NO175826B - Microbiological process for the preparation of agronomically useful active ingredients, compounds prepared, their use and biologically pure cultures of sorangium cellulosum used in the process - Google Patents

Description

The present invention relates to agrochemically applicable macro-cyclic active substances of formula I, microbiological methods for their production, agents containing the active substances, biological pure cultures and the use of the active substances in combating and preventing plant diseases.

The macrocyclic active substances are characterized by formula I

where the macrocyclic ring is either saturated or optionally contains in the 8,9-position or in the 9,10-position or in the 14,15-position a double bond after the following substituent combination, and the substituents Ri-Rs, A, B, X and Y are present in the following combined meanings:

The invention also relates to the process for preparing compounds of formula I where the macrocyclic ring is either saturated or optionally contains in the 8,9-position or in the 9,10-position or in the 14,15-position a double bond after the following substituent combination, and the substituents Rx -R5, A, B, X and Y are present in the following combined meanings: These 32 compounds are produced by microbiological cultivation as stated in claim 3's characterizing part of Myxo bacteria from the Sorangium/Polyangium group of which all or in part, depending on the respective strain, are produced in different ratios. Common to all strains is the production of compound A as one of the main products. The compounds of formula I are referred to here and in the following as "Soraphens", i.e. the compounds in the above table are referred to as "Soraphen A" to "Soraphen a".

The invention relates in particular to six new microorganism strains of the cellulose-splitting Myxo bacteria from the Sorangium/Polyangium group which, as is known, occur frequently in soil samples, plant material or in animal manure. Characteristic of this taxonomically not usually divided group is its ability to utilize cellulose or cellulose cleavage products as the only carbon source. The a-strains according to the present invention, regardless of whether they strictly belong to the Sorangium/Polyangium group or just a taxonomically related area, are compared to the most widespread representatives of this group further characterized by the fact that they produce at least one of the above-mentioned " Soraphens" of formula I and preferably at least two "Soraphens", including "Soraphen A".

The six strains here and in the following have the collective name Sorangium (Polyangium) cellulosum. They originate from soil samples that were collected at different times in different places in Europe, Africa or USA. They were deposited in the German Collection of Microorganisms (DSM) in Braunschweig, West Germany, according to the Budapest contract. They are characterized as follows. 1) Sorangium ( Polyangium) cellulosum strain "So ce 139", isolated in May 1986 from a soil sample collected in autumn 1982 at Fort Huachaca, Arizona, USA. Deposit number: DSM 5397. Deposit date: 2 June 1989. 2) Sorangium (Polyangium) cellulosum strain "So ce 170", isolated in April 1987 from a soil sample collected in May 1981 from the island of Delos, Greece. Deposit number: DSM 4795. Deposit date: 2 September 1988. 3) Sorangium (Polyangium) cellulosum strain "So ce 191", isolated in August 1987 from a soil sample collected in February 1987 from the island of Madeira, Portugal. Deposit number: DSM 4796. Deposit date: 2 September 1988. 4) Sorangium (Polyangium) cellulosum strain "So ce 192", isolated in August 1987 from a soil sample collected in March 1987 from Nigeria. Deposit number: DSM 4797. Deposit date: 2 September 1988. 5) Sorangium (Polyangium) cellulosum strain "So ce 231", isolated in April 1988 from a soil sample collected in April 1987 at Didyma, Turkey. Deposit number: DSM 5393. Deposit date: 2 June 1989. 6) Sorangium (Polyangium) cellulosum strain "So ce 242", isolated in October 1988 from a soil sample collected in April 1988 at Pozzuoli, Italy. Deposit number: DSM 5414. Date of deposit: 19 June 1989.

In what follows, the term Sorangium cellulosum is used for these strains. =

The process according to the invention for the production of a "Soraphen" of formula I is characterized by aerobic cultivation of one of the strains Sorangium cellulosum "So ce 139", "So ce 170", "So ce 191", "So ce 192", "So ce 231" or "So ce 242" or a derivable clone, mutant etc. in a suitable nutrient medium and isolation of the formed "Soraphen".

In the European patent application EP-A-282,455, another Sorangium cellulosum microorganism "Soce 26" is described, the fermentation of which yields the above-mentioned Soraphens A and B. The present invention does not relate to these two preparations and their use on their own in remedies.

The present invention further relates to valuable plant microbicide fermentation products which are produced by the micro-organisms "So ce 26" (deposition number NCIB 12 411) and/or by the microorganisms claimed here "So ce 139", "So ce 170", "So ce 191 ", "So ce 192", "So ce 231" or "So ce 242".

General production conditions for the cultivation of the six production stops

The Sorangium cellulosum strains can be cultivated in suitable nutrient media according to usual biological methods, e.g. in shaking cultures or in fermenters. The fermentation temperature is usually 10-40°C, preferably 10-35°C, and particularly

drawn approx. 30-32°C, the pH value is 6-8, preferably 7.4. The process takes place aerobically and under sterile conditions.

The composition of the nutrient medium can vary in larger areas. For essential nutrients, there must be a carbon and a nitrogen source as well as a source for inorganic mineral salts which include P, S, Mg, K, Fe, Ca.

The fermentation processes preferably use glucose, starch and cellulose as well as their cleavage products (e.g. cellobiose), as well as disaccharides, glycerin, acetic acid and others, as a C source. As an N source, e.g. NH4, N03 and also peptones. An organic compound as an N source cannot, as a rule, simultaneously be the only C source and energy source during the fermentation.

As mineral salts, chlorides, nitrates, sulphates, carbonates and phosphates of the elements Na, K, NH4, Mg, Fe and Ca come into consideration, moreover Cu, Mn, Mo, Zn, Co and others may be present as trace elements. As far as possible, such salts can also be present bound to ethylenediaminetetraacetic acid

(EDTA).

The microorganism culture is introduced into the shaking culture or into the fermenter in an inoculation amount of 0.1-20%, preferably 0.5-10%, particularly preferably 0.5-5% (v/v). The culture duration is at approx. 30°C around 2-7 days, for large volumes sometimes up to 10 days or longer. For large volumes, it is advantageous to initially ferment smaller pre-cultures. The use of Sorangium cellulosum strains is also possible in immobilized form, e.g. in the form of carrier-fixed cells on alginate.

Cultivation takes place aerobically, i.e. e.g. in a still surface culture or preferably below the surface, under shaking or stirring with air or oxygen in shaking cultures or in fermenters. Preferably, you cultivate in stages, i.e. you first prepare one or more pre-cultures in a liquid nutrient medium which are then inoculated into the actual production medium, e.g. in the ratio 1:20.

The isolation of "Soraphen" takes place in a physicochemical way using known isolation methods, such as filtration, but especially solvent extraction and chromatography, above all adsorption and distribution chromatography and also crystallization.

From the aqueous fermentation liquid, the compounds of formula I can be extracted with lipophilic organic solvents, e.g. with ketones, such as methyl ethyl ketone, cyclohexanone; with medium chain alcohols, such as iso-butanol, pentanol, hexanol; with acetic acid C 1 -C 8 alkyl esters (ethyl acetate, propyl acetate, butyl acetate, isobutyl acetate, etc.); with toluene, dichloromethane, 1,2-dichloroethane,

chlorobenzene, dichlorobenzene, etc.

From the filtered cell cake, the compounds are easily extracted with alcohols or ketones (e.g. methanol, ethanol, acetone, methyl ethyl ketone).

The compounds of formula I can then be prepared from each extract by concentration and/or precipitation, and they can be separated by fractional crystallization or other chromatographic separation methods in Soraphenene A to p (= ro).

Advantageously, the fermentation process for the preparation of the compounds of formula I is terminated by adding an adsorption resin, from which the desired products are taken up, or the fermentation is carried out from the beginning in the presence of such an adsorption resin. As adsorbing resins, above all, neutral, organic polymer substances come into question, especially non-ionic, hydrophobic adsorbing resins which are suitable for lipophilic extraction. The compound of formula I binds to these almost quantitatively. Examples of such resins are semi-polar acrylic ester resins, non-polar polystyrene/divinylbenzene resins and especially presaturated polystyrene. Such resins are added in amounts of 0.1-5% (v/v) to the fermentation volume, preferably 0.5-2% (v/v). Activated carbon also comes into play. Technically particularly advantageous are polystyrene resins, such as e.g. XAD-1180 or XAD-16

(manufacturer: Rohm and Haas) which is available in filterable form (granules or granules). After the end of the fermentation, the resin is filtered off, it is washed with water and treated with methanol or ethanol. The alcoholic extract is concentrated. By adding diethyl ether, ethyl acetate or butyl acetate, the widely crystallized compound IA (= Soraphen A) can be isolated. The filtrate is purified chromatographically to prepare the remaining Soraphen A, but above all to prepare the remaining Soraphenes B to o (sigma).

The invention relates to the compounds of formula I in pure form or in crystallized form. However, the invention also relates to biomasses, crude extracts and adsorption resins from the fermentation which contain the compound of formula I and which can be used as such or in further formulated form to combat plant diseases. Biomass can also be used further as ground or pressed dry substances ("cake") or brought into the trade.

The described production methods, including all sub-steps, are a component of the present invention.

In particular, the invention relates to a method for the production of Soraphen with formula I, characterized by aerobically cultivating one of the strains Sorangium cellulosum "So ce 139", "So ce 170", "So ce 191", "So ce 192", "So ce 231" and "So ce 242", or a microorganism containing the same structural genes that cause formation of Soraphen, in an aqueous nutrient medium containing a carbon and nitrogen source as well as inorganic salts, and isolates the Soraphenes of formula I.

In the first line, the invention relates to an embodiment of the above-mentioned method which is characterized by cultivating a Soraphen-forming microorganism from the group of cellulose-splitting Myxo bacteria.

A preferred embodiment of the above method is characterized by cultivating one of the strains Sorangium cellulosum "So ce 139", "So ce 170", "So ce 191", "So ce 192", "So ce 231", "So ce 242" or a Soraphen-forming mutant of this strain.

