Biological preparation for stimulation of growth and protection of plants from diseases, increase in productivity and soil fertility

FIELD: agriculture.

SUBSTANCE: biological preparation for stimulation of growth and protection of plants from diseases, increase in productivity and soil fertility contains biomass Bacillus amyloliquefaciens RNCIM B-11008 and humates at the following ratio of components in vol. %: biomass Bacillus amyloliquefaciens RNCIM B-11008 - vegetative cells and spores of 1.24-1.30×1010 CFU/ml of culture fluid and the spore content 94% of the total amount of CFU - 99.0, humates - 1.0.

EFFECT: biological preparation enables to protect plants from fungal and bacterial diseases, to improve the phytosanitary condition of soil and improve its fertility, to increase crop yields and improve the quality of agricultural products.

5 tbl, 8 ex

 

The invention relates to biotechnology and agricultural Microbiology, in particular for complex biological activities for crop production, and can be used for presowing treatment of seeds of agricultural crops, spraying of plants in the growing season, when preparing agricultural products for storage, for soil prior to planting and during the growing season of plants in the conditions of healthy soil and agro-ecosystem, and in terms of soil contamination with residual quantities of fungicides, herbicides, insecticides, phytopathogenic micromycetes and products of their metabolism.

Currently for agriculture known bacterial preparations on the basis of one culture of microorganisms, which either inhibit the development of pathogenic fungi and bacteria, or stimulate the growth and development of plants, or contribute to the fixation of atmospheric nitrogen or transfer of insoluble soil phosphates into soluble forms.

Known drug backoff based on Bacillus subtilis strain IPM-215 [RF Patent №2019966, IPC A01N 63/00, publ. 1994]. The disadvantages of BackOffice are limited scope, lack of accessible sources of information about health-improving effect on plants in bacterial defeat, effective, protective effect against diseases in terms of what s open ground and the capacity for nitrogen fixation and phosphatability. In addition, the composition of the drug, with many components to it, the technology for preparation of multi-stage and complex.

Known drug on the basis of Azotobacter chroococcum BH-1811 [RF Patent №2289620, IPC C12N 1/20, A01N 63/00, C12R 1/065, publ. 2006], designed to increase crop yields and improve their resistance to various diseases. The disadvantage of the drug is low effectiveness against fungal and bacterial pathogens of agricultural crops and the lack of ability to phosphatability.

There are also biological products on the basis of the Association of microorganisms of different taxonomic affiliation.

Known drug to stimulate growth and protect plants from disease - Extrogen [RF Patent №2302114, IPC A01N 63/00, C12N 1/20, C12R 1/065, C12R 1/07, publ. 2007], which contains strains of Bacillus mycoides var. B.A. ARRIAM No. D and Azotobacter vinelandii var. NP ARRIAM No. D. Known biological product for autumn, spring and summer soil, root and basal fertilizing during the growing season of plants, as well as pre-sowing treatment of seeds, which are a mixture of suspensions thirteen different strains [RF Patent №2322061, IPC A01N 63/00, C12N 1/20, publ. 2008]: Agrobacterium tumefaciens (B-4116, Agrobacterium radiobacter B-956, Azotobacter chroococcum B-2375, Bacillus thuringiensis B-2918, Bacillus subtilis B-6554, Bacillus subtilis B-4419, B. megaterium Bacillus B-4440, B. megaterium Bacillus B-200, Bradyrhizobium japonicum B-1978, Erwinia ananas B-5292,Lactobacillus casei B-3961, Pseudomonas fluorescens B-l138, Rhodopseudomonas palustris B-1620. Known biological preparation to enhance soil fertility, healing and stimulating the growth of plants on the basis of the Association strains of Bacillus subtilis K-4, Bacillus subtilis Be-12, Bacillus amyloliquefaciens 30-40 with the addition of the hydrolysate mixture of pine extract and carotene paste [RF Patent №2314693, IPC A01N 63/02, C12N 1/20, publ. 2008]. A significant drawback of drugs on the basis of several strains of bacteria of different taxonomic affiliation is the complexity of the specific and the generic structure of microorganisms with different nutritional needs that require individual culture conditions and different environmental conditions for maximum manifestation of the properties. In addition, these biological products are not effective in protecting plants against a broad spectrum of pathogens of plants.

