CN101032251A - A fungicide whose active substance is risamectin - Google Patents

Abstract

The invention relates to a bactericide with a helatin as an active substance, which is prepared by extracting the helatin with bactericidal activity through microbial fermentation. The active component of the bactericide is a material with bactericidal activity, namely the rhamnan, and the active material is extracted from soil through separation, screening and microbial fermentation.

Description

A fungicide whose active substance is risamectin

1. Technical field:

The invention relates to a pesticide, in particular to a bactericide for preventing and treating rice blast and having bactericidal active substances extracted by a microbial fermentation method.

2. Background technology:

Since the advent of organochlorine and organomercury pesticides in the 1940s, people have mainly relied on chemical pesticides as a modern weapon in the process of fighting agricultural pests and weeds. However, with the rapid increase in the variety and output of chemical pesticides, the scope of application is expanding day by day, and the pollution of pesticide residues has caused worldwide public nuisance, which has aroused people's worries. Along with the enhancement of human's awareness of environmental protection, biological pesticides that are safe for humans and animals, non-toxic, non-polluting, and non-poisonous to kill insect natural enemies have sprung up. Among biological pesticides, microbial-derived pesticides are currently the most widely used, fastest-growing, and most promising category, and are increasingly favored by people. In order to protect the ecological environment and promote the sustainable development of agriculture, the development of microbial pesticides has broad prospects and far-reaching significance. The most typical microbial pesticides are agricultural antibiotics. Agricultural antibiotics are derived from microorganisms, especially actinomycetes. According to statistics, more than 2/3 of more than 1,000 kinds of antibiotics are produced by actinomycetes, and the antibiotics produced by streptomyces actinomycetes account for more than 90% of actinomycetes; and actinomycetes are soil ecological It is one of the three major groups of microorganisms in the system and widely exists in various soils. As far as microorganisms are concerned, less than 1‰ of those that exist in nature have been studied by humans, and only 1% of these studied microorganisms have been found to produce biologically active substances, not to mention that microorganisms are variable, due to genetic recombination And chromosomal aberrations, new variants are produced every day, so the research in this field can be said to be endless. The research and application of agricultural antibiotics in my country began in the early 1950s. Since the 1960s, blasticidin, Jinggangmycin, kasugamycin, Qingfengmycin and other agricultural resistant varieties have been successively developed and applied.

3. Contents of the invention:

The object of the present invention is to provide a fungicide whose active substance is risamectin, which extracts the bactericidal substance risamectin through microbial fermentation, and a large number of drug efficacy tests show that it is effective against many plant pathogens. All have good control effects, and are compatible with the environment, and have the characteristics of low residue, high control efficiency, and wide bactericidal spectrum.

To achieve the above object, the technical solution adopted in the present invention is:

A fungicide whose active substance is relamectin, is characterized in that: the active ingredient of the fungicide is the substance relamectin with fungicidal activity:

The active substance is extracted from the soil through separation, screening and microbial fermentation.

The above-mentioned active substance relamectin is separated, screened, and the process steps of extraction are as follows:

1 Collect soil samples

2 Isolation of soil actinomycetes

3 Screening of antagonistic bacteria

3.1 Preliminary screening of antibacterial activity

3.2 Re-screening of antagonistic actinomycetes

3.3 Single spore isolation

3.4 Antagonistic screening of monospora

3.5 Optimization of fermentation conditions for antagonistic bacteria

3.5.1 Selection of fermentation medium

(1) Selection of organic nitrogen source

(2) Selection of organic carbon source

(3) Selection of inorganic ions

(4) Optimal culture medium orthogonal experiment

3.5.2 Selection of PH of fermentation medium

3.5.3 Selection of fermentation time

3.5.4 Selection of fermentation temperature

3.5.5 Selection of shaker speed

3.5.6 The relationship between the growth of actinomycetes hyphae and the production of antibiotics

3.6 Extraction of active product of highly efficient strain S-159-05

3.6.1 Pretreatment of fermentation broth

3.6.2 Rough extraction

3.6.3 Extraction

The above-mentioned bactericide includes the active ingredient relamectin, and the bactericide also includes surfactants, defoamers, synergists, hypertonic agents, emulsifiers, water, etc., and can be made into water emulsion, which consists of:

