CN106834180B - Improve enterobacteria 3bh19 and its application of Phyllosphere of cucumber Tiny ecosystem prevention and treatment downy mildew - Google Patents
Improve enterobacteria 3bh19 and its application of Phyllosphere of cucumber Tiny ecosystem prevention and treatment downy mildew Download PDFInfo
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Abstract
The invention discloses the enterobacteria 3bh19 of improvement Phyllosphere of cucumber Tiny ecosystem prevention and treatment downy mildew and its applications.Biocontrol microorganisms 3bh19 is enterobacteria (Enterobacter sp.).Concentration 1 × 10 is made in the bacterium8The microbial inoculum of CFU/mL concentrations above, in greenhouse under the conditions of normal management, enterobacteria 3bh19 microbial inoculum leaf enclose spray processing after 63.32% or more is reached to cucumber downy mildew biocontrol effect；And it the 30th day cucumber plant biomass and compares after handling and compares and increase above 32% or more；In field, to a variety of cucumber downy mildew control efficiency 53.55% or more, growth-promoting effect highly significant；And the resistance of plant can be significantly proposed, leaves of plants can be significantly improved and enclose microbial diversity, improve Tiny ecosystem.
Description
Technical field
The invention belongs to plant protection art, it is related to the enterobacteria for improvement Phyllosphere of cucumber Tiny ecosystem prevention and treatment downy mildew
3bh19 and its application.
Background technique
Cucumber (Cucums sativus L.) belongs to Curcurbitaceae (Cucurbitaceae), Cucumis in Plant Taxonomy
(Cucums), it is one of global ten big vegetable growing crops, is occupied an important position in China's the "shopping basket' project.According to the United Nations
Food and Agriculture Organization's statistics, from 2000 to 2004 year, China's cucumber cultivation area is from 116 549 hectares of (hm2) increase to 1
502 900 hectares of (hm2), yield increases to 25 558 000 million tons (Mt) from 19 869 181 million tons (Mt).With field
Between facility plastic greenhouse it is prevailing, huge change occurs for cucumber cultivation mode.According to incompletely statistics, China cucumber largely uses at present
It is planted in field facility plastic greenhouse, while bringing tremendous economic income to vegetable grower, to plant protection expert it is also proposed that being directed to disease control
High request.As China industrial crops and vegetable at protected field area extend rapidly, wherein especially cucumber is this simultaneous with vegetables
The famous bulk vegetable of fruit.In the facility plastic greenhouse green cucumber of field, the disease of Chang Fasheng has gray mold of cucumber, cucumber mosaic virus
Viral disease, powdery mildew of cucumber, cucumber bacterial angular leaf spot, cucumber downy mildew and cucumber root-knot nematode, cucumber fusarium axysporum etc., wherein especially
With cucumber downy mildew, popular fast, severe disease, difficulty of prevention and cure are big.Its leaf diseases because its condition be suitable for and due to vegetable continuous cropping it is increasingly tight
Weight, wherein downy mildew Host transfer, is 10-30% in big Tanaka's average attack rate；Facility plastic greenhouse average attack rate 50%, severe one
Total crop failure.
Cucumber downy mildew is caused by Cuba artificial downy mildew (Pseudoperonospora cubensis), is the whole world
One of main leaf diseases in cucumber producing region in range, the whole world about more than 70 country this disease successively occurs, to Huang
Melon production threatens very big.During carrying out disease investigation to Beijing periphery, Shandong, Hebei and other places, it has been found that the disease causes harm sternly
Weight, in serious greenhouse of falling ill, for sick leaf rate up to 85% or more, fruit development is slow, and yield reduces, and causes to peasant very big
Economic loss.Only Hebei Qing County spring greenhouse is investigated, and cucumber downy mildew occurring area is more than 1330hm2, is lost more than 20,000,000
First (Li Baoju, 2008)；It is investigated according to Jiangsu Tongzhou agricultural station in yellow sky brilliance village's Vegetable Base, the Huang of 2 cucumber booths
Melon downy mildew canopy disease incidence is 33.33%, and sick leaf rate average 5%, the village Xin San Yuan on March 22 investigates 10 cucumber booths, canopy
Disease incidence 50%, sick leaf rate average 10.98%, is early stage (Jiangsu Tongzhou, 2009)；In Jiangsu Huaian, cucumber is mould
White disease is to occur that most common, the duration is long, endangers most serious disease in heliogreenhouse facility plastic greenhouse, and cucumber is used at present
The maximum disease of dose causes the underproduction up to 20%-40%, or even total crop failure (Liu Feng, 2011) throughout the year.
