Actinomyces strain streptomyces lateritious 19/97-m used for stimulating growth and protecting coniferous seedling against disease pathogen caused by fungi of genus fusarium and alternaria

FIELD: microbiology, agriculture.

SUBSTANCE: actinomyces strain Streptomyces lateritius 19/97-M is isolated from tree nursery soil (56°04' North latitude; 92°42 East longitude) in Krasnoyarsk region in 1997 year and deposited in All-Russian Collection of Industrial Microorganissms, FGUPSosNIIgenetics at number VKPM Ac-1637. The actinomyces strain Streptomyces lateritius 19/97-M VKPM Ac-1637 is used for stimulation of growth and protection of coniferous seedlings against pathogens of vascular mycosis. The strain shows high competition ability and growth rate. Invention provides rapid biomass accumulation and high reproductive capacity. Invention can be used for stimulation of growth and protection coniferous seedlings against pathogens of diseases caused by fungi of genus Fusarium and Alternaria.

EFFECT: enhanced effectiveness and valuable properties of strain.

1 tbl, 6 dwg, 3 ex

 

The invention relates to the field of Microbiology, relates to a new strain actinomycete Streptomyces lateritius 19/97-M and can be used to stimulate growth and protect seedlings from agents of vascular mycoses caused by fungi of the genera Fusarium and Alternaria.

In the practice of plant protection are fungi of the genus Trichoderma [1;2]. Actinobacteria, forming antibiotic substances, recommended only for the protection of agricultural plants [3;4] and are rarely used in practice, protection of seedlings of tree species [5]. The lack of work on the selection of strains and the use of this group of microorganisms for the protection of seedlings of tree species constrains their potential in obtaining healthy nursery stock material.

Institutions and development: the Uzbekistan Academy of Sciences and Institute of Microbiology and Virology, Academy of Sciences of the Kazakh SSR recommended accordingly strains-producers of Actinomyces netropsis 1592 and 2129 and Streptomyces griseoruber 3-79 No. 314, with a broad spectrum of activity towards the pathogens of vegetable crops [3;4].

Currently microbiological industry not mastered the production of biological preparations on the basis of active strains of actinomycetes to protect seedlings from agents of vascular mycosis.

The technical result of the invention is a strain actinomycete Streptomyces lateritius 19/97-M PMBC As-1637, with high antagonistical the th activity against phytopathogenic micromycetes genera Fusarium and Alternaria - pathogens vascular lodging and stimulating the growth of coniferous seedlings.

The technical result is achieved by the use of strain actinomycete Streptomyces lateritius 19/97-M PMBC As-1637 to stimulate growth and protection of coniferous seedlings from agents of vascular mycosis. Feature of the strain is high competitiveness and growth rate. Recommended strain is characterized by a rapid accumulation of biomass and high reproductive capacity.

Offered the use of this strain as bioproducts for creation on its basis of environmentally friendly biological products.

The strain isolated from the soil of the nursery (56°04/n; 92°42/C. D.) Krasnoyarsk territory in 1997

The strain is characterized by the following features.

Morphological features: solid nutrient medium krakhmalopatochny agar (KAA) forms a richly branched mycelium, partially disintegrating into fragments. Vegetative hyphae with a diameter of 0.5-2.0 μm (figure 1). Aerial mycelium in the Mature state is a chain of three or four stationary dispute. During growth on solid nutrient media forms a relatively slow-growing colonies that reach 2-4 mm in diameter on day 5-7 (27±2 °). Colonies are thick, leathery, correctly rounded, velvety plaque on the surface appears to 12-14 with the TCI cultivation. On glycerinated agar color of colonies varies from red to dark Burgundy (figure 2). Growth in liquid nutrient medium is accompanied by the formation of pink-red globules with a diameter of 2-3 mm, gram stain positive, aerobe, chemoorganotrophs.

Cultural characteristics: under cultivation on solid nutrient medium (oatmeal agar) for 7 days at 27±2° strain produces 3900 ± 200 colony-forming elements with the area of 100 mm2(1 cm2). In submerged cultivation on chromoleana environment within 7 days strain produces 2.5 g/l of biomass. When surface cultivation on chromoleana environment at 25°With the productivity of the strain is 1.13±0.326 g/l on the 7th day, 1.75±0.118 g/l at 14 days and 2.08±0.15 g / l on the 21st day of cultivation; the biomass growth of strain 0.15±0.05 g/l/day after 7 days, 0.12±0.01 g/l/day after 14 days, 0.10±0.01 g/l/day over 21 days.