A particularly preferred embodiment of the above method is characterized by growing one of the strains "So ce 139", "So ce 170", "So ce 191", "So ce 192", "So ce 231" deposited according to the Budapest contract ", "So ce 242" .

Fermentation is preferably carried out under the conditions described in the example section.

Microorganisms that contain the same for the formation of Soraphen causal structural genes as the six strains, can e.g. is produced artificially by genetic manipulation, whereby the corresponding structural genes are isolated from the six strains and incorporated into suitable places in the genetic material of another suitable microorganism. Suitable microorganisms are those in which the relevant structural genes can not only be inserted, but in which these structural genes are also expressed and in which the formed Soraphen is not broken down again, but is preferably excreted in the fermentation liquid. Such suitable microorganisms are primarily other strains of Myxo bacteria, especially strains of the species Sorangium cellulosum. provided they do not already have the above-mentioned structural genes.

Soraphen-forming mutants can e.g. produced under the influence of ultraviolet rays or X-rays or by chemical mutagens, e.g. N-methyl-N'-nitro-N-nitroso-guanidine, and is isolated by selection according to its specific properties in a manner known per se. Further process features in the production of mutants and recombinants of a microorganism are known and routine to the person skilled in the art.

The invention also relates to a method for the production of a fermentation product which contains Soraphen in a detectable amount and which is characterized by growing one of the strains Sorangium cellulosum "So ce 139", "So ce 170", "So ce 191", "So ce 192", "So ce 231", "So ce 242", or a clone derivable from these strains in a nutrient medium, recovering Soraphen in suitable form. Such culture solutions can be used as such or in concentrated form, possibly with the addition of further carrier substances and/or distribution substances as a means against plant pathogenic microorganisms, and they are therefore an important part of the present invention.

In a narrower sense, the method is characterized by cultivating one of the microorganisms Sorangium cellulosum "So ce 139", "So ce 170", "So ce 191", "So ce 192", "So ce 231" or "So ce 242" in a culture medium containing at least one assimilable C source and N source, as well as corresponding inorganic salts, at 10-40°C, preferably at 10-35°C, in the presence or absence of an adsorption resin, then extracting the culture solution or the filtered off adsorption resin with a suitable solvent phase, concentrates the solution obtained and purifies the remaining residue, if desired by chromatography and/or recrystallization.

Strain cultures and morphological description

Stock cultures are obtained as dish cultures on VY/2 agar (0.5% baker's yeast according to fresh weight; 0.1% CaCl2; 1.5% agar; pH 7.2) or on filter paper (= cellulose source) over ST 21 agar (0 .1% KN03; 0.1% MgS04-7 H20; 0.1% CaCl2; 0.1% K2HP04; 0.01% MnS04-7 H20; 0.02% FeCl3; 0.002% yeast extract; standard trace element solution<1> ; 1% agar). The plates are incubated at 30°C.

On both media, the organisms form swarm colonies which slowly spread over the substrate. In some cases, clear differences are observed between the a-stems.

<1>0.02-0.5 mg Mn, Mo, Cu, Co and/or Zn salt per liter [R.Y. Stanier et al. "General Microbiology", 4th ed., p. 36

(1976)]; or 500 mg EDTA, 300 mg FeS04-7 H20, 3 mg MnCl2-4

H20, 5 mg CoCl2- 6 H20, 1 mg CuCl2- 2 H20, 2 mg NiCl2- 6 H20, 3 mg Na2Mo04-2 H20, 5 mg ZnS04-7 H20, 2 mg H3B03 per liter of distilled water (approx. pH 4) [G. Drew's "Microbiologist. Praktikum", 4th edition, p. 11, Springer forlag, Berlin, Heidelberg, New York, Tokyo

(1983)].

1) " So ce 139" ( DSM 5397): On filter paper over ST 21 agar, the swarm colony remains substantially restricted on the filter paper. In older colonies, the filter edge of the agar is torn, and the microorganisms and the agar roll in. The filter paper is completely broken down, in places you will find a thick layer of black-brown fruiting bodies. These are composed of small sporangioles with a cross-section of 15-30 µm, which in turn are bundled together into small packets or elongated, unarticulated masses. The vegetative cells are slender, cylindrical small rods with bluntly rounded ends and measure 0.7-0.9 x 2.5-4 µm. In a phase contrast microscope, they appear dark, often with bright small polar grains. On VY/2 agar, very large swarm colonies are formed with colorless to orange radial veins and many black-brown fruiting bodies of a similar structure as described above. The yeast cells in the nutrient medium are hardly attacked. The vegetative cells appear somewhat firmer and are approx. 1 um thicker. 2) " So ce 170" ( DSM 4795): On filter paper over ST 21 agar, the swarm penetrates widely over the filter onto the agar surface. The agar is later deeply shaken, and the swarm colony rolls in. On the filter paper, fruiting bodies can develop: These consist of small, dark brown, thick-walled sporangioles, with mostly 20-30 um 0, are tightly compressed in mostly sharply demarcated, elongated packages with approx. 50-200 um 0. In places, coherent elongated aggregates of fruiting bodies may occur. On VY/2 agar, medium-sized, thick-mucous, intensively orange swarming colonies are formed sparingly, mostly 1-2 cm 0. The yeast cells are slowly degraded. Here, too, the agar is torn up deeply in places, so that the swarm can fall completely out of the dish. On the agar surface, orange, light brown to dark brown fruiting bodies with a similar structure as above are often formed. The vegetative cells are solid, in phase contrast microscope dark small rods with broadly rounded ends and usually measure 0.8-1.0 x 2-5 pm. The strain also breaks down chitin very efficiently. 3) "So ce 191" (DSM 4796) grows on filter paper over ST 21 agar without transferring to the agar surface. In this case too, the agar can be torn up deeply around the filter paper, and the swarm colony can roll in. On VY/2 agar, large swarms of 6-8 cm 0 with fine to strong radial veins develop. The yeast cells are hardly attacked. The cylindrical, vegetative small rods have broadly rounded ends, are in phase- contrast microscope dark, often quite long and measuring mostly 0.7-0.9 x 3-8 um. The strain forms no fruiting bodies in pure culture under the mentioned culture conditions. The strain breaks down chitin efficiently. 4) "So ce 192" (DSM 4797) spreads when cultivated on filter paper over ST 21 agar widely beyond the agar surface. The agar can be torn up deeply, the swarm colony can roll in. On the agar surface, and especially on the filter paper, many luminous, orange-coloured fruiting bodies develop, consisting of very small sporangioles with mostly 10-20 pm 0 in sharply limited packets with a spread of mostly 50-500 pm. On VY/2 agar medium-sized, powerful swarms of 2-3 cm 0 develop with radial veins and many intensively brown-orange colored fruiting bodies in and on the agar of a similar structure as described above. The vegetative cells are cylindrical, dark small rods with broad ends and measure mostly 0.7-0.9 x 3-5 pm. The yeast cells are not broken down, but the strain breaks down chitin very efficiently. 5) "So ce 231" (DSM 5393) is cultured on filter paper over ST 21 agar, not on the agar plate. The agar can be torn up at the edge of the filter, but the colonies do not roll in. The filter paper is completely destroyed, in its places you will later find a very thick layer of intensively brown-orange fruiting bodies. These are similarly structured as in other strains, but the sporangioles are often arranged in bands. On VY/2 agar, very large swarms develop with radial bundles of fine veins. The light develops locally the, orange-brown fruiting bodies. The slender, vegetative small canes are very graceful and measure 0.6-0.8 x 2.5-6 jim. The yeast cells in the nutrient medium are slowly lysed. The strain also breaks down chitin very efficiently. 6) "So ce 242" (DSM 5414) penetrates the filter paper over ST 21 agar and forms fruit bodies there as well. The agar is torn up at the edge of the filter paper deeply. The filter paper is completely broken down and replaced by a thick mass of black-brown fruiting bodies. In these, the sporangioles are often arranged in chains. On VY/2 agar, large swarms develop with fine radial veins. The agar is clawed up in the swarm area in several places. Furthermore, abundant fruiting bodies occur. The yeast cells in the substrate slowly break down. The vegetative cells measure 0.7-1.0 nm x 2-4 ym. The strain is a very effective chitin degrader.

The strains produce substances that inhibit the growth of numerous yeast cells and hyphae. Chemically, the inhibitors are not only Soraphens, whose structure has already been elucidated, as indicated below. These mixtures of macrocyclic compounds of the Soraphen type appear in liquid cultures in the culture fluid, but can also be isolated from the cells. The production of antibiotics occurs during the logarithmic and also during the stationary growth phase.

All six strains must be adapted to growth in the liquid medium. They first grow in the form of small, firm, orange-coloured small tubers and only form, after many transfer steps from one liquid medium to another, more or less homogeneous cell suspensions.

Biological characterization of the "Soce 139" strains. "So ce 170". "So ce 191". "So ce 192", "So ce 231" and "So ce 242"

Cellulose degradation: positive

Glucose breakdown: positive

Starch breakdown: positive

NH4 as N source: positive

N03 as N source: positive

The invention relates to Soraphenenes C to o of formula I in pure form. However, the invention also relates to biomasses, crude extracts or adsorption resins from the fermentation which contain Soraphenenes C to o with formula I, and as such or in further formulated form can be used to combat plant diseases. Biomass can also be used as ground or pressed dry substances ("cake") and brought to the market.

The described manufacturing methods, including all sub-steps, form part of the present invention.

Surprisingly, it has now been found that the compounds of formula I exhibit a very favorable biocidal spectrum against phytopathogenic microorganisms, in particular against fungi. They have very beneficial curative, systemic and especially preventive properties and can be used to protect numerous cultivated plants. With the active ingredients of formula I, the parasites appearing on plants or plant parts (fruits, flowers, foliage, stems, tubers, roots) on various useful crops can be controlled or destroyed, whereby even later growing plant parts remain spared from the phytopathogens microorganisms.