Rilamectin 12 parts by weight

Surfactant 3 parts by weight

Defoamer 2 parts by weight

Synergist 3 parts by weight

Auxiliary 2 parts by weight

Emulsifier 2 parts by weight

Hypertonic agent 2 parts by weight

the rest is water

The above-mentioned surfactant is sodium lauryl sulfate, the defoamer is dimethyl polysiloxane, and the synergist is a synergist made of tea saponin 15% aliphatic amine 3% and water 72%, hypertonic The agent is fatty alcohol polyoxyethylene (6-7) ether, the emulsifier is sodium maleic acid monoester sulfonate, and the auxiliary agent is sodium tripolyphosphate.

The above-mentioned fungicides include various pesticide formulations used for crop sterilization.

The above-mentioned bactericide is an aqueous emulsion.

Compared with prior art, the advantages and effects that the present invention has are as follows:

The invention has the characteristics of low residue and high control effect, wide bactericidal spectrum and good control effect on various bacteria.

4. Description of drawings:

Fig. 1 is an overall process diagram of the present invention;

Fig. 2 is the concrete process flowchart of the present invention.

5. Specific implementation methods:

The invention belongs to a kind of pesticide, which is a new type of agricultural antibiotic prepared by fermentation of Streptomyces venezuela var. Rice blast.

Example:

Specific process steps and process parameters for the separation, screening and extraction of the active substance relamectin

1. Collect soil samples

Select a representative location, according to the method of mixing multi-point sampling (different locations, different soil depths), shovel off the surface soil, use a simple hole puncher, take a soil sample with a depth of 3-15cm, and put it into a sterile Kraft paper bag or aluminum box, keep records. The collected soil samples should have no large particles, and the actinomycetes should be isolated immediately after being brought back to the laboratory to dry in the shade.

2. Isolation of soil actinomycetes

(1) Preparation of soil suspension: Take a certain amount (about 10g) of shade-dried soil, put it in a sterilized mortar, and grind it on a sterile operating table until the soil particles are relatively uniform and stop when there are no large particles. Weigh 1g of the ground soil sample and put it into a sterilized test tube, add 10ml of sterile water and shake it for 30 minutes to make a 1:10 soil dilution. Take 1ml out of the above solution, add 9ml of sterile water and shake fully to make a 1: ¹⁰² dilution, and so on, make 1: ¹⁰³ , 1: ¹⁰⁴ , 1: ¹⁰⁵ soil dilutions for later use .

(2) Plate inoculation: use smear method and mixed bacteria method to inoculate

Smearing method: take 0.05ml of the above-mentioned 5 concentrations of soil dilutions, add them to the prepared Gaoshi No. 1 plate, and spread the dilutions evenly with a sterilized glass spatula. Place in a 28°C incubator for cultivation. Set up three repetitions.

Mixed bacteria method: Take 1ml of the soil dilution solution of the above 5 concentrations, add it to a petri dish, pour the melted 45-50°C Gaoshi No. 1 medium, shake gently to mix the sample and the medium evenly, After the medium solidified, put it into the incubator at 28°C for constant temperature cultivation. Set up three repetitions.

(3) Purification: Observe and record the original culture results of the strains at any time, and number them uniformly. According to the characteristics of actinomycetes, a single actinomycete colony was picked in time and placed on Gaoshi No. 1 plate, and purified by streak separation.

3. Screening of antagonistic bacteria

3.1 Preliminary screening of antibacterial activity

Activate the actinomycetes strains preserved in 2.2.2, and use the cross method to directly measure with the agar plate in the dish. The specific steps are: draw a vertical cross line on the PDA plate with the origin as the center, connect the pathogenic bacteria at the center point first, connect the same kind of actinomycetes at the two ends of one line of the cross at the same distance from the center, and connect the two ends of the other line with the other An actinomycete. Pathogenic bacteria and actinomycetes with similar growth rate can be inoculated at the same time, and slow growth of pathogenic bacteria should be delayed for 1-2 days. Cultivate for a certain period of time to see if there is a zone of inhibition. If there is a bacteriostatic zone, measure the relative radius of the bacteriostatic zone (the radius of the bacteriostatic zone—the radius of the pathogenic bacteria colony). Select the actinomycetes with good antagonistic ability to enter the re-screening.

