CN1023323C - Preparations of new antibiotics SP-120, SP-120A, SF-120B and pesticide for agricultural horticulture - Google Patents

Abstract

The present invention provides antibiotics SP-120, SP-120A and SP-120B and a preparation method thereof, and also provides a pesticide whose effective components are antibiotics SP-120, SP-120A and SP-120B or the compounds of the antibiotics SP-120, SP-120A and SP-120B for agricultural horticulture. The antibiotics of the present invention have the advantages of strong selectivity, high safety, no influence on users, and no medicinal damage. The antibiotics of the present invention are produced by cultivation of streptomyces_SP. SP-120.

Description

Preparations of new antibiotics SP-120, SP-120A, SF-120B and pesticide for agricultural horticulture

The invention relates to new antibiotic SP-120, SP-120A, SP-120B and its preparation method, and a kind of effective ingredient is SP-120, SP-120A, the pesticide for agricultural horticulture of SP-120B or its complex body.

In the past, pesticide for agricultural horticulture was to use as copper always, mercury, arsenic and so on heavy metal compound preparation and organochlorine, organophosphorus medicament.But these medicaments all are harmful to animal and human's body, and soil is had contamination, and the objectionable impurities that remains in occurring in nature acts on animal and plant again chronically.The environmental pollution that causes has like this become current important social issue, thereby has caused the present this situation that is under an embargo or limits use.In addition, along with the minimizing of selectivity medicament and the appearance of anti-medicine bacterium, various Plant diseasess-especially the main disease of paddy rice has the trend that obviously increases the weight of.For this reason, people urgently wish to develop some existing strong selectivity effects, and the novel agrochemical of tight security is arranged again.

The object of the present invention is to provide a kind of to all effective microbiotic SP-120 of various Plant diseasess, SP-120A, SP-120B and preparation method thereof.And the present invention also aims to provide more than one to state microbiotic SP-120, and SP-120A, SP-120B or its complex body are the pesticide for agricultural horticulture of effective ingredient.

Microbiotic SP-120 of the present invention, SP-120A, SP-120B is not for seeing the new antibiotic of bibliographical information, it has physico-chemical property described later.Simultaneously, in spraying described later test, to main disease such as the rice sheath blight disease of paddy rice, rice blast and other be as gray mold of cucumber, cucumber anthracnose, and the eggplant black spot, various Plant diseasess such as glomerella leaf spot and die back of grape have all shown outstanding prevention effect.And both not had any poisoning, and again the personnel of using had been had no effect, is a kind of good pesticide for agricultural horticulture.

Below the present invention will be described in detail.

Microbiotic SP-120, SP-120A, the preparation of SP-120B

The microorganism of using

Be used for producing microbiotic SP-120 of the present invention, the microorganism of SP-120A and SP-120B is a kind of bacterial classification that belongs to streptomyces, and it has the microbiotic of production SP-120, the ability of SP-120A and SP-120B.

Streptomycete SP.SP-120(Streptomyces SP.SP-120) (the preserving number CGMCC0022 at China Committee for Culture Collection of Microorganisms common micro-organisms preservation center) (be designated hereinafter simply as " SP-120 bacterial strain) is exactly an example wherein.This bacterial strain with the corresponding Japanese patent application of the application in be named as " streptomycete SP.RK-120.This microorganism has above-mentioned characteristic, is suitable for producing microbiotic SP-120 of the present invention, SP-120A, and SP-120B, thereby can be used in effectively among the preparation method of the present invention.

Above-mentioned " SP-120 bacterial strain " is to separate in the soil of Sanming City, People's Republic of China (PRC) Fujian Province area and get.Separate and adopt conventional soil dilution method to carry out.Promptly cultivated 6 days in 28 ℃ on asparagine-agar glucose earlier, picking list bacterium colony moves on the inclined-plane of this substratum.Method for screening is after the fermentation of SP-120 bacterium, gets fermented liquid and is sprayed on the plant, inserts pathogenic bacterium then on plant, places check result after 2～4 days in the greenhouse, finds to have remarkable prevention effect.Separation screening " the SP-120 bacterial strain " that come out comes down to pure like this.