As microbicides, the active substances with formula I are effective, e.g. against phytopathogenic fungi belonging to the following classes: Fungi imperfecti (e.g. particularly Botrytis, further Pyricularia, Helminthosporium, Fusarium, Septoria, Cercospora and Alternaria); Basidiomycetes (eg Rhizoctonia, Hemileia, Puccinia). Furthermore, they work against the classes Ascomycetes (e.g. especially Venturia and Erysiphe, also Podosphaera, Monilinia, Uncinula) and Oomycetes (e.g. Phytophthora, Plasmopara). The compounds of formula I can also be used as a mordant when treating seeds (fruits, tubers, grains) and plant cuttings to protect against fungal infections and against phytopathogenic fungi that occur in the soil.

The invention also relates to the agents which contain as an active substance component one or more of the Soraphenenes C to o of formula I, in particular pesticides and their use in the agricultural sector or related areas.

Also included is a method for treating plants, which method is distinguished by the application of the new compounds Soraphen C to a with formula I or of the corresponding new funds.

As target crops for the plant protection application developed here, the framework for this invention applies e.g. the following plant species: cereals (wheat, barley, rye, oats, rice, maize, sorghum and related species); beet (sugar beet and beet); stone, stone and berry fruits (apples, pears, plums, peaches, almonds, cherries, strawberries, raspberries and blackberries); legumes (beans, lentils, peas, soy); oil crops (rapeseed, mustard, poppy, olive, sunflower, coconut, castor, cocoa, peanut); cucurbits (pumpkins, cucumbers, melons); fiber crops (cotton, flax, hemp, jute); citrus fruits (oranges, lemons, grapefruit, tangerines); vegetable varieties (spinach, lettuce, asparagus, cabbage varieties, carrots, onions, tomatoes, potatoes, peppers); laurel plants (avocado, cinnamon, camphor) or plants such as tobacco, nuts, coffee, pineapple, sugar cane, tea, pepper, vines, hops, banana and natural rubber plants as well as ornamental plants. This enumeration does not constitute a limitation.

Active substances with formula I are usually used in the form of compositions and can be added simultaneously or one after the other with additional active substances on the surface or the plants to be treated. These additional active substances can be both fertilisers, trace element mediators or other preparations that have an impact on plant growth. Also selective herbicides as well as insecticides, fungicides, bactericides, nematicides, molluscicides or mixtures of several of these preparations, together with possibly additional carrier substances common in the formulation technique, surfactants or other application-promoting additives are thereby used.

Suitable carriers and additives can be solid or liquid and correspond to substances suitable in the formulation technique, such as e.g. natural or regenerated mineral substances, solvents, dispersants, cross-linkers, adhesives, thickeners, binders or fertilisers.

A preferred method for placing an active substance of formula I or an agrochemical that contains at least one of these active substances is to apply it to the foliage (foliar application). Application frequency and quantity are therefore based on the attack pressure from the parasite in question. However, the active substances with formula I can also penetrate into the root system of the plant through the soil (systemic effect), by soaking the plant's stand with a liquid preparation or adding the substances in solid form to the field, e.g. in the form of a granule (soil application). This granulate or a corresponding powder can also be dry mass from the biomass that precipitates from the fermenter or the adsorption resin that is filtered out of the fermentation liquid and contains the active substances of formula I. Compounds of formula I can. also applied to the seed grains ("coating"), by soaking the grains either in a liquid preparation of the active substance or coating them with a solid preparation.

The compounds of formula I are thereby used in unchanged form or preferably together with auxiliaries which are common in the formulation technique. Thereby, they are processed appropriately, e.g. for emulsion concentrates, spreadable pastes, directly sprayable or dilutable solutions, diluted emulsions, sprayable powders, soluble powders, nebulizers, granules, by encapsulation in e.g. polymeric substances in a known manner. Applications such as spraying, atomisation, fine distribution, spreading, coating or watering are chosen according to the nature of the agent, according to the desired goals and the given conditions. Favorable application quantities are usually 10 g to 2 kg of active substance (AS) per hectare, preferably at 50-500 g AS/ha.

The formulations, i.e. the agents containing the active substances of formula I and a solid or liquid additive, are prepared in a known manner.

Suitable solvents include: aromatic and aliphatic hydrocarbons, such as e.g. xylene mixtures, cyclohexane or paraffins; further alcohols and glycols and their ethers and esters, such as ethanol, ethylene glycol, ethylene glycol monomethyl or ethyl ether, acetic acid ester; ketones, such as cyclohexanone, strongly polar solvents, such as N-methyl-2-pyrrolidone, dimethylsulfoxide or dimethylformamide, as well as optionally epoxidized vegetable oils, such as epoxidized coconut oil or soybean oil; or water.

As solid carriers, e.g. for nebulizers and dispersible powders, natural stone flours, such as calcite, talc, kaolin, montmorillonite or attapulgite, are usually used. To improve the physical properties, highly dispersed silicic acid or highly dispersed absorbent polymers can also be added. Porous types such as e.g. pumice stone, broken brick, sepio-lite or bentonite, as non-sorptive carrier materials e.g. calcite or sand. Furthermore, a large number of pre-granulated materials of an inorganic nature can be used, such as especially dolomite or finely divided plant residues.

As surface-active compounds, non-ionic, cationic and/or anionic surfactants with good emulsifying, dispersing and cross-linking properties come into consideration, depending on the nature of the active substance of formula I to be formulated. Surfactants also mean surfactant mixtures.

Suitable anionic surfactants can be both so-called water-soluble soaps and also water-soluble, synthetic surface-active compounds.

More often, however, so-called synthetic surfactants are used, especially alkane sulphonates, fatty alcohol sulphates, sulphonated benzimidazole derivatives or alkyl sulphonates.

Relevant as non-ionic surfactants are polyglycol ether derivatives of aliphatic or cycloaliphatic alcohols, saturated or unsaturated fatty acids and alkylphenols, which may contain 3-30 glycol ether groups and 8-20 carbon atoms in the (aliphatic) hydrocarbon residue and 6-18 carbon atoms in alkyl -the alkyl residue of the phenols.

As further examples of nonionic surfactants, nonylphenol polyethoxyethanols, castor oil polyglycol ethers, polypropylene-polyethylene oxide adducts, tributylphenoxypolyethyleneethanol, polyethyleneglycol and octylphenoxypolyethoxyethanol can be mentioned.

Furthermore, fatty acid esters of polyoxyethylene sorbitan, such as the polyoxyethylene sorbitan trioleate, also come into consideration.

The surfactants most widely used in the formulation technique are described, among others, in the following publications: "Mc Cutche-on's Detergents and Emulsifiers Annual", MC Publishing Corp., Ridgewood New Jersey, 1980; Sisley and Wood, "Encyclopedia of Surface Active Agents", Chemical Publishing Co., Inc. New York, 1980.

Particularly advantageous, application-promoting additives which can lead to a strong reduction of the amount used are further natural (animal or plant-like) or synthetic phospholipids from the range of kephalins and lecithins, such as e.g. phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerine or lysolecithin.

The agrochemical preparations usually contain 0.1-95% active substance with formula I, 99.9-5% of a solid or liquid additive and 0-25% of a surfactant.

While rather concentrated means are preferred as commercial goods, the end consumer usually uses diluted means.

The agents may also contain further additives, such as stabilisers, anti-foaming agents, viscosity regulators, binders, adhesives as well as fertilizers and other active substances to achieve special effects.

1. Production examples

Example H-1: Preparation of the compounds of formula I in the presence of an adsorption resin

The procedure is carried out in a fermentation volume of 1000 1 with the addition of 0.5% (v/v) adsorption resin XAD-1180 (Rohm and Haas). The procedure conditions for this rate correspond to those for the following example H-2. Instead of the strain Soce 26 from EP-A-282.-455 used here, one of the other six mentioned strains from the present invention can also be used.

After completion of the fermentation, the polymer carrier is filtered, it is flushed into a glass column, washed with 3 bed volumes of water and eluted with 4 bed volumes of methanol. The eluate is concentrated in vacuo to dryness. Crude extract weight: 165 g.

The further chromatographic purification follows the scheme for the attached figure, where each individual separation step takes place under the following conditions. The yields are indicated in brackets (h = hour).

Separation procedure for isolation of Soraphen A to 0

1. Silica gel separation; column: 100 mm 0; 45 cm length.

Adsorbent: Lichroprep Si 100; 40-63 pm (manufacturer: Merck).

Lubricant: dichloromethane/acetone in steps 98/2, 95/5, 93/7, 90/10, 50/50; each step 2 1.

5 fractions are taken. Fraction 3 contains Soraphen A, fraction 4 (25 g) is further purified. 2. Gel chromatography; column: 60 mm 0; 100 cm length. Adsorbent: Sephadex LH-20 (manufacturer: Pharmacia LKB GmbH).

Solvent: methanol.

5 fractions are taken. Fraction 3 (17 g) is further purified. 3. Reversed phase separation; column: 76 mm 0.70 cm length.

Adsorbent: HDSIL RP-18; 18-60-60 35-70 pm (producer: Labomatic).

Solvent: methanol/water 40/60 lin. gradient for 2 h after methanol.

9 fractions are taken. Fractions 3 (1.3 g), 4 (1.5

g), 5 (3.5 g) and 6 (1.6 g) are further purified.

4. Silica gel separation; column: 40 mm 0; 30 cm length.

Adsorbent: Lichrosorb Si 100; 7 pm (manufacturer: Merck).

Solvent: dichloromethane/hexane 1/1 + 2% methanol.

6 fractions are taken. Fractions 2 (500 mg) and 4 (88 mg) are further purified. 5. Silica gel separation; column: 40 mm 0; 30 cm length. Adsorbent: Lichroprep Si 100; 7 pm (manufacturer : Merck).

Solvent: dichloromethane/hexane 1/1 + 2% methanol.