The closest to the achieved result, taken as a prototype, is the drug Integral [RF Patent №2216173, IPC A01N 63/00, AS 1/06, C12N 1/20, C12N 1/20, C12R 1:065, publ. 2003]. In the preparation contains a mixture of the culture fluid containing the bacterial strain Bacillus subtilis 24D (ARRIAM 129) at a concentration of 5×109cells/ml, humates and trace elements in the following ratio, wt.%: the culture fluid is 70-75, humates - 20, microelements - 5-10. Significant disadvantages of the prototype are long the process of cultivation of the producer strain of Bacillus subtilis 24D (ARRIAM 129) (48 h); low concentration of microbial cells in the preparation of (5×109cells/ml); the need for more inclusion in the drug composition of trace elements (5-10% of the total weight of the preparation) and a large number of humates (20% of total weight), which reduces the titer of cells of the producer strain in the structure of the biological product and increases its cost; low and insufficient spectrum fungicidal activity; lack of data about the intensity of sporoobrazovanie culture depends on the ability of bacteria to survive in the environment; the lack of antibacterial activity and comprehensive actions on soil agro-ecosystem (the drug is intended only for presowing treatment of seeds and vegetative plant crops, fruit trees and berry bushes).

Preparations on the basis of one of the living culture of bacteria of the genus Bacillus, which had both high levels of a number of activities (antifungal, antibacterial, stimulating the growth and development of plants, increasing their productivity and level of soil fertility), available to the applicant the information sources are not identified.

The task of the invention is the creation of a biological product of complex action (antifungal, antibacterial, stimulating the growth and development of plants, increasing the yield of the plant is the first and the level of soil fertility) with high quantitative characteristics of biomass-based one strain of bacteria.

The technical result is the development of a biological product of complex action with fungicidal, bactericidal, growth-stimulating effect, increasing the yield of plants and the level of soil fertility, high quantitative characteristics of biomass-based one strain of bacteria of the genus Bacillus.

The technical result in the claimed biological product is achieved by the fact that, as the biomass of vegetative cells and spores of a strain of bacteria of the genus Bacillus, it contains biomass of Bacillus amyloliquefaciens In PMBC-11008 with the title of vegetative cells and spores 1,24÷1,30×1010CFU/ml and a content dispute 94% of the total number of CFU and humates in the following ratio of components,%:

biomass B.amyloliquefaciens VKPM B-11008 - vegetative cells and spores 1,24÷1,30×1010CFU/ml of culture fluid and the content of the dispute 94% of the total number of CFU - 99,0;

humates to 1.0.

Strain B.amyloliquefaciens VKPM B-11008 selected from wheat seeds and ofselection by multistage trait selection: a wide range of high antagonistic activity to various phytopathogenic micromycetes and bacteria; no toxic effects on biological (plant and animal) genetic and organismal levels; high productivity of culture and technology in a production environment; high level of the activity of secreted enzymes (amylase, protease, cellulase, phosphatase), contributing to the fertility level of the soil; the ability to fix atmospheric nitrogen in quantities comparable with nitrogen-fixing activity effective symbiotic strains-producers of nitrogen-fixing preparations; can be used in tank mixtures with chemical fungicides, herbicides, insecticides, agricultural chemicals and biological agents and the ability to detoxify the pesticide.

Taxonomic affiliation of strain determined using analysis signatory nucleotide sequences of 16S rRNA genes gyrA and in GenBank/EMBL/DDBJ (http://www.ncbi.nlm.nih.gov) using the multiple alignment program BLASTn (http://www.ncbi.nlm.nih.gov/BLAST/). The strain is deposited in Russian national Collection of Industrial Microorganisms at FSUE gosniigenetika and has a collection number VKPM B-11008.

Culture B.amyloliquefaciens VKPM B-11008 has the following phenotypic characteristics. Straight rod-shaped cells with rounded ends on the 1.8×0.5 µm, arranged singly, in pairs, rarely in chains; movable with the help of flagella. With growth MPA at a temperature of 28°C for 17-18 h incubation form ellipsoidal endospores that are located in the cell subterminale. Have gram-positive morphotype. Chemoorganotrophic with respiratory type of metabolism, what timolol pH of 7.2 and a temperature of 28°C. Utilizing glucose, arabinose, maltose, galactose, lactose, mannitol, sucrose, starch, cellulose, gelatine, casein; use urea, citrate, but do not use propionate; do not form indole and hydrogen sulfide; do not have lecithinase, coagulase, hyaluronidase activities. Catalanopolonesa, reaction Voges-Proskauer positive. On MPA at a temperature of 28°C for 17-18 h in culture increases with the formation of strongly folded colonies with a domed centre and jagged edge, viscous consistency of the skin color. Capable of growing in a medium with a content of 7.0% NaCl in the range of pH 4.5...9,5.

