CN1281135C - Mixture of bacterial medicine for controlling vegetable epidemic disease and preparation method - Google Patents
Mixture of bacterial medicine for controlling vegetable epidemic disease and preparation method Download PDFInfo
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Abstract
The present invention belongs to the field of chemical and biological soil treating agents and the preparation methods thereof, particularly to the field of a bacterial medicine mixture for preventing and controlling vegetable epidemic disease and a preparation method thereof. The steps of the method can be orderly carried out: (a) fermenting and culturing biologic strains with antagonistic effect on pathogenic bacteria of the vegetable epidemic disease, and depositing bacteria obtained by centrifuging the fermentation liquid, and mixing with calcium carbonate micropowder according to a mass ratio of 1: 1 to obtain a biologic bacterium; (b) compounding the biologic bacterium formed by the step (a) with a germicide whose content of effective components is 50% according to a mass ratio of 1: 1. The application rate of chemical pesticides on vegetables can be decreased by more than 50%, the bacterial medicine mixture can partially or entirely substitute for traditional high-poison high-residue pesticides, the quantity of pesticide residues on vegetables can be ensured to be lower than permissible criteria, and production requirements for pollution-free vegetables can be satisfied.
Description
Technical field
The invention belongs to chemistry, biological soil inorganic agent and preparation method thereof field, particularly a kind of bacteria medicament mixture that vegetable epidemic disease is had prevention and control action and preparation method thereof field.
Background technology
Eqpidemic disease is the great disease of a class on vegetables produce, and common eqpidemic disease mainly contains capsicum epidemic disease, tomato late blight and cucumber eqpidemic disease etc.In China, the generation of capsicum epidemic disease is the most serious.In recent years, because the change of kind, cropping system, fertilizer and water condition, disease increases the weight of year by year, and the lesion is expansion year by year from south to north, and the withered rate in grave illness district can reach more than 80%, and production loss is very remarkable.Capsicum epidemic disease all can take place in the whole growing of capsicum, especially takes place serious when capsicum buddings and bear fruit.Under the suitable situation of weather conditions, this disease can endanger root, stem stalk, branch, fruit and blade, can cause dead seedling, decayed fruit, the underproduction even total crop failure when serious.In the production practices, the comprehensive regulation of eqpidemic disease mainly is to rely on to select disease-resistant variety, efficent rotation and use systemic fungicide for use.Upward applicable antilemic vegetable variety is limited but produce at present, because the eqpidemic disease pathogen is a soil inhabitant, crop rotation also is difficult to control effectively this disease simultaneously.Therefore chemical control is one of the most effective means of prevention of this disease always since the long term, mainly prevents and treats capsicum epidemic disease with reagent spray and filling root in the production.The existing systemic fungicides such as the main medicament metalaxyl of eqpidemic disease, white urea cyanogen, aliette of preventing and treating, this series bactericidal agent belongs to specificity site inhibitor, action site to pathogen is single, only the single metabolism link to pathogen works, in case undergo mutation in this site, medicament promptly can not with its generation effect, thereby cause germ to develop immunity to drugs, the effect of chemical control disease is reduced.Along with the rapid raising of socio-economic development and people's living standard, " green vegetables " and " pollution-free vegetable " demand is also increasing, and crop safety production and agricultural product security are extremely paid attention to.Therefore, the strategy and the practice of single utilization chemical pesticide control eqpidemic disease have been subjected to challenge, and exploring new controlling way is the major issue that present vegetable epidemic disease control must solve.Development biological control technology comes controlling plant diseases more and more to receive national governments, scientific worker and the common people's concern, according to the knowledge of the applicant, studies the existing many reports of biological control measure control phytophthora root rot evil both at home and abroad.But because the single biological and ecological methods to prevent plant disease, pests, and erosion factor is difficult to the preventive effect that obtains and use chemical bactericide to be complementary under the varying environment condition; And the fluctuation ratio of biological and ecological methods to prevent plant disease, pests, and erosion factor pair soil or crop rhizosperic environmental condition is responsive, exists in the field under the ecological condition of pathogen, and the colonization ability of the biological and ecological methods to prevent plant disease, pests, and erosion factor has very big-difference.The research of biological control control vegetable epidemic disease is many at present to be carried out under laboratory and green-house's condition, and large tracts of land is not used for production practices as yet.
