CN101182483B - Streptomycete S1-5 and uses thereof - Google Patents
Streptomycete S1-5 and uses thereof Download PDFInfo
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Abstract
The invention provides a novel strain of Streptomyces sp. S1-5 which is capable of controlling and preventing plant pathogenic fungi and killing mite and the application in the anti-plant pathogenic fungi and mite killing. The Streptomyces sp. S1-5 is stored in China Center for Type Culture Collection on 24 May, 2007. The preservation number of the strain is CCTCC No: M207073. Only one time application of the pesticide made by the invention can reach the effect of anti-plant pathogenic fungi and anti-mite; the invention decreases agriculture cost and reduces loads of peasant; the duration period against mites is 30 days; the invention also has the advantages that the strain is safe to human beings and animals which does not kill the natural enemy and the beneficial insect; the strain is difficult to develop resistance for the control object; the strain does not pollute the environment which is degraded by microorganism in the soil rapidly; the strain does not affect soil fertility and microbial flora; the strain does not generate bioconcentration phenomenon which is very safe for the ecology. The invention is a key enzyme inhibitor type of pesticide, which has no cross resistance with other insect resistant pesticides.
Description
(1) technical field
The present invention relates to the new bacterial strain that a strain can prevent and treat plant pathogenic fungi and kill mite---streptomycete S 1-5-5 (Streptomyces sp.S1-5) and at anti-plant pathogenic fungi with kill application in the mite, and can be used for anti-plant pathogenic fungi and the emulsion of mite extremely with what streptomycete S 1-5-5 made.
(2) background technology
Biological pesticide with its low toxicity, noresidue, pollution-free, raw material is easy to get etc., and characteristic is known as by people is the environment friendly agricultural of 21 century, desirable agricultural chemicals.The development of new biological pesticide is undoubtedly a kind of leading behavior that has social benefit and economic benefit concurrently.Mite class and plant pathogenic fungi are the main harm that causes diversified economy crops such as vegetables, tealeaves, fruit tree and cotton, and according to statistics, the whole nation only mite evil needs the control area to be no less than hundred million mu, causes production loss 12%～20% throughout the year, reaches more than 25% when serious.The loss that the harm of plant pathogenic fungi causes is difficult to estimate especially.Traditional biological pesticide, no matter be that thuringiensis cladosporioides bacillus insecticide or other microbial source sterilant all have a common drawback, that is exactly that insecticidal spectrum is narrow, the lasting period is short, drug effect is slow.Avrmectin is active high to the mite class, but not ideal enough to a lot of lepidoptera pest virulence.Polyoxin and nikemycin are the antimycotic pyrimidine nucleoside peptide antibioticses of class energy, the both mainly is used as the agricultural antibiotic of anti-plant pathogenic fungi now, but the ability that Polyoxin penetrates in the fungi born of the same parents is relatively poor, active relatively poor undesirable, along with the life-time service of these medicines, the resistance of fungi to such medicine appears again.Therefore, developing safely and efficiently the novel anti fungi, to kill the acarid biological pesticide very urgent.
Still there is not streptomycete (Streptomyces sp.) can possess control plant pathogenic fungi and the report that kills the acarid double effects simultaneously so far.
(3) summary of the invention
New bacterial strain---streptomycete S 1-5-5 (Streptomyces sp.S1-5) in order to provide a strain can prevent and treat plant pathogenic fungi and kill mite is provided, and in anti-plant pathogenic fungi and the application of killing in the mite.
For reaching goal of the invention the technical solution used in the present invention be:
Streptomycete S 1-5-5 (Streptomyces sp.S1-5) is preserved in Chinese typical culture collection center, preservation date on May 24th, 2007, deposit number CCTCC No:M207073.
The acquisition of bacterial strain of the present invention: from scenic spot in all parts of the country, gather 55 parts of pedotheques as Mt. Huang in Anhui, Mount Lushan, Jiangxi and Guilin, Sanya, Hainan, by the sample primary dcreening operation, multiple sieve, acquisition has the pure growth of anti-plant pathogenic fungi and acaricidal ability.This pure growth is carried out form and Physiology and biochemistry evaluation, it has been accredited as streptomycete (Streptomyces sp.), called after streptomycete S 1-5-5 (Streptomyces sp.S1-5).