6 fractions are taken. Fraction 3 (500 mg) is further purified.

6. Silica gel separation; column: 35 mm 0; 45 cm length. Adsorbent: Lichroprep Si 60; 40-63 pm (pro-

ducent: Merck).

Lubricant: dichloromethane/hexane lin. gradient for 1 h after methanol.

6 fractions are taken. Fractions 1 (2.4 g) and 2 (400 mg) are further purified. 7. Reversed phase separation; column: 35 mm 0.45 cm length.

Adsorbent: HDSIL RP-18; 18-30-60 25-40 pm (producer: Labomatic).

Solvent: methanol/water 80/20.

9 fractions are taken. Fraction 7 (1.4 g) is further purified. 8. Reversed phase separation; column: 20.5 mm 0.25 cm length.

Adsorbent: Nucleosil 100-7 C18; 7 pm (producer : Macherey Nagel).

Solvent: methanol/water 70/30.

7 fractions are taken. Fraction 3 (15 mg) is further purified. Fraction 7 contains Soraphen N (18 mg). 9. Reversed phase separation; column: 20.5 mm 0.25 cm length.

Adsorbent: Nucleosil 100-7 C18; 7 pm (producer : Macherey Nagel).

Solvent: methanol/water 65/35.

7 fractions are taken. Fraction 3 contains Soraphen H (7 mg). 10. Reversed phase separation; column: 20.5 mm 0.25 cm length.

Adsorbent: HDSIL 100-C18; 10 pm (producer:

Labomatic).

Solvent: methanol/water 65/35.

4 fractions are taken. Fraction 2 contains Soraphen D (108 mg). 11. Silica gel separation; column: 40 mm 0; 30 cm length. Adsorbent: Lichrosorb Si 100; 7 µm (manufacturer: Merck).

Solvent: dichloromethane/hexane 1/1 + 1% methanol. 9 fractions are taken. Fractions 4 (530 mg) and 6 (280 mg) are further purified. 12. Silica gel separation; column: 40 mm 0; 30 cm length. Adsorbent: Lichrosorb Si 100; 7 µm (manufacturer: Merck).

Solvent: dichloromethane/hexane 1/1 + 1% methanol. 8 fractions are taken. Fraction 7 (290 mg) is further purified. 13. Silica gel separation; column: 20.5 mm 0.25 cm length. Adsorbent: Nucleosil 100-7; 7 um (manufacturer:

Macherey Nagel).

Gluing agent: tert-butyl methyl ether/hexane 1/2 + 1% methanol.

2 fractions are taken. Fraction 1 (325 mg) contains Soraphen F. 14. Reversed phase separation; column: 20.5 mm 0.25 cm length.

Adsorbent: Nucleosil 100-7 C18; 7 um (producer : Macherey Nagel).

Solvent: methanol/water 68/32.

2 fractions are taken. Fraction 1 contains Soraphen 0 (3 mg). 15. Reversed phase separation; column: 20.5 mm 0.25 cm length.

Adsorbent: Nucleosil 100-7 C18; 7 um (producer : Macherey Nagel).

Solvent: methanol/water 66/34.

4 fractions are taken. Fraction 1 contains Soraphen C

(21mg); fraction 2 contains Soraphen B (230 mg). 16. Reversed phase separation; column: 20.5 mm 0.25 cm length.

Adsorbent: Nucleosil 100-7 C18; 7 um (producer : Macherey Nagel).

Solvent: methanol/water 65/35.

10 fractions are taken. Fraction 4 contains Soraphen J<_> (18 mg), fraction 9 contains Soraphen E (96 mg). 17. Reversed phase separation; column: 20.5 mm 0.25 cm length.

Adsorbent: Nucleosil 100-7 C18; 7 um (producer : Macherey Nagel).

Solvent: methanol/water 70/30.

10 fractions are taken. Fraction 6 contains Soraphen M (98 mg).

Physicochemical characterization of Soraphenene C to 0

Soraphen C

<C>28<H>42°8

MZ 506

IR (film):

3411; 2939; 2831; 1725; 1461; 1382; 1266; 1230; 1187;

1152; 1098; 1068; 1023; 988; 975.

<13>C-NMR shifts (CDC13; 6 in ppm):

10.3; 11.7; 12.5; 23.0; 26.0; 29.4; 35.2; 35.6; 35.8;

46.2; 57.3; 57.6; 68.8; 72.5; 74.6; 74.9; 76.1; 83.7;

99.4; 125.0; 126.2; 126.2; 128.2; 128.6; 137.3; 141.0;

170.6.

HPLC Rt = 8.6 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen D

<C>27<H>42<0>8

Mz 494

IR (film):

3411; 2937; 2831; 1733; 1461; 1432; 1382; 1359; 1328;

1268; 1208; 1195; 1096; 1050; 988; 933.

<13>C-NMR shifts (CDC13; 6 in ppm):

10.7; 14.4; 22.8; 24.3; 25.0; 27.5; 27.7; 31.6; 35.3;

35.6; 43.2; 57.5; 58.6; 69.1; 70.8; 71.2; 74.8; 80.7;

82.6; 97.7; 126.4; 126.4; 128.0; 128.6; 141.0; 168.6.

HPLC Rt = 7.4 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen E

<C>29<H>46°9

Mz 538

IR (film):

3398; 2939; 2829; 1725; 1461; 1430; 1380; 1266; 1235;

1191; 1154; 1102; 1044; 971.

<13>C-NMR shifts (CDC13; 6 in ppm):

10.3; 10.5; 11.5; 23.0; 25.0; 29.5; 30.0; 35.3; 35.5;

40.2; 46.3; 56.4; 57.4; 58.0; 69.0; 71.3; 71.7; 75.0;

76.3; 80.2; 81.1; 99.9; 126.5; 126.5; 128.1; 128.6;

140.9; 170.9.

HPLC Rt = 11.8 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen F

<C>29<H>46<0>8

Mz 522

IR (film):

3394; 2937; 2827; 1729; 1710; 1461; 1382; 1326; 1314;

1266; 1253; 1232; 1189; 1154; 1104; 1075; 1048; 1021;

994; 971; 907.

<13>C-NMR shifts (CDC13; 6 in ppm):

10.4; 11.7; 14.1; 23.0; 24.2; 28.3; 28.8; 33.3; 34.1;

35.2; 45.7; 57.4; 57.4; 57.9; 69.0; 70.1; 75.9; 76.3;

80.8; 82.1; 99.6; 126.6; 126.6; 128.2; 128.6; 128.6;

140.4; 171.8.

HPLC Rt = 8.6 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 75/25.

Detector: UV 210 nm.

Soraphen H

<C>28<H>42°9

Mz 522

IR (film):

3444; 2935; 2867; 2833; 1723; 1459; 1409; 1382; 1334;

1270; 1230; 1179; 1156; 1104; 1069; 996; 971.

<13>C-NMR shifts (CDC13; 6 in ppm):

10.9; 11.6; 14.4; 23.4; 25.0; 28.6; 35.6; 36.3; 45.6;

57.7; 57.8; 67.5; 68.7; 69.2; 73.9; 73.9; 76.1; 81.7;

100.1; 122.9; 126.0; 126.0; 128.2; 128.7; 128.7;

138.0; 140.6; 171.5.

HPLC Rt = 7.0 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 65/35.

Detector: UV 210 nm.

Soraphen J

<C>28<H>44°8

Mz 508

IR (film):

3394; 2939; 2829; 1729; 1461; 1380; 1313; 1270; 1187;

1158; 1100; 1042; 987.

<13>C-NMR shifts (CDC13; 6 in ppm):

11.5; 13.8; 14.3; 22.5; 24.1; 25.6; 25.8; 27.7; 31.6;

35.2; 36.3; 49.3; 57.3; 59.4; 69.6; 70.5; 72.6; 75.7;<* >81.0; 84.6; 98.2; 126.3; 126.3; 128.1; 128.7; 128.7;

141.3; 172.5.

HPLC Rt = 8.1 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen M

<C>28<H>44°9

Mz 524

IR (film):

3396; 2939; 2831; 1725; 1461; 1380; 1270; 1235; 1191;. 1154; 1075; 1048; 994; 971; 898; 850.

<13>C-NMR shifts (CDC13; 6 in ppm):

9.0; 10.4; 11.5; 23.1; 25.3; 28.3; 29.3; 35.1; 35.7;

40.3; 46.5; 57.2; 57.9; 68.8; 68.8; 71.9; 72.9; 74.4;

76.3; 83.8; 99.6; 126.3; 126.3; 128.0; 128.5; 128.5;

141.2; 171.0.

HPLC Rt = 8.5 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen N

<C>29<H>44°9

Mz 536

IR (film):

3444; 2966; 2939; 1731; 1461; 1380; 1309; 1268; 1224;

1164; 1098.

<13>C-NMR shifts (CDC13; 6 in ppm):

13.8; 13.9; 23.3; 23.4; 24.2; 26.3; 26.7; 29.5; 32.1;

36.3; 50.3; 54.5; 57.6; 68.1; 75.2; 78.4; 79.9; 105.7;

125.8; 125.8; 127.5; 128.4; 128.4.

<1>H-NMR shifts (CDC13; 6 in ppm):

0.9d; 1.05 d; 1.24 d; 5.23 d; 5.85 dd.

HPLC Rt = 13.4 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen O

^28^44^<8>

MZ 508

IR (film):

3448; 2939; 2831; 1733; 1459; 1382; 1330; 1270; 1203;

1098; 1048; 987.

<13>C-NMR shifts (CDC13; 6 in ppm):

11.5; 14.5; 23.1; 24.9; 26.2; 27.1; 27.6; 31.7; 35.5;

35.9; 39.5; 50.7; 57.4; 58.7; 69.5; 71.3; 71.6; 74.6;

79.4; 81.9; 102.2; 126.6; 126.6; 127.9; 128.4; 128.4;

137.9; 168.0.