3.2 Re-screening of antagonistic actinomycetes

Select the actinomycetes with good antagonistic properties in the preliminary screening, propagate them by means of liquid fermentation culture, and measure the antagonistic properties of the fermentation filtrate to pathogenic bacteria. The method is as follows:

(1) Actinomycetes were inserted into the fermentation medium, cultured on a constant temperature water bath shaker, and cultured at 28° C. for 72 hours.

(2) Fermentation broth pretreatment Take out the actinomycetes liquid after culturing for 72 hours, adjust the pH to 4 with oxalic acid (0.5m/l), let it stand for 1 hour, and filter under sterile conditions to obtain the fermentation filtrate.

(3) Determination of antagonism by growth rate method: 3ml of the fermentation filtrate is injected into a petri dish, then the melted PDA medium (about 25ml) is poured into the dish and mixed evenly with the fermentation filtrate, after cooling, the bacterium cake of pathogenic bacteria is connected. Distilled water and oxalic acid were added as controls. Measure the colony diameter after 24h, 48h, and 72h, and calculate the inhibition rate.

(4) Select the strain with the best antagonistic properties, and use the auxin rate method to measure the effect of different concentrations of fermentation filtrate on the growth of pathogenic fungus mycelia.

3.3 Single spore isolation

The strains with broad antibacterial spectrum and good antagonisticity in the double screening were selected for single spore isolation. Specific steps are as follows:

(1) Make Gao's No. 1 plate.

(2) Add spores of actinomycetes (without culture medium) to 10ml of sterile water to make a 1:10 spore suspension, shake fully for 10 minutes, then take out 1ml and add it to 9ml of sterile water to make a 10-2 Spore suspensions, and so on, to make spore suspensions of 10-3, 10-4, 10-5, 10-6.

(3) Use a pipette gun to take 0.5ml of the diluted spore suspension and add it to the Gaoshi No. 1 plate, spread it evenly with a spatula, and set 3 repetitions for each treatment. Incubate at 28°C for 7h.

(4) Select a plate with 5-20 colonies, number the colonies, pick the single-spore colonies into the Gaoshi No. 1 slant, culture them for 7 days, and store them in a 4°C refrigerator.

3.4 Antagonistic screening of monospora

The antagonism of the fermentation filtrate of each monospora was determined by the auxin ratio method. The method is the same as that of 3.2 Actinomycetes re-screening. Select the most antagonistic strain as the target actinomycete strain.

3.5 Optimization of fermentation conditions for antagonistic bacteria

The optimization of following fermentation medium, the optimization of substratum pH, fermentation time, fermentation temperature, shaking table rotating speed all measure with the inhibition rate index of the fermentation filtrate (50ml/l) of growth rate method, for testing pathogenic bacteria is Botrytis cinerea, The culture temperature was 28°C, and the measurement time was 48 hours after inoculation with pathogenic bacteria. Set up three repetitions. The preparation of fermentation filtrate and the calculation formula of inhibition rate are the same as 3.2.

3.5.1 Selection of fermentation medium

(1) Selection of organic nitrogen source

On the basis of the fermentation medium used in the original screening test, the carbon source content was kept unchanged, and the fermentation medium only changed the composition of the nitrogen source. The inhibition rate of the fermentation filtrate was measured by the growth rate method, and the antibacterial activity of the fermentation filtrate was compared. The selected nitrogen source combinations are: peptone, yeast powder, soybean meal, soybean meal + corn flour, soybean meal + KNO3, peptone + corn steep liquor, yeast powder + NH4SO4, wheat bran + yeast + soybean meal.

(2) Selection of organic carbon source

The screened soybean cake powder + KNO3 was used as nitrogen source, the composition of carbon source was changed, the inhibition rate of fermentation filtrate was measured, and the antibacterial activity of fermentation filtrate was compared. The selected carbon sources are: starch, dextrin, glucose, sucrose, glucose+dextrin, glucose+starch, starch+sucrose, glucose+corn oil.

