CN105309082B - A kind of tobacco bacterial wilt Synthetical prevention method - Google Patents
A kind of tobacco bacterial wilt Synthetical prevention method Download PDFInfo
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- CN105309082B CN105309082B CN201510682109.2A CN201510682109A CN105309082B CN 105309082 B CN105309082 B CN 105309082B CN 201510682109 A CN201510682109 A CN 201510682109A CN 105309082 B CN105309082 B CN 105309082B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01B—SOIL WORKING IN AGRICULTURE OR FORESTRY; PARTS, DETAILS, OR ACCESSORIES OF AGRICULTURAL MACHINES OR IMPLEMENTS, IN GENERAL
- A01B79/00—Methods for working soil
- A01B79/02—Methods for working soil combined with other agricultural processing, e.g. fertilising, planting
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05C—NITROGENOUS FERTILISERS
- C05C11/00—Other nitrogenous fertilisers
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
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Abstract
The invention discloses a kind of tobacco bacterial wilt Synthetical prevention method, carries out following handle to soil before tobacco is planted：1）Soil pretreatment：Within 10~November after upper 1 year tobacco harvests, with quick lime 90~100kg/ mus, upper soll layer is uniformly sprinkling upon；2）Green manure crop is sowed on soil, and makees green manuring processing in March in the coming year 2；3）Soil is reprocessed：Upper soll layer is uniformly sprinkling upon again with the kg/ mus of quick lime 90~100 after green manuring；4）Soil is ploughed deeply；5）Apply biological organic fertilizer.Comprehensive measures for the prevention and control of the present invention can effectively reduce the bacterial wilt incidence of disease, increase yield of tobacco, the output value, reduce cause of disease bacteria concentration in soil and then reduction pathogen is infected to root system, the ecosystem for being advantageous to disease soil converts to the direction of sustainable health development, reach the control effect good to grave illness cigarette district tobacco bacterial wilt, can solve the problem that the problem of existing single measure is limited to bacterial wilt control effect.
Description
Technical field
The present invention relates to a kind of tobacco bacterial wilt Synthetical prevention method, belongs to tobacco bacterial wilt Control Technology field.
Background technology
Tobacco bacterial wilt is by blue or green withered Raul Salmonella（Ralstonia solanacearum）Caused one kind is passed with soil
Crushing bacillary tobacco diseases based on broadcasting.Tobacco bacterial wilt is the most important bacterial disease for influenceing Tobacco Leaves in Guizhou production
One of.In recent years, as scientific and technological progress and the reinforcement of people's environmental consciousness, biological preventing control method gradually cause the weight of people
The flat board antagonism of Antagonistic Fungi in the lab to pathogen is all stopped depending on, prevention and control to Stalk Rot at present, directly
Control effect applied to greenhouse test or field test is little, the Pseudomonas bar that Anuratha etc. obtains plate screening
Bacterium is directly used in field test, and its prevention and control rate to bacterial wilt of tomato is only 36 % or so.
Antagonistic Fungi is not easy to colonize after being individually manured into soil, and improves Antagonistic Fungi colonizing as biological prevention and control in rhizosphere soil
Research emphasis, use to apply Antagonistic Fungi combination organic fertilizer or biological organic fertilizer disease is made by 2 fermentations at present and apply regulation soil
Tiny ecosystem, improve diversity of soil microorganism, suppress the growth of pathogen or improve plant self resistance, so as to suppress bacterial wilt
Generation.Because the complexity of environment and the limitation of single prevention and control measure serious field of being fallen ill to disease use individual event prevention and control
The effect of measure is unsatisfactory, therefore it is significant to explore a kind of measure of effective Synthetical prevention bacterial wilt.
The content of the invention
The technical problem to be solved in the present invention is：The vega progress three that inventor falls ill serious for tobacco bacterial wilt is older
Phase positioning is repaired, and by studying the growth and decline relation and soil microflora variation characteristic of pathogen and Antagonistic Fungi in soil,
A kind of tobacco bacterial wilt Synthetical prevention method is proposed, this method effectively cooperates with various preventions, can solve the problem that existing list
The problem of one measure is limited to bacterial wilt control effect.