HPLC Rt = 7.7 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Example H-2: Preparation of the compounds of formula I

a) Preculture:

The pre-culture is grown in 2 liter flasks with 500 ml of a

culture medium (containing 0.1% peptone of casein, 0.5% glucose, 0.05% CaCl2-H20, 0.05% MgSO4-7 H20) at pH 7.4 without buffer (or with 50 mM HEPES buffer2) at 160 rpm and 30°C on a shaking table. The most favorable time to transfer to the fermenter culture is achieved after 2-3 days (upper log phase). Instead of the strain Soce 192 used here, one can also use one of the other five strains in the present invention or the strain Soce 26 according to EP-A-282.-455.

b) Fermentation:

A 70 liter fermenter from the company Giovanola Fréres, Monthey,

Switzerland, with 60 1 of the same culture medium inoculated with 10 1 of pre-culture. Fermentation takes place at 30-32°C. The stirring speed is 500 rpm, the aeration speed 0.12 1 per liters of medium and hour. The fermentation duration is 7-14 days. It is ensured that the pH value does not fall below 7.0, especially not below 6.2. The macrocyclic compounds of formula I formed are partly in the culture liquid, partly in the cells. They can be extracted from the cells with alcohols or

<2>HEPES is the K and/or Na salt of 4-(2-hydroxyethyl)-piperazine-1-ethanesulfonic acid.

ketones (e.g. acetone) from the culture liquid with ethyl acetate or butyl acetate. The fermentation liquid is then shaken, e.g. five times with 2 1 ethyl acetate for 5 min. These combined extracts are washed twice with water and concentrated in vacuo. The remaining dark oil can be separated according to the method shown in Figure 2.

It is more advantageous to remove the formed compounds of formula I after completion of the fermentation with adsorption resin from the fermentation flask. In this way, 0.5% v/v fine-grained resin (eg XAD-1180 or XAD-16) is added to the fermentation solution and stirred for 4 hours, after which the compounds of formula I are completely bound to the resin. After separation of the fermenter solution by filtration, the resin is flushed into a glass column, washed with 3 bed volumes of water and eluted with 4 bed volumes of methanol. The eluate is concentrated to dryness in vacuo. Such an eluate or such a crude extract can be formulated with suitable dispersants and/or carriers for professionally applicable pesticides, but the crude extract can also, as shown below, and in the attached Figure 2, be separated to isolate the individual Soraphens A to a. The separation described here was carried out with 65 g of crude extract from a total of 400 1 fermentation solution.

Chromatographic purification (see: separation method II for isolation of Sora<p>henes) 1. Silica gel separation; column: 200 mm 0; 200 cm length. Adsorbent: Lichroprep Si 100; 25-40 µm (manufacturer: Merck).

Fluid: gradient in steps; 1) dichloromethane; 2-9) dichloromethane/acetone 98/2, 96/4, 95/5, 90/10, 85/15, 80/20, 50/50, 25/75; 10-11) dichloromethane/methanol 75/25, 50/50. step 2 1.

14 fractions are taken. Fractions 2, 3, 5, 6, 7, 8, 9, 10, 11 are further purified. 2. Gel chromatography; column: 60 mm 0; 100 cm length. Adsorbent: Sephadex LH-20 (manufacturer: Pharmacia LKB GmbH).

Solvent: methanol.

7 fractions are taken. Fraction 3 contains Soraphen A (2.5 g). 3. Gel chromatography; column: 30 mm 0; 100 cm length. Adsorbent: Sephadex LH-20 (manufacturer: Pharmacia LKB GmbH).

Solvent: methanol.

5 fractions are taken. Fraction 2 contains Soraphen C (6.6 g). 4. Gel chromatography; column: 30 mm 0; 100 cm length. Adsorbent: Sephadex LH-20 (manufacturer: Phar-<* >macia LKB GmbH).

Solvent: methanol.

6 fractions are taken. Fraction 2 (1 g) is further purified. 5. Reversed phase separation; column: 37 mm 0.34 cm length.

Adsorbent: HDSIL RP-18; 18-20-60 15-25 pm (producer: Labomatic).

Solvent: methanol/water 72/28.

9 fractions are taken. Fraction 7 contains Soraphen V (1.1 g). 6. Gel chromatography; column: 30 mm 0; 100 cm length. Adsorbent: Sephadex LH-20 (manufacturer: Pharmacia LKB GmbH).

Solvent: methanol.

4 fractions are taken. Fraction 2 (1.1 g) is further purified.

7. Gel chromatography; column: 30 mm 0; 100 cm length. Adsorbent: Sephadex LH-20 (manufacturer: Phar-

macia LKB GmbH).

Solvent: methanol.

6 fractions are taken. Fractions 2 (0.54 g) and 3 (0.18 g) are further purified. 8. Gel chromatography; column: 30 mm 0; 100 cm length. Adsorbent: Sephadex LH-20 (manufacturer: Pharmacia LKB GmbH).

Solvent: methanol.

6 fractions are taken. Fractions 2 (0.3 g) and 3 (1.3 g) are further purified. 9. Gel chromatography; column: 60 mm 0; 100 cm length. Adsorbent: Sephadex LH-20 (manufacturer: Pharmacia LKB GmbH).

Solvent: methanol.

7 fractions are taken. Fraction 2 (3.3 g) is further purified. 10. Gel chromatography; column: 60 mm 0; 100 cm length. Adsorbent: Sephadex LH-20 (manufacturer: Pharmacia LKB GmbH).

Solvent: methanol.

6 fractions are taken. Fraction 2 (6.1 g) is further purified. 11. Reversed phase separation; column: 37 mm 0.34 cm length.

Adsorbent: HDSIL RP-18; 18-20-60 15-25 um (manufacturer: Labomatic).

Solvent: methanol/water 75/25.

8 fractions are taken. Fraction 5 contains Soraphen D (40 mg), fraction 7 contains Soraphen B (0.5 g). 12. Reversed phase separation; column: 35 mm 0.45 cm length.

Adsorbent: HDSIL RP-18; 18-30-60 25-40 um (manufacturer: Labomatic).

Solvent: methanol/water 80/20.

10 fractions are taken. Fraction 8 contains Soraphen E (30 mg), fraction 4 is further purified. 13. Reversed phase separation; column: 37 mm 0.34 cm length.

Adsorbent: HDSIL RP-18; 18-20-60 15-25 pm (producer: Labomatic).

Solvent: methanol/water 75/25.

10 fractions are taken. Fraction 6 (80 mg) is further purified, fraction 8 (20 mg) is further purified. 14. Reversed phase separation; column: 37 mm 0.34 cm length.

Adsorbent: HDSIL RP-18; 18-20-60 15-25 pm (producer: Labomatic).

Solvent: methanol/water 75/25.

11 fractions are taken. Fraction 5 contains Soraphen X (16 mg). 15. Reversed phase separation; column: 35 mm 0.45 cm length.

Adsorbent: HDSIL RP-18; 18-30-60 25-40 pm (producer: Labomatic).

Solvent: methanol/water 70/30 lin. gradient for 2 h after methanol.

8 fractions are taken. Fraction 5 contains Soraphen U (150 mg). 16. Reversed phase separation; column: 35 mm 0.45 cm length.

Adsorbent: HDSIL RP-18; 18-30-60 25-40 pm (producer: Labomatic).

Solvent: methanol/water 70/30 lin. gradient for 1 h after 90/10.

8 fractions are taken. Faction 8 contains Soraphen M

(0.93 g), fraction 7 (250 mg) is further purified.

17. Reversed phase separation; column: 37 mm 0.34 cm length.

Adsorbent: HDSIL RP-18; 18-20-60 15-25 ym (manufacturer: Labomatic).

Solvent: methanol/water 70/30.

8 fractions are taken. Fraction 4 (105 mg) is further purified, fraction 6 contains Soraphen R (20 mg), fraction 7 contains Soraphen S (490 mg), fraction 8 contains Soraphen T (80 mg), fraction 9 (80 mg) is further purified. 18. Reversed phase separation; column: 37 mm 0.34 cm length.

Adsorbent: HDSIL RP-18; 18-20-60 15-25 ym (manufacturer: Labomatic).

Solvent: methanol/water 7 0/30.

14 fractions are taken. Fraction 7 (700 mg) is further purified, fraction 8 (570 mg) is further purified, fraction 9 contains Soraphen 6 (120 mg). 19. Reversed phase separation; column: 20.5 mm 0.25 cm length.

Adsorbent: RP-18 Nucleosil 100-7 C18; 7 \ im (Producer: Macherey Nagel).

Solvent: methanol/water 67/33.

6 fractions are taken. Fraction 5 contains Soraphen y (11 mg). 20. Silica gel separation; column: 20.5 mm 0.25 cm length. Adsorbent: Si 100 Nucleosil 100-7; 7 jim (Producer: Macherey Nagel).

Gluing agent: tert-butyl methyl ether/n-hexane/+ methanol 1/2/ + 1%.

3 fractions are taken. Fraction 2 contains Soraphen v (8 mg). 21. Silica gel separation; column: 20.5 mm 0.25 cm length. Adsorbent: Si 100 Nucleosil 100-7; 7 nm (manufacturer: Macherey Nagel).

Gluing agent: tert-butyl methyl ether/n-hexane/+ methanol 1/2/ + 1%.

2 fractions are taken. Fraction 2 contains Soraphen £>

(8 mg).

22. Silica gel separation; column: 20.5 mm 0.25 cm length. Adsorbent: Si 100 Nucleosil 100-7; 7 ym (producer: Macherey Nagel).

Gluing agent: tert-butyl methyl ether/n-hexane/+ methanol 1/2/ + 5%.

7 fractions are taken. Fraction 1 contains Soraphen Y (64 mg), fraction 4 contains Soraphen o (5 mg), fraction 6 contains Soraphen £ (7 mg). 23. Reversed phase separation; column: 20.5 mm 0.25 cm length.

Adsorbent: RP-18 Nucleosil 100-7 C18; 7 jim (Producer: Macherey Nagel).