Chromoleana environment, g/l: (NH4)2SO4- 1; K2HPO4- 1; NaCl - 1; MgSO4- 1; starch - 1, agar-agar - 15, water - 1000.

According to the experts of the Krasnoyarsk regional veterinary laboratory, this strain of avirulent, non-toxic, netoxygen in relation to warm-blooded organisms and does not cause infectious diseases.

Figure 1-5 presents the following is e illustrations:

Figure 1. The morphology of cells of the strain Streptomyces lateritius 19/97 - M on chromoleana agar.

Figure 2. The morphology of colonies of the strain Streptomyces lateritius 19/97 - M on glycerinated agar.

Figure 3. The growth inhibition of Fusarium lateritium (a) and Fusarium avenaceum (B) metabolites of Streptomyces lateritius strain 19/97)

Fig 4. Antagonistic activity of Streptomyces lateritius strain 19/97)

1 - Fusarium moniliforme, 2 - F.avenaceum, 3 - F.oxysporum, 4 - F.solani, 5 - F.nivale, 6 - F.gibbosum, 1 - F.semitectum, 8 - F.sambucinum, 9-F.sporotrichioides, 10 - F.heterosporum, 11 - F.lateritium, 12 - F.chlamydosporum, 13 - Alternaria alternata.

Figure 5. Stimulating growth of seedlings of Larix sibirica L. 10 day 1 - control; 2 - processing Streptomyces lateritius strain 19/97)

Implementation using Streptomyces lateritius strain 19/97-M as the product of a biological product to promote growth and protection of coniferous seedlings from lodging.

Example 1. Technology in-depth periodic cultivation involves several steps:

Sowing Streptomyces lateritius strain 19/97-M produce culture at the rate of 1.5-2×106propagules per 1 ml of culture medium. As inoculum using a culture of the strain grown on stubble chromoleana agar. From tubes do flush in flasks with sterile water and then determine the titer of propagules in the resulting suspension method brida and the method of conversion for the entire volume of the nutrient medium using the required amount of suspension to be planted. Sowing produced in the bulb it is aera volume of 250 ml per 100 ml of culture medium. For maximum sporulation growing produce for 7 days in liquid chromoleana medium of the following composition: (NH4)2SO4- 1; K2HPO4- 1; NaCl - 1; MgSO4- 1; starch - 1, tap water is 1000.

After the cultivation, the biomass is separated from the nutrient medium, dried to a constant weight and ground in a homogenizer; in this form of biological product is missing nutrient medium, on which grew actinomycetes.

The resulting biological product is a dry powder light grey with a title propagules of 5.5-6×1012propagules/g

Example 2. A culture of Streptomyces lateritius strain 19/97-M solid media inoculated in liquid chromoleana nutrient medium and cultured at 27±2°C. After 7 days of cultivation take samples of the culture fluid, which is filtered through membrane filters with a pore diameter of not more than 0.5-1 μm. Then the culture fluid moistened with sterile paper disks with a diameter of 5 mm, Dried disks are laid on a nutrient medium in Petri dishes, pre-seeded "lawn" of test organisms: Fusarium moniliforme, F.avenaceum, F.oxysporum, F.solani, F.nivale, F.gibbosum, F.semitectum, F.sambucinum, F.sporotrichioides, F.heterosporum, F.lateritium, F.chlamydosporum, Alternaria alternata. Cup incubated in a thermostat at 27±2°C for 5-7 days. Control are paper disks, see the Chennai sterile water.

The lack of growth of the test organism around the paper disk indicates the presence of the antibiotic properties actinomycete Streptomyces lateritius strain 19/97-M (figure 3). The ordinate axis lay a zone of no growth of the test organism in mm. So, all of the investigated pathogens vascular mycosis of coniferous seedlings had medium and high sensitivity to the action of metabolites of strain (figure 4).

