CN109526986A - A kind of tobacco diseases prevention biological prevention and control agent and preparation method thereof - Google Patents
A kind of tobacco diseases prevention biological prevention and control agent and preparation method thereof Download PDFInfo
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- 235000002637 Nicotiana tabacum Nutrition 0.000 title claims abstract description 43
- 241000208125 Nicotiana Species 0.000 title claims abstract description 38
- 201000010099 disease Diseases 0.000 title claims abstract description 37
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title claims abstract description 37
- 238000002360 preparation method Methods 0.000 title claims abstract description 27
- 230000002265 prevention Effects 0.000 title description 49
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- 230000000443 biocontrol Effects 0.000 claims abstract 13
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- 241000187747 Streptomyces Species 0.000 claims abstract 3
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- 235000017173 flavonoids Nutrition 0.000 claims abstract 3
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- 239000000203 mixture Substances 0.000 claims description 4
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- YWILDZMVBIMXID-KGHBUBBXSA-N (4s,4as,5as,6s,12ar)-n-[[4-[n-(diaminomethylidene)carbamimidoyl]piperazin-1-yl]methyl]-4-(dimethylamino)-1,6,10,11,12a-pentahydroxy-6-methyl-3,12-dioxo-4,4a,5,5a-tetrahydrotetracene-2-carboxamide;dihydrochloride Chemical compound Cl.Cl.OC([C@@]1(O)C(=O)C=2[C@@H]([C@](C3=CC=CC(O)=C3C=2O)(C)O)C[C@H]1[C@@H](C1=O)N(C)C)=C1C(=O)NCN1CCN(C(=N)N=C(N)N)CC1 YWILDZMVBIMXID-KGHBUBBXSA-N 0.000 description 1
- WLJVXDMOQOGPHL-PPJXEINESA-N 2-phenylacetic acid Chemical compound O[14C](=O)CC1=CC=CC=C1 WLJVXDMOQOGPHL-PPJXEINESA-N 0.000 description 1
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- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
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- 229910000403 monosodium phosphate Inorganic materials 0.000 description 1
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Classifications
-
- 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
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
-
- 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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- Biotechnology (AREA)
- Virology (AREA)
- Agronomy & Crop Science (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Environmental Sciences (AREA)
- Mycology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
本发明涉及植物生防制剂技术领域，具体涉及一种烟草防病生防制剂及其制备方法。本发明的目的在于提供一种烟草防病生防制剂及其制备方法，将混合培养基冻干粉、甘蔗粉和草木灰按一定比例混匀，即得烟草防病生防制剂粉剂，将混合培养基冻干粉和一些无机物混合加水溶解，即得烟草防病生防制剂粉剂。所述混合培养基冻干粉包括枯草芽孢杆菌、哈茨木霉菌和细黄链霉菌的培养基冻干粉，所得烟草防病生防制剂能防治烟草的一些常见且危害较大的病害。The invention relates to the technical field of plant biocontrol preparations, in particular to a tobacco disease prevention biocontrol preparation and a preparation method thereof. The object of the present invention is to provide a kind of tobacco disease prevention and biocontrol preparation and preparation method thereof. The mixed medium freeze-dried powder, sugarcane powder and plant ash are mixed in a certain proportion to obtain tobacco disease prevention and biocontrol preparation powder. The base freeze-dried powder and some inorganic substances are mixed with water to dissolve, and the tobacco disease prevention and biocontrol preparation powder is obtained. The mixed medium freeze-dried powder includes the medium freeze-dried powder of Bacillus subtilis, Trichoderma harzianum and Streptomyces flavonoids, and the obtained tobacco disease prevention and biocontrol preparation can prevent and treat some common and more harmful diseases of tobacco.
Description
Technical field
The present invention relates to plant biological prevention and control agent technical fields, and in particular to a kind of tobacco diseases prevention biological prevention and control agent and its preparation side
Method.
Background technique
Tobacco is the higher crop of economic value, and China's tobacco planting area is big, and plantation is with a long history, but planting process
In tobacco disease pest problem and traditional chemical sterilizing insecticide it is increasingly severe to the pollution problem of environment.Root rot, arteries and veins
Pinta, bacterial wilt and balck shank etc. are the infectious diseases that tobacco plants disease incidence is higher, harm is more serious, and circulation way is with soil
Based on earth is propagated, traditional chemical prevention and control method is difficult to obtain apparent control efficiency, and easily causes environmental pollution.In current state
Family proposes under the strategy of sustainable development overall background of pollution-less agriculture, environment protective plant protecting, is carried out using soil beneficial microbe resource
Disease control because of its no pollution to the environment, suitable for controlling disease and shows good development and application prospect.