Technical scheme：A kind of tobacco bacterial wilt Synthetical prevention method, before tobacco is planted to soil carry out with
Lower processing： 1）Soil pretreatment：Within 10~November after upper 1 year tobacco harvests, with quick lime 90~100kg/ mus,
It is even to be sprinkling upon upper soll layer；2）Green manure crop is sowed on soil, and makees green manuring processing in month in coming year 2-3；3）Soil is reprocessed：
Upper soll layer is uniformly sprinkling upon again with the kg/ mus of quick lime 90~100 after green manuring；4）Soil is ploughed deeply；5）Applying biology has
Machine fertilizer.
The preparation method of the biological organic fertilizer is：Antagonistic Fungi L-25 is inoculated in beef extract-peptone fluid nutrient medium,
Triangular flask liquid amount is 1/5,30 DEG C, 170 r/m vibrate 48 h, and 6000 × g of zymotic fluid then is centrifuged into 10 min, collects thalline
The organic fertilizer carrier after sterilizing is inoculated in by 5% inoculum concentration, regulation organic fertilizer water content is 40%, and ferment 5~6 d, periodically turns over throwing
Obtain the biological organic fertilizer of L-25 secondary fermentations.
The preparation method of the biological organic fertilizer is：L-9 is inoculated in Gause I fluid nutrient medium, triangular flask liquid amount
For 1/5,30 DEG C, 170 r/m vibrate 48 h, 6000 × g of zymotic fluid is then centrifuged into 10 min, thalline is collected and is connect by 5% inoculum concentration
Organic fertilizer carrier of the kind after sterilizing, regulation organic fertilizer water content are 40%, and ferment 5~6 d, periodically turns over and throws to obtain L-9 secondary fermentations
Biological organic fertilizer.Organic fertilizer is applied after Antagonistic Fungi is combined and microbial organic fertilizer is made in the present invention, to tobacco bacterial wilt
Harmful prevention and control have good effect.
The biological organic fertilizer is that foregoing L-25 and L-9 is distinguished into the biological organic fertilizer of secondary fermentation with mass ratio 1:1 is mixed
Manufactured biological organic fertilizer after conjunction.L-25 has the characteristics of cost is low and control effect is poor in the present invention, L-9 have cost high and
The characteristics of control effect is good, the two is combined, and can be played a part of Synthetical prevention effect and be taken into account production cost.
The organic fertilizer carrier is the amino acid fertilizer that rapeseed meal digests and cow dung with mass ratio 1:1 mixing, containing organic matter
33.8%th, amino acid 4.3%, N 4.20%, P2O5 2.26%、K2O 1.08%。
The green manure plant is late-maturing rape, wheat or clover.Such green manure plant is easy to plant, and cost is relatively low.
The L-25 be using early stage from health tobacco rhizosphere soil separation screening to Antagonistic Fungi, the L-9 for use
Early stage from health tobacco rhizosphere soil separation screening to Antagonistic Fungi.The antagonistic microbe used in the present invention is from tobacco rhizosphere
Soil screening obtains, and has very strong colonization ability in tobacco rhizosphere；Antagonistic Fungi is using organic fertilizer as carrier simultaneously, by secondary solid
In being manured into soil after body fermentation, the colonization ability in soil is stronger, good inhibiting effect is played to pathogen, and then postpone
Tobacco plant disease time and significantly reduce bacterial wilt disease index.