Be subjected to increasing query in chemical control, under the also incomplete condition of biological control system, the strategy of development biological-chemical Collaborative Control vegetable epidemic disease causes national governments and scientific research personnel's attention gradually.
Summary of the invention
The object of the present invention is to provide a kind of efficient, low toxicity, with environmentally compatible and the bacteria medicament mixture and preparation method thereof that vegetable epidemic disease had the biological-chemical Collaborative Control of prevention and control action.
The object of the present invention is achieved like this: provide a kind of control vegetable epidemic disease bacteria medicament mixture to contain biocontrol fungicide and bactericide, described biocontrol fungicide and bactericide amount ratio by mass are 1: 1.Preferred bactericide is selected from that the Amici reaches, in the tpn, mancozeb, metalaxyl, green Supreme Being, flumorph, dimethomorph, aliette, silver-colored Thailand one or more.Further preferred fungicide is a dimethomorph, further the wetting powder of preferred dimethomorph active constituent content 25%-90%, most preferably 50% peace gram wetting powder (dimethomorph preparation).The biocontrol bacterial strain that contains in the preferred biocontrol fungicide is selected from one or more among bacillus subtilis BS, bacillus subtilis GF1, pseudomonas fluorescens LX1, the pseudomonas fluorescens BCA1.The biocontrol bacterial strain that contains in the further preferred biocontrol fungicide is bacillus subtilis BS.
The present invention also provides a kind of method for preparing above-mentioned control vegetable epidemic disease bacteria medicament mixture, and it comprises the steps:
A) will carry out fermented and cultured to the biocontrol bacterial strain that the vegetable epidemic disease germ has an antagonism, again with the bacterial sediment of the centrifugal acquisition of zymotic fluid by 1: 1 mass ratio and micro mist calcium carbonate are mixed must biocontrol fungicide;
Preferred biocontrol bacterial strain is selected from one or more among bacillus subtilis BS, bacillus subtilis GF1, pseudomonas fluorescens LX1, the pseudomonas fluorescens BCA1.Further preferred biocontrol bacterial strain is bacillus subtilis BS.
B) biocontrol fungicide that step a is formed and bactericide are composite by 1: 1 mass ratio.
Preferred bactericide is the preparation of active constituent content 25%-90%, and further preferred bactericide is the preparation of active constituent content 50%.
Preferred bactericide is selected from that the Amici reaches, in the tpn, mancozeb, metalaxyl, green Supreme Being, flumorph, dimethomorph, aliette, silver-colored Thailand one or more.Further preferred fungicide is a dimethomorph, further the wetting powder of preferred dimethomorph active constituent content 25%-90%, most preferably 50% peace gram wetting powder (dimethomorph preparation).
The present invention is based on the bacteria medicament mixture of biological-chemical Collaborative Control principle, synergistic function by biotic factor and chemokines, can reduce the chemical pesticide amount of application on the facilities vegetable more than 50%, substitute traditional high poison, persistent pesticide fully, the persticide residue of guaranteeing vegetables is below the mark in permission, meets the pollution-free vegetable production requirement.Thereby save production cost and improved quality of vegetable.
Description of drawings
Fig. 1 is a process flow diagram of the present invention.
Embodiment
Below in conjunction with the described embodiment of accompanying drawing the present invention is described in further detail.
Control vegetable epidemic disease bacteria medicament mixture, it contains biocontrol fungicide and 50% peace gram wetting powder; The mass ratio of described biocontrol fungicide and 50% peace gram wetting powder is 1: 1.
As shown in the figure, control vegetable epidemic disease bacteria medicament mixture preparation method, can carry out as follows successively:
A) will carry out fermented and cultured to the biocontrol bacterial strain bacillus subtilis BS that the capsicum epidemic disease germ has an antagonism, again with the bacterial sediment of the centrifugal acquisition of zymotic fluid by 1: 1 mass ratio and micro mist calcium carbonate are mixed must biocontrol fungicide;
B) to be that 50% peace restrains wetting powder composite by 1: 1 mass ratio for the biocontrol fungicide that step a is formed and active constituent content.