The evaluation of bacterial strain:
1) cultural characteristic of bacterial strain: fibrillae of spores is straight or gentle bent, no spiral.Spore surface is smooth.Sucrose Cha Shi agar colony circle, gas velvet shape, ripple is curved, grows thickly, the fibrillae of spores merogenesis, light yellow to green grey.In glucose asparagine agar colony canescence, circle has concentric ring, and fibrillae of spores is mixed and disorderly, grows thickly, and nearly loose spiral end type is viewed as left-handedly counterclockwise from the top, and spore is spherical in shape or avette, and size is uneven, and spore surface is smooth.In Gause I agar colony circle, concentric annular is arranged, there is corrugation protuberance on the surface, neat in edge to around be radial diffusion, powdery slightly is light yellow or light grey, nearly loose spiral end type of fibrillae of spores or ripple song are grown thickly, and are seen as left-handed counterclockwise from the top.In medium oatmeal bacterium colony circle, concentric circles is arranged, uneven, canescence is concentric circles, upward gives birth to the dark circles pearl.In agar colony grey of Ke Shi or canescence, circle is upward given birth to pore, and concentric(al) circles is arranged, and nearly loose spiral end type of fibrillae of spores or ripple song are grown thickly.Bacterial strain microscopic features (15 * 100) and JEOL-1230 electromicroscopic photograph are seen Fig. 1.
2) physio-biochemical characteristics of bacterial strain: produce light yellow soluble pigment.Bacterial strain can make liquefy gelatin, reddish brown pigment, and hydrolyzed starch peptonizes cow's milk, produces H
2S produces melanochrome, can grow on Mierocrystalline cellulose.12 kinds of sugar alcohols such as raffinose, D-wood sugar, L-arabinose, semi-lactosi, cellobiose, L-rhamnosyl, maltose, seminose, D-fructose, inositol, N.F,USP MANNITOL all there is in various degree the ability of utilizing.The result shows that the physiological characteristic of bacterial strain S1-5 is very similar to the streptomyces (Streptomyces) in the actinomycetes (Actinomycete).
3) determining of bacterial strain: according to thalli morphology, the base silk of S1-5 bacterial strain do not have every, do not rupture, fibrillae of spores is the chain raw silk rings, determines that S1-5 belongs to streptomyces.S1-5 cultivates the bacterium colony circle by the standard of cultivating the macroscopical identification kind, and concentric annular is arranged, and there is the corrugation protuberance on the surface, neat in edge to around be radial diffusion, powdery; Many single tines of aerial hyphae branch, the curved or near loose spiral end type of fibrillae of spores ripple; Gelatine liquefication, litmus milk peptonize after coagulation earlier; The starch hydrolysis ability is stronger, grows on the Mierocrystalline cellulose, produces hydrogen sulfide, has melanochrome to produce.Consult document according to this and determine that tentatively S1-5 belongs to streptomycete (Streptomyces sp.).
Streptomycete S 1-5-5 can be used for preventing and treating in the crop pest that plant pathogenic fungi and/or mite pest cause.Described plant pathogenic fungi is one of following: 1. tomato leaf mold germ (brown cladosporium Fulriafulva), 2. Phytophthora nicotianae bacterium (Phytophthora nicotianae), 3. tobacco brown spot pathogen (long handle lattice spore bacterium Alternaria alternata), 4. Rhizoctonia solani Kuhn (dry thread Pyrenomycetes Rhizotoniasolani).Mite pest is representative with cotton, fruit tree red spider.
Concrete, described being applied as: streptomycete S 1-5-5 is seeded to fermention medium, and the supernatant liquor dilution back after fermented liquid that obtains with fermentation culture or fermented liquid are centrifugal is sprayed on the crop plant as soup.The filter cake that obtains after fermented liquid is centrifugal, available distilled water washing back is centrifugal, and the filter cake that obtains adds dehydrated alcohol, and is centrifugal after soaking, and gets also to can be used as medicine liquid spray on crop plant after supernatant liquor dilutes.
Perhaps, described being applied as: streptomycete S 1-5-5 is seeded to fermention medium, fermentation culture, get supernatant liquor after fermented liquid is centrifugal, use ethyl acetate extraction, lower floor's liquid gets parting liquid through column chromatography for separation, and parting liquid is through concentrated, purifying, obtain the biologically active substance crystallization and can be mixed with aqua or emulsion according to a conventional method, be used for the control of crop pest.
Described biologically active substance crystalline virtual mass concentration is 2-3% (refer to the concentration in aqua or the emulsion mother liquor, can dilute 1000～5000 times as required during actual the use).