Solvent: methanol/water 60/40.

6 fractions are taken. Fraction 2 (35 mg) is further purified. 24. Silica gel separation; column: 20.5 mm 0.25 cm length. Adsorbent: Si 100 Nucleosil 100-7; 7 ym (producer: Macherey Nagel).

Gluing agent: tert-butyl methyl ether/n-hexane/+ methanol 1/2/ + 10%.

5 fractions are taken. Fraction 2 contains Soraphen o (20 mg). 25. Reversed phase separation; column: 20.5 mm 0.25 cm length.

Adsorbent: RP-18 Nucleosil 100-7 C18; 7 nm (manufacturer: Macherey Nagel).

Solvent: methanol/water 60/40.

9 fractions are taken. Fraction 4 (194 mg) is further purified, fraction 5 (130 mg) is further purified, fraction 6 (50 mg) is further purified. 26. Silica gel separation; column: 20.5 mm©, 25 cm length. Adsorbent: Si 100 Nucleosil 100-7; 7 ym (producer: Macherey Nagel).

Gluing agent: tert-butyl methyl ether/n-hexane/+ methanol 1/2/ + 5%.

9 fractions are taken. Fraction 4 contains Soraphen p (45 mg). 27. Silica gel separation; column: 20.5 mm 0.25 cm length. Adsorbent: Si 100 Nucleosil 100-7; 7 \ im (Producer: Macherey Nagel).

Gluing agent: tert-butyl methyl ether/n-hexane/+ methanol 1/2/ + 10%.

4 fractions are taken. Fraction 2 contains Soraphen

(26 mg).

28. Silica gel separation; column: 20.5 mm 0.25 cm length. Adsorbent: Si 100 Nucleosil 100-7; 7 jim (Producer: Macherey Nagel).

Gluing agent: tert-butyl methyl ether/n-hexane/+ methanol 1/2/ + 5%.

4 fractions are taken. Fraction 2 contains Soraphen \ i (18 mg), fraction 3 contains Soraphen ti (69 mg). 29. Silica gel separation; column: 20.5 mm 0.25 cm length. Adsorbent: Si 100 Nucleosil 100-7; 7 nm (manufacturer: Macherey Nagel).

Gluing agent: tert-butyl methyl ether/n-hexane/+ methanol 1/2/ + 5%.

5 fractions are taken. Fraction 4 contains Soraphen k (5.5 mg). 30. Silica gel separation; column: 20.5 mm 0.25 cm length. Adsorbent: Si 100 Nucleosil 100-7; 7 ym (producer: Macherey Nagel).

Gluing agent: tert-butyl methyl ether/n-hexane/+ methanol 1/2/ + 5%.

4 fractions are taken. Fraction 4 (10 mg) is further purified. 31. Silica gel separation; column: 20.5 mm 0.25 cm length. Adsorbent: Si 100 Nucleosil 100-7; 7 jim (Producer: Macherey Nagel).

Gluing agent: tert-butyl methyl ether/n-hexane/+ methanol 1/2/ + 5%.

2 fractions are taken. Fraction 1 contains Soraphen rc (2.2 mg), fraction 2 contains Soraphen Z (6.5 mg).

Physicochemical characterization of the further produced Soraphenes R to o

Soraphen V

<C>28<H>42°8

Mz 506

IR (film, v in cm'1) :

3394; 2942; 2900; 2829; 1723; 1461; 1380; 1270; 1233;

1189; 1068; 988; 898; 851.

<13>C-NMR shifts (CDC13; 6 in ppm):

10.3; 11.7; 12.6; 23.2; 26.0; 32.7; 35.3; 35.6; 36.2;

46.3; 55.8; 57.3; 68.9; 72.5; 72.9; 74.6; 76.3; 84.5;

99.5; 122.1; 126.2; 126.2; 128.1; 128.5; 128.5; 139.9;

141.2; 170.9.

HPLC Rt = 9.2 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen X

<C>28<H>42°8

Mz 522

IR (film, v in cm"1) :

3404; 2941; 1716; 1598; 1461; 1380; 1272; 1233; 1189;

1156; 1100; 1069; 988.

<13>C-NMR shifts (CDC13; in ppm) :

10.3; 11.7; 12.5; 23.1; 25.9; 29.4; 35.2; 35.6; 35.7;

46.2; 57.3; 57.6; 68.9; 72.5; 74.5; 75.0; 76.1; 83.8;

99.5; 113.4; 115.2; 118.0; 124.9; 129.8; 137.4; 142.7;

156.2; 170.9.

HPLC Rt = 4.4 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen U

<C>29<H>44°9

Mz 536

IR (film, v in cm"1) :

3386; 3361; 2977; 2939; 2892; 2823; 1698; 1461; 1380;

1268; 1185; 1152; 1096; 1064; 1023; 975; 900; 840.

<13>C-NMR shifts (CDC13; 6 in ppm) :

10.1; 11.4; 12.2; 31.8; 32.9; 35.2; 35.2; 38.6; 46.3;

56.0; 57.2; 57.9; 68.7; 72.2; 73.3; 76.2; 80.0; 84.5;

99.3; 122.2; 126.1; 126.1; 127.9; 128.4; 128.4; 140.6;

141.4; 171.7.

HPLC Rt = 5.2 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen R

<C>26<H>40°8

Mz 480

IR (film, v in cm'1) :

3404; 2937; 1731; 1459; 1430; 1384; 1355; 1330; 1270;

1212; 1110; 1083; 1033; 988.

<13>C-NMR shifts (CDC13; 5 in ppm) :

10.7; 14.5; 22.7; 23.3; 24.8; 27.9; 30.9; 31.8; 35.5;

43.4; 58.6; 68.9; 71.0; 72.9; 73.3; 74.6; 80.5; 97.6;

126.5; 128.0; 128.6; 128.6; 141.1; 168.8.

HPLC Rt = 5.1 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel.<* >Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen S

<C>27<H>40°8

Mz 492

IR (film, v in cm"1) :

3431; 2941; 1720; 1604; 1459; 1426; 1380; 1264; 1187;

1150; 1104; 1062; 977.

<13>C-NMR shifts (CDC13; 6 in ppm) :

10.5; 11.9; 12.7; 24.9; 26.2; 35.5; 36.2; 36.3; 37.9;

47.5; 57.6; 69.5; 73.5; 74.1; 75.8; 76.3; 77.9; 100.6;

125.1; 127.1; 128.7; 129.4; 129.4; 138.8; 143.3;

173.8.

HPLC Rt = 5.9 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen T

<C>27<H>42°8

Mz 494

IR (film, v in cm'<1>) :

3427; 2941; 1720; 1461; 1382; 1268; 1191; 1154; 1106;

1068; 994; 971.

<13>C-NMR shifts (CDC13; 6 in ppm) :

10.4; 11.7; 14.2; 22.7; 24.8; 25.3; 28.3; 3 0.6; 34.6;

35.2; 45.8; 57.3; 69.1; 69.8; 73.0; 73.3; 76.0; 76.4;

99.6; 126.5; 126.5; 128.1; 128.5; 128.5; 140.6; 171.9.

HPLC Rt = 6.5 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen 6 (delta)

<C>27<H>42°8

Mz 506

IR (film, v in cm'1) :

3440; 3311; 2937; 1731; 1600; 1461; 1380; 1340; 1185;

1096; 988; 850.

<13>C-NMR shifts (CDC13; 6 in ppm) :

10.2; 12.0; 12.7; 23.2; 25.6; 29.6; 34.9; 36.0; 37.3;

45.3; 56.2; 57.6; 70.3; 72.1; 72.6; 74.3; 74.9; 84.2;

99.5; 124.9; 126.3; 126.3; 128.0; 128.6; 128.6; 131.6;

141.2; 171.2.

HPLC Rt = 7.1 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/3 0.

Detector: UV 210 nm.

Soraphen y (gamma)

<C>28<H>42°9

Mz 522

IR (film, v in cm'1) :

3398; 2939; 1723; 1461; 1382; 1264; 1189; 1152; 1102;

1066; 975; 894.

<13>C-NMR shifts (CDC13; 6 in ppm) :

8.6; 10.3; 11.5; 23.6; 25.8; 28.8; 33.9; 35.4; 36.3;

46.6; 54.7; 57.3; 58.1; 59.2; 68.8; 69.3; 73.5; 74.3;

76.2; 83.5; 99.7; 126.1; 126.1; 128.0; 128.6; 128.6;

141.4; 170.4.

HPLC Rt = 8.4 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Sora<p>hen v (new)

<C>28<H>42°9

Mz 522

IR (film, v in cm'1) :

3402; 2971; 2935; 2827; 1733; 1459; 1367; 1181; 1095;

1050; 990; 860.

<13>C-NMR shifts (CDC13; 6 in ppm) :

8.7; 10.3; 11.5; 26.3; 33.4; 35.2; 40.2; 40.8; 46.2;

57.4; 58.1; 67.8; 68.9; 71.5; 73.5; 76.3; 78.9; 82.6;

99.4; 125.9; 128.3; 128.7; 128.7; 128.8; 129.5; 140.7;

170.3.

HPLC Rt = 8.5 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel.

Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen R (beta)

<C>29<H>44°8

Mz 520

IR (film, v in cm'1) :

3408; 2971; 2935; 2825; 1733; 1459; 1365; 1343; 1181;

1095; 1050; 990; 958; 738.

13C-NMR shifts (CDC13; 5 in ppm) :

10.1; 12.1; 12.6; 23.4; 25.5; 30.4; 35.0; 35.9; 37.2;

45.3; 56.2; 56.3; 57.9; 70.3; 72.1; 72.7; 74.2; 83.1;

84.8; 99.4; 122.6; 126.3; 126.3; 128.0; 128.6; 128.6;

140.0; 141.3; 171.3.

HPLC Rt = 16.8 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen Y

<C>27<H>40°9

Mz 508

IR (film, v in cm"1) :

3436; 2937; 1721; 1459; 1376; 1268; 1199; 1114; 1048;

988; 961.