It is in recent years that the generation and induction that disease is controlled using environmental beneficial microorganism, which improve crop anti-adversity ability,
The hot spot of research, especially these microorganism no pollution to the environment, cause scientists using biological control means control disease,
And the interest of reply poor environment.Enterobacteria (Enterobacter sp.), can produce in the growth course of its own
A series of metabolite, these metabolites enable enterobacteria to have the activity for widely inhibiting fungi and bacterium, or
Person's inducing plant resistance.It can in different ways or many metabolic processes of approach influence plant, increase plant root long change and plant
Object Root morphology is built up, and absorption of the root system to moisture content and nutrient is promoted, and enterobacteria can be improved the disease resistance of plant, support
Anti-microbial pathogen infects.
Enterobacteria is distributed very wide in nature, while can generate inhibiting effect to a variety of fungies and bacterium.Cause
This, has broad application prospects in terms of biological control, can be appropriate instead of traditional chemical pesticide.Meanwhile with point
The gene function of the raising of sub- investigative technique, enterobacteria is more appreciated that, increases applicable feasibility.And fermentation condition
Optimization so that the large-scale application of enterobacteria also has objective condition.
Summary of the invention
The purpose of the present invention is providing for the existing deficiency for preventing and treating prevention and treatment cucumber downy mildew, one kind is environmentally protective to be mentioned
The enterobacteria 3bh19 that high broad-spectrum plant diseases prevention, growth-promoting, Tiny ecosystem are improved.
It is a further object of the present invention to provide the applications of the enterobacteria.
The purpose of the present invention can be achieved through the following technical solutions:
Biocontrol microorganisms 3bh19 is enterobacteria (Enterobacter sp.).For a variety of cucumber diseases preventions, growth-promoting, degeneration-resistant, micro- life
The biocontrol microorganisms 3bh19 of state improvement, which was named as enterobacteria (Enterobacter sp.), on 06 13rd, 2016
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, culture presevation number is CGMCC NO.12608.
The enterobacteria 3bh19 preferably in prevention and treatment cucumber downy mildew, promotes cucumber overground part or underground part to grow and mention
High-quality, improve its resistance, Tiny ecosystem improvement in application；The resistance selected from drought resisting, cold-resistant, it is heat-resisting, waterlogging in
It is one or more.
The microbial inoculum of enterobacteria 3bh19 preparation of the present invention.
Microbial inoculum of the present invention is preferably prepared by the following method to obtain: by enterobacteria 3bh19 described in claim 1
28 DEG C of 180rpm shaken cultivation 12-16h in LB culture solution, then 6000rpm be centrifuged 10min obtain thallus, with aqua sterilisa into
Microbial inoculum is made in row dilution, and viable bacteria total concentration is 1 × 10 in microbial inoculum finished product9-1×1010CFU/mL。
The microbial inoculum is enclosed by pouring root or leaf sprays processing.
Beneficial effect
The present invention, which is utilized, prevents and treats cucumber downy mildew to the microbial bacterial agent of environment nonhazardous, provides a kind of environmentally protective
Enterobacteria microbial inoculum promotes the growth of cucumber plant and improves quality and to improve its resistance (including drought resisting, cold-resistant, heat-resisting, waterlogging
Deng the bad climates such as resistance cold spell in later spring).It is ineffective or have the problems such as residual to solve other methods such as chemical agent, protects
Retaining ring border promotes agricultural sustainable development.
Have after the enterobacteria 3bh19 preparation root irrigation to the plant height and root long of cucumber and significantly improve, has to plant
There are certain growth-promoting functions, compared with the control group, the 30th day biomass compares increasing with control to the cucumber handled through 3bh19 after processing
Add more than 32%.Enterobacteria microbial inoculum, can be room temperature preservation -1 year 6 months.The plant greenhouse diseases prevention of enterobacteria 3bh19 microbial inoculum processing promotees
It gives birth to test result to show to be up to diseases control 62.32% or more, the plant field diseases prevention of enterobacteria 3bh19 microbial inoculum processing promotees
Raw test result shows to be up to diseases control 53.55% or more, and the quality of cucumber can be improved, and improves leaf and encloses micro- life
The diversity of object.
