CN102550607A - Bacillus subtilis PTS-394 for preventing and controlling soil-borne diseases of solanaceous fruit vegetables and application thereof - Google Patents
Bacillus subtilis PTS-394 for preventing and controlling soil-borne diseases of solanaceous fruit vegetables and application thereof Download PDFInfo
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Abstract
The invention relates to a biocontrol bacterial agent. An effective active constituent consists of bacillus subtilis PTS-394. The biocontrol bacterial agent is characterized in that: the biocontrol bacterial agent is aqueous solution or wettable powder. The aqueous solution comprises the following components in percentage by weight: bacillus subtilis PTS-394 with 2 *1,010 cfu/mL of content of bacteria, 0.5 percent of NNO (sodium methylene naphthalene sulfonate), 6 percent of SDBS (sodium dodecyl benzene sulfonate) and 0.1 percent of sodium benzoate. The wettable powder comprises the following components in percentage by weight: 40 percent of bacillus subtilis PTS-394 viable bacterium raw powder with 5 *1,010 cfu/g of content of bacteria, 7 percent of NNO (sodium methylene naphthalene sulfonate), 3 percent of polyethylene glycol, 5 percent of SDBS (sodium dodecyl benzene sulfonate) and 45 percent of light calcium carbonate. The biocontrol bacterial agent is poured into vegetable roots when applied and is used for preventing and controlling vegetable soil-borne diseases such as solanaceous fruit vegetable blight and bacterial wilt mainly; and the content of effective agent components is between 104 and 108 cfu/mL or between 104 and 108 cfu/g after finished product agents are diluted. The biocontrol bacterial agent has the advantages that: the aims of preventing and controlling two kinds of soil-borne diseases by means of one kind of biocontrol bacterial agent are fulfilled by introducing the bacillus subtilis PTS-394 which is safe to people and livestock and has better inhibition effect on the solanaceous fruit vegetable blight and the bacterial wilt; the using amount of chemical agents is reduced, and the agent cost is reduced; and the ecological environment is improved, and the residues of the chemical agents in the vegetables are reduced and the same time.
Description
Technical field
The present invention relates to a kind of biocontrol fungicide and application thereof, especially a bacillus subtilis PTS-394 and the application in control two kinds of important soil-borne diseases of solanaceous vegetables (fusarium wilt and bacterial wilt) thereof.
Background technology
Along with the adjustment of crop mix, the facilities vegetable cultivated area increases year by year, and high yield, continuous cropping, diversified facilities vegetable planting structure distribution are that damage by disease and insect provides abundant nutrition and breeding condition.Therefore, the great disease of facilities vegetable is frequently broken out in recent years.At present, prevent and treat disease in the facilities vegetable production and mainly rely on chemical pesticide.This not only causes the ecotope severe contamination, and directly increases the residual of toxic chemical substance in the vegetables, and human health is brought serious harm.In order to solve contradiction between yield of vegetables and the quality, reach the purpose of the usage amount of controlling plant diseases, minimizing chemical pesticide, use the method for biological control controlling plant disease more and more to receive the national governments and the people's concern.
Biological control mainly is to utilize bio-diversity, through discharge or the introduction beneficial organism sustainable Control plant disease at natural world.All carry out the research that utilizes the biocontrol fungicide controlling plant diseases both at home and abroad since nineteen fifty, obtained first-stage success.As: utilize bacillus subtilis (Bacillus subtilis) to prevent and treat rice sheath blight disease, soybean rust, soft rot of Chinese cabbage etc.; Utilize the fusarium wilt of crops such as Pseudomonas fluorescence (Fluorescent pseudomonads) control cotton, flax, clover, cucumber; Utilize saccharomycete (Candida saitoana), pseudomonas syringae (Pseudomonas syringae) and the multiple fruit postharvest diseases of pantoea agglomerans (Pantoea agglomerans) control or the like.