(3) Selection of inorganic ions

According to the optimal carbon source and nitrogen source that have been screened, dipotassium hydrogen phosphate, ferrous sulfate, copper sulfate, and zinc sulfate were added respectively, and the L9 (34) orthogonal experiment was used to investigate the effect of the three levels of each inorganic salt on the potency. Influence. The orthogonal experiment scheme is as follows:

Table 1 L9(34) Orthogonal experiment factor level table factor I II III IV Level 1(%) Level 2(%) Level 3(%) 00.050.1 00.050.1 00.010.1 00.010.1

Note: I dipotassium hydrogen phosphate, II copper sulfate, III ferrous sulfate, IV zinc sulfate

(4) Optimal culture medium orthogonal experiment

From the research on carbon and nitrogen sources mentioned above, the optimal types of carbon and nitrogen sources and the appropriate concentrations of carbon and nitrogen sources were determined. In addition, since calcium carbonate plays a role in regulating pH in the medium, a certain amount is often added to the liquid fermentation medium. In order to further study the correlation between the relevant nutrient source factors, four factors including soybean flour cake, glucose, starch and calcium carbonate were selected to conduct an orthogonal experiment. Take 3 levels for each factor, and arrange experiments according to the orthogonal table L9(34). As in Table 2.

Table 2 L9(34) Orthogonal experiment factor level table factor A B C D. Level 1(%) Level 2(%) Level 3(%) 1.52.02.5 0.51.01.5 0.51.01.5 0.10.250.5

Note: A soybean cake powder, B glucose, C starch, D calcium carbonate

3.5.2 Selection of PH of fermentation medium

Use NaOH or HCl to adjust to different pH values: 5.0, 6.0, 7.0, 8.0, and 9.0, and select the optimum pH value of the medium according to the inhibition rate of the fermentation broth to the tested bacteria.

3.5.3 Selection of fermentation time

Measure the inhibition rate of 24, 36, 48, 60, 72, 84, 96, 120h fermentation broth respectively, and choose the best fermentation time.

3.5.4 Selection of fermentation temperature

The actinomycetes were cultured in a constant temperature shaking incubator at 26°C, 28°C, 30°C, 32°C, 34°C, 36°C and 38°C, and the inhibition rate of the fermentation broth was calculated to determine the optimum fermentation temperature.

3.5.5 Selection of shaking table speed

Test the influence of 100r/min, 150r/min, 200r/min, 250r/min shaker speed on fermentation respectively. Determine the optimum shaker speed.

3.5.6 The relationship between the growth of actinomycetes hyphae and the production of antibiotics

(1) Determination of pH changes during actinomycetes metabolism

The actinomycetes were cultured under the optimized shaker fermentation conditions, samples were taken every 24 hours, the pH value of the fermentation broth was measured, and the pH change curve was drawn.

(2) Drawing of the curve of antibiotic production by actinomycetes

The actinomycetes were cultured under the optimized shaker fermentation conditions, samples were taken every 12 hours, the mycelium morphology was observed, the inhibition rate of the fermentation filtrate was measured, and the antibiotic production curve of the actinomycetes was drawn.

(3) Drawing of actinomycetes growth curve

The growth curve of actinomycetes can be measured by cell accumulation volume method: get fermented liquid 10ml every 24h and be packed in the graduated centrifuge test tube, centrifuge 5min under the rotating speed of 3500rpm, take the sediment volume percentage content as relative hyphae concentration (weight). The growth curve of actinomycetes is drawn with time as the abscissa and the percentage of sediment volume as the ordinate.

According to the above determination results, the relationship between the growth of actinomycetes mycelium and metabolism was analyzed.

3.6 Extraction of active product of highly efficient strain S-159-05

3.6.1 Pretreatment of fermentation broth

The screened high-yield strain S-159-05 was selected from the medium optimized by the orthogonal test, and cultured for 72 hours under the conditions of pH 8.0, 30-34°C, and shaker speed 150r/min, and then taken out. Use 1mol/L oxalic acid to adjust the pH of the fermentation broth, stir while adding oxalic acid, and measure the pH value of the fermentation broth with a pH meter. When the pH is 4.0, let stand for 0.5h, then adjust the pH to 4.0, let stand for half an hour, filter and collect the filtrate.