Example 3. Sowing Streptomyces lateritius strain 19/97-M produce culture at the rate of 1.5-2×106propagules per 1 ml of culture medium. As inoculum using a culture of the strain grown on stubble chromoleana agar. From tubes do flush in flasks with sterile water and then determine the titer of propagules in the resulting suspension method brida and the method of conversion for the entire volume of the nutrient medium using the required amount of suspension to be planted. Sowing produced in the Erlenmeyer flask with a volume of 250 ml per 100 ml of culture medium. For maximum sporulation growing produce for 7 days in liquid chromoleana medium of the following composition: (NH4)2SO4- 1; K2HPO4- 1; NaCl - 1; MgSO4- 1; starch - 1, tap water is 1000.

After the cultivation, the biomass is separated from the nutrient medium, dried to a constant weight and ground in a homogenizer; when TA is th form of a biological product is missing nutrient medium, which grew actinomycetes. Crops Larix sibirica L., Pinus sylvestris L., Picea obovata L. treated with the suspension of the strain containing 10 mg of biomass per 1 ml at a rate of 10 ml per 100 seeds. Seedlings grown in the laboratory under continuous illumination 32 LIGHTS in the clay within 21 days of the growing season. Conduct evaluation of seed vigor on the 7th day of the growing season, seed germination at 21 days of vegetation.

The increase in energy of germination and germination of seeds indicates the allocation of strain metabolites that stimulate the growth processes in seedlings of Larix sibirica L. (5), Pinus sylvestris L., Picea obovata L. (table).

Thus, the strain Streptomyces lateritius 19/97-M can be used as a producer of a biological product for the biological control of pathogens vascular mycosis of the genera Fusarium and Alternaria and growth of coniferous seedlings.

Sources of information

1. Tretyakov A.P. Shcherbakov GY, Shirmanova NI Strain micromycete Trichoderma lignorum for the production of drug against phytopathogenic fungi: Pat. 2034468 Russia, MKI6A 01 N 63104, C 12 N 1/14 - No. 93028450/13. Appl. 21.06.93, epubl. Bulletin no.13.

2. Thunderous TI, Shmarlovskaya SV, Tyulpanov, VA, Thunderous B.C. Strain of the fungus Trichoderma sp. MG-97 used to protect seedlings from coniferous fusariosis. Patent for invention No. 2171580. M.10.08.2001, - 12 C.

3. Askarov C. Actinobacteria-EN the antagonists of phytopathogenic microorganisms and their role in the resistance of cotton to Verticillium wilt. Autoreplies.... Prof. biolay, Tashkent, 1970, - 45 C.

4. Tulemisova K.A. Igina E.F., Nikitina ET Strain Streptomyces griseoruber 3-79 No. 314 producer of the antibiotic to fight with the mucous bacteriosis of cabbage / Auth. mon. The USSR №1042.349. - Inst. of Microbiology and virus. The Academy of Sciences of the Kazakh SSR. 16.05.1983, - 8 C.

5. R.K. Dumroese, R.L. James, Wenny D.L. Interaction among Streptomyces griseoviridis, Fusarium root disease and Douglas-fir seedlings // New Forest, 1998. - 15. - P.181-191.

Strain actinomycete Streptomyces lateritius PMBC Ac-1637 used for growth promotion and protection of coniferous seedlings from diseases caused by fungi of the genera Fusarium and Alternaria.



 

Same patents:

FIELD: food processing industry, in particular cannery industry.

SUBSTANCE: claimed method includes potato washing, inspection, peeling, post-peeling, cutting, and blanching to obtain half-finished product, pre-packing in vacuum into packets from duplex or laminated film followed by sealing and pasteurization. Prior post-peeling potato is treated with preparation, obtained from biomass of Pythium catenulatum micromycete by sequential extraction with non-polar extractant in above-critical state, water, alkali, water, acid, water, alkali, and water and blending of the first extract with solid precipitate in amount of 0.7-1.105 mg/t and conditioned by approximately 5 hours.

EFFECT: garnish potato of improved organoleptic properties.

FIELD: food processing industry, in particular cannery industry.

SUBSTANCE: claimed method includes potato washing, inspection, peeling, post-peeling, cutting, and blanching to obtain half-finished product, pre-packing in vacuum followed by sealing and pasteurization. Prior post-peeling potato is treated with preparation, obtained from biomass of Mortierella hygrophila micromycete by sequential extraction with non-polar extractant in above-critical state, water, alkali, water, acid, water, alkali, and water and blending of the first extract with solid precipitate in amount of 0.7-1.105 mg/t and conditioned by approximately 5 hours.