Detailed description of the invention
Fig. 1, enterobacteria 3bh19 microbial inoculum handle the biocontrol effect to cucumber downy mildew
The influence of Fig. 2, the processing of enterobacteria 3bh19 microbial inoculum to cucumber biomass
The influence of Fig. 3, the processing of enterobacteria 3bh19 microbial inoculum to cucumber resistance
Phyllosphere of cucumber sample DGGE is analyzed in Fig. 4, the processing of enterobacteria 3bh19 microbial inoculum
1-3 swimming lane: 3bh19 bacterial strain 0d Phyllosphere of cucumber bacteria samples (3 repeat)；4-6 swimming lane: 3bh19 bacterial strain 30d cucumber leaf
Enclose bacteria samples (3 repeat)；7-9 swimming lane: Control 0d Phyllosphere of cucumber bacteria samples (3 repeat)；10-12 swimming lane: Control
30d Phyllosphere of cucumber bacteria samples (3 repeat)；Arrow is new band.
Biomaterial preservation information
Enterobacteria 3bh19, for enterobacteria (Enterobacter sp.).The micro- life of China was preserved on 06 13rd, 2016
Object culture presevation administration committee common micro-organisms center, culture presevation number are CGMCC NO.12608, address: Beijing's southern exposure
The institute of microbiology, the Chinese Academy of Sciences, institute of area North Star West Road 1.
Specific embodiment
The molecules of 1 enterobacteria 3bh19 of embodiment is identified
After bacterial genomes kit (SBS Genetech company) extracts bacterial gene DNA, with gyrB universal primer gyrB1 (5'-
GAAGTCATCATGACCGTTCTGCAYGCNGGNGGNAARTTYGA-3')；gyrB2R(5'–
AGCAGGATACGGATGTGCGAGC CRTCNACRTCNGCRTCNGTCAT-3') gyrB gene is amplified from genomic DNA
Segment.Specific amplification system and program (25ul) are as follows: each 1 μ l of primer 1,2 (10 μM)；Mix:12.5μl；Taq:0.5μl.
ddH2O:10μl；Template: 1 μ l.94 DEG C of initial denaturation 5min, 94 DEG C of denaturation 1min, 56 DEG C of annealing 2min, 72 DEG C of extension 2m in, 35
A circulation, 72 DEG C of extension 10min, 4 DEG C of preservations.Pcr amplification product is detected with 1% agarose gel electrophoresis, 90V,
30min；EB dyes 30min, detects amplified production.Amplified production is connected to pMD19-T carrier after purification, conversion in
Escherichia coli DH5 α, extracts positive colony plasmid, and NCBI (Natio nal is submitted in the sequencing of commission TaKaRa company
Center for Biotechnology Information) it is compared.Enterobacteria (E is accredited as according to comparison result
nterobacter sp.)。
Embodiment 2
It is a kind of to improve cucumber downy mildew biological control effect and growth-promoting, degeneration-resistant, Tiny ecosystem improvement enterobacteria 3bh19,
Containing microbe composition: enterobacteria 3bh19 (deposit number is CGMCC NO.12608).By being separated in the Cucumber rhizosphere soil of Huaian
It arrives.
Enterobacteria 3bh19 microbial inoculum the preparation method is as follows:
Enterobacteria 3b,h19 28 DEG C of 180rpm shaken cultivation 12-16h in LB culture solution, then 6000rpm is centrifuged 10min
Thallus is obtained, thallus is diluted with aqua sterilisa, microbial inoculum is made, viable bacteria total concentration is 1 × 10 in microbial inoculum finished product9-1×
1010CFU/mL。
3 enterobacteria 3bh19 of embodiment tests the greenhouse growth-promoting of cucumber
Method: will participate in the experiment cucumber seeds 10%H2O2Surface sterilization 10min, sterile water dry spare after cleaning.Each battalion
It supports alms bowl and sows 10 seeds.After normal watering management 4 weeks, by the consistent transplantation of seedlings of growing way into 330ml plastic cup, when transplanting, will
The enterobacteria 3bh19 microbial inoculum to have suspended is with 1.0 × 107The concentration pouring root of CFU/mL is applied to crop root, every plant of 30mL amount.So
It is placed in greenhouse and cultivates, 28 DEG C, 70% relative humidity.Using clear water processing as control, every group of 48 young plant of processing is put at random
It sets, altogether in triplicate.The biomass such as cucumber plant plant height, stem thickness, fresh weight, dry weight are measured after 30d.
