CN1281135C - Mixture of bacterial medicine for controlling vegetable epidemic disease and preparation method - Google Patents

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

Bacteria-drug mixture for preventing and controlling vegetable blight and preparation method thereof

technical field

The invention belongs to the field of chemical and biological soil treatment agents and preparation methods thereof, in particular to the field of a fungus-drug mixture capable of preventing and controlling vegetable blight and the preparation method thereof.

Background technique

Blight is a serious disease in vegetable production. Common blight mainly includes pepper blight, tomato late blight and cucumber blight. In my country, pepper blight is the most serious. In recent years, due to changes in varieties, cultivation systems, and fertilizer and water conditions, the disease has increased year by year, and the diseased area has expanded from south to north year by year. The death rate in severely diseased areas can reach more than 80%, and the yield loss is very significant. Capsicum blight can occur throughout the growth period of peppers, especially when peppers are budding and fruit-setting. Under favorable weather conditions, the disease can harm roots, stems, branches, fruits and leaves, and in severe cases, it can cause dead seedlings, rotten fruits, reduced production or even extinction. In production practice, the comprehensive management of epidemic diseases mainly relies on the selection of disease-resistant varieties, reasonable crop rotation and the use of systemic fungicides. However, there are limited varieties of anti-blight vegetables that can be used in production. At the same time, because the pathogenic bacteria of blight are soil-inhabiting bacteria, it is difficult to effectively control the disease by crop rotation. Therefore, chemical control has been one of the most effective means of preventing and controlling the disease for a long time. In production, chemical spraying and root irrigation are mainly used to prevent and control pepper blight. Existing systemic fungicides such as metalaxyl, cymoxanil, and fofosalum, the main agents for disease prevention and control, are specific site inhibitors, which have a single site of action on pathogenic bacteria, and are only effective on a single site of pathogenic bacteria. The metabolic link plays a role. Once this site is mutated, the medicine will not be able to act on it, which will lead to the resistance of the bacteria and reduce the effect of the medicine on disease control. With the development of social economy and the rapid improvement of people's material living standards, the demand for "green vegetables" and "pollution-free vegetables" is also increasing, and the safety of crop production and agricultural product safety has received much attention. Therefore, the strategy and practice of solely using chemical pesticides to control diseases has been challenged. Exploring new control methods is an important problem that must be solved in the prevention and control of vegetable diseases. The development of biological control technology to prevent and treat plant diseases has attracted more and more attention from governments, scientific and technological workers and the public. As far as the applicant knows, there have been many reports on the research of biological measures to prevent and control Phytophthora disease at home and abroad. However, it is difficult for a single biocontrol factor to achieve the same control effect as the use of chemical fungicides under different environmental conditions; moreover, biocontrol factors are sensitive to fluctuations in the environmental conditions of the soil or the root circle of crops. , the colonization ability of biological control factors is very different. At present, research on biological control of vegetable diseases is mostly carried out in laboratories and greenhouses, and has not been used in large-scale production practice.

Under the condition that chemical control has been questioned more and more and the biological control system is still not perfect, the development of bio-chemical synergistic control strategy of vegetable diseases has gradually attracted the attention of governments and researchers in various countries.

Contents of the invention

The object of the present invention is to provide a bio-chemical synergistically controlled bacterial-drug mixture that is highly efficient, low-toxic, compatible with the environment and has the effect of preventing and controlling vegetable blight and its preparation method.

The object of the present invention is achieved in the following way: provide a kind of anti-vegetable blight bacteria medicine mixture containing biocontrol bacteria agent and bactericide, the dosage ratio of biocontrol bacteria agent and bactericide is 1:1 by mass. The preferred fungicides are selected from one or more of amida, chlorothalonil, mancozeb, metalaxyl, lvdi, flumorph, dimethomorph, phosphonium, and Intime. A further preferred bactericide is dimethomorph, more preferably a wettable powder with a dimethomorph active ingredient content of 25%-90%, most preferably a 50% anke wettable powder (dimethomorph preparation). The biocontrol bacterial strain contained in the preferred biocontrol agent is selected from one or more of Bacillus subtilis BS, Bacillus subtilis GF1, Pseudomonas fluorescens LX1, and Pseudomonas fluorescens BCA1. The biocontrol bacterial strain contained in the further preferred biocontrol agent is Bacillus subtilis BS.