Several antagonistic microbes that the present invention is based on having obtained, prevent and treat eqpidemic disease several bactericide commonly used in biocontrol strains of having registered and the production both at home and abroad, with the capsicum epidemic disease germ is the target bacterium, by exsomatizing and the live body screening, obtain a strain respectively Phytophthora capsici is had the biocontrol bacterial strain of strong antagonism and a kind of efficient, low toxicity, eco-friendly chemical pesticide, by exsomatizing and live test mensuration biopesticide and chemical pesticide virulence coefficient, the specific synergistic effect of specialization between clear and definite biotic factor and the chemical agent, the synergistic combination of screening high-efficiency broad spectrum biological bactericide, be developed into bacteria medicament mixture, and its method of application is studied.
Concrete operations step of the present invention is as follows:
(1) antagonistic microbe screening on the basis of Preliminary screening, select bacillus subtilis BS, pseudomonas fluorescens LX1, bacillus subtilis GF1, pseudomonas fluorescens BCA1 four strain biocontrol bacterial strains to carry out stripped bacteriostatic experiment and the screening of live body plant respectively, find that bacillus subtilis (Bacillus subtilis) BS has stronger antagonism to the capsicum epidemic disease germ, have higher preventive effect in the greenhouse diseases prevention test.Concrete data see Table 1 and table 2.Therefore select the biological components of bacillus subtilis BS as the development bacteria medicament mixture.
(2) 9 kinds of dissimilar bactericide commonly used during the screening of bactericide is adopted and produced, measured inhibitory action respectively to the different thalli morphologies of capsicum epidemic disease germ, the result is as shown in table 3, and two kinds of bactericide of dimethomorph and flumorph all have significant inhibitory effect to the various thalli morphologies of being measured.On the basis of vitro Screening, be contrast with the typical medicament metalaxyl of preventing and treating eqpidemic disease again, carried out the greenhouse diseases prevention test of dimethomorph and flumorph, find that the control efficiency of dimethomorph is better than flumorph (seeing Table 4).Therefore select dimethomorph as the chemical constituent that makes up bacteria medicament mixture.
(3) development of bacteria medicament mixture is pressed the bacterial sediment of the centrifugal acquisition of zymotic fluid 1: 1 mass ratio and the mixed biocontrol fungicide that gets of micro mist calcium carbonate again with bacillus subtilis BS fermented and cultured, and its bacterium amount is greater than 10
8Cfu/g.Prevent and treat the bacteria medicament mixture of vegetable epidemic disease then by the composite acquisition the present invention of 1: 1 mass ratio with 50% peace gram wetting powder (dimethomorph preparation).
(4) content's index of the detection dimethomorph of bacteria medicament mixture quality index and bacillus subtilis BS is the gross index of bacteria medicament mixture, the dimethomorph Determination on content adopts high performance liquid chromatography to carry out (〉=25%), and the bacterium number of bacillus subtilis adopts dilution-plate method to measure (viable bacteria quantity 〉=10 that fall
7Cfu/mL).Show that through heat storage experiment bacillus subtilis does not have metabolism to dimethomorph, dimethomorph does not have interference effect to the clump count formation of bacillus subtilis yet.
Embodiment 1:
Adopt the main capsicum variety capital that plants, Beijing area to grind capsicum No. 3, the mode of infection of capsicum epidemic disease under the simulating natural condition, adopt the mode of root-pouring method and spray-on process inoculation germ to set up pathogenic system respectively, studied of the influence of different insecticide-applying way bacteria medicament mixture DMBS control capsicum epidemic disease effect.The result is shown in table 5, table 6, under different pathogenic systems, the control efficiency of bacteria medicament mixture is relevant with the occupation mode of medicament, and the root-pouring method dispenser is better for the disease expansion effect of control underground part, and the spray-on process dispenser is better for the communication effect of control overground part germ.But which kind of no matter adopts connect the bacterium mode can be found, in the agent of bactericide list, the agent of biocontrol microorganisms list and three kinds of processing of bacteria medicament mixture for examination, single biological prevention and control agent preventive effect performance is understable, bacteria medicament mixture then shows the trend identical with single bactericide, with the rising of drug concentration, control efficiency improves; The more important thing is, reduced a half-value dose bacteria medicament mixture control efficiency with can obtain roughly suitable control efficiency with the single bactericide of concentration, under individual concentrations, even be higher than bactericide and handle.
