-mentioned complex microbial inoculum for preventing vegetables root knot, are used for
The usage amount for preventing the complex microbial inoculum of vegetables root knot is 450-550ml/ mus, in use, for preventing vegetables root nodule
The complex microbial inoculum of disease dilutes 200 times.
Compared with prior art, the present invention can be obtained including following technique effect：
The present invention will apply after multifunctional microbial compound bacteria and cow dung mixture and fermentation, compound by multifunctional microbial
The interaction of bacterium and cow dung, soil and root nodule germ can kill vegetables root nodule germ, improve soil texture, improve and plant
Nutrient needed for object growth, improves the yield and quality of agricultural product.The present invention has very high economic benefit, ecological benefits and society
Benefit, using having a bright future.
Certainly, it implements any of the products of the present invention it is not absolutely required to while reaching all the above technique effect.
Description of the drawings
Attached drawing described herein is used to provide further understanding of the present invention, and constitutes the part of the present invention, this hair
Bright illustrative embodiments and their description are not constituted improper limitations of the present invention for explaining the present invention.In the accompanying drawings：
Fig. 1 is influence of the complex microbial inoculum of the present invention to wild cabbage, wherein (a) is not plus composite bacteria agent is by root knot
The wild cabbage root of bacterium infection；(b) it is not add composite bacteria agent processing, control group；(c) it is plus composite bacteria agent is handled.
Specific implementation mode
Carry out the embodiment that the present invention will be described in detail below in conjunction with embodiment, thereby to the present invention how application technology hand
Section solves technical problem and reaches the realization process of technical effect to fully understand and implement.
It is including following the invention discloses a kind of preparation method for preventing the complex microbial inoculum of vegetables root knot
Step：
Step 1, selection fertile soil, scalp topsoil, fetch earth earth, spare, and the soil for scalping topsoil is put into nothing
In bacterium water and triangular flask with bead, setting shaking table oscillation 3-8min, so that soil sample is dispersed in dilution outstanding as soil
The mass fraction of liquid, soil supension is 0.5%-1.5%；
Step 2 is drawn in the test tube that soil supension injection fills sterile water, and pressure-vaccum 3 times vibrates mixing, it is dense that volume is made
Degree is 10-1-10-5Soil dilution liquid；Soil dilution liquid is put on meat soup peptone culture medium tablet, is applied with sterile glass
Stick is lightly coated with uniformly in media surface；Ware bottom certain angle is rotated, continues to be coated with, until uniformly；Tablet is inverted in
45-50h is cultivated in 37 DEG C of incubators；The microorganism that culture obtains is handled and is sealed with desivac, micro- life is prepared
Object compound bacteria；
The microbial composite bacteria being prepared and water, culture medium according to mass ratio are 1 by step 3:1000:100-3:
1000:100 are mixed and are cultivated 8-12 days, are added cow dung and are carried out mixed fermentation；The quality of cow dung and microbial composite bacteria
It is 1000 than (kg/g):5-1000:15, the water content of cow dung is 25%-35%；During the fermentation, the water content of fermentate
It is maintained at 60%-70%；Urea is added in fermentate and supplements nitrogen source, it is 25: 1 to keep carbon-nitrogen ratio；And suitable lime is added
Or calacareous soil, keep neutral or alkalescence environment；The complex microbial inoculum for preventing vegetables root knot is prepared.
Embodiment 1
A kind of preparation method for preventing the complex microbial inoculum of vegetables root knot includes the following steps：
Step 1, selection scalp topsoil compared with fertile soil, and fetch earth earth 10g, spare, weighs soil sample 1.0g and is put into Sheng
99mL sterile waters and with bead triangular flask in, set shaking table oscillation 5min make soil sample be dispersed in dilution become soil
Earth suspension；
Step 2 is therefrom drawn in the test tube that the injection of 0.5mL soil supensions fills 4.5mL sterile waters, pressure-vaccum with sterile pipette tip
3 times, mixing is vibrated, is made a concentration of 10-1The soil liquid；Soil dilution liquid is put on meat soup peptone culture medium tablet,
Stick is applied with sterile glass to be lightly coated with uniformly in media surface；Ware bottom certain angle is rotated, continues to be coated with, until uniformly；
Tablet is inverted in 37 DEG C of incubators and cultivates 48h；The microorganism that culture obtains is handled and is sealed with desivac, is prepared
Obtain microbial composite bacteria；
Step 3,
10 grams of microbial composite bacteria being prepared is taken, adds water 5000ml, is placed 10 days at 500 grams, 25 DEG C of culture medium；Again
It is mixed with 1000kg cow dungs (water content 30%).Fermentation condition is：1,65% water content is kept；2, it keeps having in mixture suitable
When air；3, lime or calacareous soil can be added in right amount, keep neutral or alkalescence environment；4, urine is added in fermentate
Element supplement nitrogen source, it is 25: 1 to keep carbon-nitrogen ratio；The complex microbial inoculum for preventing vegetables root knot is prepared.