Solanaceous vegetables fusarium wilt (pathogen is Fusarium oxysporum Fusarium oxysporum f.sp.lycopersici) and bacterial wilt (pathogen is the blue or green withered Raul Salmonella Ralstonia solanacearum of eggplant) are two kinds of main soil-borne diseases during vegetables produce.Bacterial wilt is claimed bacterialo wilt disease again, and field disease symptom and fusarium wilt are closely similar: how take place at the tomato duration of flowering, along with bearing fruit and fruit expands, the state of an illness increases the weight of gradually.The pathogen infection vascular bundle blocks it and carries nutriment, sick stem vertical section vascular bundle browning.Similar illness has brought very hang-up for the diagnosis and the control of two kinds of diseases.The fusarium wilt biocontrol fungicide of having reported mainly contains Trichoderma, non-pathogenic Fusarium oxysporum, pseudomonas and bacillus etc.; The bacterial wilt biocontrol bacterium agent mainly contains had no pathogenicity Ralstonia solanacearum and bacillus etc.But these biocontrol fungicides all are to be action target to single disease at present, because two kinds of disease illnesss are similar, and frequent generation inappropriate medication productions in, thus cause preventive effect not good.
One of approach that addresses this problem is exactly: with solanaceous vegetables wilt and ralstonia solanacearum is the target bacterium, screening, identifies two kinds of pathogens are all had the biocontrol fungicide and a field application of antagonism.
The academy of agricultural sciences, Jiangsu Province is from crop rhizosphere soils such as the tomato on ground such as Yinchuan, Ningxia, Shuyang, Jiangsu, Xiamen, Fujian and Kunming, Yunnan, capsicum, watermelon, and separation and purification obtains 1239 bacterial isolates.Screening obtains multiple fungal disease is had bacillus subtilis (Bacillus subtilis) PTS-394 of fine preventive effect.Its characteristic is recited as:
A, morphological feature: bacillus subtilis PTS-394 bacterial strain trophosome is shaft-like, and two ends are oval, peritrichous.Size is (0.7～0.8) micron * (2.0～3.0) microns.No pod membrane.Give birth in the gemma, ellipse, size is (0.6～0.9) micron * (1.0～1.5) microns.
B, colony morphology characteristic: when bacillus subtilis PTS-394 cultivates on beef broth peptone agar plate, the colony edge waveform, cream-coloured, there is gauffer on the surface.Quality is opaque shape, not chromogenesis.
C, physio-biochemical characteristics: bacillus subtilis PTS-394 Gram-reaction is positive.H
2O
2Enzyme reaction is positive, the V.P. reacting positive.L-arabinose, D-glucose, D-wood sugar, D-mannitol are utilized as the positive.Starch hydrolysis, gelatin liquefaction, sodium citrate utilization, dihydroxyacetone (DHA) form and react also all positive.30 ℃ of optimum growth temps are not grown more than 55 ℃.
D, kind are identified: the DNA extraction of bacillus subtilis PTS-394 adopts conventional method, adopts 1% agarose gel electrophoresis to detect its purity and concentration.Genomic DNA to extract is a template, with bacterium universal primer amplification 16S rRNA sequence.Primer BSF (27f): AGAGTTTGATCCTGGCTCAG; BSR (1541r): AAGGAGGTGATCCAGCCGCA.PCR reaction condition: 95 ℃ of 4min; 94 ℃ of 40s, 55 ℃ of 45s, 72 ℃ of 1min, 25 circulations; 72 ℃ of 5min, PCR product serve the sea and give birth to the order-checking of worker's biotechnology Services Co., Ltd after the checking of 1% agarose gel electrophoresis, on the NCBI nucleic acid database, adopt the Blast program to carry out the autoploidy search 16S rRNA sequence that records.Sequencing result shows: bacillus subtilis PTS-39416S rRNA sequence and bacillus subtilis Bacillus subtilis have very high autoploidy, and submitting GenBank, to obtain the gene accession number be JF419551.
E, bacillus subtilis PTS-394 also have many premium properties, as: fast growth; Accommodative ability of environment is strong; Can grow surely at soil and plant surface, and form dominant population; Induce the host to produce disease resistance; Secretion produces multiple antibacterial material.
At present, biocontrol fungicide (bacillus subtilis PTS-394) is used to prevent and treat two kinds of important soil-borne diseases of solanaceous vegetables (fusarium wilt and bacterial wilt) and is not seen in report.
Summary of the invention
The purpose of this invention is to provide a kind of bacillus subtilis (Bacillus subtilis) PTS-394, this bacterial strain obtained gene accession number JF419551 on NCBI nucleic acid database GenBank on February 24th, 2011.