3.6.2 Rough extraction

(1) Concentrate the filtrate to a small volume under reduced pressure at 70°C in a rotary thin film evaporator, adjust the pH to 7.0 with 6mol/L NaOH, centrifuge at 5000rpm for 30min, discard the precipitate, and take the supernatant. The supernatant was precipitated with 10 times the volume of frozen acetone, the supernatant was discarded, and the precipitate was collected and dried.

(2) 732 cation exchange resin adsorption

After soaking 732 cationic resin in water for 2 hours, remove the air bubbles under reduced pressure, pour off the water, wash with a large amount of ion-free water until clarified, add 4 times the amount of 2mol/l HCl after removing the water, stir for 4 hours, remove the acid solution, and wash with water until medium Add 4 times the amount of 2mol/L NaOH, stir for 4 hours, remove the lye, wash with water to neutrality, soak in 4 times the amount of 2mol/L NaOH for 2 hours, and convert to Na ⁺ type column. Fully dissolve the acetone precipitate with redistilled water, add the sample, rinse with distilled water until the effluent is colorless and clear, then elute with 3% ammonia water, and collect the eluate in sections. The activity of the eluate from each section was detected by the spore germination method. The active fraction was concentrated under reduced pressure to obtain a crude extract.

3.6.3 Refining Extraction

(1) Macroporous resin column chromatography

The new resin was first soaked in absolute ethanol for 3-4 days, then loaded into a chromatography column, rinsed with absolute ethanol and allowed to stand for 6 hours. After the macroporous adsorption resin settles and compacts, wash the column with 6 times the column volume of distilled water. Then rinse the resin with 3-4 times the volume of 2% HCl solution and soak it for 2-4 hours, and then wash it with distilled water until the pH of the effluent is neutral. Rinse the resin with 3-4 times the volume of 2% NaOH solution, soak it for 2-4 hours, and then wash it with distilled water until the pH of the effluent is neutral. The active eluent extracted by the ion exchange resin is statically adsorbed on the macroporous resin, and distilled water, 40% ethanol, 60% ethanol, and 100% ethanol are used as the eluent respectively, and 20ml of the eluent is collected in each tube, concentrated under reduced pressure, The activity was measured by the spore germination method, and the active eluent was dried to obtain the fine extract.

(2) The fine extract was further cut and separated on the reversed-phase preparative column high-performance liquid chromatography, and the activity of each peak of the preparative chromatogram was measured by the spore germination method, and the single component with strong antibacterial effect was separated and prepared, and it was subjected to physical and chemical analysis Property and structure analysis.

The specific preparation process of fungicide (water emulsion) pesticide:

1. The original drug is dissolved in water

Rilamectin is easily soluble in water, and the water-emulsion has stable quality and is easy to store and transport, which is beneficial to crop absorption and environmental protection.

2. Determine the content of fermentation

It has been proved by repeated fermentation experiments that under normal fermentation conditions, most batches of rilamectin-producing bacteria have a relamectin content ≥ 1.2%. In order to determine the product quality, commercial rilamectin A content of 1.2% is a qualified product standard.

Table 3 Determination table of ten batches of relamectin-producing bacteria fermentation broth samples serial number Sample lot number Sample determination content (%) serial number Sample lot number Sample determination content (%) 1 20030407 1.201 6 20030429 1.212 2 20030411 1.225 7 20030512 1.257 3 20030415 1.209 8 20030516 1.248 4 20030420 1.215 9 20030520 1.238 5 20030424 1.198 10 20030526 1.279 average value 1.228

3. Add emulsifier, defoamer, etc.

Add surfactant sodium lauryl sulfate, antifoaming agent dimethyl polysiloxane, synergist made of tea saponin 15% fatty amine 3% and water 72%, hypertonic agent fatty alcohol polyoxyethylene (6-7) Ether, emulsifier sodium maleic acid monoester sulfonate, auxiliary agent is sodium tripolyphosphate. Its composition is:

Rilamectin 12 parts by weight

Surfactant 3 parts by weight

Defoamer 2 parts by weight

Synergist 3 parts by weight

Auxiliary 2 parts by weight

Emulsifier 2 parts by weight

Hypertonic agent 2 parts by weight

Water 74 parts by weight

4. Stir

5. Packaging

Field efficacy test of active substance relamectin aqueous emulsion of the present invention for controlling rice blast