Beneficial effects of the present invention：Pre-processed and reprocessed with quick lime in the present invention to adjust soil pH, energy
Enough growth and breedings for suppressing blue or green withered Raul Salmonella；Green manure crop is sowed, other types of microorganism can be cultivated, it is blue or green withered to tie up
The existence resource of Raul Salmonella；Soil is ploughed deeply and using biological organic fertilizer, also can further suppress blue or green withered Raul
The breeding of Salmonella.The present invention can be effectively improved edaphon balance by measures first mentioned, recover destroyed soil ecology
System, so as to set up complicated and health edaphon system.Antagonistic Fungi forms " matrix-flora " under organic fertilizer assistance
The ecosystem, it is more beneficial for adjusting soil microenvironment, rhizosphere soil microorganism ecological characteristic can be changed and physical chemistry is special
Sign, so as to play diseases prevention, the effect of suppression disease.
The present invention, which applies, suppresses tobacco bacterial wilt type biological organic fertilizer cooperation comprehensive measures for the prevention and control, can effectively reduce blue or green withered
The sick incidence of disease, increase yield of tobacco, the output value, reduce cause of disease bacteria concentration in soil and then reduction pathogen is infected to root system, have
Converted beneficial to the ecosystem of disease soil to the direction of sustainable health development, reach good to grave illness cigarette district tobacco bacterial wilt
Control effect.
Brief description of the drawings
Fig. 1 is the growth and decline relation figure of pathogen and Antagonistic Fungi in soil between different years；
Fig. 2 is different disposal Beta diversity PCoA analysis charts；
Fig. 3 is different disposal UPGMA cluster tree graphs；
Fig. 4 is that pedotheque prepares the procedure chart being sequenced with upper machine.
Embodiment
To make the object, technical solutions and advantages of the present invention clearer, the present invention is made into one below in conjunction with accompanying drawing
It is described in detail on step ground.
Embodiment 1：
1st, materials and methods
1.1 material to be tested
Tobacco bred uses Yun yan85, the susceptible bacterial wilt of the kind.Antagonistic Fungi uses early stage from health tobacco rhizosphere soil
The L-25 that separation screening arrives（Brevibacillus brevis,Brevibacillus brevis）And L-9（Lou Che Shi streptomycetes,Streptomyces rochei）, wherein, L-25 protects through China Committee for Culture Collection of Microorganisms's common micro-organisms center
Hide（CGMCC No.3175）.Organic fertilizer carrier is the amino acid fertilizer that rapeseed meal digests and cow dung with 1:1（Mass ratio）Mixing,
Containing organic matter 33.8%, amino acid 4.3%, N 4.20%, P2O5 2.26%、K2O 1.08%。
1.2 suppress the preparation of tobacco bacterial wilt type biological organic fertilizer
Antagonistic Fungi L-25 is inoculated in beef extract-peptone fluid nutrient medium, L-9 is inoculated in Gause I fluid nutrient medium,
Triangular flask liquid amount is 1/5,30 DEG C, 170 r/m vibrate 48 h.Above-mentioned 6000 × g of zymotic fluid is centrifuged into 10 min, collects thalline
The organic fertilizer carrier after sterilizing is inoculated in by 5% inoculum concentration.It is 40% to adjust organic fertilizer water content, and ferment 5~6 d, periodically turns over throwing.
After fermentation ends, counted by dilution plate gradient and quantitative fluorescent PCR determines L-9 and L-25 biological organic fertilizer living bacteria counts
And Antagonistic Fungi quantity（Table 1）.The biological organic fertilizer of L-25 and L-9 difference secondary fermentations is with 1:1（w:w）Manufactured biology after mixing
Organic fertilizer is referred to as BOF.
Micro organism quantity before and after the antagonism microzyme secondary fermentation of table 1（cfu/g）
1.3 field experimental design
Respectively at 2012-2014 in Guizhou Province In Qiannan Changshun County Guang Shun towns pit hole village（26.03 ° of N, 106.45 ° of E）Even
It is continuous to carry out 3 years long-term tillage repairing tests.The bacterial wilt incidence of disease reaches 100 before selected long-term tillage repairing test experimental tests
%, decide not to plant tobacco then.Field test sets four processing altogether：T1（CK）：Only apply 975 kg/hm2Tobacco
Special complex fertilizer（N:P2O5:K2O=10:15:25）；T2：Biological organic fertilizer processing（BOF）, applying 817.5 kg/hm2Cigarette
375 kg/hm are affixed by the basis of careless special complex fertilizer2BOF, phosphorus and potassium deficiency part use calcium superphosphate and sulfuric acid respectively
Potassium polishing；T3：Soil deep tillage is taken on the basis of T2（Break plough sole）, depth is more than 30cm.T4：T3 processing on the basis of
Apply 600 kg/hm within 20 days before transplanting2Quick lime adjusts soil pH.