Embodiment 2:
Adopt the general susceptible variety eggplant door pimento of capsicum epidemic disease, the mode of infection of capsicum epidemic disease under the simulating natural condition, adopt the mode of root-pouring method and spray-on process inoculation germ to set up pathogenic system respectively, studied of the influence of different insecticide-applying way bacteria medicament mixture DMBS control capsicum epidemic disease effect.The result is shown in table 7, table 8, and it is similar that capsicum No. 3 is ground in its rule and capital.
Table 1,4 strain biocontrol bacterial strains are measured the bacteriostasis of 7 strain capsicum epidemic disease germs
| Strains tested (eqpidemic disease germ different strains) | The biological and ecological methods to prevent plant disease, pests, and erosion factor | |||
| BS | GF1 | LX1 | BCA1 | |
| SD21-3 GZ01 Pc112 Pc33E Pc1E-8 P1319 P1314 | 35.60 38.15 40.80 59.43 55.01 44.53 33.15 | 27.72 26.70 34.81 41.32 40.49 41.30 21.10 | 24.73 28.07 38.36 43.02 47.65 41.21 35.07 | 32.34 29.16 39.69 44.15 47.85 42.91 34.25 |
Table 2, the greenhouse test of biological and ecological methods to prevent plant disease, pests, and erosion factor control capsicum epidemic disease
| Prevent and treat method | BCAs | Spray-on process inoculation germ | Root-pouring method inoculation germ | ||
| Disease index (%) | Control efficiency (%) | Disease index (%) | Control efficiency (%) | ||
| The root control is irritated in the spraying control | CK BS GF1 LX1 BCA1 BS GF1 LX1 BCA1 | 19.91ABa 13.61DCbc 18.00ABCab 17.87ABCab 14.00BDCbc 13.08DCc 18.96ABCa 21.69Aa 8.40Dd | 31.64 9.60 10.27 29.69 34.32 4.79 -8.93 57.81 | 14.80BAba 6.39Dd 14.28BACba 17.76Aa 14.00BACba 11.60BDCbc 12.80BACb 13.56BACba 8.40DCdc | 56.83 3.53 -19.97 5.41 21.62 13.51 8.41 43.24 |
Table 3,9 kinds of bactericide are to the inhibitory action of the different thalli morphologies of capsicum epidemic disease germ
| Bactericide | Mycelial growth | Sporangium produces | The spore that stops is sprouted | Sporangium is sprouted | ||||
| EC 50 (μg/mL) | EC 95 (μg/mL) | EC 50(μg/mL) | EC 95(μg/mL) | EC 50(μg/mL) | EC 95(μg/mL) | EC 50(μg/mL) | EC 95(μg/mL) | |
| It is safe that the Amici reaches the green Supreme Being's dimethomorph of Bravo Mancozeb flumorph metalaxyl aliette silver | ＞100 ＞100 ＞100 0.18 0.01 55.46 0.14 ＞100 28.82 | ＞1000 ＞1000 ＞1000 0.67 0.03 ＞1000 0.45 ＞1000 102.35 | 6.15 1.97 8.70 0.68 2.75 20.16 0.61 25.21 62.66 | 244.61 24.13 114.32 2.57 5.97 57.66 2.44 466.98 460.43 | 0.094 0.211 18.70 0.30 ＞100 70.24 0.12 61.34 39.49 | 0.409 1.116 116.72 0.78 ＞1000 147.60 0.52 178.50 155.71 | 1.27 0.99 3.82 0.64 155.53 41.11 0.25 78.56 26.09 | 139.69 21.89 22.66 1.48 490.06 222.54 0.99 390.19 130.88 |
Table 4, the greenhouse test of bactericidal agent for preventing and treating capsicum epidemic disease
| Handle | Drug concentration (μ g/mL) | Capsicum variety | |||
| Capsicum is ground No. 3 in the capital | Eggplant door pimento | ||||
| Disease index (%) | Control efficiency (%) | Disease index (%) | Control efficiency (%) | ||