1. Crops and targets

Test crop: rice variety "Sanlicun"

Prevention object: rice blast (Pyricularia oryzae Cav.e)

2. Environmental conditions

The test was carried out in the farmer's responsibility field in the suburbs of Shahe Town, Yunmeng City, Xiaogan City, Hubei Province. Rice blast occurs moderately and severely all year round. The test subjects are all rice leaf blast. The soil type of the test site is loam, and the cultivation conditions of each test plot are uniform.

3. Experiment design and arrangement

3.1 Pharmacy

3.1.1 Test drug and dosage

1.2% relamectin aqueous emulsion (provided by Xi'an Hailang Chemical Co., Ltd.), with three dosages: 15 grams, 16.5 grams and 18 grams of active ingredients per hectare.

3.1.2 Control drug and dosage

75% tricyclazole wettable powder (manufactured by Xiaoyang Agricultural Chemical Factory, Guanghan City, Sichuan Province), the active ingredient consumption per hectare is 225 grams. In addition, a clear water blank control was set up.

3.2 Community arrangement

In this experiment, there were five treatments and four repetitions, with a total of 20 plots, and the plots were randomly arranged in blocks, each plot was 35 square meters.

3.3 Application method

3.3.1 Application time and frequency

When it is found that there are sporadic rice blast spots on the leaves of rice, and when there is an obvious upward trend, according to the above 3.1 test design, the amount of liquid medicine in each plot is converted into the amount of liquid medicine of 75 kilograms per mu and sprayed (June 12), after 10 days (June 22) Spray the second medicine, spraying 2 times in total. The spraying equipment is Gongnong-16 manual sprayer.

3.3.2 Meteorological data

During the application period, the daily average temperature was between 24.5°C and 32.0°C, and there were light to moderate rains or showers on June 18 and 20 respectively, which was conducive to the development of the disease.

4. Survey

4.1 Survey methods and grading standards

Each community adopts the diagonal five-point sampling method, surveying 5 clusters at each point, a total of 25 clusters. Using grading standards of 0-9 grades, the final disease result investigation was carried out 14 days after the second application; the disease index and drug efficacy were calculated respectively, and biostatistical analysis (DMRT method) was carried out. Observe the injury situation.

Grading standards for rice leaf blast:

Level 0: no disease;

Grade 1: There are less than 5 lesion spots on the leaves, and the length is less than 1cm;

Grade 3: 6-10 lesions on leaves, some of which are longer than 1cm;

Grade 5: 11-25 lesions on leaves, some of which are contiguous, accounting for 10%-25% of the leaf area;

Level 7: There are more than 26 diseased spots on the leaves, and the diseased spots are connected into pieces, accounting for 26%-50% of the leaf area;

Grade 9: Lesions are contiguous into sheets, accounting for more than 50% of the leaf area.

4.2 Calculation method of efficacy

Disease index = ∑ (number of diseased leaves at all levels × relative level value) / (total number of leaves under investigation × 9) × 100

Control effect = (blank control disease index-drug treatment disease index)/blank control disease index×100

5. Analysis and discussion of test results

5.1 Test results and analysis

As can be seen from Table 4 of the test results, the test agent 1.2% relamectin aqueous emulsion has a relatively ideal control effect on rice blast, and with the increase of the dosage, the control effect increases. . The control effect of 225 grams of powder is 86.3%.

Statistical analysis results show that the control effect of 1.2% relamectin water agent 18 grams of the test agent is significantly higher than that of 16.5 grams of treatment, and the difference between the control effect of 16.5 grams of treatment and the control effect of 15 grams of treatment is not significant; Compared with the control effect of the control drug, the control effect of 18 grams of the test drug 1.2% rilamectin aqueous solution is equivalent to the control effect of the control drug 75% tricyclazole wettable powder 225 grams. No adverse effects of the test agents on rice were found in the whole test.