For trying yellow earth pH 5.45, organic matter 48.6 g/kg, the full g/kg of nitrogen 2.29, the mg/kg of alkali-hydrolyzable nitrogen 158, rapid available phosphorus
48.9 mg/kg, the mg/kg of available potassium 543.4 repetition district's groups are often handled, per 120 plants of cigarette strains of district's groups, add protection row etc. to take up an area altogether
0.05 hm2.The administration of field management and top dressing is identical with other local vegas.
1.4 field test disease survey and correlated traits measurement
Every 10 days, observation tobacco pathogenesis situation simultaneously determined rhizosphere soil microorganism within 40 to 90 days after tobacco transplant
Pathogen and Antagonistic Fungi quantity and community structure and function diversity, tobacco leaf are plucked and each processing yield are calculated after toasting.
1.4.1 disease index and control effect
Bacterial wilt investigates grade scale（In units of strain）：Disease index and control effect are investigated, the methods of using Liu
It is measured.Disease index and control effect are calculated as follows：
1.4.2 tobacco production determines
About 60~70 days after tobacco transplant, blade start it is gradually ripe from bottom to top, according to tobacco leaf mature condition by it is lower extremely
Upper Several harvests and being put in barn are toasted, and its Output value is calculated after baking.
1.5 soil surface characters and Antagonistic Fungi Fluctuation of The Number relation
Soil genomic DNA is extracted using Soil DNA Isolation Kit (Omega), using 1% Ago-Gel
Electrophoresis detection DNA purity and concentration.Using sample DNA as amplification template, pathogen is used respectively（Ralstonia solanacearum）With Antagonistic Fungi（Brevibacillus brevis）Specific primer in quantitative real time PCR Instrument
（StepOne Plus Real-time PCR System, ABI）Upper carry out amplified reaction, reaction confirm amplification curve after terminating
And melt curve analysis, the Ct values of each sample are recorded, and Ct values are substituted into calibration curve equation, calculate the first primordium of sample template
Because of copy number, the Ralstonia solanacearum of every gram of dry ground or the gene copy number of Brevibacillus brevis are finally conversed.
Fluorescent quantitative PCR result shows that the amplification efficiency of Ralstonia solanacearum reaches 91%, the amplification of Brevibacillus brevis
Efficiency reaches 100%, standard curve R2>0.99.Ralstonia solanacearum calibration curve equation is CtR=- 3.564C0R+38.744,
Brevibacillus brevis calibration curve equation is：CtB=-3.322C0B+37.209.
L-9 is counted and is used Lou Che Shi streptomycete selective mediums.Short gemma in fluorescence quantifying PCR method measure soil
Bacillus L-25 and pathogen Ralstonia solanacearum（Ralstonia solanacearum）Quantity.Antagonistic Fungi quantity is L-9 and L-
25 quantity sums.
1.6 rhizosphere soil microorganism faunas detect
Transplanting gathers rhizosphere soil after 90 days, and cigarette strain is slowly uprooped, and the soil block of attachment is fallen in gently concussion, and root is connected
It is placed in the soil that does not fall is shaken on root in 10mL aqua sterilisas, the soil obtained after the min of ultrasonic oscillation 15 is Rhizosphere Soil
Earth.