Another object of the present invention is: upward two kinds of important soil-borne diseases of solanaceous vegetables (fusarium wilt and bacterial wilt) illness is similar to producing at present; Be difficult to diagnose and prevent and treat not good problem, screening obtains a strain all has good control efficiency to fusarium wilt and bacterial wilt biocontrol fungicide (bacillus subtilis PTS-394) and application thereof.Bacillus subtilis PTS-394 can grow rapidly at plant rhizosphere surely, forms dominant population; Secrete antibiotic related substances; The inducing plant disease resistance; Thereby realize utilizing this biocontrol fungicide to control the popular purpose of solanaceous vegetables soil-borne disease lastingly.
The invention provides a kind of biocontrol fungicide, wherein the effective active composition is made up of bacillus subtilis (Bacillus subtilis) PTS-394.
Biocontrol fungicide of the present invention wherein comprises protease and has a liking for the plain two kinds of antibiotic related substanceses of iron in the metabolite of bacillus subtilis PTS-394 secretion.
Biocontrol fungicide of the present invention, it is aqua or wetting powder.
The percentage by weight of each component of aqua is:
The percentage by weight of each component of wetting powder is:
Biocontrol fungicide of the present invention, wherein auxiliary agent can be chosen any one kind of them or multiple following material: NNO (sodium metnylene bis-naphthalene sulfonate), azone, thiophene ketone; Adhesive can be chosen any one kind of them or multiple following material: polyvinylpyrrolidone, polyethylene glycol, polyvinyl alcohol (aggregate number 12000-20000); Surfactant can be chosen any one kind of them or multiple following material: dodecyl sodium sulfate, neopelex, sodium lignin sulfonate, the withered powder of tea, spaonin powder; Preservative can be chosen any one kind of them or multiple following material: Sodium Benzoate, potassium sorbate, dehydro sodium acetate, calcium propionate, sodium Diacetate.Filler can be chosen any one kind of them or multiple following material: precipitated calcium carbonate, diatomite, kaolin, sericite in powder and white carbon black.
The purposes of biocontrol fungicide of the present invention is: be used to prevent and treat multiple soil-borne disease with pouring into the vegetables root after the described biocontrol fungicide dilution; Its characteristic comprises: described soil-borne disease is solanaceous vegetables fusarium wilt and bacterial wilt.
The present invention further provides the application of said biocontrol fungicide, it is characterized in that: the active drug component content 10 behind the finished medicines dilution agent
4-10
8Cfu/mL or 10
4-10
8Cfu/g.
The invention has the advantages that:, reach and utilize a kind of biocontrol fungicide to prevent and treat the purpose of two kinds of important soil-borne diseases of solanaceous vegetables simultaneously through introducing the person poultry safety, solanaceous vegetables fusarium wilt, bacterial wilt all being had stronger inhibiting bacillus subtilis PTS-394; Reduce the chemical agent dosage, reduce drug cost; Improved the ecological environment, reduced the residual quantity of chemical agent in vegetables simultaneously.
Description of drawings
Below in conjunction with accompanying drawing and embodiment the present invention is further specified.
Fig. 1 is bacillus subtilis PTS-394 and dilution thereof the inhibitory action to tomato wilt bacterium mycelial growth.
Fig. 2 is bacillus subtilis PTS-394 and dilution thereof the inhibitory action to the bacterial wilt of tomato bacterium.
Fig. 3 is that bacillus subtilis PTS-394 can produce and has a liking for the iron element.
Fig. 4 is that bacillus subtilis PTS-394 can produce protease.
A among the figure: bacillus subtilis PTS-394 stoste; A-1: 100 times of dilutions of bacillus subtilis PTS-394; A-2: 500 times of dilutions of bacillus subtilis PTS-394; B: tomato wilt bacterium.
Embodiment:
Bacillus subtilis PTS-394 is to the inhibitory action of wilt and ralstonia solanacearum.
Supply examination pathogen bacterial strain:
The tomato wilt bacterium is provided by Institute of Plant Protection, academy of agricultural sciences, Jiangsu Province.Cultivate 5d for 28 ℃ with PSA medium (200g potato, 20g sucrose, 20g agar-agar and 1000mL distilled water, pH 5.6～6.6), subsequent use.
The bacterial wilt of tomato bacterium is provided by Institute of Plant Protection, academy of agricultural sciences, Jiangsu Province.Cultivate 2d with 28 ℃ of vibrations of NA medium (beef extract 3.0g, yeast extract 1g, peptone 5g, glucose 10g, agar 20g, distilled water 1000mL) (150rpm/min), subsequent use.