As nutrient medium to obtain the claimed biological product using low cost environment based on waste grain production of the following composition, g/l: wheat bran - 40,0; corn extract to 0.5; MgSO4- 0,9; KH2PO4- 0,5; caso3to 1.0; pH 7.0 ą 0.2. To obtain a seed culture B.amyloliquefaciens VKPM B-11008 scatter in test tubes with beveled environment MPA, incubated in the conditions of thermostat at a temperature of 28°C for 24 h, and then make a tube 6-7 ml of 0.85% NaCl solution, bacteriological loop is removed from the surface of the agar bacterial mass, shaken for 3-4 min obtained seed material containing cells and spores 1,0÷2,0×108CFU/ml contribute in a liquid nutrient medium in alyemeni less than 5%, giving the initial optical density of cell suspension in 0.1% (λ=590 nm, l=10 mm) and grown at 28°C for 24 h the composition environment provides a high level of accumulation of biomass B.amyloliquefaciens VKPM B-11008 (5,6÷5.9 g/l), high titer of viable cells and spores (1,24÷1,28×1010CFU/ml) and the intensity of sporoobrazovanie (content dispute - 94,0% of total CFU). Nutrients maximally utilized by the crop (residual sugar and amino nitrogen in the environment by the end of the growing amounts of 14.0% and 2.0% respectively from baseline). Maximum antagonistic and plant growth activity, the biomass yield and the intensity of sporoobrazovanie culture B.amyloliquefaciens VKPM B-11008 achieved at the temperature of cultivation 28°C. the Culture fluid B.amyloliquefaciens VKPM B-11008 contains metabolites, resulting in high levels of prolonged antagonistic activity to a wide range of pathogens of various crops: Fusarium frost, Fusarium oxysporum, Fusarium sambucinum, Bipolaris sorokiniana, Helminthosporium solani, Alternaria alternata, Alternaria solani, Alternaria brassicae, Rhizoctonia solani, Botrytis cinerea, Gliocladium roseum, Phytophthora infestans, Verticillium dahliae, Colletotrichum coccodes, Cladosporium herbarum, Oospora pustulans. The advantage of a biological product in contrast to the prototype is bactericidal effect on pitapata the i.i.d. bacteria and fungicidal effect on micromycetes, causing plesnevelae seeds. Unlike the prototype, after completion of the cultivation process to culture B.amyloliquefaciens VKPM B-11008 add only the sodium and potassium salts of humic acids and a lower final concentration of 1.0 vol.%), than the biological prototype (20,0%). In this case, the humates are used to preserve the viability and stabilization of microbial biomass and increase the adsorption capacity of cells and spores in the processing of seeds and vegetative plants, unlike the prototype, in which the humates function of promoting the growth and development of plants. In the invention stimulating properties due to their own ability culture B.amyloliquefaciens VKPM B-11008 to synthesize substances with regulatory activity and to allocate them in the culture fluid when growing culture and the environment in the introduction. This biological product stores at least 12 months from the date of manufacture in the temperature range from +4 to +28°C to preserve the entire set of properties.

A high level of profitability of the production process of the proposed biological product according to the formula of the invention provides a low duration of the process, low cost of raw materials, high productivity and intensity of sporoobrazovanie culture B.amyloliquefaciens VKPM B-11008, from which the head is sit further the ability of the strain to survive under adverse environmental conditions (drought, low temperatures, acidification of soils and so on).

The advantage of the strain B.amyloliquefaciens In-PMBC 11008 is a growth activity in the pH range from 4.5 to 9.5, with a maximum fungistatic/fungicidal activity of the active acidity in the range of 4.5...of 8.5. Unlike the prototype, it is the predominant characteristic of the requested drug is practically widespread acidification of soils and sharing in agriculture mixtures with fungicides, herbicides, insecticides, having an acidic pH (up to pH 5.0).

The invention is illustrated below by examples.

Example 1 demonstrating suppressed spectrum of phytopathogenic micromycetes and the level of antagonistic activity of the proposed biological product compared to the prototype. Biologic receive according to the invention. Antagonistic activity against the test cultures phytopathogenic micromycetes detected by the method of the hole. To obtain a suspension of the test culture micromycete fungal culture grown lawn on the surface of the Petri dishes with agar medium of čapek [see Semenov S.M. Laboratory environment for actinomycetes and fungi: a Handbook / Samsonov. - M.: Agropromizdat, 1990. - P.172] within 14 days, add 10 ml of sterile tap water, soskrebajut grown spore-mycelial mass from p the surface environment loop, transferred into a test tube. The resulting suspension from the tube introduced into the flask with 100 ml of molten and cooled to 60°With potato-glucose medium [see Semenov S.M. Laboratory environment for actinomycetes and fungi: a Handbook / Samsonov. - M.: Agropromizdat, 1990. - P.á192] thus, to obtain a final concentration of 109CFU/ml In Petri dishes poured into 15 ml of potato-glucose medium, receiving the bottom layer of agar. The environment made by the suspension of the test culture poured into 8 ml in Petri dishes on a frozen bottom layer, thus obtaining the top layer of agar. After solidification of the medium in the center of the cups in the top layer of agar doing 1 round hole borer. In wells contribute to 0.1 ml and incubated in a thermostat at a temperature of 28°C for 7 days. In a variant of "integral" in the wells contribute to 0.1 ml with a titer of 109CFU/ml In the control Petri dishes in wells contribute to 0.1 ml of sterile water. Assessment of the level of antagonistic activity carried out by the diameter sterile zones (in mm) in mushroom lawn, formed around the holes. Spectrum suppress phytopathogenic micromycetes and the level of antagonistic activity of the proposed biological product in comparison with the prototype shown in table 1.