| CK metalaxyl dimethomorph flumorph | 0 5 10 25 50 100 25 50 100 250 500 25 50 100 250 500 | 70 90 82 78 56 50 48 32 20 16 6 62 56 52 18 6 | - -28.57 -17.14 -11.43 20.00 28.57 31.43 54.29 71.43 77.14 91.43 11.43 20.00 25.71 74.29 91.43 | 78 62 48 40 22 6 70 50 38 12 0 74 56 44 24 10 | - 20.51 38.46 48.72 71.79 92.31 10.26 35.90 51.28 84.62 100.00 5.43 28.21 43.59 69.23 87.18 |
Table 5, bacteria medicament mixture of the present invention grind the control efficiency of No. 3 root-pouring method inoculations of capsicum capsicum epidemic disease germ to the capital
| Handle | Spray pesticide | Irritate the root dispenser | |||
| Disease index (%) | Control efficiency (%) | Disease index (%) | Control efficiency (%) | ||
| CK | 37.50Aa | ||||
| DMM (μg/mL) BS (cfu/mL) DMBS (μg/mL) | 50 100 250 10 7 10 8 10 9 25 50 125 | 18.00DEFdef 12.44GFgh 9.33Hgh 35.60Aab 31.59Aab 35.20Aab 22.40CDcd 20.40CDEde 16.71DEFefg | 52.00 66.82 75.11 5.07 15.76 6.13 40.27 45.60 55.43 | 26.00BCc 5.11HIi 0.00Ij 34.50Aab 15.33EFfg 35.38Aab 16.00EFefg 2.00Iij 1.20Iij | 30.67 86.37 100.00 8.00 59.11 5.66 57.33 94.67 96.80 |
Annotate: DMM is a dimethomorph, and BS is a bacillus subtilis, and DMBS is a bacteria medicament mixture.Down together.
Table 6, bacteria medicament mixture of the present invention grind the control efficiency of No. 3 spray-on process inoculations of capsicum capsicum epidemic disease germ to the capital
| Handle | Spray pesticide | Irritate the root dispenser | |||
| Disease index (%) | Control efficiency (%) | Disease index (%) | Control efficiency (%) | ||
| CK | 44.40Aa | ||||
| DMM (μg/mL) BS (cfu/mL) DMBS (μg/mL) | 50 100 250 10 7 10 8 10 9 25 50 125 | 5.60HIg 4.17Ig 4.00Ig 33.50BCc 26.80CDd 40.67ABab 6.40HIg 4.00Ig 3.34Ig | 87.39 90.62 90.99 24.55 39.64 8.41 85.59 90.99 92.47 | 24.67DEd 18.40EFe 12.58FGHef 27.91CDd 35.50Bbc 40.45ABb 15.50FGe 13.20FGHef 9.02GHIgf | 44.44 58.56 71.67 37.14 20.05 8.91 65.09 70.27 79.68 |
Table 7, bacteria medicament mixture of the present invention are inoculated the control efficiency of capsicum epidemic disease germ to eggplant door pimento root-pouring method
| Handle | Spray pesticide | Irritate the root dispenser | |||
| Disease index (%) | Control efficiency (%) | Disease index (%) | Control efficiency (%) | ||
| CK | 45.20Aa | ||||
| DMM (μg/mL) BS (cfu/mL) DMBS (μg/mL) | 50 100 250 10 7 10 8 10 9 25 50 125 | 23.60CDcd 21.20CDEcde 14.50EFfg 22.80CDcd 26.13Cc 34.80Bb 24.84Cc 14.50EFfg 16.67DEFef | 47.78 53.10 67.92 49.56 42.19 23.01 45.04 67.92 63.13 | 3.50GHij 2.40GHij 1.60Hj 21.00CDEcde 16.40DEFef 38.50ABb 18.80CDEdef 9.60GFgh 7.20GHih | 92.26 94.69 96.46 53.54 63.72 14.82 58.41 78.76 84.07 |
Table 8, bacteria medicament mixture of the present invention are inoculated the control efficiency of capsicum epidemic disease germ to eggplant door pimento spray-on process
| Handle | Spray pesticide | Irritate the root dispenser | |||
| Disease index (%) | Control efficiency (%) | Disease index (%) | Control efficiency (%) | ||
| CK | 23.33Aa | ||||