EFFECT: garnish potato of improved organoleptic properties.

FIELD: food processing industry, in particular cannery industry.

SUBSTANCE: claimed method includes potato washing, inspection, peeling, post-peeling, cutting, and blanching to obtain half-finished product, pre-packing in vacuum into packets from duplex or laminated film followed by sealing and pasteurization. Prior post-peeling potato is treated with preparation, obtained from biomass of Mortierella minutissima micromycete by sequential extraction with non-polar extractant in above-critical state, water, alkali, water, acid, water, alkali, and water and blending of the first extract with solid precipitate in amount of 0.7-1.105 mg/t and conditioned by approximately 5 hours.

EFFECT: garnish potato of improved organoleptic properties.

FIELD: immunology, proteins.

SUBSTANCE: invention proposes a method for preparing affinity surfaces based on chemically cross-linked staphylococcus protein A. Method involves applying a layer of staphylococcus protein A on hydrophobic surface followed by chemical cross-linkage of protein A. Also, method involves additional applying specific antibodies on the prepared affinity surface with a layer of chemically cross-linked staphylococcus protein A. This method provides preparing the affinity surface eliciting the enhanced strength that can be separated from a backing if necessary and with retaining its integrity. Also, the flat and uniform surface of layer of crossed-linked staphylococcus protein A provides low level of interferences in carrying out study by different methods of microscopy. Invention can be used in investigation of biological objects by different methods, among them by method of atomic-force microscopy. Invention can be used in immunological researches of biological objects.

EFFECT: improved preparing method.

3 dwg, 3 ex

FIELD: veterinary virology.

SUBSTANCE: the virus strain of bursal infectious disease (IBB) "52/70-M" is isolated from bursas of Fabricius of sick and killed chickens. The strain shows the expressed precipitating and infectious activity. Invention can be used for producing inactivated vaccine against IBB virus and for evaluation of properties of vaccines against IBB virus.

EFFECT: valuable properties of strain.

4 tbl, 3 ex

FIELD: biotechnology, microbiology, vitamins.

SUBSTANCE: method relates to a method for preparing riboflavin by culturing the microorganism Bacillus subtilis as a producer in the nutrient medium containing rib-operon from Bacillus amyloliquefaciens, or microorganism able for utilization of glycerophosphate as a single carbon source, or eliciting the resistance against inhibition of growth by glyoxylate, and extraction of riboflavin prepared. Invention uses the following strains as producers of riboflavin: B. subtilis GM51/pMX45, B. subtilis GM41/pMX45, B. subtilis GM44/pMX45. Invention provides preparing riboflavin of the high degree of effectiveness.

EFFECT: improved preparing method.

5 cl, 4 ex

FIELD: biotechnology, microbiology, amino acids.

SUBSTANCE: invention proposes the strain Enterobacter agglomerans/FERM BP-7207 that is able to metabolize the carbon source at pH value when L-glutamic acid precipitates in liquid cultural medium containing L-glutamic acid in the saturation concentration and the carbon source. Also, the strain is able to accumulate L-glutamic acid in the amount exceeding the amount that corresponds to its saturation concentration in liquid cultural medium at this pH value. Also, invention relates to a method for preparing L-glutamic acid by fermentation that involves culturing this microorganism in liquid cultural medium wherein pH value is brought about to the value when L-glutamic acid precipitates to prepare and accumulate L-glutamic acid and to precipitate L-glutamic acid in this cultural medium. The proposed microorganism is prepared by addition of a sample comprising microorganisms to acid cultural medium containing L-glutamic acid in the saturation concentration and the carbon source followed by removal of strain that is able to metabolize this carbon source. Invention provides preparing L-glutamic acid with the high degree of effectiveness.

EFFECT: improved preparing method, valuable properties of strain.

20 cl, 9 dwg, 3 tbl, 8 ex

FIELD: food-processing industry.