Table 1
Antagonistic activity of the proposed biological product and prototype against phytopathogenic micromycetes
The test-cultureZones of growth inhibition, mm
The proposed biological productThe placeholder
Fusarium frost41,3±1,313,4±1,0
Fusarium solani36,7±1,912,6±0,9
Fusarium oxysporum35,2±1,411,1±0,8
Fusarium sambucinum38,3±0,812,3±1,1
Bipolaris sorokiniana40,6±1,517,5±1,1
Helminthosporium solani39,1±1,016,4±1,3
Alternaria alternata39,4±2,020,7±1,3
Alternaria solani*18,5±1,5
Alternaria brassicae41,5±2,119,7±1,2
Rhizoctonia solani39,1±1,315,7±1,4
Botrytis cinerea39,0±1,118,0±1,2
Gliocladium roseum36,4±0,810,5±0,7
Phytophthora infestans28,6±0,98,0±0,6
Verticillium dahliae36,1±1,511,5±1,0
Colletotrichum coccodes*11,1±1,0
Botrytis cinerea39,2±1,718,5±0,8
Cladosporium herbarum36,7±1,116,3±0,3
Oospora pustulans26,2±0,58,4±1,8
Mucor racemosus*nd
Mucor racemosus
Penicillium spp
Aspergillus spp.
Trichotecium roseum
Nigrospora spp
Note: * - the fungicide is th activity (no growth of the test culture); nd - no data

Example 2, demonstrating the level of antagonistic activity of the proposed biological product against phytopathogenic bacteria compared to the prototype. Antagonistic activity against the test cultures phytopathogenic bacteria evaluate method deferred antagonism. To do this, get a biological product according to the formula of the invention and the loop is sown in the centre of Petri dishes with medium Gause [see Semenov S.M. Laboratory environment for actinomycetes and fungi: a Handbook / Samsonov. - M.: Agropromizdat, 1990. - P.148]. Crops incubated at 28°C for 3 days. Grown culture B.amyloliquefaciens VKPM B-11008 podshuveyt stroke DSA suspension of the test cultures of phytopathogenic bacteria (titer suspensions 108CFU/ml). Then Cup incubated at 28°C for 24 h assessment of the level of antagonistic activity of the claimed drug against phytopathogenic bacteria carry out the largest zones of suppression (in mm) of growth of the test cultures phytopathogenic bacteria formed around colonies of culture B.amyloliquefaciens VKPM B-11008. Spectrum suppress phytopathogenic bacteria and the level of antagonistic activity of the proposed biological product in comparison with the prototype shown in table 2.

Table 2
Antagonistic activity of the proposed biological product and prototype against phytopathogenic bacteria
The test-cultureZones of growth inhibition, mm
The claimed preparationThe placeholder
Pseudomonas atrofaciens*nd
Erwinia carotovora subsp.carotovora*10,2±0,4
Erwinia carotovora subsp.atroseptica*14,1±1,1
Xanthomonas campestris*the 15.6±1,0
Xanthomonas translucens*10,3±0,9
Note: * - bactericidal activity (complete inhibition of growth of test cultures); nd - no data

Example 3, demonstrating the level of antagonistic activity of the proposed biological product against the test cultures phytopathogenic micromycetes active at different pH of the medium. The drug is produced according to the invention, while the pH of the medium for culturing lead to values of 4.5, 5,5, 6,5, 7,, to 8.5, and 9.5 by adding 1M HCl solution. The test culture phytopathogenic micromycetes are grown separately in Petri dishes with the environment of čapek. Antagonistic activity against the test cultures phytopathogenic micromycetes detected by the method of the hole (as in example 1). Strain B.amyloliquefaciens VKPM B-11008 able to grow with the active acidity in the range of pH from 4.5 to 9.5, with a maximum fungistatic/fungicidal activity are in the range of pH 4.5 to 8.5. This is illustrated by data shown in table 3.