Described fermention medium can be the common substratum that is applicable to streptomycete in this area, prepares by following composition among the present invention: 1% soybean cake powder immersion liquid 20g, glucose 20g, lime carbonate 2.0g, sodium-chlor 2.5g, distilled water 1000mL.
Lepidoptera pest can be used and kill to streptomycete S 1-5-5 also.
The invention still further relates to the emulsion of a kind of usefulness described streptomycete S 1-5-5 (Streptomyces sp.S1-5) preparation, described emulsion is prepared by following method: streptomycete S 1-5-5 is seeded to fermention medium, fermentation culture, get supernatant liquor after fermented liquid is centrifugal, use ethyl acetate extraction, lower floor's liquid gets parting liquid through column chromatography for separation, parting liquid is through concentrated, purifying, obtain the biologically active substance crystallization, ratio by biologically active substance crystallization, water, dimethylbenzene, washing powder quality is 0.1: 2: 2 again: 0.03 prepares, and obtains described emulsion.
Described fermention medium is prepared by following composition: 1% soybean cake powder immersion liquid 20g, glucose 20g, lime carbonate 2.0g, sodium-chlor 2.5g, distilled water 1000mL.
The present invention obtains a streptomycete (Streptomyces sp.) from the nature screening, antimycotic and the insecticidal constituent of a kind of New-type wide-spectrum of its generation, it not only has the high activity of killing to multiple piercing-sucking mouthparts acarid that harms the crops and worm's ovum thereof evidence, also to a lot of lepidoptera pest virulence very high (Avrmectin is not ideal enough to a lot of lepidoptera pest virulence), and to plant pathogenic fungi good prevention effect is arranged also.Therefore its advantage is embodied in: 1. antibiotic desinsection double effects.The traditional biological agricultural chemicals will use 2 times, utilizes the present invention to make agricultural chemicals and uses the effect that can reach fungicidal for 1 time and kill acarid, reduces agricultural cost greatly, reduces farmer's burdens.2. drug effect is fast.Lasting period reaches 30 days, also possess simultaneously to person poultry safety, non-killed natural enemies and useful insect, to controlling object be difficult for developing immunity to drugs, free from environmental pollution, can by soil microorganisms rapidly degraded do not influence soil fertility and micro-flora again, the lifeless matter enrichment, to advantage such as ecology is fool proof, be a kind of key enzyme inhibitor class agricultural chemicals, do not have cross resistance with other pest-resistant medicines.
(4) description of drawings
Fig. 1 be bacterial strain microscopic features (15 * 100) (A) and JEOL-1230 electromicroscopic photograph (B);
Fig. 2 is a long handle lattice spore bacterium inhibition zone;
Fig. 3 is the dry thread Pyrenomycetes inhibition zone;
The crystallization HPLC color atlas of Fig. 4 for obtaining after concentrating.
(5) embodiment
The present invention is described further below in conjunction with specific embodiment, but protection scope of the present invention is not limited in this:
Embodiment 1: the acquisition of fermented liquid
(1) slant culture: adopt the PDA substratum, inclined-plane coating streptomycete S 1-5-5 bacterial classification was cultivated 2～3 days down, is obtained slant strains for 30 ℃;
(2) seed culture: No. 1 substratum of slant strains inoculation Zhi Gaoshi, cultivated 2～3 days down, obtain streptomycete S 1-5-5 bacterium colony for 30 ℃;
(3) fermentation culture: original fermention medium preparation: 1% soybean cake powder immersion liquid 20mL, glucose 20g, peptone 3.0g, lime carbonate 2.0g, sodium-chlor 2.5g, distilled water 1000ml.Scrape from streptomycete S 1-5-5 bacterium colony of No. 1 culture medium culturing of Gao Shi and to get spore and place sterilized water, vibration 10min is made into concentration 1 * 10
8The spore suspension of individual/ml inserts in the fermention medium with 10% (V/V) inoculum size, and 250ml Erlenmeyer flask on shaking table places the constant temperature shaking table to cultivate 5d, gets fermented liquid, and is standby.