The bacillus subtilis PTS-394 that the Jiangsu Province Agriculture Science Institute screening is obtained carries out activation on the NA medium, move into then in the 50mL NA culture fluid, cultivates 2d at 28 ℃ vibrate down (150rpm/min), and is subsequent use.Tomato Fusarium oxysporum cake (diameter is 7mm) places the dull and stereotyped side (anomaly panel edges 30mm) of blank PDA.Parallel opposite side (anomaly panel edges 30mm) is placed 5mm sterilization filter paper (anomaly plate center 30mm), and filter paper adds the bacillus subtilis PTS-394 bacterium liquid 5 μ L of different extension rates.Every processing repetition 3 wares.If clear water contrast.28 ℃ of constant temperature culture are measured investigation pathogen colony diameter, and are calculated inhibiting rate when flat-plate bacterial colony to be contrasted covers with.Inhibiting rate=(contrast pathogen colony diameter-processing pathogen colony diameter)/contrast pathogen colony diameter * 100%.
Draw Ralstonia solanacearum liquid coating NA dull and stereotyped (0.3mL bacterium liquid/flat board).Culture dish becomes crux to place 4 sterilization filter papers, adds the bacillus subtilis PTS-394 bacterium liquid 5 μ L of different extension rates on the filter paper: establish the clear water contrast.Every processing repetition 3 wares.28 ℃ of constant temperature culture 2d measure antibacterial circle diameter.
Result of the test is seen table 1, Fig. 1-2.Result of the test shows that bacillus subtilis PTS-394 stoste, 100 times of dilutions and 500 times of dilutions all have stronger inhibitory action to tomato wilt bacterium and ralstonia solanacearum.
Table 1 bacillus subtilis PTS-394 is to the inhibitory action of tomato wilt bacterium and ralstonia solanacearum
As depicted in figs. 1 and 2.
The generation of bacillus subtilis PTS-394 metabolic secretion thing-antibiotic related substances
Supply the examination material: protease detects medium: skim milk powder 100g, agar 20g is settled to 1000mL.Chitinase detects medium: gluey chitin 15g, MgSO47H
2O 0.5g, FeSO
47H
2O 0.01g, K
2HPO
40.7g, KH
2PO
40.3g, agar 20g, be settled to 1000mL, pH7.0～7.2.Cellulase detects medium: peptone 10g, dusty yeast 10g, sodium carboxymethylcellulose 10g, NaCl 5g, KH
2PO
41g, agar 20g is settled to 1000mL, pH7.0.Have a liking for the plain medium that detects of iron: CAS 60.5g, 10mL ferric iron solution (1mmol.L
-1FeCl
36H
2O), HDTMA 72.9g, agar 20g is settled to 1000mL, pH7.0.
The bacillus subtilis PTS-394 that the Jiangsu Province Agriculture Science Institute screening is obtained carries out activation on the NA medium, move into then in the 50mL NA culture fluid, cultivates 2d at 28 ℃ vibrate down (150rpm/min), and is subsequent use.
Protease detects: protease detects culture medium flat plate central authorities and places 5mm sterilization filter paper, drips on every filter paper and picks up antimicrobial fluid 5 μ L.Every processing repetition 4 wares.Cultivate 2d for 28 ℃, observation has or not transparent circle to produce.
Chitinase detects: chitinase detects medium central authorities and places 5mm sterilization filter paper, drips on every filter paper and picks up antimicrobial fluid 5 μ L.Every processing repetition 4 wares.Cultivate 3～7d for 28 ℃, observation has or not transparent circle to produce.
Cellulase detects: cellulase detects culture medium flat plate central authorities and places 5mm sterilization filter paper, drips antimicrobial fluid 5 μ L short of money on every filter paper.Every processing repetition 4 wares.Cultivate 3～7d for 28 ℃, observation has or not transparent circle to produce.
Having a liking for the iron element detects: have a liking for the plain placement 5mm of the culture medium flat plate central authorities sterilization filter paper that detects of iron, drip inspection antimicrobial fluid 5 μ L short of money on every filter paper.Every processing repetition 4 wares.Cultivate 3～7d for 28 ℃ and observe change color,, then have the iron of having a liking for element to produce if there is the orange colour haloing to produce.