1.6.1 PCR reacts
The soil DNA of said extracted, the sample of vector is taken in centrifuge tube, 1 ng/ μ L are diluted to using sterilized water.With dilute
Genomic DNA after releasing is template, according to the selection in sequencing region, uses the special primer with Barcode；Use New
The Phusion High-Fidelity PCR Master Mix with GC Buffer of England Biolabs companies.Make
Enter performing PCR with efficient and high-fidelity enzyme, it is ensured that amplification efficiency and accuracy.Primer corresponding region：16S V4 areas primer is
515F-806R.PCR primer carries out electrophoresis detection using the Ago-Gel of 2 % concentration；Carried out etc. according to PCR primer concentration dense
Sample mixing is spent, PCR primer is detected using 2% agarose gel electrophoresis after fully mixing, uses Thermo Scientific companies
GeneJET glue reclaim kit recovery products.
1.62 library constructions, sequencing and information analysis
Use the NEB Next Ultra DNA Library Prep Kit of New England Biolabs companies
For Illumina build the structure that storehouse kit carries out library, and the library built is quantified by Qubit and library detection, qualified
Afterwards, upper machine sequencing is carried out using MiSeq.Based on Illumina MiSeq microarray datasets, double end sequencings are utilized（Paired-
End）Method, structure small fragment library carries out double end sequencings.Prepared by pedotheque sees Fig. 4 with the process of upper machine sequencing.
Obtained initial data is sequenced, a certain proportion of interference data be present, in order that the result of information analysis is more accurate
Really, reliably, initial data is spliced first, filtered, obtain valid data.It is soft using Uparse to be then based on valid data
Part (Uparse v7.0.1001) is to whole Effective Tags Sequence clusterings of all samples, with 95% uniformity
(Identity) carry out OTUs (Operational Taxonomic Units) clusters and species taxonomy analysis, and by OTU and
Species annotation combines, so as to obtain the OTUs of each sample and pedigree of classifying fundamental analysis result.Again to OTUs carry out abundance,
Diversity indices etc. is analyzed, while carries out the statistical analysis of structure of community in each categorization levels to species annotation.Finally exist
On the basis of analyzing above, it can carry out a series of based on OTUs, Shannon, the cluster analysis of species composition, principal component point
Analysis（Principal Component Analysis, PCoA）Deng.
2nd, result and analysis
The Synthetical prevention effect of 2.1 tobacco bacterial wilts
After biological prevention and control aggregate measures, each processing cigarette strain all breaks out bacterial wilt after transplanting 53d in 2012, each processing
Between control effect without significant difference.Each processing in 2013 is considerably better than 2012 to the control effect of tobacco bacterial wilt（Table 2）,
Control effect of the T4 processing in the tobacco time of infertility is significantly higher than T2 and T3 processing, after tobacco gives birth to 90d still respectively than T2 with
T3 high 78.1% and 18.2%.The bacterial wilt of T4 in 2014 and T3 processing occurs to postpone 40d, T4 processing prevention and control after 60d than control
Significant effect is higher than T3 and T2 processing, and control effect is 5.25 and 0.99 times of T2 and T3 respectively after 90d.
The Synthetical prevention tobacco bacterial wilt control effect dynamic change of table 2
2.2 yield and output value
Because each processing cigarette strain all breaks out bacterial wilt after transplanting 60d in 2012, the Output value between each processing is zero（Table
3）.T4 processing yield in 2013 is significantly higher than other each processing, respectively 3.71,1.61 and 1.13 times of T1, T2 and T3 processing,
The output value of T4, T3 and T2 processing is the 3.44 of T1,2.62 and 1.87 times respectively.T4 processing yield in 2014 and the output value are significantly higher than
Other processing, T3 processing is substantially less than between T1 and T2 yield and the output value without significant difference bacterium.