Influence of the 1 enterobacteria 3bh19 of table to cucumber biomass
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
As seen from the above table, said preparation has significantly the plant height and root long of cucumber to after enterobacteria 3bh19 root irrigation
Improve, to plant have certain growth-promoting functions, through 3bh19 handle cucumber compared with the control group, the 30th day biology after processing
Amount is compared with control and increases above 32%.
4 enterobacteria 3bh19 of embodiment tests the cucumber downy mildew greenhouse diseases prevention of cucumber
To participate in the experiment cucumber seeds 10%H2O2Surface sterilization 10min, sterile water dry spare after cleaning.Each nutritive cube
Sow 10 seeds.After normal watering management 4 weeks, by the consistent transplantation of seedlings of growing way into 330ml plastic cup, 10 are grown to it
True leaf carries out spraying treatment.Every processing sets 4 repetitions, 48 seedlings of each repetition.The every young plant of processing group sprays what 30ml had suspended
1.0×107The enterobacteria 3bh19 aqueous solution of CFU/mL concentration, control group are spraying in the same way with equivalent aqua sterilisa；It is above each
Final concentration of 0.01% surfactant Tween-20 is added in a processing group when spraying.5d is sprayed in a manner of spraying treatment
Inoculum density is 105The cucumber downy mildew pathogen of a sporangium/ml.Disease severity is investigated after inoculation pathogen 15d and is counted
Calculate biocontrol effect.Disease survey method: 0 grade: without illness；1 grade: vaccination has slight scab, and diameter is less than 0.5cm；3 grades:
Lesion diameter 0.5-1.3cm；5 grades: vaccination yellow area accounts for cotyledon area 1/2 hereinafter, necrosis area accounts for 1/3 or less；7 grades:
Necrotic plaque area accounts for leaf area 1/3-2/3；9 grades: necrotic plaque area accounts for 2/3 or more leaf area or complete stool is withered.
Disease index (%)=[∑ (the sick series × disease grade plant number)/(highest disease grade × total plant number)] × 100；
Biocontrol effect (%)=[(control disease severity-processing group disease severity)/control disease severity] × 100
Influence of the 9 enterobacteria 3bh19 of table to cucumber plant
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
Test result shows that enterobacteria 3bh19 can significantly reduce the disease severity of greenhouse cucumber downy mildew, to disease
Preventive effect is up to 62.32%, and can significantly improve yield and quality.
The prevention and treatment fruit cucumber downy mildew of 5 enterobacteria 3bh19 microbial inoculum of embodiment processing and raising fruit yield test
To participate in the experiment cucumber seeds 10%H2O2Surface sterilization 10min, sterile water dry spare after cleaning.
Cucumber seeds after vernalization are seeded in equipped in basin Portland, one, every basin keeps soil before sowing in sterile soil
It is more dry.When tomato grew to for 2 leaf phase, it is transplanted to field, every group of 48 young plant of processing randomly places, the intestines bar that will have been suspended
Bacterium 3bh19 biocontrol agent is with 1.0 × 107The concentration of CFU/mL is sprayed for crop leaf portion, and cucumber growth-promoting, diseases prevention are counted after 30d
And disease control result.Disease survey method: 0 grade: without illness；1 grade: vaccination has slight scab, and diameter is less than 0.5cm；3
Grade: lesion diameter 0.5-1.3cm；5 grades: vaccination yellow area accounts for cotyledon area 1/2 hereinafter, necrosis area accounts for 1/3 or less；7
Grade: necrotic plaque area accounts for leaf area 1/3-2/3；9 grades: necrotic plaque area accounts for 2/3 or more leaf area or complete stool is withered.
Disease index (%)=[∑ (the sick series × disease grade plant number)/(highest disease grade × total plant number)] × 100；
Biocontrol effect (%)=[(control disease severity-processing group disease severity)/control disease severity] × 100
Test result shows that the disease severity of cucumber downy mildew can significantly drop in enterobacteria 3bh19, and average preventive effect reaches
59.96%；And yield and quality can be significantly improved.