Result of the test shows: bacillus subtilis PTS-394 can not secrete chitinase and cellulase; The ability extracellular proteinase; Also can secrete and have a liking for the iron element, ability chelating ferric ion forms faint yellow haloing (Fig. 3 and Fig. 4) on test slab.
Like Fig. 3 and shown in Figure 4.
Embodiment 3
The preparation of 20,000,000,000 bacillus subtilis PTS-394 aquas
A, bacterial classification are prepared and activation.
NA medium (with embodiment 1) is by conventional method sterilization, beveling, cultivate 2 days in advance after, inoculation bacillus subtilis PTS-394, cultivation temperature: 28 ℃; Incubation time: 2 days.
B, liquid fermentation.
One grade fermemtation: with above-mentioned cultured test tube slant, move a ring lawn with oese and go in the 500mLNA culture fluid (with embodiment 1), move 2 bottles altogether.Cultivate 2d at 28 ℃ vibrate down (150rpm/min), subsequent use.
Second order fermentation: culture medium prescription: soybean cake powder 1.0%, soybean meal 0.5%%, dusty yeast 0.15%, corn flour 2.0%, fish meal 1.0%, NaCl 0.05%, CaCO
30.3%.PH 7.5 before disappearing.Feed intake 1 ton in 2 tons of fermentation tanks.Sterilized 30 minutes for 121 ℃.Inoculate above-mentioned one grade fermemtation strain liquid 1000mL when being cooled to 30 ℃ of left and right sides.30 ℃ of liquid fermentation temperature, speed of agitator 250rpm/min.Throughput 1: 1.5.Fermentation period is 24 hours.
Three grade fermemtation: the same second order fermentation of culture medium prescription.Feed intake 10 tons in 15 tons of fermentation tanks.Sterilized 30 minutes for 121 ℃.Inoculate 1 ton of above-mentioned second order fermentation strain liquid when being cooled to 30 ℃ of left and right sides.33 ℃ of liquid fermentation temperature, speed of agitator 300rpm/min.Throughput 1: 1.5.Fermentation period is about 36 hours, puts jar when 90% thalline forms gemma in the zymotic fluid.
C, aqua configuration.
Zymotic fluid is put into storage tank, adds NNO (sodium metnylene bis-naphthalene sulfonate) 0.5%, neopelex 6%, Sodium Benzoate 0.1%, stirs.Recording zymotic fluid pH value is 5.4, and the living spores number is 2 * 10
10Cfu/mL.
Embodiment 4
The preparation of 20,000,000,000 bacillus subtilis PTS-394 wetting powders
A, bacterial classification are prepared and activation.With embodiment 3.
B, liquid fermentation.With embodiment 3.
C, the former powder of preparation viable bacteria.
Zymotic fluid is put into settling tank, utilize the isoelectric point sedimentation after 3 days, anhydrate through disc centrifuge is centrifugal, stay the bacterium slurry.The precipitated calcium carbonate (detailed catalogue 325 orders) that by volume adds 10% (w/v) in the bacterium slurry is starched bacterium in the former powder of gemma that sprays the dried bacillus subtilis PTS-394 of one-tenth with spray drying process.Inlet temperature is 175 ℃ during atomized drying, 72 ℃ of outlet wind-warm syndrome.Record living spores several 5 * 10 in the former powder
10Cfu/g.
D, preparation wetting powder
Auxiliary agent is selected NNO (sodium metnylene bis-naphthalene sulfonate); Adhesive is a polyethylene glycol; Surfactant is a neopelex; Filler is a precipitated calcium carbonate.
Each component is got the raw materials ready according to following percentage by weight:
The former powder of bacillus subtilis PTS-394 viable bacteria: 40%; NNO (sodium metnylene bis-naphthalene sulfonate): 7%; Polyethylene glycol: 3%; Neopelex: 5%; Precipitated calcium carbonate: 45%.
The preparation process: the former powder of bacillus subtilis PTS-394 viable bacteria etc. is placed blender according to the above ratio, after mixing, is crushed to 325 orders through airslide disintegrating mill to sieve, packing, 20,000,000,000 bacillus subtilis PTS-394 wetting powders.
Embodiment 5
20,000,000,000 bacillus subtilis PTS-394 aquas are to the potted plant control test of tomato wilt and bacterial wilt.