The complex microbial inoculum for preventing vegetables root knot being prepared, fusobacterium (Clostridium_
Sensu_stricto_1) 42.69%；Pseudomonas (Pseudomonas) 27.40%；Azotobacter (Azotobacter)
3.78%；Blattabacterium (Blattabacterium) 1.56%；Fusobacterium (Clostridium_sensu_stricto_
11) 1.39%.
The usage amount of complex microbial inoculum for preventing vegetables root knot is 450-550ml/ mus, in use, being used for
The complex microbial inoculum for preventing vegetables root knot dilutes 200 times.
Embodiment 2
A kind of preparation method for preventing the complex microbial inoculum of vegetables root knot includes the following steps：
Step 1, selection fertile soil, scalp topsoil, fetch earth earth, spare, and the soil for scalping topsoil is put into nothing
In bacterium water and triangular flask with bead, setting shaking table oscillation 8min, so that soil sample is dispersed in dilution outstanding as soil
The mass fraction of liquid, soil supension is 0.5%；
Step 2 is drawn in the test tube that soil supension injection fills sterile water, and pressure-vaccum 3 times vibrates mixing, it is dense that volume is made
Degree is 10-3Soil dilution liquid；Soil dilution liquid is put on meat soup peptone culture medium tablet, applying stick with sterile glass is training
Foster primary surface is lightly coated with uniformly；Ware bottom certain angle is rotated, continues to be coated with, until uniformly；Tablet is inverted in 37 DEG C of guarantors
50h is cultivated in incubator；The microorganism that culture obtains is handled and is sealed with desivac, microbial composite bacteria is prepared；
The microbial composite bacteria being prepared and water, culture medium according to mass ratio are 3 by step 3:1000:100 are mixed
Merge culture 12 days, adds cow dung and carry out mixed fermentation；The mass ratio (kg/g) of cow dung and microbial composite bacteria is 1000:5,
The water content of cow dung is 35%；During the fermentation, the water content of fermentate is maintained at 60%；Urea is added in fermentate to mend
Nitrogen charging source, it is 25: 1 to keep carbon-nitrogen ratio；And suitable lime or calacareous soil is added, keep neutral or alkalescence environment；System
It is standby to obtain the complex microbial inoculum for preventing vegetables root knot.
The usage amount of complex microbial inoculum for preventing vegetables root knot is 450-550ml/ mus, in use, being used for
The complex microbial inoculum for preventing vegetables root knot dilutes 200 times.
Embodiment 3
A kind of preparation method for preventing the complex microbial inoculum of vegetables root knot includes the following steps：
Step 1, selection fertile soil, scalp topsoil, fetch earth earth, spare, and the soil for scalping topsoil is put into nothing
In bacterium water and triangular flask with bead, setting shaking table oscillation 3min, so that soil sample is dispersed in dilution outstanding as soil
The mass fraction of liquid, soil supension is 1.5%；
Step 2 is drawn in the test tube that soil supension injection fills sterile water, and pressure-vaccum 3 times vibrates mixing, it is dense that volume is made
Degree is 10-5Soil dilution liquid；Soil dilution liquid is put on meat soup peptone culture medium tablet, applying stick with sterile glass is training
Foster primary surface is lightly coated with uniformly；Ware bottom certain angle is rotated, continues to be coated with, until uniformly；Tablet is inverted in 37 DEG C of guarantors
45h is cultivated in incubator；The microorganism that culture obtains is handled and is sealed with desivac, microbial composite bacteria is prepared；
The microbial composite bacteria being prepared and water, culture medium according to mass ratio are 1 by step 3:1000:100 are mixed
Merge culture 8 days, adds cow dung and carry out mixed fermentation；The mass ratio (kg/g) of cow dung and microbial composite bacteria is 1000:15,
The water content of cow dung is 25%；During the fermentation, the water content of fermentate is maintained at 70%；Urea is added in fermentate to mend
Nitrogen charging source, it is 25: 1 to keep carbon-nitrogen ratio；And suitable lime or calacareous soil is added, keep neutral or alkalescence environment；System
It is standby to obtain the complex microbial inoculum for preventing vegetables root knot.
The usage amount of complex microbial inoculum for preventing vegetables root knot is 450-550ml/ mus, in use, being used for
The complex microbial inoculum for preventing vegetables root knot dilutes 200 times.
Began the microbial composite bacteria developed in the application in continuous 2 years of the Wenchuan County in Sichuan Province townshiies Cao Po wild cabbage vegetable field from 2016
Agent simultaneously does control experiment (being illustrated in fig. 1 shown below), achieves good result.Use complex microbial inoculum 500ml, when use dilute per acre
Release 200 times.The results show complex microbial inoculum can fertile soil, sufficient nutrition is provided for wild cabbage fruit tree, and can kill
It goes out root nodule germ, prevention plant root nodule disease.