42,9±1,0
Table 3
The active influence of the pH on the level of antagonistic activity of the proposed biological product against the test cultures phytopathogenic micromycetes
The pH value of the environmentThe diameter of the zones of inhibition of growth of test cultures mm
Bipolaris sorokinianaAlternaria alternataFusarium frost
4,520,0±0,829,3±0,734,2±0,9
5,525,2±0,735,8±1,0
6,526,1±1,036,5±1,143,7±1,2
7,524,9±0,935,6±0,842,3±0,9
8,522,8±0,733,4±1,340,7±1,3
9,519,9±0,828,2±1,233,2±1,3

Example 4 demonstrating the effectiveness of the proposed drug for the treatment of seeds against phytopathogenic micromycetes and bacteria. The claimed biological receive according to the invention. Obtained by the biological product to handle the seeds of spring wheat varieties Omsk 32 and spring barley varieties Raushan semi-dry method with the consumption rate of the product of 1.5 l/t of grain (fluid flow to 10 l/t). Seeds of control variant treated with sterile water at a rate of 10 l/I. Evaluate the efficacy of seed treatment by the claimed biological product compared to the prototype (at the dose of 1.5 l/t seed) and a chemical fungicide, Kaluga super (at a dose of 2.0 l/t seed) [see List of pesticides and agrochemicals permitted for use on the territory of the Russian Federation. 2010: the reference and the creation, Supplement to the journal "Protection and quarantine of plants" No. 6, 2010. - M., 2010. - P.150]. Evaluation of effectiveness is carried out by analysis of the seeds in a moist chamber and taking into account the development of diseases of seedlings.

The results are presented in tables 4 and 5. Processing grains declare a biological product leads to a more significant reduction in the infestation of seed pathogens (by 95.7 100.0%of cases)than the prototype (49.1-68,8%) and the chemical fungicide (49.5-78.5 per cent) relative to the control, depending on the culture of grain and the pathogen. While the fungicidal effect of the proposed biological product is maintained over long-term (over 30 days) compared with the action of the prototype and chemical fungicide (up to 7 days after treatment). In addition, the claimed biological product, unlike the prototype and chemical fungicide, eliminates pathogens of placevine seeds and bacterial diseases. The proposed biological product reduces the level of development of diseases of seedlings, depending on the cultivar and crop processing by 94.6 100.0%of cases, the integral on 56,0 of 62.3%, Kaluga super - 64,5-to 78.7%.

Table 4
Prevalence of seeds of spring wheat varieties Omsk 32 phytopathogenic micromycetes in the processing of grains declare a biological product,prototype and chemical fungicide
Phytopathogen-
nye
the micromycetes
Prevalence of seeds, %
Control (without treatment)The proposed biological productThe placeholderChemical fungicide
Bipolaris sorokiniana82,52,528,526,0
Fusarium spp46,52,014,513,5
Alternaria alternata93,03,030,527,0
Penicillium spp28,5014,510,5
Aspergillus spp.8,003,53,0
Mucor spp.13,506,05,0
Cladosporium spp. 7,003,02,5

Table 5
Prevalence of seeds of spring barley varieties Raushan phytopathogenic micromycetes in the processing of grains declare a biological product, prototype and chemical fungicide
Phytopathogen-
nye
the micromycetes
Prevalence of seeds, %
Control (without treatment)The proposed biological productThe placeholderChemical fungicide
Bipolaris sorokiniana84,03,033,524,5
Fusarium spp.39,51,519,513,0
Alternaria alternata81,52,032,0of 17.5
Penicillium spp.29,5 014,011,0
Aspergillus spp.25,0011,59,0
Mucor spp.10.505,03,5
Cladosporium spp.3,001,51,0

Thus, the claimed biological product significantly improves the phytosanitary condition of seed, and is substantially outweighed by the effectiveness of the prototype.

Example 5, showing the presence of the stimulating action of the proposed biological product on the processes of seed germination, growth and development of plants.

The stimulating effect of the proposed biological product show on the seeds of spring wheat, grade Fret. Biologic receive according to the invention. Wheat seeds in quantities of 100 pieces soaked for 24 h in 15 ml of inoculant control seeds soaked in water. After soaking the seeds are laid on moist filter paper, rolled up and germinated at 25°C. Germination was determined on 4 days by the number of germinated seed (expressed in percentage of the total number of treated seeds), vigor - 3 day ground biomass and root parts of seedlings 14 days.

The difference in germination between the experimental (treatment claimed by the biological product) and control (water treatment) seeds is 19%, vigor - 10%, in the terrestrial biomass and root parts of seedlings - 29 and 47%, respectively. Thus, the claimed biological product has a significant stimulatory effect on the processes of seed germination, growth and development of plants.