Subtilin, polymyxins, system are generated during bacillus subtilis (Bacillussubtilis) thalli growth
Moldin, gramicidins isoreactivity substance, these active materials have obviously the conditioned pathogen of pathogenic bacteria or autogenous infection
Inhibiting effect, protease, the cellulase etc. that thallus generates be conducive to other strain growths in soil environment；Most of trichodermas
Bacterium can produce the bioactive substance that a variety of pairs of plant pathogenic fungis, bacterium and insect have antagonism, such as cell wall drop
Enzyme and secondary metabolite are solved, and the resistance of crops can be improved, promote plant growth and improves agricultural output, therefore
It is widely used in biological control, bio-feritlizer and soil conditioner, since negative effect of the chemical pesticide to environment is more serious,
So more environmentally friendly biological pesticide Trichoderma has received widespread attention；Streptomyces microflavus (Streptomyces
Microflavus contain the plant growths such as auxin, antibiotic, phenylacetic acid, succinic acid and the basic element of cell division in metabolite)
Necessary growth regulator ingredient has conversion the soil nitrogen, phosphorus phosphorus, potassium, increases soil fertility, reduce fertilizer amount, inhibits
Germ reproduction, diseases prevention are kept a full stand of seedings, and cell division is stimulated, and promote crop take root, germinate, is mature, improving crop yield, promoting effective substance
The effects of matter synthesizes, and significantly improves the quality of agricultural product reaches " increasing both production and income is retained and supports field, inhibit disease, improve quality " four
The effect of effect unification.
In conclusion there are many research shows that the metabolite of certain micro-organisms is conducive to control of crop disease and rush at present
Into the effect of plant growth, these metabolites include reactive protein or carbohydrate etc..There are many microbial bacterial agents to use at present
In the edaphon environment for adjusting crop, achieve the purpose that reducing pathogenic microorganisms grows and promote plant growth, it is still, raw
Production person needs to guarantee that the microorganism in microbial inoculum is state living, and can live and reproduce in severe soil environment, practical raw
It is competition existent relationship if adding several different microorganisms in crop soil simultaneously, between them in production, it will certainly
The growth vigor for leading to certain microorganism, the growth and effect for being unfavorable for other useful microorganisms play；Original crop soil is micro-
In the case that biotic environment is unknown, the addition of external microorganism can affect to original edaphon environment,
The quantity kind of microorganism is all difficult to control.It is by its active generation during the growth process that beneficial microorganism effect is mostly
It thanks to product and plays, " can generally live with peacefully " between metabolite, be the technological means to solve the above problems.
Summary of the invention
In view of the above shortcomings of the prior art, the purpose of the present invention is to provide a kind of tobacco diseases prevention biological prevention and control agent and its systems
Mixed culture medium freeze-dried powder, cane powder and plant ash are mixed in a certain ratio to get tobacco diseases prevention biological prevention and control agent powder by Preparation Method
Agent, the mixed culture medium freeze-dried powder include the culture medium freeze-dried powder of bacillus subtilis, trichoderma harzianum and streptomyces microflavus,
Gained tobacco diseases prevention biological prevention and control agent can prevent and treat some common of tobacco and the biggish disease of harm.
A kind of tobacco diseases prevention biological prevention and control agent, including mixed culture medium freeze-dried powder, mixed culture medium freeze-drying meal component and
Weight ratio are as follows: bacillus subtilis bacteria culture fluid freeze-dried powder: Trichoderma harzianum bacteria culture fluid freeze-dried powder: streptomyces microflavus culture solution freeze-drying
Powder=3:2:3.
The preparation method of above-mentioned tobacco diseases prevention biological prevention and control agent comprising the steps of:
(1) microculture
Bacillus subtilis, trichoderma harzianum, streptomyces microflavus are expanded culture respectively, culture to bacterial concentration reaches
107CFU/mL；
(2) culture solution is collected
Above-mentioned inoculum is centrifuged, 8000 × g of revolving speed, the time 1 hour respectively, collects the culture for being free of bacterium
Liquid supernatant, freeze-drying process, until water content is 10% hereinafter, the culture solution freeze-dried powder of three kinds of bacterium of mixing, each freeze-dried powder mixing
Weight ratio are as follows: bacillus subtilis: trichoderma harzianum: streptomyces microflavus=3:2:3 is to get mixed culture medium freeze-dried powder.
Above-mentioned tobacco diseases prevention biological prevention and control agent can be applied to prevention and treatment tobacco head mold disease, arteries and veins pinta, bacterial wilt and balck shank.