Above description has shown and described several preferred embodiments of invention, but as previously described, it should be understood that invention is not
It is confined to form disclosed herein, is not to be taken as excluding other embodiments, and can be used for various other combinations, modification
And environment, and can be carried out by the above teachings or related fields of technology or knowledge in the scope of the invention is set forth herein
Change.And changes and modifications made by those skilled in the art do not depart from the spirit and scope of invention, then should all be weighed appended by invention
In the protection domain that profit requires.
Claims (9)
1. a kind of preparation method for preventing the complex microbial inoculum of vegetables root knot, which is characterized in that including following step
Suddenly：
Step 1, selection fertile soil, scalp topsoil, fetch earth earth, spare, and the soil for scalping topsoil is put into sterile water
And in the triangular flask with bead, setting shaking table oscillation 3-8min makes soil sample be dispersed in dilution as soil supension；
Step 2 is drawn in the test tube that soil supension injection fills sterile water, and pressure-vaccum 3 times vibrates mixing, soil dilution liquid is made；
Soil dilution liquid is put on meat soup peptone culture medium tablet, apply stick with sterile glass is lightly coated in media surface
It is even；Ware bottom certain angle is rotated, continues to be coated with, until uniformly；Tablet is inverted in incubator and is cultivated；Culture is obtained micro-
Biology desivac is handled and is sealed, and microbial composite bacteria is prepared；
The microbial composite bacteria being prepared is mixed and is cultivated with water, culture medium by step 3, is added cow dung and is mixed
Close fermentation；The complex microbial inoculum for preventing vegetables root knot is prepared.
2. preparation method according to claim 1, which is characterized in that the mass fraction of the soil supension is 0.5%-
1.5%.
3. preparation method according to claim 1, which is characterized in that the volumetric concentration of the soil dilution liquid is 10-1-
10-5。
4. preparation method according to claim 1, which is characterized in that the cultivation temperature in the step 2 is 37 DEG C, culture
Time is 45-50h.
5. preparation method according to claim 1, which is characterized in that the microbial composite bacteria in the step 3 and water, training
The mass ratio for supporting base is 1:1000:100-3:1000:100, incubation time is 8-12 days, and cultivation temperature is 8-12 days.
6. preparation method according to claim 1, which is characterized in that the cow dung in the step 3 and microbial composite bacteria
Mass ratio (kg/g) be 1000:5-1000:15, the water content of cow dung is 25%-35%.
7. preparation method according to claim 1, which is characterized in that in the step 3 during the fermentation, fermentate
Water content be maintained at 60%-70%；Urea is added in fermentate and supplements nitrogen source, it is 25: 1 to keep carbon-nitrogen ratio；And it is added suitable
The lime or calacareous soil of amount keep neutral or alkalescence environment.
8. what the preparation method in a kind of 1-7 by claim described in any claim was prepared is used to prevent vegetables root nodule
The complex microbial inoculum of disease.
9. a kind of by the application according to any one of claims 8 for preventing the complex microbial inoculum of vegetables root knot, feature exists
In the usage amount of the complex microbial inoculum for preventing vegetables root knot is 450-550ml/ mus, in use, for preventing vegetable
The complex microbial inoculum of dish root knot dilutes 200 times.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109362790A (en) * | 2018-12-10 | 2019-02-22 | 河北省农林科学院石家庄果树研究所 | It is a kind of for preventing and treating the complex microbial inoculum and preparation method thereof of pear tree root knot |
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| CN101514331A (en) * | 2009-01-05 | 2009-08-26 | 中国农业大学 | Pseudomonas. chlororaphis subsp. Aurantiaca Pa40 and application thereof |
| JP4625626B2 (en) * | 2003-11-27 | 2011-02-02 | 片倉チッカリン株式会社 | Method for suppressing plant parasitic nematode damage using nematode damage-controlling microorganisms and nematode damage-controlling microorganism materials |
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| JP4625626B2 (en) * | 2003-11-27 | 2011-02-02 | 片倉チッカリン株式会社 | Method for suppressing plant parasitic nematode damage using nematode damage-controlling microorganisms and nematode damage-controlling microorganism materials |
| CN1946297A (en) * | 2004-02-27 | 2007-04-11 | 株式会社树 | Method of controlling plant disease damage by using bacillus and controlling agent |
| CN1847388A (en) * | 2006-04-21 | 2006-10-18 | 贵州大学 | Aspergillus niger strain with Agrobacterium tumefaciens inhibiting effect and its use |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109362790A (en) * | 2018-12-10 | 2019-02-22 | 河北省农林科学院石家庄果树研究所 | It is a kind of for preventing and treating the complex microbial inoculum and preparation method thereof of pear tree root knot |
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