Influence of the 9 enterobacteria 3bh19 of table to cucumber plant
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
Influence of 10 3bh19 of table to cucumber quality
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
The induced resistance test of 6 enterobacteria 3bh19 microbial inoculum of embodiment processing
Method: will participate in the experiment cucumber seeds 10%H2O2Surface sterilization 10min, sterile water dry spare after cleaning.
Cucumber seeds after vernalization are seeded in equipped in basin Portland, one, every basin keeps soil before sowing in sterile soil
It is more dry.When cucumber grew to for 6 leaf phase, by the enterobacteria 3bh19 biocontrol agent to have suspended with 1.0 × 107CFU/mL's is dense
Degree pouring root is applied to crop root, is subsequently placed in greenhouse and cultivates, arid of cutting off the water supply is carried out after 1 month, and drought stress continues 15d,
It normally waters after drought stress, observes the restoration ecosystem situation of cucumber seedling.Every group of 48 young plant of processing, randomly places, and altogether repeatedly 3
It is secondary.It is put in the greenhouse according to randomized complete-block design.
Watering carry out restoration ecosystem 1d after, enterobacteria 3bh19 processing group plant restore normal growth, and and control group plant
It is all dead.
Influence of the 11 enterobacteria 3bh19 of table to drought resistance in plants
Note: different lowercase persons indicate in 0.05 horizontal upper significant difference.
The Tiny ecosystem improved test of 7 enterobacteria 3bh19 microbial inoculum of embodiment processing
Method: will participate in the experiment cucumber seeds 10%H2O2Surface sterilization 10min, sterile water dry spare after cleaning.
Cucumber seeds after vernalization are seeded in equipped in basin Portland, one, every basin keeps soil before sowing in sterile soil
It is more dry.When tomato grew to for 2 leaf phase, it is transplanted to field, every group of 48 young plant of processing is randomly placed, be repeated 3 times altogether.To 30d
After (10 leaf), by the enterobacteria 3bh19 biocontrol agent to have suspended with 1.0 × 107The concentration of CFU/mL is sprayed for crop leaf portion,
The sampling of 30d rear blade (marks, the leaf to prevent newly growing influences) in advance, and the leaf of biocontrol microorganisms is sprayed with sterilizing scissors clip
Son.The leaf sample fetched respectively claims 10g, be transferred under aseptic condition 100ml eluent (0.1mol/L kaliumphosphate buffer,
PH7.0), 15 DEG C of 200r/min shake 30min, are ultrasonically treated 5min in ultrasonic washing instrument.Collect eluent, 4 DEG C of 10000r/
M in is centrifuged 20min, collects precipitating, is used as microbial DNA and extracts.Plant phyllospheric is directly extracted using improved chemical cleavage method
Genomic DNA, the method is as follows: take the plant phyllospheric microorganism of the above-mentioned collection of 0.2g to be suspended in 800 μ L lysates, 5 μ are added
The Proteinase K of L20mg/ml, 37 DEG C of warm bath 30min；Add 20% SDS solution make its final concentration of 2%, mix, 60 DEG C
Warm bath 1h takes out concussion 4-5 times during warm bath；Isometric chloroform is added, mixes, 1000r/min is centrifuged 8min；In supernatant
The isopropanol of 0.6 times of volume is added, is stood overnight under room temperature, 12000r/min is centrifuged 20min；Supernatant is abandoned, is resuspended in after dry
In 30 μ L TE buffers.Bacterium is expanded using 16SrDNA sequence V3 section Specific PCR primers F338-GC and R518:
F338-GC:5-CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGCA CGGGGGGACTCCTACGGGAGGCAGCAG-3′,
R518:5′-GCGTGTGTACAAGACCC-3′.Primer is synthesized by Beijing SBS Genetech gene technology Co., Ltd.PCR product is about
230bp.PCR response procedures are as follows: 94 DEG C, 7min；94℃,30s；55℃,30s；72 DEG C, 30s, 35 circulations；72℃,7min；4
DEG C save.PCR reaction system are as follows: 50 μ l systems, Buffer5 μ l；1 μ l of DNA profiling；Mg2+4μl；dNTP 4μl；Each 0.8 μ of primer
l；0.8 μ l of Taq DNA polymerase；33.6 μ l of deionized water.Primer is synthesized by Beijing SBS Genetech gene technology Co., Ltd.PCR is produced
Object is about 260bp.PCR response procedures are as follows: 94 DEG C, 5min；94℃,30s；56℃,30s；72 DEG C, 90s, 35 circulations；72℃,
10min；4 DEG C of preservations.Denaturing gradient gel electrophoresis (DGGE) analysis that PCR product carries out, DGGE is in DCode abrupt climatic change system
Uniting, (Dcode Universal Mutation Detection System) (B io-Rad) is middle to be completed.It is concentrated with the PCR of 25 μ L
Product carry out DGGE, use denatured gradient be 30%~60% 8% acrylamide gel (chemical denaturant be 100% urinate
The deionized formamide of plain 7mol L-1 and 40% (v/v)) in 1 × TAE electrophoretic buffer, 130V voltage, electrophoresis at 60 DEG C
7h.10~15min of Silver stain is used after electrophoresis, is then taken pictures with Al phimager system.DGGE map is analyzed and processed point
Analysis, using Quantity One analysis software (Bio-Rad) determine sample electrophoresis band number and band brightness peak value, obtain
To the dendrogram of DGGE map, the variation of cucumber different growing rhizosphere Tiny ecosystem is determined.