The SP-120 bacterial strain belongs to streptomyces, and its development condition and physiological property on substratum is as follows:

1. morphological specificity:

The SP-120 bacterial strain is at oatmeal-nutrient agar, and starch can be grown on the yeast nutrient agar (its component is as described below) extremely well, aerial hyphae, and adhering to of spore is very abundant, the aerial hyphae light gray that is white in color.Adhering to of its development condition on sucrose nitrate nutrient agar, yeast malt agar and aerial hyphae, spore is also all fine.At other substratum, as at glucose-l-asparagine nutrient agar, glycerine-l-asparagine nutrient agar, starch-inorganic salt nutrient agar, tyrosine nutrient agar, nutrient agar, also all can grow on peptone-yeast iron nutrient agar, but insufficient.And at glucose-l-asparagine nutrient agar, nutrient agar, and on peptone-yeast iron nutrient agar, the growth of the aerial hyphae of this bacterium is faint or be difficult to confirm.Generally speaking, the generation of soluble pigment is very faint, only can produce little yellow class pigment.Rely on tyrosine nutrient agar and peptone-yeast iron nutrient agar, then be difficult to generate melanochrome.Substrate mycelium does not present on the surface and has distinctive tone and be little yellow or little brown.Aerial hyphae is the straight chain shape and grows, and goes out to be strong spiral helicine coil by these straight chain shape mycelia upper branch again, and this has just formed long spore lock.Spore is similar to sphere, and length is in 1 micron, and width is about 0.5 micron, the surface smoothing of head.

In the utilization of carbon source test of carrying out with carbohydrate, this bacterial strain can utilize L-arabinose well, D-fructose, and raffinose, lactose, close disaccharides, it physically well develops.At the sweet sugar of D-glucose, inositol, in the D-N.F,USP MANNITOL, though also can grow, it is relatively poor to have the part strain growth.At the D-wood sugar, the growth in the rhamnosyl is bad, particularly difficult generation aerial hyphae in the L-rhamnosyl.

This bacterial strain can be grown on glucose-peptone-gelatine culture, but can not make gelatine liquefication; Can peptonize skimming milk formation fully and be bordering on transparent liquid; On starch-inorganic salt nutrient agar, can impel the starch intense hydrolysis.

With the hydrochloric acid hydrolysis cell walls time, with the amino acid composition of Paper Chromatography detection cell, can find the L-diaminopimelic acid, this has just shown that this bacterial strain is the bacterial classification that belongs to streptomyces.

2. the character on various substratum

The 3rd week after inoculation, to observe bacterial strain and write down the result as follows, color is recorded and narrated according to the color name mark of " color designation is described dictionary " (Descriptive color names dictionary) the 4th edition.

1) sucrose nitrate nutrient agar

Well-grown

Adhering to of aerial hyphae is good

The color 2ba canescence (pearl) of aerial hyphae

The color 3lc of substrate mycelium amber (amber)

Soluble pigment 3ga pawpaw Huang (melon yellow)

2) glucose-l-asparagine nutrient agar

Poor growth

Aerial hyphae adhere to nothing

The color of aerial hyphae is not clear

The color 5ca light red of substrate mycelium (shell pink)

Soluble pigment 6ca light red (shell pink)

3) glycerine-l-asparagine nutrient agar

It is common to grow

Adhering to of aerial hyphae is bad

The color 7cb of aerial hyphae secretly pale red (cloud pink)

The color 5pe russet of substrate mycelium (meaning terra cotta)

Soluble pigment 61/2gc gray coral look (dust coral)

4) starch-inorganic salt nutrient agar

It is common to grow

Adhering to of aerial hyphae is common

The color a white (white) of aerial hyphae

The color 3ca deep pink of substrate mycelium (pearl pink)

Soluble pigment does not have

5) tyrosine nutrient agar

It is common to grow

Adhering to of aerial hyphae is common

The bright grey of the color b of aerial hyphae (light grey)

The color 4pi Mongolian oak brown (oak brown) of substrate mycelium

Soluble pigment 5ie copper brown (copper tan)

6) nutrient agar

It is common to grow

Aerial hyphae adhere to nothing

The color of aerial hyphae is not clear

The color 2ea glassy yellow of substrate mycelium (light maize)

Soluble pigment 2ea glassy yellow (light maize)

7) yeast-malt agar

Well-grown

Adhering to of aerial hyphae is good

The color a white (white) of aerial hyphae

The color 2ea glassy yellow of substrate mycelium (light maize)

Soluble pigment does not have

8) oatmeal nutrient agar

Well-grown

Adhering to of aerial hyphae is abundant

The bright grey of the color c of aerial hyphae (light grey)