The present invention also provides a method for preparing the above-mentioned mixture of medicines for preventing and treating vegetable blight, which comprises the following steps:

a) Fermenting and culturing biocontrol strains that have antagonistic effects on vegetable blight pathogens, and then centrifuging the fermented liquid to obtain a bacterial cell precipitate that is mixed with micropowder calcium carbonate at a mass ratio of 1:1 to prepare a biocontrol agent;

The preferred biocontrol strain is selected from one or more of Bacillus subtilis BS, Bacillus subtilis GF1, Pseudomonas fluorescens LX1, and Pseudomonas fluorescens BCA1. A further preferred biocontrol strain is Bacillus subtilis BS.

b) Compounding the biocontrol agent and fungicide formed in step a at a mass ratio of 1:1.

A preferred fungicide is a preparation with an active ingredient content of 25%-90%, and a further preferred fungicide is a preparation with an active ingredient content of 50%.

The preferred fungicides are selected from one or more of amida, chlorothalonil, mancozeb, metalaxyl, lvdi, flumorph, dimethomorph, phosphonium, and Intime. A further preferred bactericide is dimethomorph, more preferably a wettable powder with a dimethomorph active ingredient content of 25%-90%, most preferably a 50% anke wettable powder (dimethomorph preparation).

The bacterial-drug mixture based on the principle of bio-chemical synergistic control, through the synergistic effect of biological factors and chemical factors, can reduce the amount of chemical pesticide application on facility vegetables by more than 50%, completely replacing traditional high-toxicity and high-residue pesticides , to ensure that the pesticide residues in vegetables are below the allowable standards and meet the requirements for pollution-free vegetable production. Thereby saving production cost and improving vegetable quality.

Description of drawings

Fig. 1 is a process flow block diagram of the present invention.

Detailed ways

The present invention will be further described in detail below with reference to the embodiments described in the accompanying drawings.

The mixture of bacteria and medicines for preventing and controlling vegetable blight contains biocontrol bacteria agent and 50% Anke wettable powder; the mass ratio of the biocontrol bacteria agent to 50% Anke wettable powder is 1:1.

As shown in the figure, the preparation method of the bacterial medicine mixture for preventing and treating vegetable blight can be carried out in sequence as follows:

a) Ferment and cultivate the biocontrol strain Bacillus subtilis BS which has an antagonistic effect on the pathogen of Phytophthora capsici, and then centrifuge the fermented broth to obtain the bacterial precipitate and mix it with micropowder calcium carbonate at a mass ratio of 1:1 to prepare a biocontrol agent ;

b) compound the biocontrol agent formed in step a with the Anke wettable powder with an active ingredient content of 50% in a mass ratio of 1:1.

The present invention is based on the obtained several antagonistic microorganisms, the registered biological control strains and several fungicides commonly used in domestic and foreign production to prevent and control epidemic diseases, and takes the pepper blight pathogen as the target bacteria, and obtains a A biocontrol strain with strong antagonistic effect on Phytophthora capsici and a chemical pesticide with high efficiency, low toxicity and environmental friendliness. Through in vitro and in vivo tests, the toxicity coefficient of biological pesticides and chemical pesticides was determined, and the relationship between biological factors and chemical agents was determined. The synergistic effect of specialization and specificity among them, the synergistic combination of high-efficiency broad-spectrum biofungicides were screened, the fungicide mixture was developed, and its application method was studied.

Concrete operation steps of the present invention are as follows:

(1) Screening of antagonistic microorganisms On the basis of preliminary screening, four biocontrol strains of Bacillus subtilis BS, Pseudomonas fluorescens LX1, Bacillus subtilis GF1 and Pseudomonas fluorescens BCA1 were selected for in vitro inhibition respectively. Bacillus subtilis (Bacillus subtilis) BS has a strong antagonistic effect on the pathogen of pepper blight through bacterial experiments and live plant screening, and has a high control effect in the greenhouse disease control test. See Table 1 and Table 2 for specific data. Therefore, Bacillus subtilis BS was selected as the biological component for the development of bacterial-drug mixture.

(2) The screening of fungicides adopts 9 kinds of different types of fungicides commonly used in production to measure the inhibitory effect on the different thalline forms of Phytophthora capsici respectively, and the results are as shown in table 3. Dimethomorph and flumorph The two fungicides had significant inhibitory effects on the various cell forms measured. On the basis of in vitro screening, and using metalaxyl, a typical drug for epidemic prevention and control, as a control, the greenhouse disease prevention test of dimethomorph and flumorph was carried out, and it was found that the control effect of dimethomorph was better than that of flumorph. morphine (see Table 4). Therefore, dimethomorph was selected as the chemical component for constructing the bacteria-drug mixture.