Test period: (tomato falls to planting) carried out in greenhouse, academy of agricultural sciences, Jiangsu Province about 1 day in October, 2010.
Test kind: Jiangshu 1 (providing) by academy of agricultural sciences, Jiangsu Province vegetables.
Supply examination pathogen bacterial strain:
The preparation of bacterial wilt of tomato bacterium is with embodiment 1.
The preparation of tomato wilt bacterium spore liquid is following: the flat board among the embodiment 1 is washed with sterile water, break up, make it into uniform spore suspension.Microscopically is counted with blood counting chamber, transfers to concentration 106 spores/mL with sterile water.
Test medicine and contrast agents:
20,000,000,000 bacillus subtilis PTS-394 aquas are provided by embodiment 3.The contrast medicament is 50% carbendazim (being provided by Huifeng Agricultural Chemistry Co., Ltd., Jiangsu) and 72% agricultural streptomycin sulphate (effluent Bei Sheng Shijiazhuang dawn pharmaceutical factory provides), and they all are the chemical agents commonly used of preventing and treating fusarium wilt and bacterial wilt on producing.
Field control effectiveness test: No. 1 tomato species of river vegetables is sowed in the sterile soil, when tomato seedling grows 6 true leaves, transplants seedlings.
Fusarium wilt control test: with shoot transplanting equipment to the basin alms bowl that the 100g sterile soil is housed, every basin 1 strain.Every strain tomato seedling is irritated root 20mL wilt spore liquid, and (concentration is 10
6Spore/mL), (bacterial content is 10 to irritate root 20mL bacillus subtilis PTS-394 aqua behind the 6h
8Cfu/mL).Every processing 20 basins, Routine Management.Clear water and chemical agent commonly used 50% carbendazim are contrast.The growth 30d " Invest, Then Investigate " state of an illness is calculated preventive effect.
Bacterial wilt control test: with shoot transplanting equipment to the basin alms bowl that the 100g sterile soil is housed, every basin 1 strain.Every strain tomato seedling is irritated root 20mL bacillus subtilis PTS-394 aqua, and (bacterial content is 10
8Cfu/mL), (bacterial content is 10 to irritate root 20mL Ralstonia solanacearum liquid behind the 6h
8Cfu/mL).Every processing 20 basins, Routine Management.Clear water and 72% agricultural streptomycin sulphate are contrast.The growth 30d " Invest, Then Investigate " state of an illness is calculated preventive effect.
Tomato wilt and bacterial wilt disease symptom are closely similar, and state of an illness grade scale is following: 0 grade is that vascular bundle is normal in the stem, outside asymptomatic; 1 grade is vascular bundle 1/4 following variable color in the stem; 3 grades is vascular bundle 1/4～1/2 variable color in the stem; 5 grades is vascular bundle 1/2～3/4 variable color in the stem; 7 grades is vascular bundle 3/4 above variable color in the stem, and partial blade is wilted; 9 grades withered for whole strain.
Disease index=[(morbidity numbers at different levels * typical values at different levels)/(investigating total strain number * highest typical value)] * 100
Control efficiency (%)=[disease index of (disease index of the disease index-control of contrast)/contrast] * 100%
Result of the test is seen table (table 2).After the artificial infection, the disease index of tomato wilt is 48.54%, and the disease index of bacterial wilt of tomato is 78.39%.Bacillus subtilis PTS-394 aqua is higher to the preventive effect of tomato wilt and bacterial wilt, is respectively 77.40% and 74.49%.Conventional chemical medicament carbendazim and the potted plant preventive effect of agricultural streptomycin sulphate are all undesirable, are respectively 46.09% and 55.35%, are starkly lower than the control efficiency of bacillus subtilis PTS-394 aqua.
Table 2 bacillus subtilis PTS-394 aqua is to the potted plant control efficiency of tomato wilt bacterium and ralstonia solanacearum
Embodiment 6
20,000,000,000 bacillus subtilis PTS-394 wetting powders are to the field controling test of eggplant fusarium wilt and bacterial wilt.
Test period: in the facility plastic greenhouse of modern agriculture industrial zone, city, place, Shuyang County, Jiangsu Province, carried out about 1 day in October, 2010.
No. one, test kind: Su Qi.