The color 3ba canescence (pearl) of substrate mycelium

9) peptone-yeast iron nutrient agar

It is common to grow

Aerial hyphae adhere to nothing

The color of aerial hyphae is not clear

The color 2ea glassy yellow of substrate mycelium (light maize)

Soluble pigment does not have

10) starch-yeast nutrient agar *

Well-grown

Adhering to of aerial hyphae is abundant

The color a white (white) of aerial hyphae

The color 3pc of substrate mycelium amber (amber)

Soluble pigment 3ca wind rose (pearl pink)

* the component of starch-yeast nutrient agar

Zulkovsky starch 10 grams

Yeast extract paste 1 gram

NZ amino acid A 1 gram

Agar 15 grams

1 liter of PH7.4 of water

3. the utilization of carbon source (general dagger-axe nutrient agar)

Carbon source development condition *

L-arabinose ++

The D-wood sugar ±

D-glucose+

D-fructose ++

Sucrose+

Inositol+

Rhamnosyl ±

Raffinose ++

D-N.F,USP MANNITOL+

Lactose +++

Close disaccharides ++

* development condition

+++grow very good

++ physically well develop

+ grow

± grow slightly

-aplasia

4. other physiological property

25～30 ℃ of suitable temps

Gelatin does not liquefy

(glucose-peptone-gelatine culture)

Starch hydrolysis hydrolysis

(starch-inorganic salt nutrient agar)

The liquefaction of skimming milk is liquefied fully

Melanic generation (tyrosine agar

Substratum and peptone-yeast iron agar training does not generate

Support base)

The retrieval of streptomyces bacterial classification with above-mentioned each character is from the report of the wild Cun Shi in open country, promptly by " zymotechnique magazine " (Joural of FermentationTechnology) the 5th No. 2 contained actinomycetes ISP458 bacterium classification methods of volume that wild Cun Shi showed and put down in writing.The name of this classification is called " 458 kinds of streptomycete kind classification and the clue of differentiating " Key for Classificat ion and Identificat ion of 458 Species of the Streptomyces Included in ISP).

This Pseudomonas belongs in white or grey colour cell, and its aerial hyphae is shape in the shape of a spiral, the surface smoothing of spore and do not form melanochrome.Its inside and soluble pigment all do not present significant tone simultaneously.Relatively poor to utilizing of rhamnosyl in the carbohydrate and D-wood sugar, but for other carbohydrate, then number average can utilize mostly.The bacterial classification that can satisfy above condition has living streptomyces albus (Streptomyces albofacience), and therefore, this Pseudomonas gives birth near a kind of streptomyces albus in S..But novel bacterial SP-120(Streptomyces SP.SP-120) difference with known living streptomyces albus is:

(1) on glycerine-asparagine substratum:

The back side of SP-120 bacterium is russet, and the beige soluble pigment is arranged;

And the back side of the living streptomyces albus of S. does not have characteristic color, no soluble pigment.

(2) to the utilization of carbohydrate:

The SP-120 bacterium can utilize rhamnosyl slightly, can utilize sucrose, and can utilize pectinose, raffinose well;

And the living streptomyces albus of S. can not utilize rhamnosyl, can utilize sucrose slightly, can utilize pectinose, raffinose.

Cultivation of SP-120 bacterium and SP-120, SP-120A, SP-120B producing and purifying

With the above-mentioned production of antibiotics bacterium of streptomyces,, can cultivate the SP-120 bacterium and make microbiotic SP-120 of the present invention, SP-120A, SP-120B by the production of antibiotics method of routine.Training method both can be liquid culture and also can be solid culture.But the cultivation when producing in order to help industrialness, the spore suspension of above-mentioned production bacterium or nutrient solution should being inoculated into goes forward side by side on the substratum, and cultivation is mixed in the gas mixing that works.

Nutrition source in the substratum be there is no special regulation, can contain the carbon source that is usually used in microorganism culturing, nitrogenous source and other nutrition source.Wherein carbon source can be starch, dextrin, glycerine, glucose, sucrose, lactose, inositol, N.F,USP MANNITOL etc.Nitrogenous source can be peptone, soyflour, meat extract, rice sugar, wheat skin, urea, corn steep liquor, ammonium salt, nitrate, and other organic or inorganic nitrogenous compound.For other nutrition source, can suitably add some inorganic salts, for example, and salt, phosphoric acid salt and potassium, calcium, zinc, manganese, the metal-salt of iron and so on also can add a little moving in case of necessity, plants, and the mineral wet goods is as defoamer.