SUBSTANCE: method involves preparing and rubbing beet; sterilizing resultant pulp and fermenting while providing combined cultivation thereon of mycelium fungi of Trichoderma kind and of Aspergillus kind of citric acid fermentation; separating cultural liquid and concentrating to dry substance content of 58-60%; introducing resultant concentrate into prepared raw materials and boiling out; introducing into cultural liquid solid residue produced after sequential extracting of Mortierella zychae micromycet biomass with the use of non-polar extractant in above-critical state, water, alkaline, water, acid, water, alkaline, and water, said solid residue being introduced in an amount of 6-8% by weight of dry substances; introducing into concentrate extract produced after extracting of equivalent amounts of the same biomass with the use of non-polar extractant in above critical state.

EFFECT: provision for obtaining of base product with improved organoleptical properties.

5 ex

FIELD: microbiology, agriculture.

SUBSTANCE: invention proposes the strain Pseudomonas aureofaciens IB-6 that exceeds known strains by the level of accumulation of cytokinines. The strain is isolated from sewage waters from aerotank of biological cleaning units and maintained in the Collection of microorganisms of Biology Institute UNTS RAN at number IB-6. The strain in the growth on LB medium forms cytokinines in the amount 680 ng-equiv. of zeatine per 1 ml of medium. Invention can be used for preparing the growth-stimulating preparations used in plant growing by microbiological synthesis.

EFFECT: improved preparing method, high yield of cytokinines.

1 tbl, 2 ex

FIELD: biotechnology, medicine industry.

SUBSTANCE: Alkaloid agroclavine is produced by coltivation of fungi Claviceps sp. c106 under aeration conditions and with agitation in aqueous broth containing sucrose, citric acid, yeast extract and mineral salts. Additionally broth contains 1.0-20.0 mol of tryptophan. Medium pH is adjusted with 25 % solution of NH4OH to 4.5-5.4. Target product is isolated from culture liquid by conventional methods.

EFFECT: increased content of agroclavine in culture liquid and in alkaloid mixture.

10 ex

FIELD: medicinal microbiology.

SUBSTANCE: method involves stage-by-stage immunization of rabbits with lipase from Pseudomonas sp. and preparing anti-lipase antibodies that are immobilized on polymeric microspheres. Then in the reaction of volume agglomeration in analyzed hemolytic strains of choleraic vibrios the property to produce lipase in interaction with anti-lipase antibodies immobilized on polymeric carrier is detected and the positive reaction is found confirming atoxigenic property of the analyzed strain being toxigenic - non-hemolytic strains don't show such ability but show the negative response reaction. Method provides reducing time for detection. Invention can be used in laboratory diagnosis of extreme danger infections.

EFFECT: improved method for detection.

4 cl, 1 tbl, 3 ex

FIELD: biotechnology, microbiology.

SUBSTANCE: nutrient medium comprises the following components per 1 l of distilled water: enzymatic hydrolyzate of soybeans, sodium hydrogen phosphate, sodium chloride, sodium sulfite, lipoic acid and 20% sodium hydroxide solution. Invention provides preparing the qualitative and inexpensive nutrient medium for culturing plague microorganism.

EFFECT: improved properties of nutrient medium.

3 ex

FIELD: biotechnology, animal breeding, in particular probiotic production.

SUBSTANCE: claimed method includes production of nutrient medium, its fermentation with strain consortium of Clostridium cellobioparum, Ruminococcus flavefaciens, Lactobacillus acidophilus, Propionibacterium acnes and collection of biomass. Said biomass is mixed with protective medium and dried.

EFFECT: cow milk productivity.

2 cl, 1 tbl, 3 ex

FIELD: biotechnology, food processing and medicine industry , in particular production of bacterium preparations.

SUBSTANCE: Strain Bifidobacterium bifidum is obtained by selection without using of genetic modification methods and is capable to utilize of inulin and cellobiose. Strain accumulates biomass on plant-origin substrates and artificial broth for short period with high bifidobacterium concentration. Strain has acid-forming properties and represents antagonist of pathogenic and saprogenic microflora. Bacterium preparations, bioactive food supplements, ferments, fermented milk products, fermented and non-fermented foodstuffs, hygienic and cosmetic products containing the same have probiotic effect and provide normalization of microbiocenose in human organism, including gastrointestinal and urogenital tracts, skin integument and mucilaginous.

EFFECT: new strain Bifidobacterium bifidum with probiotic and acid-forming properties.

2 tbl, 5 ex

FIELD: biotechnology, in particular food processing, dairy and medicine industry.