Embodiment 2: the acquisition of fermentation broth extract and mensuration
Get fermented liquid 50ml, 5000r/min, centrifugal 10min, supernatant liquor add the equal-volume dehydrated alcohol and are biologically active substance supernatant liquor crude extract.Filter cake is washed two times centrifugal with distilled water, abandoning supernatant, filter cake adds dehydrated alcohol to reverting to fermentating liquid volume, soak 1h after, the centrifugal 10min of 5000r/min, getting supernatant liquor is biologically active substance mycelium crude extract.These two kinds of crude extracts are carried out bacteriostatic action with the supernatant liquor of fermented liquid after centrifugal and kill the mensuration of mite test, compare with identical aqueous ethanolic solution simultaneously.
Bacteriostatic action measuring method: (four kind of plant pathogenic fungies are so kind as to give for plant protection system of Zhejiang University, and other test microorganism is the preservation of Microbiological Lab of Zhejiang Polytechnical University)
Method with reference to summer Chen En etc. adopts bacterium cake method, filter paper method and cylinder plate method respectively, and 3 repetitions all are set.Leach a small amount of centrifugal back supernatant liquor, supernatant liquor crude extract and biologically active substance mycelium extract respectively with filter paper and be put into containing on the bacterium flat board of brown cladosporium, Phytophthora nicotianae bacterium, long handle lattice spore bacterium, main withered silk kernel fungus, streptococcus aureus and subtilis.Flat board was cultivated 2-3 days under 28 ℃ of conditions, measured its antibacterial circle diameter size with vernier callipers respectively.The results are shown in Table 1:
Table 1:S1-5 biologically active substance crude extract is to the bacteriostatic action of plant pathogenic fungi
| The strains tested title | Antibacterial circle diameter/mm | ||
| Centrifugal back supernatant liquor | The supernatant liquor crude extract | The mycelium crude extract | |
| The leaf muld of tomato bacterium | 14.00 | 6.62 | 6.80 |
| Tobacco brown spot pathogen | 15.00 | 6.45 | 7.33 |
| Rhizoctonia solani Kuhn | 12.00 | 6.43 | 11.56 |
| The Phytophthora nicotianae bacterium | 16.00 | 6.00 | 7.00 |
| Streptococcus aureus | 13.0 | Do not survey | 6.54 |
| Subtilis | 12.5 | Do not survey | 6.81 |
| Physiological saline compares | - | - | - |
| Equivalent physiological saline+ethanol compares | - | - | - |
The acaricidal action measuring method:
Centrifugal back supernatant liquor is supplied examination with stoste and 10 times of liquid of dilution, 100 times of liquid of dilution respectively, distilled water contrast, every concentration are handled and are established 3 repetitions, gather the female one-tenth mite of citrus red mite (crm) from the citrus garden in the indoor mite mensuration of killing, observe once every 6h, acaricidal rate=killed mite number former mite number.The results are shown in Table 2:
Table 2:S1-5 biologically active substance kills mite test determination result
| Extension rate | Acaricidal rate (acaricidal rate=kill mite number former mite number) | |||
| Behind the 6h | Behind the 12h | Behind the 18h | Behind the 24h | |
| Centrifugal back supernatant liquor stoste | 94％ | 100％ | ||
| Dilute 10 times of liquid | 87％ | 100％ | 100％ | |
| Dilute 100 times of liquid | 88％ | 98％ | 100％ | |
| The distilled water contrast | 0 | 0 | 0 | 0 |
Conclusion: to killing cotton red spider good killing effect just being arranged behind the S1-5 biologically active substance supernatant liquor stoste 6h, can kill cotton red spider substantially behind 10 times of diluent 12h, is 100% to the cotton red spider killing rate behind 100 times of diluent 18h.
Embodiment 3: the acquisition of biologically active substance and mensuration
After streptomycete S 1-5-5 fermented liquid is centrifugal, get supernatant liquor, add isopyknic ethyl acetate, the upper strata concentrating under reduced pressure is collected in extraction.Bacteria inhibition assay is done in lower floor's collection, walked pillar with ethyl acetate as eluent, eluent is a petrol ether/ethyl acetate, column chromatography silica gel (200～300 order) is a stationary phase, uses dry column-packing, collects elutriant with test tube in the lower end, every pipe is collected 10mL, collects 10 pipes altogether.Biological activity is followed the tracks of: the 10 pipe elutriants that will collect and lower floor's solution of extraction use the filter paper method to measure it respectively to the fungistatic effect (result data is seen Fig. 2, Fig. 3) for examination bacterium (long handle lattice spore bacterium and dry thread Pyrenomycetes, plant protection system of Zhejiang University is so kind as to give).HPLC test column chromatography effect, when sample is analyzed on Carbohydrate Analysis 3.9 * 300mm Column analytical column, the effluent liquid of its antibacterial substance main peak correspondence is simultaneously in long (the enterprising line scanning of 190～400nm) detectors of Water 2996 Photodiode diode array all-waves, measure the wavelength at its maximum absorption band place, and draw ultra-violet absorption spectrum.Column temperature: 25 ℃; Flow velocity: 1.0ml/min; Sample size: 10 μ L; Moving phase: methyl alcohol: water (30: 70 volume ratios).