Temperature during cultivation, culture condition such as time there is no strict restriction, being suitable for the using growth of bacterium to be as the criterion, and selecting SP-120, SP-120A, the highest condition of the output of SP-120B is for well.For example, the PH scope of substratum is 4～9, with near neutral for well.Culture temperature is 25 ℃ to 35 ℃.Certainly, comprise the cultivation constituent, hydrionic concentration in the substratum, culture temperature stirs or the like all these culture condition, all should carry out suitable adjusting according to the kind of employed bacterial strain and external conditions etc., to obtain best effect.

Culture by above-mentioned gained sets out, and just can make SP-120 by some appropriate means, SP-120A and SP-120B.These methods are often to be used in the method for extracting metabolite, for example, can utilize SP-120, and SP-120A, SP-120B and impurity be in solubleness, ionic bond power, and the difference of adsorbing aspects such as avidity and molecular weight is extracted.These methods can be used separately, also can suitably cooperate or use repeatedly.Specifically, SP-120 comprises SP-120A and SP-120B, and their major parts are present in the culturing filtrate.Extract nutrient solution repeatedly with butanols, or adsorb, and then just can obtain SP-120(with the washing of 50% propionyl and wherein contain SP-120A and SP-120B with synthetic adsorbent as Diaion HP10 and so on) enriched material.The enriched material of gained carries out column chromatography repeatedly with silica gel, and solvent is selected the mixing solutions of propyl carbinol-methanol-water for use, regulates their component ratio, can reach the purpose of chromatographic separation.So just obtain two differentiations, one of them is distinguished and mainly contains SP-120A, and another is distinguished and mainly contains SP-120B.To these two differentiations, carry out column chromatography with biogel such as glucose gel LH-20 etc. respectively, this moment used solvent with use with the same class of above-mentioned solvent system for suitable.It is further refining to use gel-filtration to carry out, and can obtain the SP-120A of white powder and the highly finished product of SP-120B respectively.

At last, adopt high-efficient liquid phase color to compose and SP-120A and SP-120B can be separated fully.Filling agent is selected Nucleosil 5C for use ₁₈, solvent is Virahol-methanol-water, and is added with little acetic acid.

So new antibiotic SP-120A and the SP-120B that is produced has following physico-chemical property respectively.

The physico-chemical property of SP-120A and SP-120B.

1. fusing point:

A:235～255 a ℃ decomposition is brown.

B:235～265 a ℃ decomposition is brown.

2. ultimate analysis:

A: carbon 50.98%, hydrogen 7.11%, nitrogen 16.42%, sulphur 3.55%,

B: carbon 50.91%, hydrogen 6.95%, nitrogen 16.12%, sulphur 3.39%,

3. specific optical rotation

A:[α] ²⁵ _D-90 ° (C=0.05, methyl-sulphoxide)

B:[α] ²⁵ _D-59 ° (C=0.03, methyl-sulphoxide)

4. ultra-violet absorption spectrum:

A：λ ^{0.1N-HCl-MeOH} _max

220nm(E ^1% _1cm186) peak shoulder (referring to Fig. 1)

B：

λ ^{0.1N-Hcl-MeOH} _max

220nm(E ^1% _1cm293) peak shoulder (referring to Fig. 2)

5. infrared absorption spectrum: press the KBr method

A：3300，2903，2850，1655，1520，1460，1395，1382，1320，1280，1250，1215，1163，1100，1020，1002，900，840，760cm

(referring to Fig. 3)

B：3300，2900，2850，1650，1520，1435，1392，1370，1342，

1322，1285，1212，1185，1168，1100，1024，1004，958，903，846，825，765cm ^-1

(referring to Fig. 4)

6. solvability:

SP-120A and SP-120B all are insoluble to methyl alcohol, ethanol, acetone, ethyl acetate, chloroform, benzene, normal hexane, organic solvents such as ether; Be insoluble in water; Dissolve in the mixed solution of propyl carbinol-methanol-water.

7. molecular weight: press the FAB mass spectrum

A：911

B：923

8. color reaction:

Equal and the potassium permanganate of SP-120A and SP-120B, aubepine-sulfuric acid, iodine is positive.

9. material color

SP-120A and SP-120B are white powder.