Example 6 showing the effectiveness of the proposed biological product at pre-sowing seed treatment against diseases and impact on yield in pot experiment. Biologic receive according to the invention. The growing experience is performed on the plots with the area of 0,5 ha for spring barley Hordeum vulgare L. varieties Raushan according to the standard technique of field and vegetation experiments [see Armor B.A. Methods of field experience / Bagishbekov. - M.: Agropromizdat, 1985. - 351 S.]. Seed processing is performed by using dry etching at a rate of seeding of viable seeds 5.5 million/ha and the rate of application of the proposed biological product 1.5 l/t of grain (fluid flow to 10 l/t), the integral and Kaluga super (chemical standard) - 1.5, and 2.0 l/t, respectively; in the control variant, the seeds treated with water. Predecessor - spring barley. The soil is leached Chernozem. The yield is of cause standard 14%humidity and 100%purity.

The proposed biological product contributes to a more significant reduction in the development of root rot compared with the prototype. Biological efficiency (defined as the difference between the percentage of infected plants in the control and experience, as a percent of control) of a biological product at the end of the growing season is 67%more than the effectiveness of the prototype of 6.9% and a chemical fungicide to 19.7%.

The experience carried out at a high infectious background: the number of conidia of Bipolaris sorokiniana and Fusarium is before sowing 270 and 490 1 g of air-dry soil, respectively. By the end of vegetation the proposed biological product reduces the number of conidia B.sorokiniana to 26 and Fusarium - up to 42 in 1 g of air-dry soil; in the variant with integral and chemical fungicide significant differences from control were not found. Thus, the claimed biological product not only protects plants from root rot pathogens transmitted through seeds, but also improves the phytosanitary condition of the soil by reducing soil infection.

Biological efficacy of the proposed drug against Fusarium head blight, mobiles ear and Helminthosporium leaf spot is 85.9, 90,3, 87,2%, respectively, and exceeds the efficiency of the integral 13.9, 12.4, and 11.9 percent, respectively, of the effectiveness of Kaluga super - 19.5, 13,2 and 23.5%, respectively. Thus, the proposed biological product is effektivnym remedy not only from root rot, but from leaf-steam diseases and diseases of the ear.

The positive impact of the proposed drug on the yield and quality of grain recorded in terms of patterns of grain yield and protein content. The proposed biological product increases the mass of 1000 grains by 14.2%, the number of productive stems by 11.3% and the grain number in ear ears by 6.6% compared with control, while increasing the protein content in grain by 11.4% compared with the control. Grain yield when using the proposed biological product exceeds the level of control by 29.8%. Significant differences between the control, option "integral" and "Kaluga super" elements of yield structure and yield was not detected.

Example 7 showing nitrogenase activity of the proposed biological product without paying and when introduced into the soil.

Biologic receive according to the invention. Nitrogenase activity of the proposed biological product determine the acetylene method using a gas chromatograph [see Kalinin T.A. application of the acetylene method for quantitative estimation of different groups of nitrogen-fixing method of limiting dilutions / Taskalinsky, Tvedten, Umelb, Ltiplied, Avemano // Microbiology. - 1981. - T, Iss.5. - S-927]. Nitrogenase activity of the proposed biological product without soil composition is yet of 0.18 µg N 2/ml/h

The activity of nitrogen fixation in the soil with the introduction of the proposed biological product determined by acetylene method according to [see Zvyagintsev became popular Methods of soil biochemistry and Microbiology: a manual / D.g.zvjagintsev. - M.: Moscow state University, 1991. - 303 S.]. For this purpose, 0.5 kg of soil distributed in a plastic cuvette with a size of 60×40 cm, evenly irrigate the claimed product with the help of a spray, soil thoroughly mixed, transferred into sterile jars with a capacity of 750 ml, closed with a thin plastic film and incubated at 28°C, maintaining a constant humidity in the soil samples within 60% of full capacity. Control samples treated with water.

The difference in nitrogen-fixing activity of soil experienced (processing of the claimed preparation) and control (water treatment) samples was 256%. Thus, the claimed biological product increases the natural level of soil fertility. High nitrogen-fixing activity of the soil due to both its own nitrogenase activity of the proposed biological product, and by enhancing soil microflora under the influence of stimulating metabolites producer strain B.amyloliquefaciens VKPM B-11008.

Example 8 showing dephosphorylates activity of the proposed biological product.