<13>C-NMR shifts (CDC13; in ppm) :

10.8; 11.2; 18.8; 23.0; 23.5; 31.7; 33.4; 36.2; 40.6;

41.1; 51.1; 61.1; 70.8; 74.6; 76.4; 77.1; 78.7; 86.4;

99.9; 125.5; 125.5; 127.4; 128.4; 128.4; 142.5; 171.1;

212.4.

HPLC Rt = 6.1 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen o (omicron)

<C>29<H>44°9

Mz 536

IR (film, v in cm"1) :

3402; 3361; 2933; 2894; 1708; 1457; 1360; 1269; 1187;

1150; 1096; 1062; 975; 898; 838; 765.

<13>C-NMR shifts (CDC13; 6 in ppm) :

10.4; 11.6; 12.6; 31.7; 33.4; 35.3; 35.7; 38.7; 46.3;

56.3; 57.4; 58.3; 65.7; 69.0; 72.3; 74.2; 76.2; 80.0;

84.8; 99.5; 122.9; 126.3; 126.3; 128.2; 128.6; 128.6;

140.1; 170.8.

HPLC Rt = 4.8 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen C (ksi)

<C>27<H>40°8

Mz 492

IR (film, v in cm'<1>) :

3402; 2969; 2933; 1731; 1459; 1367; 1179; 1095; 1050;

990.

<13>C-NMR shifts (CDC13; 6 in ppm) :

10.4; 11.7; 12.5; 20.1; 26.2; 31.8; 35.2; 35.7; 36.2;

46.4; 57.4; 68.9; 72.3; 73.5; 75.0; 76.1; 76.3; 99.6;

126.2; 126.2; 127.0; 128.1; 128.6; 128.6; 138.3;

141.2; 170.7.

HPLC Rt = 4.1 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Sora<p>hen o (sigma)

<C>29<H>44°9

Mz 536

IR (film, v in cm"1) :

3398; 3363; 3301; 2971; 2933; 2894; 1702; 1453; 1378;

1262; 1167; 1150; 1100; 979; 838.

<13>C-NMR shifts (CDC13; 6 in ppm) :

10.3; 11.5; 12.4; 31.6; 34.2; 35.0; 35.5; 38.9; 46.2;

56.1; 57.5; 58.5; 64.3; 69.1; 73.1; 76.1; 77.2; 79.8;

85.2; 99.7; 121.8; 126.0; 126.0; 128.0; 128.5; 128.5;

141.9; 142.0; 171.4.

HPLC Rt = 6.2 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen p (calm)

<C>27<H>42°9

Mz 510

IR (film, v in cm"1) :

3398; 3363; 3301; 2971; 2933; 2894; 1700; 1453; 1378;

1262; 1187; 1150; 1100; 1064; 979; 898; 838; 765.

13C-NMR shifts (CDC13; 6 in ppm):

8.7; 10.3; 11.4; 23.1; 25.5; 26.4; 32.3; 35.0; 36.1;

40.5; 46.6; 57.2; 68.5; 68.8; 71.8; 73.1; 73.7; 74.3;

76.4; 99.7; 126.3; 127.9; 128.5; 128.5; 141.4; 171.4.

HPLC Rt = 4.7 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen S (zeta)

<C>28<H>42°9

Mz 522

IR (film, v in cm'1) :

3398; 2937; 1718; 1457; 1378; 1332; 1272; 1203; 1102;

978.

<13>C-NMR shifts (CDC13; in ppm) :

11.8; 21.9; 25.9; 25.9; 28.7; 36.3; 39.9; 43.0; 45.7;

56.7; 57.4; 58.6; 70.6; 75.5; 76.2; 78.2; 78.2; 84.3; <* >98.9; 125.9; 128.0; 128.4; 128.7; 133.1; 141.2; 171.6.

HPLC Rt = 3.5 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen \ i (my)

<C>28<H>44°9

Mz 524

IR (film, v in cm"1) :

3423; 2939; 1725; 1459; 1380; 1264; 1191; 1154; 1102;

1052; 992; 971.

<13>C-NMR shifts (CDC13; 6 in ppm) :

10.4; 11.7; 14.1; 21.3; 27.7; 28.2; 30.9; 32.9; 34.1;

35.2; 45.6; 57.4; 60.7; 69.1; 70.0; 70.4; 72.1; 76.2;

76.3; 85.4; 99.6; 126.6; 126.6; 128.2; 128.6; 128.6;

140.2; 172.0.

HPLC Rt = 4.7 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen n (eta)

<C>28<H>42°9

Mz 522

IR (film, v in cm'1) :

3411; 2935; 1718; 1459; 1382; 1266; 1189; 1152; 1104;

1066; 981.

13C-NMR shifts (CDC13; 5 in ppm) :

10.2; 11.4; 12.3; 31.6; 32.9; 35.1; 35.4; 38.1; 46.2;

57.2; 57.7; 64.0; 68.8; 72.2; 73.7; 74.7; 76.2; 81.2;

99.3; 124.8; 126.1; 126.1; 127.9; 128.4; 128.4; 137.8;

141.1; 171.4.

HPLC Rt = 4.6 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/3 0.

Detector: UV 210 nm.

Soraphen k (coat)

<C>27<H>40°8

Mz 492

IR (film, v in cm'1) :

3411; 2935; 1731; 1461; 1382; 1347; 1270; 1189; 1104;

1048; 990.

<13>C-NMR shifts (CDC13; 6 in ppm) :

10.1; 12.1; 12.8; 23.4; 25.4; 33.5; 35.0; 36.1; 37.2;

45.4; 56.1; 70.3; 72.0; 72.7; 74.2; 74.2; 75.6; 99.4;

124.1; 126.2; 126.2; 127.9; 128.5; 128.5; 138.6;

141.3; 171.5.

HPLC Rt = 4.7 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen ti (pi)

<C>27<H>40°8

Mz 492

IR (film, v in cm'1) :

3396; 3363; 3299; 2933; 2694; 1702; 1451; 1378; 1262;

1167; 1150; 1100; 979.

<13>C-NMR shifts (CDC13; 6 in ppm) :

10.3; 12.3; 14.6; 23.4; 25.0; 32.2; 35.6; 36.0; 36.8;

50.4; 55.9; 70.2; 71.2; 73.9; 75.0; 75.5; 98.3; 125.2;

126.2; 126.2; 128.1; 128.6; 128.6; 137.8; 141.0;

171.5.

HPLC Rt = 4.9 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

Soraphen Z

<C>28<H>42°9

Mz 522

IR (film, v in cm'1) :

3405; 2937; 1718; 1459; 1380; 1266; 1189; 1152; 1104;

1069; 987; 902; 850; 813; 738.

<13>C-NMR shifts (CDC13; 5 in ppm) :

10.5; 11.7; 12.6; 31.8; 33.5; 35.3; 35.7; 41.1; 46.3;

56.0; 57.3; 66.0; 69.0; 70.5; 72.5; 74.3; 76.2; 84.3;

99.5; 122.4; 126.2; 126.2; 128.2; 128.6; 128.6; 140.4;

140.9; 170.9.

HPLC Rt = 7.7 min.

Column: 4 x 250 mm Nucleosil 100-7 C18, Macherey Nagel. Flow rate: 1.5 ml/min.

Solvent: methanol/water 70/30.

Detector: UV 210 nm.

On the basis of X-ray structure analysis of Soraphen A and the data obtained above, it is assumed that the Soraphenes C to Q have the following spatial structures IC to IQ<1>. It is assumed that the remaining connections IR<1>, IS<1>, IT<1>, IU<1>, IV, IX', IY', IZ<1>, IB1 , ly', I5<1>, IC <1>, ItT, Ik • , I|i'f 1$' and lp<1> have similar structures.

For Soraphen Iv 1, the following structure is probable:

For the soraphenes o, u and o, the structure is not certain. Furthermore, it is clear that some Soraphenes are mutually epimeric. Soraphen 6 is epimeric with Soraphen C; Soraphenes k and £ are mutually epimeric and with Soraphen S, so are Soraphenes U, o and o. Binding in each case are the measurement values obtained for a particular compound of formula I, which currently does not allow any string to - scheme to a specific structure, which the person skilled in the art knows.

As mentioned, the invention encompasses all stereoisomeric forms of the individual listed compounds with formula I as well as their preparation and their use for combating or prevention of plant diseases. Furthermore, however, the invention in particular also covers the compounds IC<*> to Io<1> on the basis of their physicochemical data, for which the above-mentioned structure is assumed.

It is worth noting that the macrocyclic soraphenes of formula I usually exist in the indicated hemiacetal form, however, this form can undergo reversible ring-opening according to the scheme

depending on the manufacturing or processing methodology, depending on pH value and solvent, Soraphenene occurs in one form or another or as a mixture of both forms. Characteristic of the ring opening is the shift of the <13>C-NMR signal in the 3-position and the shift of the <1>H-NMR signals in certain other positions. At Soraphen A, e.g. following changes: <13>C-NMR (CDC13, 6 in ppm) : 99.5-203.1 (3-C). <1>H-NMR (CDCl 3 , 6 in ppm): 3.14-3.72 (2-H); 3.18-4.5 (4-H); 3.83-3.16 (7-H); 5.86-5.7 (17-H). Similar shifts are also observed with Soraphenene B to p. Formula I in the present invention in principle comprises both the 3-hemiacetal form preferred at lower pH values and also the opened 3-keto-7-hydroxy form.

2. Formulation examples for one or more of the active substances - with formula I (% = percentage by weight)

In the following, active substance means one of the Soraphenes in pure form or several of the Soraphenes C to o or an eluate obtained from the microbiological preparation, in liquid form or as a dry concentrate containing the unseparated total amount of Soraphenes.

2.1 Emulsion concentration methods

Such concentrates can be diluted with water to produce emulsions of any desired concentration.