As a result (Fig. 4) is shown: enterobacteria 3bh19 can significantly improve the diversity of Phyllosphere of cucumber microorganism.
Claims (7)
1. one plant of enterobacteria 3bh19, which was named as enterobacteria (Enterobacter sp.), in 06 month 2016 13
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center day, culture presevation number is CGMCC
NO.12608。
2. enterobacteria 3bh19 described in claim 1 preparation promote cucumber growth and improve quality, improve drought resistance, change
Good leaf encloses the application in the microbial inoculum of Tiny ecosystem.
3. application according to claim 2, it is characterised in that the enterobacteria 3bh19 is in preparation promotion cucumber plant
Top or underground part grow and improve quality, improve its drought resistance, Tiny ecosystem improvement microbial inoculum in application.
4. the microbial inoculum prepared with enterobacteria 3bh19 described in claim 1.
5. microbial inoculum according to claim 4, it is characterised in that be prepared by the following method to obtain: will be described in claim 1
Enterobacteria 3b,h19 28 DEG C of 180 rpm shaken cultivation 12-16 h in LB culture solution, then 6000 rpm are centrifuged 10 min and obtain
Thallus is diluted with aqua sterilisa and microbial inoculum is made, and viable bacteria total concentration is 1 × 10 in microbial inoculum finished product9 -1×1010 CFU/mL。
6. the microbial inoculum of claim 4 or 5 is promoting cucumber overground part or underground part to grow and improve quality, is improving its drought resisting
Property, Tiny ecosystem improvement in application.
7. application according to claim 6, it is characterised in that the microbial inoculum sprays processing by pouring root and leaf portion.
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| CN102321561A (en) * | 2011-10-12 | 2012-01-18 | 淮阴师范学院 | Biocontrol strain CYW72 for controlling greenhouse cucumber downy mildew and application thereof |
| CN102428964A (en) * | 2011-09-26 | 2012-05-02 | 南京农业大学 | Biocontrol compound microbial inoculum CB28 mixture for preventing and treating cucumber downy mildew |
| CN102533610A (en) * | 2012-02-09 | 2012-07-04 | 河北省农林科学院植物保护研究所 | Bacillus subtilis for preventing and treating cucumber ampho disease, and microbial agent thereof |
| CN105907689A (en) * | 2016-07-01 | 2016-08-31 | 淮阴工学院 | Biocontrol strain for preventing and controlling cucumber downy mildew and biocontrol microbial agent prepared by biocontrol strain |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102428964A (en) * | 2011-09-26 | 2012-05-02 | 南京农业大学 | Biocontrol compound microbial inoculum CB28 mixture for preventing and treating cucumber downy mildew |
| CN102321561A (en) * | 2011-10-12 | 2012-01-18 | 淮阴师范学院 | Biocontrol strain CYW72 for controlling greenhouse cucumber downy mildew and application thereof |
| CN102533610A (en) * | 2012-02-09 | 2012-07-04 | 河北省农林科学院植物保护研究所 | Bacillus subtilis for preventing and treating cucumber ampho disease, and microbial agent thereof |
| CN105907689A (en) * | 2016-07-01 | 2016-08-31 | 淮阴工学院 | Biocontrol strain for preventing and controlling cucumber downy mildew and biocontrol microbial agent prepared by biocontrol strain |
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