10. thin-layer chromatography (U.S. Merck corporate system thin layer plate 0.25m/m):

Mierocrystalline cellulose

(solvent is pressed propyl carbinol: methyl alcohol: water=4: 1: 2)

The Rf value

A：0.60

B：0.66

Silica gel

(solvent is pressed propyl carbinol: methyl alcohol: water=4: 1: 2)

The Rf value

A：0.51

B：0.51

11. acidic hydrolysis

In 105 ℃ of heating hydrolysis 20 hours, in SP-120A, can detect glycine in 6N-HCl, leucic each amino acid can detect glycine and Xie Ansuan in SP-120B.

12. antimicrobial spectrum:

Adopt common agar plate dilution method, (MIC) represents with minimum inhibitory concentration.Bacterium is used broth agar culture medium, and mould is used the potato sucrose nutrient agar

MIC（mcg/ml）

A B

Gold-coloured staphylococci 12.5 12.5

（Staphylococcus aureus）

Mycobacterium phlei 12.5 12.5

（Mycobacterium phlei）

Pseudomonas pseudomonas bacteria＞100＞100

（Pseudomonas aeruginosa）

Bacillus subtilus 25 12.5

（Bacilus subtilis）

Lattice pink mold 12.5 12.5

（Alternaria mali）

Rice blast 25 12.5

（Pyricularia oryzae）

Cucumber anthracnose 12.5 12.5

（Collectotrichum lagenarium）

Botrytis cinerea pers 12.5 12.5

（Botoryt is cinerea）

Sickle spore bacterium point spore bacterium 50 25

（Fusarium oxysporum）

Glomerella leaf spot and die back of grape bacterium 25 25

（Glomeralla cingurata）

Revolve spore bacterium 25 25

（Cochliobolus miyabeanus）

Anti-rhizoctonia eggplant fusarium 12.5 25

（Rhizoctonia solani）

Aspergillus tubigensis 50 50

（Aspergillus oryzae）

As what compare with microbiotic SP-120A of the present invention and SP-120B, the known peptide antibiotics that contains 3～4% sulphur in molecule has: ripple monomycin (Bottormycin), (Enshumycin), actinoleukin (Actinoleukin), kobenomycin (Kobenomycin), zorbonomycin (Zorbomycin) and phleomycin (Phleomycin) or the like.But SP-120A and SP-120B and above-mentioned microbiotic are having evident difference aspect ultra-violet absorption spectrum and the anti-microbial activity.Therefore can reach a conclusion, microbiotic SP-120A promptly of the present invention and SP-120B belong to the new antibiotic that is not seen in the document record.

SP-120, SP-120A, the pesticide for agricultural horticulture of SP-120B or their complex body.

Here the complex body of said SP-120A and SP-120B is meant the enriched material that how much contains some SP-120A and SP-120B.This enriched material is after adsorbing above-mentioned culturing filtrate by synthetic adsorbent, to make with the aqueous acetone extracting again.These two kinds of materials are owing to making simultaneously from a culture, so can use as effective constituent with the form of complex body.This is very favourable at economic aspect, thereby noticeable.Can certainly use SP-120 separately, SP-120A or SP-120B are as effective constituent.

When effective constituent of the present invention is used for preparations such as pesticide for agricultural horticulture, can use equally known agricultural chemicals in this technical field general solid phase carrier, preparation such as liquid phase carrier and emulsifying dispersant, and can be mixed with granule, pulvis, emulsifying agent, wettable powder, tablet, finish, sprays, smoke substance etc. are formulation arbitrarily.Solid phase carrier can be a clay in these carriers, kaolin, bentonite, acidic white earth, diatomite, lime carbonate, soluble cotton, starch, gum arabic etc.; The phase of wave carrier can be methyl alcohol, ethanol, acetone, dimethyl formamide, ethylene glycol etc.Simultaneously, in preparation, generally can suitably add a little auxiliarys, as the sulfuric ester of higher alcohols, polyoxyethylene, alkyl-allyl ethers, alkyl allyl polyglycol ether, the anhydrous sorbitol laurate of alkyl allyl group, alkylallyl sulfonate, quaternary amine, polyalkylene oxide etc.

Generally speaking, the proportioning of effective constituent in each preparation is 10～90% in emulsion and wettable powder; In pulvis and finish etc. is 0.1～10%; Certainly, also can suitably increase and decrease according to different application targets.

Preparation of the present invention also can with other sterilant, weedicide, sterilant, the fertilizer material, soil improvement agents etc. are used.

Below with regard to manufacturing of the present invention, implement, the test example is made specific description, but and does not mean that qualification the present invention.