Changshun Synthetical prevention experiment Output value of table 3
The growth and decline relation of 2.3 soil surface characters and Antagonistic Fungi
From figure 1 it appears that when repairing 1 year（2012）Each processing pathogen quantity occupies height after tobacco transplant 30d
Under not, it is always held at more than 107 cfu/g soil, and the Antagonistic Fungi quantity time of infertility of T2, T3 and T4 processing is substantially linear
Decline and be finally reduced to 104 cfu/g soil；After 2 years repair（2013）, the pathogen quantity of T1 processing is to reach after 30d
To 107 cfu/g soil, T2, T3 and the T4 handled by biological organic fertilizer handles pathogen quantity and the trend slowly risen is presented,
But the Antagonistic Fungi that biological organic fertilizer is respectively handled is all remarkably higher than each self-corresponding pathogen quantity before 70d, in 80d, T3 and
Without significant difference between the pathogen quantity and Antagonistic Fungi quantity of T4 processing, and the pathogen quantity of T2 processing is Antagonistic Fungi
1.49 times into significant difference；Slow ascendant trend is presented in T1 in 2014 and other processing pathogen quantity before 50d, in 50d
T1 handles pathogen quantity and risen rapidly afterwards, and T2 processing pathogen quantity is just presented ascendant trend after 70d, and T4 cause of disease bacterium numbers
Amount is then all the time below the 106 cfu/g soil orders of magnitude, after the d of tobacco transplant 90, respectively handles in addition to T4 cause of disease bacterium number in soil
Amount rises to more than 107 cfu/g soil.The trend gradually reduced, T2 processing cause of diseases are presented in the time of infertility for the quantity of Antagonistic Fungi
The quantity of bacterium and Antagonistic Fungi crosses the time in 72d, and the time that crosses of T3 processing is 78d, and T4 Antagonistic Fungi quantity after 90d still
So it is higher than pathogen quantity, reaches 1.23 × 106 cfu/g.soil, with the extension Antagonistic Fungi of biological prosthetic prevention and control time
Quantity, which reduces speed, tends to be slow.
2.4 rhizosphere soil microorganism Floristics
2.4.1 different disposal rhizosphere soil microorganism species abundance is horizontal
The microorganism category horizontal distribution significant difference handled through analysis, T1 and T4, T1 processingPlanctomyces, Sphingobium, Rhodanobacter, Gemmata, Candidatus_NitrososphaeraWithStenotrophomonas
Six category are the higher population of abundance level, andChitinophaga、Niastelia、MethyloteneraDeng category relative abundance
It is horizontal relatively low, and T4 handles the higher population of abundance level and only hadNitrospiraWithNovosphingobium, relatively low only hasPhenylobacteriumCategory, abundance difference is smaller between other microorganisms belong to, and level distribution is close, at overall microbiologic population
In balance, uniform state.
2.4.2 different disposal rhizosphere soil microorganism door is horizontal relatively
Through analyzing, T4 processingCyanobacteria（Cyanobacteria door）Relative amount is significantly larger than T1 processing.Cyanobacteria is
A kind of large-scale prokaryotic micro-organisms that can carry out acting on entirely from oxygen light, can improve rice field and other Soil Nitrogen water by fixed nitrogen
It is flat, and in T1Acidobacteria（Acidfast bacilli door）WithCrenarchaeota（Spring Gu bacterium）Relative amount is significantly larger than T4.
2.4.3 different disposal rhizosphere soil microorganism alpha diversity indices
As can be seen from Table 4, T2, T3 and T4 after Synthetical prevention, its OTUs quantity are significantly higher than T1, diversity
For index Shannon four in gradually rising state from T1 to T4, T4 diversity indices is higher than T1.
The Alpha diversity of the different disposal of table 4
Note：Alpha diversity indices under different disposal edaphon kind is horizontal is obtained based on 97% sequence similarity
Arrive.
2.4.4 different disposal rhizosphere soil microorganism Community composition PCoA is analyzed
PCoA is a kind of multidimensional variable extracting data primary variables from complexity, and carries out visualization method.It is right
After Beta diversity carries out PCoA analyses, as shown in Fig. 2 T3 and T4 processing diversity of soil microorganism relation is nearer, it is in
Lower-left half portion, the diversity of soil microorganism of T2 processing close in the upper left quarter as limit between T1 processing and T2, T3 and T4 processing
It is farther out, in the upper right quarter as limit.