2.2 Solutions

The solutions are suitable for application in the form of very small drops.

2.3 Granules

The active substance is dissolved in methylene chloride, sprayed onto the carrier substance, and the solvent is then evaporated in a vacuum.

2.4 Atomizing agent

By thoroughly mixing the carrier substances with the active substance, ready-to-use nebulizers are obtained. These can be ground to a ready-to-use dust with 0.001% active ingredient by further addition of the three carrier substances.

2.5 S spray powder

The active substance is mixed well with the additives and ground in a suitable mill. Spray powder is obtained which can be diluted with water to form suspensions of any desired concentration.

tration.

2.6 Encapsulation granules

The finely ground active substance is applied evenly distributed in a mixer onto the kaolin which is moistened with polyethylene glycol. In this way, dust-free encapsulation granules are obtained.

2.7 Suspension concentrate

The finely ground active substance is mixed well with the additives. A suspension concentrate is thus obtained from which, by diluting with water, suspensions of any desired concentration can be prepared.

2.8 Biomass concentrate

The biomass that falls out in the production culture is dried, ground and mixed in a weight ratio of 80:20 with ethylene glycol monomethyl ether. Such a concentrate can be diluted arbitrarily with water for spray suspensions. 3. Biological examples of plants (In the following, "active ingredient" means one of the Soraphenenes C to a according to the present invention).

Example 3. 1: Effect against Puccinia graminis on wheat

a) Post-protective effect

Wheat plants are sprayed 6 days after sowing with a

spray solution made from spray powder with active substance (0.02% active substance). After 24 hours, the treated plants are infected with a uredospore suspension of the fungus. After incubation for 48 hours at 95-100% relative humidity and approx. 20°C, the infected plants are placed in a greenhouse at approx. 22°C. The assessment of rust blister development takes place 12 days after infection.

b) Systemic effect

For wheat plants, 5 days after sowing one is poured

spray solution which is made from spray powder with active substance (0.002% active substance with regard to the soil volume). After 48 hours, the treated plants are infected with a uredospore suspension of the fungus. After incubation for 48 hours at 95-100% relative humidity and approx. 20°C, the infected plants are placed in a greenhouse at approx. 22°C. The assessment of rust blister development takes place 12 days after infection.

In both experiments, the fungal attack was completely inhibited by the active substance.

Untreated but infected control plants, on the other hand, showed a Puccinia attack of 100%.

Example 3. 2: Effect against Phytophthora on tomato plants

a) Post-protective effect

Tomato plants were sprayed after cultivation for 3 weeks with a

spray solution that was made from spray powder with active substance (0.02% active substance). After 24 hours, the treated plants were infected with a sporangia suspension of the fungus. The assessment of the fungal attack took place after incubation of the infected plants for 5 days at 90-100% relative humidity and 20°C.

b) Systemic effect

For tomato plants, after 3 weeks of cultivation, it became one

spray solution which was prepared from spray powder with active substance (0.006% active substance with regard to the soil volume). This ensured that the spray solution did not come into contact with the plant parts that were above ground. After 48 hours, the treated plants were infected with a sporangia suspension of the fungus. The assessment of the fungal attack took place after incubation of the infected plants for 5 days at 90-100% relative humidity and 20°C.

In both trials, no fungal attack was observed during the assessment, while the infected control plants were completely attacked.

Example 3. 3: Effect against Plasmopara viticola on vines Post-protective effect

At the 4-5 leaf stage, vine plants are sprayed with a spray solution made from spray powder with active substance (0.006% active substance). After 24 hours, the treated plants are infected with a sporangia suspension of the fungus. After incubation for 6 days at 95-100% relative humidity and 20°C, the fungal attack is assessed.

In contrast to the untreated but infected control plants with 100% fungal attack, the plants treated with active substance I were attack-free.

Example 3. 4: Effect against Cercospora arachidicola on groundnut plants

Post-protective effect

10-15 cm tall peanut plants are sprayed with a spray solution made from spray powder with active substance (0.006% active substance) and are infected 48 hours later with a conidia suspension of the fungus. The infected plants are incubated for 72 hours at approx. 21°C and high humidity and then placed in a greenhouse until typical leaf spots appear. The assessment of the fungicidal effect takes place 12 days after the infection on the basis of the number and size of the appearing spots.

The plants that had been treated with active substance I were free of attack. The plants that had been treated with Soraphen A, also using a spray concentration of 0.002% at the end of the assessment, did not show any attack. Untreated but infected control plants, on the other hand, showed a Cercospora attack of 100%.

Example 3. 5: Effect against Venturia inaequalis on apple shoots Post-protective effect

Apple cuttings with 10-20 cm long fresh shoots are sprayed with a spray solution made from spray powder with active substance (0.02% active substance). After 24 hours, the treated plants are infected with a conidia suspension of the fungus. The plants are then incubated for 5 days at 90-100% relative humidity and placed for a further 10 days in a greenhouse at 20-24°C. Scabies infestation is assessed 15 days after infection.

The cuttings that had been treated with active substance I were free of attack, the control plants, on the other hand, were completely attacked.

Example 3. 6: Effect against Botrytis cinerea on apple fruits Post-protective effect

Artificially damaged apples are treated as a spray solution made from spray powder with active substance (0.02% active substance) is added drop by drop to the damaged areas. The treated fruits are then inoculated with a spore suspension of the fungus and incubated for 1 week at high humidity and approx. 20°C. During the evaluation, the rotted damage sites are counted, and the fungicidal effect of the test substance is derived from this.

The active ingredient completely inhibited fungal growth. Untreated but infected control plants, on the other hand, showed 100% Botrytis attack.

Example 3. 7: Effect against Erysiphae graminis on barley

a) Post-protective effect

Barley plants approximately 8 cm high are sprayed with a spray solution

which is made from spray powder with active substance (0.006% active substance). After 3-4 hours, the treated plants are pollinated with conidia from the fungus. The infected barley plants are placed in a greenhouse at approx. 22°C, and the fungal attack is assessed after 10 days.

b) Systemic effect

To approx. 8 cm tall barley plants are poured with a spray solution which

is produced from spray powder with active substance (0.002% active substance with regard to the j word volume). This ensured that the spray solution did not come into contact with the plant parts that were above ground. After 48 hours, the treated plants are pollinated with conidia from the fungus. The infected barley plants are placed in a greenhouse at approx. 22°C, and the fungal attack is assessed after 10 days.

In both trials, the plants were attack-free, the control plants completely attacked.

Example 3. 8: Effect against Rhizoctonia solani (earthworm on rice plants)

Local protective soil application

12-day-old rice plants are watered with a spray solution made from a formulation of the active substance (0.02% active substance), without contaminating plant parts that are above ground. To infect the treated plants, a suspension of mycelium and sclerotia of R. solani is added to the soil surface. After 6 days of incubation at 27"C (day) or 23°C (night) and 100% relative humidity (humidity box) in the climate chamber, the fungal attack on the leaf sheath, leaf and stem is assessed.

After treatment with the active ingredient, no attack occurred, while the infected control plants showed complete Rhizoctonia attack.

Claims (9)

1. Macrocyclic compound characterized by formula I where the macrocyclic ring is either saturated or optionally contains in the 8,9-position or in the 9,10-position or in the 14,15-position a double bond after the following substituent combination, and the substituents R!-R5, A, B, X and Y exists in the following combined meanings: 2. Compound according to claim 1, characterized in that it is compound IY and the compound lp (=ro) in pure form. 3. Procedure for the preparation of compounds of formula I where the macrocyclic ring is either saturated or optionally contains in the 8,9-position or in the 9,10-position or in the 14,15-position a double bond after the following substituent combination, and the substituents R!-R5, A, B, X and Y exists in the following combined meanings: including their stereoisomers, characterized by aerobic cultivation of one of the strains of choice: Sorangium cellulosum "So ce 26" (NCIB 12411), Sorangium cellulosum "So ce 139" (DSM 5397), Sorangium cellulosum "So ce 170" (DSM 4795) , Sorangium cellulosum "So ce 191" (DSM 4796), Sorangium cellulosum "So ce 192" (DSM 4797), Sorangium cellulosum "So ce 231" (DSM 5393) or Sorangium cellulosum "So ce 242" (DSM 5414) in a suitable nutrient medium containing at least one assimilable C source and N source as well as corresponding inorganic salts, at 10-40°C in the presence or absence of an absorption resin, subsequent absorption of the culture solution or the filtered off absorption resin with a suitable solvent phase, concentration of the obtained solution, if desired purification by chromatography or recrystallization and if desired subsequent separation of the obtained compounds of formula I, with the proviso that the strain "So ce 26" (NCIB 12411) not used for the production of compounds IA and IB. 4. Method according to claim 3, characterized in that the fermentation is carried out at 10-35°C with one of the strains "So ce 170" (DSM 4795), "So ce 191" (DSM 4796), "So ce 192" (DSM 4797). 5. Means for combating or prevention of plant diseases characterized by the fact that they consist of culture media for agrochemical purposes containing process products according to the method in claim 3. 6. Means for combating or prevention of plant diseases, characterized by the fact that they consist of agrochemically usable raw extracts from the process according to claim 3. 7. Means for combating or prevention of plant diseases, characterized in that it contains as active substance a process product of formula I from the method according to claim 3, with the exception of compounds IA and IB. 8. Biological pure culture characterized by the fact that it is chosen from among Sorangium cellulosum "So ce 139" (DSM 5397), Sorangium cellulosum "So ce 170" (DSM 4795), Sorangium cellulosum "So ce 191" (DSM 4796), Sorangium cellulosum "So ce 192" (DSM 4797), Sorangium cellulosum "So ce 231" (DSM 5393) and Sorangium cellulosum "So ce 242" (DSM 5414), which are capable of producing at least one compound of formula I according to claim 1. 9. Use of the general process products obtained according to the method according to claim 3 or their stereoisomers for combating or prevention of plant diseases, with the exception of compounds IA and IB.