Wherein, " ratio " in an embodiment is weight ratio.

Production Example:

With glucose 2%, Zulkovsky starch 1%, gravy medicinal extract 0.1%, dry yeast 0.4%, soyflour 2.5%, salt 0.2%, the proportioning of potassium primary phosphate 0.005% is made substratum, above-mentioned SP-120 bacterial strain (microorganism store number CGMCC0022) is inoculated in this substratum, 28 ℃ of shaking culture 96 hours.Get 25 liters of its culturing filtrates, change into (strain) system in 4 liters of formaldehyde resin HP-10(Mitsubishis) adsorb activeconstituents in the chromatography column, with 40 liter 50% acetone extraction, extracting solution under reduced pressure is concentrated into 2 liters, and filtering-depositing is also washed.Be that diameter of solvent balance is 50mm with propyl carbinol-methanol-water (8: 1: 1) then, length is the silicagel column of 40cm.Use 0.1 NHCl-MeOH(1: agent dissolves 8) precipitates, and lysate is added on capital, launches then.Be 8: 1: 1 when the proportioning of propyl carbinol-methanol-water begins, change 5: 1: 1 then into, adopted at last 4: 1: 1.With every 55ml is a metering, mainly obtains SP-120B initial No57～100 li, after this, li mainly obtains SP-120A in No101～123.

With these two the active concentrating under reduced pressure respectively of distinguishing, obtain product 990mg and 170mg after the lyophilize respectively, this is rough powder.

The rough powder that contains SP-120A is packed in sephadex lh-20 (Sweden, the pharmaceuticals corporate system) chromatography column, and the diameter of this post is 50mm, and length is 90cm, is to get with propyl carbinol-methanol-water (4: 1: 2) balance.Then, launching with above-mentioned same solvent, is a pipe with every 10ml, at No28～35 pipes, mainly contains SP-120A, with these effluent liquid concentrating under reduced pressure, and obtains the 76mg white powder through lyophilize.After this, it is separated, obtain SP-120A and SP-120B with high performance liquid chromatography.Chromatographic column is selected nucleosides 5C for use ₁₈, diameter is 20mm, length is 30cm.Solution adopts Virahol-methanol-water (2: 2: 6) and adds 0.15% acetic acid.Flow velocity is the 6.0ml/ branch, and pressure is 180kg/cm ², SP-120B appears in developing time when being 30 minutes, occur SP-120A in the time of 42 minutes.Collect the part of SP-120A, concentrating under reduced pressure when eliminating Virahol and methyl alcohol till, lyophilize then obtains white powder 27mg.

The rough powder of SP-120B equally also can adopt the sephadex lh-20 column chromatography and select nucleosides 5C for use ₁₈High performance liquid phase separate and make SP-120B.Collect the part of SP-120B, under reduced pressure concentrate to remove and desolvate, after lyophilize, obtain the white powder of 14.8mgSP-120B.

Adopt following method to carry out desalination then.

SP-120A is dissolved in propyl carbinol-methanol-water (2: 1: 2) and is added with in the solvent of 0.15% acetic acid, remove insolubles.The limit adds the waterside underpressure distillation and removes propyl carbinol and water, makes the concentration of acetic acid remain on below 0.15% lyophilize then.After this, 200ml makes its dissolving with propyl carbinol-methanol-water (2: 1: 2), and concentrating under reduced pressure most about 10ml, filters and tells SP-120A precipitation and fully washing, be placed on drying in the moisture eliminator with acetone and ether washes clean again, promptly obtain the pure SP-120A powder of 12.8mg.

In kind carry out the desalting treatment of SP-120B, also can get 4.2mg powdered pure product.

Embodiment 1 wettable powder

With 10 parts of SP-120A(or SP-120, SP-120B or complex body) and 5 parts of sodium laurylsulfonates, 2 parts of ditane-sodium disulfonate-yubans and 83 parts of potter's clay mix, and pulverize, and promptly obtain 100 parts of preparations.

Embodiment 2 emulsions

With 8 parts of SP-120A(or SP-120, SP-120B or complex body) and 10 parts of ethylene glycol, 20 parts of dimethyl formamides, 10 parts of alkyl xylene ammonia chlorides and 52 parts of methanol mixed dissolvings promptly obtain 100 parts of emulsions.

Embodiment 3 pulvis

With 0.2 part of SP-120A(or SP-120, SP-120B or complex body) and 0.5 part of calcium stearate, 50 parts of talcum powder and 49.3 parts of potter's clay mix to be pulverized, and promptly gets 100 parts of pulvis.