The powder form of above-mentioned tobacco diseases prevention biological prevention and control agent, the ingredient and weight ratio of the powder form are as follows: mixing training
Support base freeze-dried powder: cane powder: plant ash=5~7:15~20:42~72.
Further, the powder form of above-mentioned tobacco diseases prevention biological prevention and control agent, the cane powder are by sugarcane by being dehydrated
To water content, rear crush is made below 10%.
Aqua form can be made in above-mentioned tobacco diseases prevention biological prevention and control agent, and the every 100mL of aqua includes mixed culture medium
Freeze-dried powder 10-50g, disodium hydrogen phosphate 2-4g.
Further, the aqua form of the tobacco diseases prevention biological prevention and control agent when in use with distilled water dilute 100 times, pour in
The tobacco plant root of morbidity.
The utility model has the advantages that
1, the Common Diseases of tobacco can be prevented and treated without changing the original edaphon environment of tobacco, it is external removes addition from
The risk for causing original edaphon environmental ecology unbalance after flora；
2, the main active in gained biological prevention and control agent is bacillus subtilis, trichoderma harzianum, streptomyces microflavus three
Kind has the culture medium lyophilized products of the active microorganism of diseases prevention through studying, and the metabolism for including medium component and microorganism secretion produces
Object, the metabolite mixing that microorganism generates will not generate uncontrollable existence reaction, can play in plant growth respectively
Effect；
3, the present invention carries out crop disease using the metabolite of bacillus subtilis, trichoderma harzianum, streptomyces microflavus
Prevention and treatment, the microorganism cultivated can also continue to carry out fermentating metabolism, saved living resources, saved production cost.
Specific embodiment
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with implementation of the invention
Example, technical scheme in the embodiment of the invention is clearly and completely described.Based on the embodiments of the present invention, this field
Those of ordinary skill's every other embodiment obtained without making creative work, belongs to protection of the present invention
Range.
The preparation of one tobacco biological prevention and control agent of embodiment
1 material source
Bacillus subtilis, trichoderma harzianum, thin xanthomycin are bought from commercial channel.
Plant ash is the ashes left after plant fuel combustion power generation.
Sugarcane waste residue of the cane powder after sugar refinery's juicing, is broken into powder.
2 microcultures
2.1 bacillus subtilis
(1) culture medium prescription
Seed culture medium: peptone 1%, yeast extract 0.5%, sodium chloride 1%, natural pH, sterilizing；
Basal fermentation medium: sucrose 1%, peptone 1%, disodium hydrogen phosphate 0.2%, sodium dihydrogen phosphate 0.2%, pH
7.0, sterilizing.
(2) cultural method
A. actication of culture
In gnotobasis, by the strain transfer of purchase to slant medium, 37 DEG C of cultures are for 24 hours, spare.
B. the preparation of seed liquor
Under aseptic condition, the strain of activation is taken with oese, access loading amount is the 250mL triangular flask of 50mL seed culture medium
In, 37 DEG C, 180r/min cultivates 18h.
C. shaking flask culture
20mL seed liquor is taken respectively, and access fills in the bottle of 1000mL fermentation medium (inoculum concentration 2%, V/V).It sets and shakes
In bed, 35 DEG C of shaken cultivation 12h, revolving speed 160r/min.
D. culture solution is collected
Bacterium solution after will be enlarged by culture is centrifuged, and revolving speed is 8000 × g, and the time is half an hour, is collected clear without thin
Bacteria culture fluid, frozen drying to water content is 10% hereinafter, culture medium at this time is powdery.
2.2 trichoderma harzianum
(1) culture medium prescription
Solid medium: PDA culture medium (200g potato is weighed, peeling slice is cleaned, water 1000mL is added to boil 15min,
After filtered through gauze plus water complements to 1000mL, then plus 12g glucose, 20g agar boil to solid and dispense after completely dissolution.Point
Culture medium after dress should sterilize in 1h, using high steam moist heat sterilization, under 0.1MPa vapour pressure, temperature up to 121 DEG C,
20min is maintained, sterile coated plate obtains solid medium after cooling)；
Fluid nutrient medium: the same solid medium of method, but agar is not added, loaded on sterilizing in culture bottle.