With above parting liquid centrifugal 10min under 15000r/min with bacteriostatic activity, get supernatant liquor, add equal-volume dimethylbenzene, organic phase is got in extraction, is condensed into thick liquid with rotatory evaporator again, places one day under room temperature.Crystallization is washed with methyl alcohol, the centrifugal precipitation that discards, rotation concentrates once more, in kept at room temperature overnight, obtains the biologically active substance crystallization.Crystallization is washed (chromatogram alcohol) with methyl alcohol, collect and be HPLC, condition the same (the HPLC color atlas is seen Fig. 4, table 3).
Table 3:HPLC result data
| Title | The treatment channel explanation | Retention time (branch) | Area (microvolt second) | The % area | Highly (microvolt) |
| Peak 1 | The PDA270.0 micron | 3.360 | 17992 | 4.71 | 1321 |
| Peak 3 | The PDA270.0 micron | 4.382 | 364283 | 95.28 | 16887 |
Embodiment 4: emulsion preparation and mensuration
Get 3 method gained biologically active substance crystallizations by embodiment, press biologically active substance crystallization, colloid sulphur (the high brilliant Fine Chemical Co., Ltd in Hangzhou), water, dimethylbenzene, washing powder is (commercial, the common washing powder of carving board) ratio of quality is 0.1: 1: 2: prepare at 2: 0.03, earlier that colloid sulphur is soluble in water, add washing powder and stir, pour dimethylbenzene afterwards at leisure into, the limit bevelling stirs, continue 5～10min, add biologically active substance crystallization mixing at last, obtain described emulsion.
Indoorly kill mite, kill the ovum effect measuring:
Gather female one-tenth mite of citrus red mite (crm) and ovum kills mite and kills ovum mensuration from 2500 times of liquid of antigalactics in indoor carrying out from the citrus garden, and contrast with 1500 times of liquid of 40% isocarbophos, 1500 times of liquid of 20% kelthane, if clear water contrast, acaricidal rate=killed mite number former mite number.The result shows that very strong to the lethality of citrus red mite (crm) female adult worm and ovum from 2500 times of liquid of antigalactics, killing rate reaches 92.1%～94.8%, and ovicidal rate reaches 89.7%～93.4%, with 40% isocarbophos and 500 times of liquid effects of 20% kelthane basically identical.
Embodiment 5: from antigalactics control cotton red spider
The preparation of emulsion:
Get the biologically active substance crystallization, press the biologically active substance crystallization: colloid sulphur: water: dimethylbenzene: washing powder=0.1: 1: 2: 2: 0.03 weight ratio preparation, earlier that colloid sulphur is soluble in water, adding washing powder stirs, pour dimethylbenzene afterwards at leisure into, the limit bevelling stirs, and continues 5～10min, add biologically active substance crystallization mixing at last, make mother liquor.Add water by the different concns dilution during use.
Field controling test:
Having carried out from the test of 3000 times of liquid field spray controls of antigalactics cotton red spider in the cotton field, is contrast with 1500 times of liquid of 40% isocarbophos, 1500 times of liquid of 20% kelthane.The result is as follows:
From 3000 times of liquid of antigalactics: the 5d preventive effect is 87.8% behind the medicine, and the 10d preventive effect is 89.3% behind the medicine, and 15d is 98.5% behind the medicine;
1500 times of liquid of 40% isocarbophos: the 5d preventive effect is 99.6% behind the medicine, and 10d is 96.1% behind the medicine.15d only 63.5% behind the medicine;
1500 times of liquid of 20% kelthane: the 5d preventive effect is 99.7% behind the medicine, and 10d is 98.7% behind the medicine, and 15d only is 56.1% behind the medicine.