Embodiment 4 granules

With 10 parts of SP-120A(or SP-120, SP-120B, or complex body) and 15 parts of starch, the sodium salt of 72 parts of powdered bentonites and 3 parts of sulfuric acid dodecyl esters mixes to be pulverized, and promptly gets 100 parts of preparations.

Test example 1

Control test to gray mold of cucumber

Will by embodiment 1 made wettable powder be diluted to fixed concentration, be sprayed onto then on cucumber (kind: horizontal mutually half the is white) seedling that after planting gives birth to 15 days, the sector-style of going forward side by side is done.

(Botorytis cinerea) places potato with the cucumber mildew bacterium, cultivates on the glucose agar medium, shines with BLB light spore is brought out.Spore after will bringing out is suspended in the yeast extracted solution of 10% glucose and 1%.

After the soup drying, cucumber is moved to inoculation tank, above-mentioned suspension is carried out spray inoculation with atomizer, per 10 cucumber seedlings spray medicine 10cc approximately.

The situation of falling ill is checked in the place that postvaccinal cucumber seedling places constant temperature to wet more after 4 days.

Prevention effect is calculated as follows

The disease index occurring degree

0 does not fall ill

1 only has scab

2 onset areas account for below 10%

3 onset areas account for more than 10% to below 20%

4 onset areas account for more than 20% to below 30%

5 onset areas account for more than 30% to below 40%

6 onset areas account for more than 40%

Prevention effect=(1-(treatment zone disease index summation)/(check plot disease index summation)) * 100

Its result is as shown in table 1

Table 1

Test compound spray concentration (ppm) prevention effect (%) poisoning

SP-120A 200 90 does not have

SP-120B 200 81 does not have

SP-120 complex body * 500 100 does not have

SP-120 complex body * 250 99 does not have

SP-120 complex body * 125 89 does not have

Benzene Lay spy 1) 250 92 do not have

ロ Block ラ-

2) 250 90 do not have

Control treatment-0-

* wherein contain effective ingredient about 30%

1) 1-butylamine formyl radical)-the 2-benzimidazole carbamate.

2) 1-sec.-propyl formamyl-3-(3, the 5-dichlorophenyl) glycolylurea (50%)

Test example 2

Control test to cucumber anthracnose

Be diluted to by embodiment 2 made emulsions fixed concentration, be sprayed onto breeding time after planting then and be on 15 days cucumber (kind: horizontal mutually half the is white) seedling, carry out air-dry.

Cucumber anthracnose (Colletotrichum lagenarium) placed on the potato Agar Plating cultivate, and the spore after will giving birth to is made suspension (spore concentration is about 200 an of visual field [microscope multiplying power * 150]).

After the soup drying, cucumber seedling is moved in the inoculation tank, carry out spray inoculation with above-mentioned suspension with atomizer, per 10 cucumber seedlings spray medicine 10cc approximately.Postvaccinal cucumber seedling was placed 24 hours in humidity is 100% inoculation tank, moved to then in the greenhouse of natural light, checked the scab number after 4 days.Each chemicals treatment district all uses 10 cucumber seedlings.

Prevention effect is calculated as follows

Prevention effect=1-(treatment zone scab sum)/(check plot scab sum) * 100

Its result is as shown in table 2

Table 2

Test compound spray concentration (ppm) prevention effect (%) poisoning

SP-120A 200 98 does not have

100 88 do not have

50 62 do not have

SP-120B 200 89 does not have

100 78 do not have

50 63 do not have

SP-120 complex body * 500 100 does not have

250 100 do not have

125 98 do not have

Bacterium clear 3) 1,000 90 do not have

Control treatment-0-

* contain effective ingredient about 30%

3) daconil M

Test example 3

Control test to the sheath and culm blight of rice

(kind: seedling ten stones) was cultured to for the 5th leaf phase in the greenhouse to prepare some potted plant (7 in every basin, every district 10 basins) paddy rice in advance.Will by the made wettable powder of embodiment 1 with methyl alcohol and water be diluted to fixed concentration, spray with the dosage of atomizer again by the about 40ml of average every basin.After 2 hours, the flora of Rhizoctonia solani Kuhn (Pellicuaria sasakii) is inserted between the new leaf sheath of paddy rice and inoculate, Ji Ye is covered with plastics film, measures the total scab length of rice pathogenesis after 7 days in greenhouse heat-insulating then.Prevention effect is calculated by following formula.