Biologic receive according to the invention. Dephosphorylates activity of the claimed drug determine in the modeling experience. Soil samples treated as in example 7. After 45 and 90 days of incubation in the control (water treatment) and experimental (treatment claimed by biological) samples to determine the content of mobile phosphorus (mg P2O5/kg soil) method Kirsanov [see Sokolov A.V. Agrochemical methods of soil testing / Aveskulov. - M.: Nauka, 1975. - 656 S.].

The difference in the content of mobile phosphorus in the soil of the experimental (treatment claimed by the biological product) and control (water treatment) samples is after 45 days of incubation 30,3%, after 90 days of 35.2%. Thus, the claimed biological product increases the natural level of soil fertility by increasing the content of mobile phosphorus.

Examples confirm the achievement of the technical result of the invention.

Thus, the full spectrum of signs combined effect of the claimed invention is as follows:

1. A wide range of high antagonistic activity to phytopathogenic micromycetes; fungicidal effect on phytopathogenic fungi that cause plesnevelae, and a bactericidal effect on phytopathogenic bacteria.

2. Stimulating effect on the growth and development of plants, which is manifested in increased germination and seed vigor, increased accumulation of root biomass and aboveground parts of plants, accelerating the passage of f the h development of plants and as a consequence, the acceleration of the maturation process agricultural products (15-20 days). It is a primary property of the proposed biological product is due both to the ability of the strain B.amyloliquefaciens VKPM B-11008 to the production of physiologically active substances (natural analogues of gibberellins, auxins, heteroauxin, cytokinins)and stimulating effect on pedieos.

3. The ability to fix atmospheric nitrogen in quantities comparable with nitrogen-fixing activity producers of nitrogen-fixing biologics.

4. The ability to synthesize lytic enzymes, which increases the availability to plants of mineral and organic compounds of the soil due to their partial or complete hydrolysis to easily assimilable compounds and increases the overall level of soil fertility.

5. Physical and chemical consistency of a biological product used in the agricultural production of chemical fungicides, herbicides, insecticides without reducing the biological activity of the components of the tank mixture. This allows to reduce to the minimum standards of consumption of chemical pesticides, the cost of processing of seeds and vegetative plants, grain processing in preparation for storage, to reduce the pesticide load on the agrophytocenoses and get environmentally friendly products, not terasul residual amounts of pesticides.

6. Compatibility with biological preparations based on bacteria of different taxonomic groups (based on Bacillus - fotosporno, integral; Pseudomonas - pseudobacteremia-2, taking into consideration that the biological product Elena; Azospirillum - asoriginal; Flavobacterium - Flavobacterium). This property is caused by resistant strains B.amyloliquefaciens VKPM B-11008 to antibacterial metabolites synthesized by bacteria-producers of various biological products. These drugs in a joint application with the declared biologic show synergistic antagonistic action against phytopathogenic micromycetes.

7. The ability to dissolve poorly soluble phosphorus-containing substances in the soil (including phosphate fertilizers) in soluble phosphorus compounds, easily involved in the metabolism of plants.

8. The improvement of the phytosanitary condition of the soil both by direct reduction of the number of phytopathogenic microorganisms in the soil, and indirectly by increasing beneficial soil microflora.

The biological product to stimulate growth and protect plants from disease, improve yield and soil fertility, including biomass of vegetative cells and spores of bacteria from the genus Bacillus and humates, characterized in that it contains biomass of Bacillus amyloliquefaciens In PMBC-11008 with the title of vegetative cells and spores 1,24÷1,30·1010 CFU/ml and a content dispute 94% of the total number of CFU and humates in the following ratio of components, vol.%:
biomass .amyloliquefaciens VKPM B-11008 - vegetative cells and spores 1,24÷1,30·1010CFU/ml of culture fluid and the content of the dispute 94% of the total number of CFU - 99,0;
humates to 1.0.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. The symbiontic strain of Corynebacterium diphtheriae tox " No.108 is isolated from a bacteria carrier - a tonsillitis patient at IKB No.1 Moscow. The strain is harmless, non-reactogenic, does not possess allergic properties, antigens of which increase resistance of the microorganism to infectious diseases. The strain of Corynebacterium diphtheriae tox" No.108 is deposited in the National Collection of Normal Microflora of the G.N. Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology under registration number 381.

EFFECT: invention enables to create non-specific resistance of farm animals to infectious bacterial and viral diseases.

4 tbl, 5 ex

FIELD: chemistry.

SUBSTANCE: invention relates to microbiology and biotechnology and can be used in food industry and medicine. The microbial strain of Lactobacillus plantarum Tensia DSM 21380 produces conjugated linoleic acid (CLA), hydrogen peroxide (H2O2), nitrogen monoxide (NO), contains plantaricin encoding genes and has antioxidant activity. This strain and its liophilised form are used to prepare an antimicrobial and hypotensive probiotic, a dairy product and a medicinal agent which lowers blood pressure. The strain and its liophilised form are also used to increase polyamine turnover in the body, to attain a dominant position among intestinal lactic bacteria in the gastrointestinal tract and to prolong the shelf life of a dairy product.