(2) cultural method
After taking Trichoderma harzianum to activate on PDA medium plate, it is inoculated in oese and is a little connected to potato, glucose fine jade
On rouge test tube slant, 26 DEG C of culture 6d are to producing spore.It is subsequently poured into 9ml sterile water, vortex oscillation, it is 10 that concentration, which is made,7A/ml
Spore suspension, take pre-fabricated spore suspension (concentration 107Cfu/ml it) is inoculated in Liquid Culture respectively by 2% inoculum concentration
Shaking table shaken cultivation in base, revolving speed 180rpm, culture are centrifuged after 7 days, and revolving speed is 10000 × g, and the time is 1 hour, are collected clear
Clear culture solution and with 0.45 μm of membrane filtration, is freeze-dried filtered culture solution, until water content is lower than 10%, obtains powder
Last shape culture medium.
2.3 streptomyces microflavus
(1) culture medium prescription
Solid medium: PDA culture medium (200g potato is weighed, peeling slice is cleaned, water 1000mL is added to boil 15min,
After filtered through gauze plus water complements to 1000mL, then plus 12g glucose, 20g agar boil to solid and dispense after completely dissolution.Point
Culture medium after dress should sterilize in 1h, using high steam moist heat sterilization, under 0.1MPa vapour pressure, temperature up to 121 DEG C,
20min is maintained, sterile coated plate obtains solid medium after cooling)；
Fluid nutrient medium: the same solid medium of method, but agar is not added, loaded on sterilizing in culture bottle.
(2) cultural method
It takes streptomyces microflavus to be inoculated in PDA solid medium to be cultivated, 25 DEG C are cultivated 3 days, with oese picking list
Clone strain, which is seeded to fluid nutrient medium by 5%, to be continued to cultivate, and revolving speed 160rpm, 28 DEG C are cultivated 5-7 days.By the mould after culture
Liquid is centrifuged, and 8000 × g is centrifuged 1 hour, collects clear supernatant, 0.45 μm of membrane filtration, by filtered culture
Liquid freeze-drying obtains powdered medium until water content is lower than 10%.
3 configuration mixed culture medium freeze-dried powders
Mix the fluid nutrient medium freeze-dried powder of three kinds of microorganisms, mixing mass ratio are as follows: bacillus subtilis: Trichoderma harzianum
Bacterium: streptomyces microflavus=3:2:3.
The preparation of 4 biological prevention and control agent pulvis
Above-mentioned mixed culture medium freeze-dried powder is uniformly mixed with cane powder, plant ash, mixing mass ratio are as follows: mixed-culture medium
Freeze-dried powder: cane powder: plant ash=5~7:15~20:42~72 gained are biological prevention and control agent pulvis.
The preparation of 5 biological prevention and control agent aquas
Mixed culture medium the freeze-dried powder 10-50g, disodium hydrogen phosphate 2-4g for taking above-mentioned steps 1-3 to prepare add distilled water
To 100mL, pH value 7.0-7.2, gained is tobacco diseases prevention biological prevention and control agent aqua.
Embodiment one illustrates method for culturing microbes involved in biological prevention and control agent of the invention and biological prevention and control agent powder
The preparation method of agent and aqua, the method are simply easily implemented, and production cost is low, can produce a large amount of biological and ecological methods to prevent plant disease, pests, and erosion system according to demand
Agent.
Application of two biological prevention and control agent of embodiment in tobacco planting
Field trial is carried out using the method for embodiment 1 complex micro organism fungicide produced, test site: Yunnan Province is high
The township Cha He, mountain county, test variety are flue-cured tobaccos, are the more serious plot of disease incidence in recent years for examination experimental plot, and apply by organic fertilizer
It spreads.2 processing of test setting, are respectively as follows: 1: without using biological prevention and control agent, as a control group；2: using biological prevention and control agent of the invention
Pulvis is spread fertilizer over the fields, and amount of application is that 5kg/ mus of each processing are repeated 4 times, plot area 100m2.In plantation 60d observation morbidity feelings
Condition, sick grade: 0 grade asymptomatic for blade face；1 grade shows wilting shape for 1/4 or less blade face on plant；Second level is 1/4~1/2 blade face
Show wilting shape；3 grades show wilting symptom for 1/2 or more blade face of plant；4 grades dead for complete stool wilting.
Disease incidence=(morbidity strain number/investigation strain number) × 100%
Disease index=∑ (diseased plant numbers at different levels × typical value at different levels)/(investigation total strain number × superlative degree typical value) ×
100%
Control effect=(control disease index-processing disease index)/control disease index × 100%
1 different disposal tobacco pathogenesis situation of table compares
Embodiment two is the result shows that diseases prevention biological prevention and control agent of the invention can play the role of preventing tobacco diseases, prevention and control effect
Fruit is between 55-63%.This biological prevention and control agent control efficiency is good, and be it is environmentally friendly, compared to existing chemical prevention side
Method more meets the demand of modern agricultural development.
Claims (7)
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