SUBSTANCE: claimed method includes broth preparation, sterilization and cooling. In prepared broth probiotic strain together with two non-lysogenic strains of thermophylic streptococcus in ratio 4:1 and 3:2, respectively, are cultivated. Mass of each probiotic strain Streptococcus salivarius subsp. Thermophilus is built up separately from one another. Then strains are mixed in equal amounts, pre-packed, dried and packed.

EFFECT: increased assortment of probiotic agents with improved properties.

4 cl, 2 tbl, 5 ex

FIELD: biotechnology.

SUBSTANCE: claimed method includes bacteria cultivation of genus Bacillus in broth followed by isolation of collected inosine from cultural liquid. As producer bacteria with resistance to growth inhibition with polymyxin B and/or colistin is used. Also disclosed are method for production of inosine-5'-monophosphate and two new strains of inosine producers: Bacillus subtilis and Bacillus amyloliquefaciens.

EFFECT: inosine production with increased yield.

8 cl, 4 tbl, 3 ex

FIELD: dairy industry.

SUBSTANCE: milk substrate is subjected to bioconversion with culture Bifidobacterium breve CNCM I-2219 having immunostimulating action. Milk substrate is brought into contact with said culture under condition wherein medium acidifying by culture is not more than 0.5 pH during incubation for 8 h and starting inoculation is 1x107 - 5x107 c/ml. Final population of Bifidobacterium breve after bioconversion is 1x105 - 1x109 c/ml. Contact time between milk substrate and strain is 6-24 h. Dairy product obtained by claimed method also is disclosed.

EFFECT: dairy product with decreased acidity useful in infant nutrition.

10 cl, 3 tbl, 3 ex

FIELD: agriculture, biotechnology, microbiology.

SUBSTANCE: invention represents the strain of bacterium Bacillus subtilis CH-13 that is used for protection of cereal agricultural crops, sunflower and grape against phytopathogenic microorganisms and vegetable crops against phytopathogenic bacteria also. The strain Bacillus subtilis CH-13 is isolated from chernozem (black) soil in Republic Moldova and deposited at registration number VNIISCHM D-606 from 10 September 1998 in the group of epiphyte microorganisms. Ability of the strain to inhibit different pathogenic microflora results to enhancing productivity of different species of agricultural crops.

EFFECT: valuable antibacterial properties of agent.

4 tbl, 2 ex

FIELD: general and medicinal microbiology.

SUBSTANCE: invention relates to preparing nutrient medium used for culturing microorganisms Haemophilus influenzae type b. The nutrient medium used for culturing microorganisms Haemophilus infleunzae type b comprises KH2PO4, K2HPO4, NaCl, MgCl2, CaCl2, glucose, NAD, hemin, thiamine, calcium pantothenate, aminopeptide and distilled water taken in the definite ratio of components. Invention provides simple method in preparing nutrient medium, reduced cost and absence of medium components interfering in preparing the pure preparation of capsule polysaccharide. The tentative cost of such nutrient medium is 20 rubles for 1 l.

EFFECT: improved preparing method, reduced cost.

1 ex

FIELD: biotechnology, medicine, microbiology.

SUBSTANCE: the strain is isolated from a sick child with meningitides and deposited in the collection GISK named for L. A. Tarasevich at № 267. The strain produces a capsule polysaccharide - polyribosyl ribitol phosphate in synthetic and semi-synthetic nutrient media.

EFFECT: valuable properties of strain.

1 ex

FIELD: biotechnology, microbiology, medicine.

SUBSTANCE: invention relates to the strain Lactobacillus paracasei CNCM I-2116 used for diarrhea prophylaxis causing by pathogenic microorganisms. Supernatant of this strain culture elicits ability to prevent colonization of intestine with pathogenic microorganisms causing diarrhea also and this strain is designated for preparing agent used for prophylaxis and/or treatment of disorders associated with diarrhea. Agent for oral administration represents therapeutically effective dose of the strain L. paracasei CNCM I-2116 or supernatant of its culture and acceptable foodstuff. Invention provides the enhanced viability of the strain in its applying and effectiveness in prophylaxis of adhesion to intestine cells and invasion to intestine cells of pathogenic microorganisms causing diarrhea.

EFFECT: valuable medicinal properties of strain.

5 cl, 8 dwg, 10 ex

Up!