In antigalactics 1000ml, add 7.5% trifoliate jewelvine missible oil 60ml mix mixed solution, get its 1500 times of liquid and use, the insect population rate of going down can reach more than 90%, during 30～60d behind the 5d, prevention effect is stabilized between 90%～99%, and isocarbophos, kelthane treatment zone only are about 40%.
This preparation raw material is easy to get, and is with low cost, prepares easyly, safe in utilization, and little to the natural enemy lethal effect, no residual hazard, residual simultaneously can doublely be controlled other insects, can extensively promote the use of on fruit tree and other crops.
Embodiment 6: prevent and treat rice sheath blight disease research
(1) colonization ability of bacterial strain S1-5 on field plant measured
At rice seedling, the bacterial strain S1-5 fermented liquid (3.6 * 10 that will make by embodiment 1 method
5Cfu/mL) spray on rice plant, from inoculating the thalline number change that the back began to observe on the record blade every square centimeter on the 1st day.
(2) field control effect is measured
Processing is respectively S1-5 fermented liquid (3.6 * 10
5Cfu/mL), 20% jinggangmeisu WP (Hunan Factory of Biologicals) is that medicament contrast and clear water contrast (CK2) (CK1).Each is handled 4 times and repeats every replicated plot area 30m
2Two ridge, each sub-district coating is isolated, and minizone distance and sub-district and guard rows spacing are 80cm, and the guard rows kind is hilllock excellent 527.Before the transplanting, the sheath blight fungus of cultivating with Semen Maydis powder mixes with the river sand mixing.Every sub-district 200g evenly spreads and plants in the sub-district, and Sheng phase and boot stage prevent and treat in tillering.According to rice leaf sheath and blade hazard symptoms grading, be 5 samplings of unit every sub-district diagonal lines with the strain, every some investigation links to each other 5 clumps, totally 25 clumps, writes down total strain number, diseased plant number and sick progression.0 grade: complete stool is anosis; 1 grade: quaterfoil and following each leaf sheath, blade morbidity (is first leaf with sword-like leave); 3 grades: the 3rd blade and following each leaf sheath, blade morbidity; 5 grades: second blade and following each leaf sheath, blade morbidity; 7 grades: sword-like leave blade and following each leaf sheath, blade morbidity; 9 grades: the complete stool morbidity, withered ahead of time.
Streptomycete S 1-5-5 fermented liquid has significant control effect to rice sheath blight disease, the multiple spot field test results showed in 4 years, streptomycete S 1-5-5 fermented liquid is prevented and treated the banded sclerotial blight effect stability between 80%～85%, and is suitable with the prevention effect of contrast medicament 20% jinggangmeisu.Survey the product result according to real cell, significant guarantor's product effect (6.91%～18.13%) is all arranged for examination streptomycete S 1-5-5 fermentation liquor treatment.
Embodiment 7: the anti-therapy assault of secondary control mulberry field lepidopteran class pest
After annual mid or late August, the lepidoptera pest in the mulberry field particularly mulberry geometrid (Hemerophila atrilineata) enters the high-incidence season, is in for 1 length of time, and the worm amount reaches 2400～62000/mu.The diaphania adult also sprouts wings successively in the mulberry field, entered the larva Sheng and incubate the phase at the beginning of 9 months.The anti-therapy assault of the secondary that present embodiment is mentioned control mulberry field lepidopteran class pest, the control time can fix on late August for the first time, and the control time can be in early September for the second time.
The preparation of emulsion: get the biologically active substance crystallization, press the biologically active substance crystallization: colloid sulphur: water: 1000 times of diluents of 80% SD-1750 (Hunan Factory of Biologicals)=0.1: 1: 2: 2 weight ratio preparation, earlier that colloid sulphur is soluble in water, add 1000 times of diluents of SD-1750, the limit bevelling stirs, continue 5～10min, add biologically active substance crystallization mixing at last, make mother liquor.Add water by the different concns dilution during use.Safety is used 5 days interlobar septum phases.