Table 4 Test results of 1.2% relamectin aqueous solution for controlling rice blast deal with Active ingredient dosage repeat 14 days after treatment Anti-efficacy (%) average(%) 0 1 3 5 7 9 Sick finger 1.2% relamectin aqueous solution 15 g/ha I 244 34 twenty three 7 0 0 4.98 72.6 71.3Bb II 273 38 20 5 0 0 4.07 74.0 III 259 46 30 4 0 0 5.11 69.8 IV 253 34 twenty two 11 2 0 5.83 68.9 16.5 g/ha I 267 39 15 5 1 0 3.94 78.3 76.3Bb II 261 28 twenty two 4 2 0 4.49 71.3 III 269 37 16 4 0 0 3.58 78.8 IV 276 30 twenty three 5 1 0 4.34 76.9 18 g/ha I 260 27 17 2 0 0 3.20 82.4 83.9Bb II 271 25 14 1 0 0 2.57 83.5 III 279 17 10 2 0 0 2.06 87.8 IV 236 twenty four 17 2 0 0 3.39 82.0 75% tricyclazole wettable powder 225 g/ha I 280 17 7 1 0 0 1.57 91.4 86.3Aa II 261 16 10 2 0 0 2.15 86.2 III 269 13 8 4 0 0 2.15 87.3 IV 229 25 17 3 0 0 3.69 80.4 blank control / I 183 69 48 37 19 7 18.18 / / II 186 54 43 28 15 4 15.62 / III 202 41 36 30 19 9 16.91 / IV 192 60 47 39 twenty two 8 18.78 /

Claims (6)

1, a kind of active substance is the bactericide of zuelaemycin, it is characterized in that: bactericide active ingredient is the material zuelaemycin with bactericidal activity:

This active substance be from soil through separation, screening, then through microbial fermentation extracted.

2, a kind of active substance according to claim 1 bactericide that is zuelaemycin is characterized in that: described active substance zuelaemycin separates, screening, the processing step that extracts are:

1, gathers soil sample;

2, the separation of soil actinomycetes;

3, the fly up and down screening of bacterium;

3.1 antibacterial activity primary dcreening operation;

The actinomycetic multiple sieve 3.2 fly up and down;

3.3 monospore separates;

3.4 the flying up and down property screening of sporangium;

The bacterium Optimizing Conditions of Fermentation 3.5 fly up and down;

3.5.1 the selection of fermentation medium;

(1) selection of organic nitrogen source;

(2) selection of organic carbon source;

(3) selection of inorganic ions;

(4) best medium orthogonal experiment;

3.5.2 the selection of fermentation medium PH;

3.5.3 the selection of fermentation time;

3.5.4 the selection of fermentation temperature;

3.5.5 the selection of shaking speed;

3.5.6 the relation between growth of actinomycetes mycelia and the antibiotic generation;

3.6 the extraction of efficient bacterial strain S-159-05 activated product;

3.6.1 the preliminary treatment of zymotic fluid;

3.6.2 slightly carry;

3.6.3 essence is carried.

3, a kind of active substance according to claim 1 bactericide that is zuelaemycin, it is characterized in that: described bactericide comprises active ingredient zuelaemycin, surfactant, defoamer, synergist, high osmotic agent, safener, auxiliary agent, impurity, filler etc., and it consists of:

Zuelaemycin 12 weight portions

Surfactant 3 weight portions

Defoamer 2 weight portions

Synergist 3 weight portions

Auxiliary agent 2 weight portions

Emulsifier 2 weight portions

High osmotic agent 2 weight portions

Water 74 weight portions.

4, a kind of active substance according to claim 3 bactericide that is zuelaemycin, it is characterized in that: described surfactant is a lauryl sodium sulfate, defoamer is a dimethyl polysiloxane, synergist is the synergist of being made by tea saponin 15% fatty amine 3% and water 72%, high osmotic agent is aliphatic alcohol polyethenoxy (6-7) ether, emulsifier is that auxiliary agent is a sodium phosphate trimer along the succinate monoester sodium sulfonate.

5, a kind of active substance according to claim 4 bactericide that is zuelaemycin, it is characterized in that: described bactericide comprises the various formulations of pesticide that are used for sterilizing.

6, a kind of active substance according to claim 5 bactericide that is zuelaemycin, it is characterized in that: described bactericide is an aqueous emulsion.

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