(3) Development of bacteria-drug mixture: Bacillus subtilis BS was fermented and cultured, and then the bacterial precipitate obtained by centrifuging the fermentation broth was mixed with micronized calcium carbonate in a mass ratio of 1:1 to prepare a biocontrol agent with a bacterial count of more than 10 ⁸ cfu/g. Then it is compounded with 50% Anke wettable powder (dimethomorph preparation) at a mass ratio of 1:1 to obtain the bacterium-drug mixture for preventing and treating vegetable blight of the present invention.

(4) The detection of dimethomorph and the content index of Bacillus subtilis BS of bacteria-drug mixture quality index is the basic quality index of bacteria-drug mixture, and the mensuration of dimethomorph content adopts high performance liquid chromatography to carry out (≥25%), The number of Bacillus subtilis was determined by the dilution plate method (number of viable colonies ≥ ^{10 7} cfu/mL). The heat storage experiment shows that Bacillus subtilis has no metabolic effect on dimethomorph, and dimethomorph has no interference effect on the colony formation of Bacillus subtilis.

Example 1:

Jingyan Pepper No. 3, the main pepper variety grown in Beijing, was used to simulate the infection mode of pepper blight under natural conditions, and the pathogenic system was established by inoculating the pathogen with root irrigation method and spraying method, and the effects of different application methods on the fungus were studied. The effect of medicine mixture DMBS on the control of capsicum blight. The results are shown in Table 5 and Table 6. Under different pathogenic systems, the control effect of the fungal-drug mixture is related to the way of use of the agent. Drugs are more effective in preventing and controlling the spread of aboveground pathogens. However, no matter which inoculation method is used, it can be found that among the three treatments of single agent of fungicide, single agent of biocontrol bacteria and mixture of bacteria and drugs, the control effect of the single agent of biocontrol is not stable, while the mixture of bacteria and drugs It showed the same trend as that of a single fungicide, and the control effect increased with the increase of the concentration of the fungicide; more importantly, the control effect of the fungicide mixture with half the dose was roughly equivalent to that of a single fungicide with the same concentration. The control effect is even higher than that of fungicide treatment at individual concentrations.

Example 2:

Using sweet pepper, a general-purpose susceptible variety of pepper blight, to simulate the infection mode of pepper blight under natural conditions, the pathogenic system was established by inoculating the pathogen by root irrigation method and spraying method, and the effects of different application methods on the bacteria were studied. The effect of medicine mixture DMBS on the control of capsicum blight. The results are shown in Table 7 and Table 8, and the rules are similar to those of Jingyan Pepper No. 3.

Table 1, Determination of the antibacterial effect of 4 strains of biocontrol bacteria on 7 strains of pepper blight Tested strains (different strains of Phytophthora bacterium) biological control factor BS GF1 LX1 BCA1 SD21-3GZ01Pc112Pc33EPc1E-8P1319P1314 35.6038.1540.8059.4355.0144.5333.15 27.7226.7034.8141.3240.4941.3021.10 24.7328.0738.3643.0247.6541.2135.07 32.3429.1639.6944.1547.8542.9134.25

Table 2. Greenhouse experiments on the control of pepper blight by biocontrol agents Prevention method BCAs spray inoculation Inoculation of pathogens by root irrigation Disease index (%) Control effect (%) Disease index (%) Control effect (%) Spray control Root irrigation control CKBSGF1LX1BCA1BSGF1LX1BCA1 19.91ABa13.61DCbc18.00ABCab17.87ABCab14.00BDCbc13.08DCc18.96ABCa21.69Aa8.40Dd 31.649.6010.2729.6934.324.79-8.9357.81 14.80BAba6.39Dd14.28BACba17.76Aa14.00BACba11.60BDCbc12.80BACb13.56BACba8.40DCdc 56.833.53-19.975.4121.6213.518.4143.24

Table 3. Inhibitory effects of 9 fungicides on different bacterial forms of Phytophthora capsici fungicide mycelial growth sporangia production germination of restospores sporangia germination _EC50 (μg/mL) _EC95 (μg/mL) _EC50 (μg/mL) _EC95 (μg/mL) _EC50 (μg/mL) _EC95 (μg/mL) _EC50 (μg/mL) _EC95 (μg/mL) Amishida 100 bacteria Qing Dai Sen mancozeb flumorph morphine metalaxyl lvdiene acyl morpholine ethyl phosphorus aluminum Yintai >100>100>1000.180.0155.460.14>10028.82 >1000>1000>10000.670.03>10000.45>1000102.35 6.151.978.700.682.7520.160.6125.2162.66 244.6124.13114.322.575.9757.662.44466.98460.43 0.0940.21118.700.30>10070.240.1261.3439.49 0.4091.116116.720.78>1000147.600.52178.50155.71 1.270.993.820.64155.5341.110.2578.5626.09 139.6921.8922.661.48490.06222.540.99390.19130.88