Test medicine and contrast agents:
20,000,000,000 bacillus subtilis PTS-394 wetting powders are provided by embodiment 4.50% carbendazim is provided by Huifeng Agricultural Chemistry Co., Ltd., Jiangsu.72% agricultural streptomycin sulphate effluent Bei Sheng Shijiazhuang dawn pharmaceutical factory provides.
Field control effectiveness test: (by the end of November) irritated root and handled during the eggplant transplantation of seedlings, and transplanting the back, every (15d) filling root is once at a distance from about two weeks.The whole growth phase needs medication 8-9 time altogether.With finished medicines dilution agent 500-1500 doubly; 2 repetitions are established in each processing, establish guard row.
State of an illness grade scale and control efficiency are calculated: with embodiment 5.
Result of the test is seen table (table 3).The disease index of (contrast) eggplant fusarium wilt and bacterial wilt is respectively 48.72 and 39.01 under the natural conditions.500-1500 times of dilution of bacillus subtilis PTS-394 wetting powder is higher to the preventive effect of eggplant fusarium wilt and bacterial wilt, is respectively 59.42%-82.78% and 57.83%-87.03%.Conventional chemical medicament carbendazim is 64.24% to the preventive effect of fusarium wilt; Agricultural streptomycin sulphate is 59.16% to the preventive effect of bacterial wilt.The effect of 500 times and 1500 times dilutions of bacillus subtilis PTS-394 wetting powder is apparently higher than the contrast medicament; The effect of 1500 times of dilutions is suitable with the contrast medicament.
Table 3 bacillus subtilis PTS-394 wetting powder is to the potted plant control efficiency of eggplant wilt and ralstonia solanacearum
Claims (9)
1. biocontrol fungicide, wherein the effective active composition is made up of bacillus subtilis (Bacillus subtilis) PTS-394.
2. biocontrol fungicide according to claim 1 wherein, comprises protease and has a liking for the plain two kinds of antibiotic related substanceses of iron in the metabolite of bacillus subtilis PTS-394 secretion.
3. according to the described biocontrol fungicide of claim 1-2, it is characterized in that it is aqua or wetting powder.
4. biocontrol fungicide according to claim 3, wherein the percentage by weight of each component of aqua is:
5. biocontrol fungicide according to claim 3, wherein the percentage by weight of each component of wetting powder is:
6. according to the described biocontrol fungicide of claim 4-5: wherein auxiliary agent can be chosen any one kind of them or multiple following material: NNO (sodium metnylene bis-naphthalene sulfonate), azone, thiophene ketone; Adhesive can be chosen any one kind of them or multiple following material: polyvinylpyrrolidone, polyethylene glycol, polyvinyl alcohol (aggregate number 12000-20000); Surfactant can be chosen any one kind of them or multiple following material: dodecyl sodium sulfate, neopelex, sodium lignin sulfonate, the withered powder of tea, spaonin powder; Preservative can be chosen any one kind of them or multiple following material: Sodium Benzoate, potassium sorbate, dehydro sodium acetate, calcium propionate, sodium Diacetate.Filler can be chosen any one kind of them or multiple following material: precipitated calcium carbonate, diatomite, kaolin, sericite in powder and white carbon black.
7. the purposes of the described biocontrol fungicide of claim 1-6 is characterized in that: be used to prevent and treat soil-borne disease with pouring into the vegetables root after the described biocontrol fungicide dilution.
8. the application of biocontrol fungicide according to claim 7 is characterized in that: described biocontrol fungicide can be prevented and treated two kinds of important soil-borne diseases of solanaceous vegetables (fusarium wilt and bacterial wilt) simultaneously.