| DMM (μg/mL) BS (cfu/mL) DMBS (μg/mL) | 50 100 250 10 710 8 10 925 50 125 | 9.70BCbc 9.60BCb c 8.80BCcbd 10.00BCbc 7.20BCcbd 11.60Bbc 6.00BCcd 6.40BCcbd 3.40BCcbd | 58.41 58.86 62.29 57.14 69.14 50.29 74.29 72.57 85.43 | 8.80BCcbd 6.90BCcbd 8.40BCcbd 8.40BCcbd 7.24BCcbd 8.80BCcbd 12.40Bb 7.20BCcbd 6.40BCcbd | 62.29 70.43 64.00 64.00 68.95 62.29 46.86 69.14 72.57 |
Claims (2)
1, a kind of control vegetable epidemic disease bacteria medicament mixture, it contains biocontrol fungicide and bactericide, and described biocontrol fungicide and bactericide amount ratio by mass are 1: 1, and wherein the biocontrol bacterial strain that contains in the biocontrol fungicide is bacillus subtilis BS, and bactericide is a dimethomorph.
2, a kind of method for preparing the described control vegetable epidemic disease of claim 1 bacteria medicament mixture, it comprises the steps:
A) will carry out fermented and cultured to the bacillus subtilis BS that the vegetable epidemic disease germ has an antagonism, again with the bacterial sediment of the centrifugal acquisition of zymotic fluid by 1: 1 mass ratio and micro mist calcium carbonate are mixed must biocontrol fungicide;
B) biocontrol fungicide that step a is formed and dimethomorph are composite by 1: 1 mass ratio.
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| AR091197A1 (en) * | 2012-05-30 | 2015-01-21 | Bayer Cropscience Ag | COMPOSITION THAT INCLUDES A BIOLOGICAL CONTROL AGENT AND A FUNGICIDE |
| CN108184905B (en) * | 2012-05-30 | 2021-12-28 | 拜尔农作物科学股份公司 | Compositions comprising biological control agents and fungicides |
| BR112014029224A2 (en) * | 2012-05-30 | 2017-06-27 | Bayer Cropscience Ag | composition comprising a biological control agent and a fungicide |
| AR091195A1 (en) * | 2012-05-30 | 2015-01-21 | Bayer Cropscience Ag | COMPOSITION THAT INCLUDES A BIOLOGICAL CONTROL AGENT AND A FUNGICIDE |
| NZ742941A (en) * | 2012-05-30 | 2019-09-27 | Bayer Cropscience Ag | Composition comprising a biological control agent and a fungicide selected from inhibitors of the mitosis and cell division or compounds having a multi-site action |
| CN102742603A (en) * | 2012-05-31 | 2012-10-24 | 青岛农业大学 | Bactericide used for controlling brassica rapa pekinensis downy mildew and prepared by complexation of bacillus subtilis and chemical bactericide |
| US9877486B2 (en) | 2014-01-31 | 2018-01-30 | AgBiome, Inc. | Methods of growing plants using modified biological control agents |
| HUE056596T2 (en) * | 2014-01-31 | 2022-02-28 | Agbiome Inc | Modified biological control agent and its uses |
| CN103975929A (en) * | 2014-06-10 | 2014-08-13 | 黑龙江大学 | Compound preparation for preventing anthracnose of grapes |
| CN105994381A (en) * | 2016-06-27 | 2016-10-12 | 中国烟草总公司郑州烟草研究院 | Bactericide composition containing carboxylic acid amide and dithiocarbamate bactericides and biocontrol bacterium, preparation and applications |
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| CN109819827B (en) * | 2019-02-12 | 2022-01-07 | 安徽农业大学 | Prevention and treatment method for pepper phytophthora blight |
| CN110249743A (en) * | 2019-07-18 | 2019-09-20 | 四川迪菲特药业有限公司 | A kind of processing method of watt of cloth fritillaria kind bulb |
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