Prevention effect (%)=1-(total scab length of treatment zone)/(total scab length of check plot)) * 100

Its result is as shown in table 3

Table 3

Test compound spray concentration (ppm) prevention effect (%) poisoning

SP-120A 200 95 does not have

SP-120B 200 92 does not have

SP-120 complex body * 500 100 does not have

250 100 do not have

150 90 do not have

Neoasozin 4) 65 90 do not have

Control treatment-0-

* contain effective ingredient about 30%

4) ammonium ferric methylarsonate (65%)

Effect of the present invention

Result according to above test can prove, pesticide for agricultural horticulture of the present invention provides a kind of new agricultural chemicals. This kind agricultural chemicals both had high prevention effect to various plant diseases, had again the security of height and did not have any poisoning.

4. the description of the drawings

Fig. 1 is the uv absorption spectra of microbiotic SP-120A of the present invention.Fig. 2 is the uv absorption spectra of SP-120B.Fig. 3 is the infrared absorpting light spectra of microbiotic SP-120A.Fig. 4 is the infrared absorpting light spectra of microbiotic SP-120B.

Claims (2)

1, a kind of agriculture and garden new antibiotic SP-120 preparation of compositions method, the effective constituent of wherein said composition is made up of new antibiotic SP-120A and new antibiotic SP-120B, and their physico-chemical property is as follows:

New antibiotic SP-120A

(1) fusing point: be brown in 235～255 ℃ of decomposition;

(2) ultimate analysis: carbon 50.98%, hydrogen 7.11%, nitrogen 16.42%, sulphur 3.55%;

(3) specific optical rotation: [α] ²⁵ _D=-90 ° (C=0.05, methyl-sulphoxide);

(4) ultra-violet absorption spectrum: λ ^0.1 _MaxN-Hcl-MeoH 220nm

(E ^1% _1cm186) peak shoulder

(5) infrared absorption spectrum: (KBr method) 3300,2903,2850,1655,1520,1460,1395,1382,1280,1250,1215,1163,1100,1020,1002,900,840,760cm ^-1

(6) solvability: be insoluble to methyl alcohol, ethanol, propyl alcohol, ethyl acetate, chloroform, benzene, normal hexane and ether; Be insoluble in water; Dissolve in the mixed solution of propyl carbinol-methanol-water;

(7) molecular weight: (pressing the FAB mass spectrum) 911;

(8) color reaction: with potassium permanganate, aubepine-sulfuric acid, iodine is positive;

(9) material color: white powder;

(10) acidic hydrolysis: acid hydrolysis gets leucine and glycine;

(11) antimicrobial spectrum: to gram-positive microorganism, the resistance to acid bacterium, negative bacterium and phytopathogen are anti-microbial activity;

2, new antibiotic SP-120B

(1) fusing point: 235～265 ℃ of decomposition are brown;

(2) ultimate analysis: carbon 50.91%, hydrogen 6.95%, nitrogen 16.12%, sulphur 3.39%;

(3) specific optical rotation: (α) ²⁵ _D=-59 ° (C=0.03, methyl-sulphoxide);

(4) ultra-violet absorption spectrum: λ ^{0.1N-Hcl-MevH} _Max220nm

(5) infrared absorption spectrum: (KBr method) 3300,2900,2850,1650,1520,1435,1392,1370,1342,1322,1285,1212,1185,1168,1100,1024,1004,958,903,846,825,765Cm ^-1;

(6) solvability: be insoluble to methyl alcohol, ethanol, propyl alcohol, ethyl acetate, chloroform, benzene, normal hexane and ether; Be insoluble in water; Dissolve in the mixed solution of propyl carbinol-methanol-water;

(7) molecular weight: (pressing the FAB mass spectrum) 923;

(8) color reaction: with potassium permanganate, aubepine-sulfuric acid, iodine is positive;

(9) material color: white powder;

(10) acidic hydrolysis: acid hydrolysis gets Xie Ansuan and glycine;

(11) antimicrobial spectrum: to gram-positive microorganism, the resistance to acid bacterium, negative bacterium and phytopathogen are anti-microbial activity; Described method comprises cultivates streptomycete SP-120(Streptomyces SP.SP-120, and CGMCC0022), the described effective constituent of separation and Extraction from culture is then mixed carrier suitable on the effective constituent that obtains and the agricultural chemicals at last.

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