9. according to the application of claim 7 or 8 described biocontrol fungicides, it is characterized in that: the active drug component content 10 behind the finished medicines dilution agent
4-10
8Cfu/mL or 10
4-10
8Cfu/g.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103146613A (en) * | 2013-03-14 | 2013-06-12 | 江苏省农业科学院 | Fermentation culture method for bacillus subtilis PTS-394 |
| CN103667134A (en) * | 2013-12-10 | 2014-03-26 | 江苏省农业科学院 | Paenibacillus polymyxa N3-4 used for preventing and treating clubroot of cruciferous crops and application thereof |
| CN104082346A (en) * | 2014-06-27 | 2014-10-08 | 江苏省农业科学院 | Chilli Lysobacter capsici NF87-2 and application thereof |
| CN104982516A (en) * | 2015-06-15 | 2015-10-21 | 珠海真绿色技术有限公司 | Environment-friendly and ecological orange preservation method and preservative |
| CN105145094A (en) * | 2015-09-23 | 2015-12-16 | 山东农业大学 | Integrated ecological tomato cultivation method for reducing pest and disease damage and improving fruit quality |
| CN105296381A (en) * | 2015-08-31 | 2016-02-03 | 哈尔滨师范大学 | Bacillus subtilis CYY-25 and application thereof |
| CN105309245A (en) * | 2015-11-10 | 2016-02-10 | 江苏省农业科学院 | Method utilizing bacillus subtilis PTS-394 wettable powder for preventing and treating crop soil-borne disease |
| CN106305792A (en) * | 2016-05-19 | 2017-01-11 | 北京启高生物科技有限公司 | Clonostachys rosea-bacillus subtilis composite wettable powder |
| CN107897179A (en) * | 2017-11-09 | 2018-04-13 | 江苏省农业科学院 | The effective use method of bacillus subtilis PTS-394 wettable powder field control Fusarium solanis |
| KR101879971B1 (en) * | 2017-01-06 | 2018-07-18 | 강원대학교산학협력단 | Bacillus subtilis ds660 strain having antimicrobial activity and uses thereof |
| CN111172056A (en) * | 2019-11-25 | 2020-05-19 | 云南大学 | Bacterial strain and application thereof in wilt prevention and control |
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| CN103146613A (en) * | 2013-03-14 | 2013-06-12 | 江苏省农业科学院 | Fermentation culture method for bacillus subtilis PTS-394 |
| CN103667134A (en) * | 2013-12-10 | 2014-03-26 | 江苏省农业科学院 | Paenibacillus polymyxa N3-4 used for preventing and treating clubroot of cruciferous crops and application thereof |
| CN103667134B (en) * | 2013-12-10 | 2016-08-17 | 江苏省农业科学院 | The Paenibacillus polymyxa N3-4 of one strain preventing and treating club-root and application thereof |
| CN104082346B (en) * | 2014-06-27 | 2016-04-13 | 江苏省农业科学院 | The molten bacillus NF87-2 of one strain capsicum and application thereof |
| CN104082346A (en) * | 2014-06-27 | 2014-10-08 | 江苏省农业科学院 | Chilli Lysobacter capsici NF87-2 and application thereof |
| CN104982516A (en) * | 2015-06-15 | 2015-10-21 | 珠海真绿色技术有限公司 | Environment-friendly and ecological orange preservation method and preservative |
| CN104982516B (en) * | 2015-06-15 | 2019-05-14 | 珠海真绿色技术有限公司 | A kind of the green ecological preservation method and antistaling agent of citrus |
| CN105296381B (en) * | 2015-08-31 | 2018-11-30 | 哈尔滨师范大学 | One bacillus subtilis CYY-25 and its application |
| CN105296381A (en) * | 2015-08-31 | 2016-02-03 | 哈尔滨师范大学 | Bacillus subtilis CYY-25 and application thereof |
| CN105145094A (en) * | 2015-09-23 | 2015-12-16 | 山东农业大学 | Integrated ecological tomato cultivation method for reducing pest and disease damage and improving fruit quality |
| CN105309245A (en) * | 2015-11-10 | 2016-02-10 | 江苏省农业科学院 | Method utilizing bacillus subtilis PTS-394 wettable powder for preventing and treating crop soil-borne disease |
| CN106305792A (en) * | 2016-05-19 | 2017-01-11 | 北京启高生物科技有限公司 | Clonostachys rosea-bacillus subtilis composite wettable powder |
| CN106305792B (en) * | 2016-05-19 | 2018-12-25 | 北京启高生物科技有限公司 | A kind of wettable powder that Gliocladium roseum is compounded with bacillus subtilis |
| KR101879971B1 (en) * | 2017-01-06 | 2018-07-18 | 강원대학교산학협력단 | Bacillus subtilis ds660 strain having antimicrobial activity and uses thereof |
| CN107897179A (en) * | 2017-11-09 | 2018-04-13 | 江苏省农业科学院 | The effective use method of bacillus subtilis PTS-394 wettable powder field control Fusarium solanis |
| CN111172056A (en) * | 2019-11-25 | 2020-05-19 | 云南大学 | Bacterial strain and application thereof in wilt prevention and control |
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