2.4.5 different disposal rhizosphere soil microorganism species similarity
With UPGMA（Unweighted PairGroup Method with Arithmetic mean）For clustering method,
By carrying out cluster analysis to sample, the similitude of species composition is studied.From figure 3, it can be seen that T3 and T4 clustering relationships
It is relatively near, and T1 and the clustering relationships of each processing are farthest.
Embodiment 2：
Following handle is carried out to soil before tobacco is planted：1）Soil pretreatment：1 month after upper 1 year tobacco harvests,
Typically within 10~November, with quick lime 100kg/ mus, upper soll layer is uniformly sprinkling upon；2）Late-maturing rape is sowed on soil,
And make green manuring processing in month in coming year 2-3；3）Soil is reprocessed：Upper soll layer is uniformly sprinkling upon again with the kg/ mus of quick lime 100；
4）Soil is ploughed deeply, depth is more than 30cm；5）Apply the biological organic fertilizer BOF in embodiment 1.
Embodiment 3：
Following handle is carried out to soil before tobacco is planted：1）Soil pretreatment：1 month after upper 1 year tobacco harvests,
Typically within 10~November, with quick lime 90kg/ mus, upper soll layer is uniformly sprinkling upon；2）Wheat is sowed on soil, and is being come
Year makees green manuring processing in 2-3 months；3）Soil is reprocessed：Upper soll layer is uniformly sprinkling upon again with the kg/ mus of quick lime 90；4）To soil
Earth is ploughed deeply, and depth is more than 30cm；5）Apply the L-25 biological organic fertilizers through secondary fermentation in embodiment 1.
Embodiment 4：
Following handle is carried out to soil before tobacco is planted：1）Soil pretreatment：1 month after upper 1 year tobacco harvests,
Typically within 10~November, with quick lime 90kg/ mus, upper soll layer is uniformly sprinkling upon；2）Sow clover on soil, and
Month in coming year 2-3 makees green manuring processing；3）Soil is reprocessed：Upper soll layer is uniformly sprinkling upon again with the kg/ mus of quick lime 100；4）It is right
Soil is ploughed deeply, and depth is more than 30cm；5）Apply the L-9 biological organic fertilizers through secondary fermentation in embodiment 1.
Claims (4)
- A kind of 1. tobacco bacterial wilt Synthetical prevention method, it is characterised in that：Following handle is carried out to soil before tobacco is planted：1） Soil pretreatment：Within 10~November after upper 1 year tobacco harvests, with quick lime 90~100kg/ mus, soil is uniformly sprinkling upon Top layer；2）Green manure crop is sowed on soil, and makees green manuring processing in month in coming year 2-3；3）Soil is reprocessed：With life after green manuring The kg/ mus of lime 90~100 are uniformly sprinkling upon upper soll layer again；4）Soil is ploughed deeply；5）Apply biological organic fertilizer；Will be short of money Antibacterial L-25 is inoculated in beef extract-peptone fluid nutrient medium, and triangular flask liquid amount is 1/5,30 DEG C, 170 r/m vibrate 48 h, Then 6000 × g of zymotic fluid is centrifuged into 10 min, collects thalline and be inoculated in the organic fertilizer carrier after sterilizing, regulation by 5% inoculum concentration Organic fertilizer water content be 40%, ferment 5~6 d, periodically turn over throw L-25 secondary fermentations biological organic fertilizer；L-9 is inoculated in height Family name's No.1 fluid nutrient medium, triangular flask liquid amount be 1/5,30 DEG C, 170 r/m vibrate 48 h, then by 6000 × g of zymotic fluid from The min of the heart 10, collect thalline and be inoculated in the organic fertilizer carrier after sterilizing by 5% inoculum concentration, regulation organic fertilizer water content is 40%, hair The d of ferment 5~6, periodically turn over throw L-9 secondary fermentations biological organic fertilizer；The biological organic fertilizer is that L-25 and L-9 difference is secondary The biological organic fertilizer of fermentation is with mass ratio 1:Manufactured biological organic fertilizer after 1 mixing；The green manure crop is late-maturing rape, wheat Or clover.