Table 4. Greenhouse test of fungicides on pepper blight deal with Drug concentration (μg/mL) Pepper Varieties Jingyan Pepper No. 3 eggplant bell pepper Disease index (%) Control effect (%) Disease index (%) Control effect (%) CK Metalaxyl Dimethomorph Flumorph 0510255010025501002505002550100250500 709082785650483220166625652186 --28.57-17.14-11.4320.0028.5731.4354.2971.4377.1491.4311.4320.0025.7174.2991.43 786248402267050381207456442410 -20.5138.4648.7271.7992.3110.2635.9051.2884.62100.005.4328.2143.5969.2387.18

Table 5, the prevention and control effect of the bacterium-drug mixture of the present invention on Jingyan No. 3 capsicum root irrigation method inoculated with capsicum blight pathogen deal with spray application Root spraying Disease index (%) Control effect (%) Disease index (%) Control effect (%) CK 37.50Aa DMM(μg/mL)BS(cfu/mL)DMBS(μg/mL) 5010025010 ⁷ 10 ⁸ 10 ⁹ 2550125 18.00DEFdef12.44GFgh9.33Hgh35.60Aab31.59Aab35.20Aab22.40CDcd20.40CDEde16.71DEFefg 52.0066.8275.115.0715.766.1340.2745.6055.43 26.00BCc5.11HIi0.00Ij34.50Aab15.33EFfg35.38Aab16.00EFefg2.00Iij1.20Iij 30.6786.37100.008.0059.115.6657.3394.6796.80

Note: DMM is dimethomorph, BS is Bacillus subtilis, and DMBS is bacterial-drug mixture. The same below.

Table 6, the prevention and control effect of the bacterium-drug mixture of the present invention on No. 3 spray method of Jingyan pepper inoculated with capsicum blight deal with spray application Root spraying Disease index (%) Control effect (%) Disease index (%) Control effect (%) CK 44.40Aa DMM(μg/mL)BS(cfu/mL)DMBS(μg/mL) 5010025010 ⁷ 10 ⁸ 10 ⁹ 2550125 5.60HIg4.17Ig4.00Ig33.50BCc26.80CDd40.67ABab6.40HIg4.00Ig3.34Ig 87.3990.6290.9924.5539.648.4185.5990.9992.47 24.67DEd18.40EFe12.58FGHef27.91CDd35.50Bbc40.45ABb15.50FGe13.20FGHef9.02GHIgf 44.4458.5671.6737.1420.058.9165.0970.2779.68

Table 7, the prevention and control effect of the bacterium-drug mixture of the present invention to the inoculation of capsicum blight pathogen by root irrigation method of eggplant sweet pepper deal with spray application Root spraying Disease index (%) Control effect (%) Disease index (%) Control effect (%) CK 45.20Aa DMM(μg/mL)BS(cfu/mL)DMBS(μg/mL) 5010025010 ⁷ 10 ⁸ 10 ⁹ 2550125 23.60CDcd21.20CDEcde14.50EFfg22.80CDcd26.13Cc34.80Bb24.84Cc14.50EFfg16.67DEFef 47.7853.1067.9249.5642.1923.0145.0467.9263.13 3.50GHij2.40GHij1.60Hj21.00CDEcde16.40DEFef38.50ABb18.80CDEdef9.60GFgh7.20GHih 92.2694.6996.4653.5463.7214.8258.4178.7684.07

Table 8, bacterium-drug mixture of the present invention is to the prevention and treatment effect of eggplant sweet pepper spraying method inoculation capsicum blight pathogen deal with spray application Root spraying Disease index (%) Control effect (%) Disease index (%) Control effect (%) CK 23.33Aa DMM(μg/mL)BS(cfu/mL)DMBS(μg/mL) 5010025010 ⁷ 10 ⁸ 10 ⁹ 2550125 9.70BCbc9.60BCbc8.80BCcbd10.00BCbc7.20BCcbd11.60BCcbc6.00BCcd6.40BCcbd3.40BCcbd 58.4158.8662.2957.1469.1450.2974.2972.5785.43 8.80BCcbd6.90BCcbd8.40BCcbd8.40BCcbd7.24BCcbd8.80BCcbd12.40Bb7.20BCcbd6.40BCcbd 62.2970.4364.0064.0068.9562.2946.8669.1472.57

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.