Field controling test:
Carried out the particularly test of mulberry geometrid (Hemerophila atrilineata) of lepidoptera pest in 2500 times of liquid field sprays of antigalactics control mulberry field with the leaf mulberry field grown silkworm, with 40% mulberry treasured (Jiangsu Province Jiangnan Farm Chemical Plant, Changzhou) 2500 extraordinarily 60% enemy horse (Jiangsu Province Jiangnan Farm Chemical Plant, Changzhou) 1500 extraordinarily 1000 times of liquid of 40% Rogor (Jiangsu Province Jiangnan Farm Chemical Plant, Changzhou) be contrast.The result is as follows:
From 2500 times of liquid of antigalactics: the 5d preventive effect is 79.4% behind the medicine, and the 10d preventive effect is 93.7% behind the medicine, and 15d is 92.3% behind the medicine;
Extraordinarily 60% oppose extraordinarily 1000 times of liquid of 40% Rogor of horse 1500 with 40% mulberry treasured 2500: the 5d preventive effect is 88.4% behind the medicine, and 10d is 96.7% behind the medicine.15d only 63.5% behind the medicine;
Phase in mid-autumn mulberry branch is long, growing way is prosperous, if add dryly, temperature is higher, and there is certain difference the mulberry shoot blade residual toxicity phase up and down, therefore tries afterwards to eat earlier with must accomplishing before the leaf, guards against the silkworm pesticide intoxication.Spray to use during medicine special-purpose atomizer, with clean water make up a prescription, concentration registration, spraying want thin, soup spray foot, to improve preventive effect and to guarantee the safety of breeding silkworms.
Claims (9)
1. streptomycete S 1-5-5 (Streptomyces sp.S1-5) is preserved in Chinese typical culture collection center, preservation date on May 24th, 2007, deposit number CCTCC No:M 207073.
2. the application of claim 1 described streptomycete S 1-5-5 in the crop pest that control plant pathogenic fungi and/or mite pest cause.
3. application as claimed in claim 2 is characterized in that described plant pathogenic fungi is one of following: 1. tomato leaf mold germ, 2. Phytophthora nicotianae bacterium, 3. tobacco brown spot pathogen, 4. Rhizoctonia solani Kuhn.
4. application as claimed in claim 2 is characterized in that described being applied as: streptomycete S 1-5-5 is seeded to fermention medium, and the supernatant liquor dilution back after fermented liquid that obtains with fermentation culture or fermented liquid are centrifugal is sprayed on the crop plant as soup.
5. application as claimed in claim 2, it is characterized in that described being applied as: streptomycete S 1-5-5 is seeded to fermention medium, fermentation culture, get supernatant liquor after fermented liquid is centrifugal, use ethyl acetate extraction, lower floor's liquid gets parting liquid through column chromatography for separation, and parting liquid obtains the control that the biologically active substance crystallization is used for crop pest through concentrated, purifying.
6. as claim 4 or 5 described application, it is characterized in that described fermention medium prepares by following composition: 1% soybean cake powder immersion liquid 20mL, glucose 20g, lime carbonate 2.0g, sodium-chlor 2.5g, distilled water 1000mL.
7. the application of claim 1 described streptomycete S 1-5-5 in killing lepidoptera pest.
8. emulsion with claim 1 described streptomycete S 1-5-5 (Streptomyces sp.S1-5) preparation, described emulsion is prepared by following method: streptomycete S 1-5-5 is seeded to fermention medium, fermentation culture, get supernatant liquor after fermented liquid is centrifugal, use ethyl acetate extraction, lower floor's liquid gets parting liquid through column chromatography for separation, parting liquid is through concentrated, purifying, obtain the biologically active substance crystallization, ratio by biologically active substance crystallization, colloid sulphur, water, dimethylbenzene, washing powder quality is 0.1: 1: 2 again: prepare at 2: 0.03, obtain described emulsion.
9. emulsion as claimed in claim 8 is characterized in that described fermention medium prepares by following composition: 1% soybean cake powder immersion liquid 20mL, glucose 20g, lime carbonate 2.0g, sodium-chlor 2.5g, distilled water 1000mL.
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| CN102250803A (en) * | 2011-06-29 | 2011-11-23 | 黑龙江大学 | Strain having high-efficiency insecticidal function |
| CN103468599B (en) * | 2013-05-22 | 2014-12-17 | 滨州学院 | Streptomycetes sp. and its application in winter jujube ring rot prevention and treatment |
| CN104357369B (en) * | 2014-11-25 | 2017-10-13 | 辽宁大学 | Biological control streptomycete and its application |
| CN109762752B (en) * | 2018-11-01 | 2020-06-30 | 福建省农业科学院植物保护研究所 | Streptomyces nodulicidal strain and application thereof |
| CN109439598B (en) * | 2018-12-24 | 2020-06-30 | 吉林省林业科学研究院 | Streptomyces and application thereof |
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