- 2. tobacco bacterial wilt Synthetical prevention method according to claim 1, it is characterised in that：The organic fertilizer carrier is dish The amino acid fertilizer of dregs of rice enzymolysis is with cow dung with mass ratio 1:1 mixing, containing organic matter 33.8%, amino acid 4.3%, N 4.20%, P2O5 2.26%、K2O 1.08%。
- 3. tobacco bacterial wilt Synthetical prevention method according to claim 1, it is characterised in that：The L-25 is using early stage From health tobacco rhizosphere soil separation screening to Antagonistic Fungi.
- 4. tobacco bacterial wilt Synthetical prevention method according to claim 1, it is characterised in that：The L-9 is using early stage From health tobacco rhizosphere soil separation screening to Antagonistic Fungi.
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| CN105777262A (en) * | 2016-03-21 | 2016-07-20 | 贵州省烟草科学研究院 | Growth-promoting bioorganic fertilizer special for tobacco and application thereof |
| CN105969686A (en) * | 2016-05-18 | 2016-09-28 | 贵州省烟草科学研究院 | Bacterial-wilt-resistant Streptomyces rochei strain and bio-organic fertilizer thereof |
| CN108660141B (en) * | 2018-05-28 | 2021-08-03 | 贵州省烟草科学研究院 | Application of NtCNGC1 gene in bacterial wilt resistance of tobacco |
| CN108586116A (en) * | 2018-08-10 | 2018-09-28 | 保康丰源肥业有限责任公司 | A kind of biological organic fertilizer and preparation method thereof of prevention pepper ralstonia solanacearum |
| CN110537470A (en) * | 2019-05-22 | 2019-12-06 | 贵州省烟草公司黔南州公司 | comprehensive prevention and control technology for tobacco rootstock diseases |
| CN110683875A (en) * | 2019-11-13 | 2020-01-14 | 贵州中烟工业有限责任公司 | Biological bacterial fertilizer for preventing bacterial wilt of flue-cured tobacco, preparation method and application |
| CN111328491A (en) * | 2020-03-12 | 2020-06-26 | 湖南农业大学 | Method for improving tobacco planting soil acidity and fertilizing by reconstructing plough layer |
| CN113186138A (en) * | 2021-06-08 | 2021-07-30 | 南京本源生态农业科技有限公司 | Biocontrol strain and application thereof in bacterial wilt prevention and control and crop growth promotion |
| CN115152568A (en) * | 2022-06-14 | 2022-10-11 | 安徽省农业科学院烟草研究所 | Method for repairing continuous cropping obstacles of tobacco field |
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| CN102154156B (en) * | 2010-12-28 | 2013-02-13 | 南京农业大学 | Antagonists capable of preventing and controlling tomato bacterial wilt or knot nematode and microbial organic fertilizer thereof |
| CN103270915B (en) * | 2012-08-22 | 2015-03-18 | 湖北省烟草科研所 | Method for preventing and treating tobacco bacterial wilt |
| CN103190222B (en) * | 2013-04-28 | 2015-01-28 | 中国烟草总公司重庆市公司烟草科学研究所 | Method for controlling tobacco bacterial wilt |
| CN103704069A (en) * | 2013-12-11 | 2014-04-09 | 南京农业大学 | Method of three-dimensionally preventing and controlling tomato bacterial wilt |
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| CN104531588A (en) * | 2015-01-07 | 2015-04-22 | 南京农业大学 | Effective antagonistic strain for preventing and controlling continuous cropping tobacco bacterial wilt disease, microorganism organic fertilizer including effective antagonistic strain, and production method and application of effective antagonistic strain |
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