EFFECT: use of the invention enables to lower blood pressure, suppress undesirable microflora and inhibit oxidative processes.

12 cl, 8 dwg, 31 tbl, 7 ex

FIELD: agriculture.

SUBSTANCE: method to stimulate growth and to protect small-fruit crops against diseases caused fungic pathogens includes treatment of plants with a biopreparation in a liquid form by soaking roots of seedlings prior to planting, or their sprinkling after planting, or sprinkling of plants in the period of vegetation and fruiting, or sprinkling of soil around plants with its further soaking with water. The biopreparation is a mixture of strains of bacteria B. subtilis VKPM B-10641, B. amyloliquefaciens VLPM B-10642, B. licheniformis VKPM B-10561 and B. licheniformis VKPM B-10562 or a mixture of strains of bacteria B. subtilis VKPM B-10641, B. amyloliquefaciens VKPM B-10642 and B. licheniformis VKPM B-10562 with a titre of each strain of not less than 105 CFU/ml in the form of a water suspension.

EFFECT: invention provides for higher protection of small-fruit crops against diseases caused by fungic pathogens, due to use of a mixture of strains of different types of bacteria of Bacillus type, having higher antagonistic activity, and also provision of the possibility of stimulation of growth of small-fruit crops.

5 cl, 10 tbl, 8 ex

FIELD: biotechnologies.

SUBSTANCE: strain Enterococcus hirae BC - 37 having high antagonistic activity is deposited in the Russian National Collection of Industrial Microorganisms (RNCIM) under the registration number VKPMV-10090 and may be used, for instance, in production of such cultured milk foods as kefir, cottage cheese or ryazhenka.

EFFECT: invention makes it possible to produce a strain having high antagonistic activity and high speed of milk turning sour.

3 ex

FIELD: biotechnologies.

SUBSTANCE: strain Lactobacillus gallinarum I-12, having high antagonistic activity is deposited in the Russian National Collection of Industrial Microorganisms (RNCIM) under the registration number VKPM V - 10134 and may be used in production, for instance, of such cultured milk foods, as kefir, acidophilic milk, acidophilic-yeast milk.

EFFECT: invention makes it possible to produce a strain having high antagonistic activity and high speed of milk turning sour.

3 ex

FIELD: biotechnology.

SUBSTANCE: method of increasing the productivity of the bacteria E.coli lies in preparation of a suspension of microorganisms, stirring it in the process of cultivation in the presence of a magnetic magnesium isotope sulfate 25Mg in an amount of 2.2 mmol/l.

EFFECT: increased stimulation of cell growth of bacteria Ecoli.

4 dwg, 4 tbl

FIELD: biotechnology.

SUBSTANCE: strain of Enterococcus hirae "И"-27 with high antagonistic activity is deposited in the Russian National Collection of Industrial Microorganisms (RNCIM) under registration number of RNCIM B-10091 and can be used in production, for example, of fermented milk products such as kefir, cottage cheese or fermented baked milk.

EFFECT: invention enables to obtain a strain with high antagonistic activity and a high rate of milk fermentation.

3 ex

FIELD: biotechnology.

SUBSTANCE: strain of Enterococcus hirae "И"-29 with high antagonistic activity, deposited in the Russian National Collection of Industrial Microorganisms (RNCIM) under registration number of RNCIM B-10088, and can be used in production, for example, of fermented milk products such as kefir, cottage cheese or fermented baked milk.

EFFECT: invention enables to obtain a strain with high antagonistic activity and a high rate of milk fermentation.

3 ex

FIELD: biotechnologies.

SUBSTANCE: invention may be used to extract carotenoids, in particular, deinoxanthine, which is used to develop new antioxidant and radioprotector preparations to increase adaptation capabilities of humans and aminals, prevention and treatment of diseases. The method provides for extraction of carotenoids from the bacterial mass Deinococcus radiodurans by the mixture acetone : ethanol (at the 1:1 ratio). Separation of carotenoids with extraction of deinoxanthine on preparative columns of a liquid low-pressure chromatograph, where the sorbent is hydroxyapatite, and the eluent - ethanol.

EFFECT: invention makes it possible to increase quantity and quality of extracted deinoxanthine.

2 dwg, 1 tbl, 4 ex

FIELD: biotechnologies.

SUBSTANCE: probiotic lacto-amylovorin is produced by means of depth growth of the strain Lactobacillus amylovorus RNCIM B-6253 on the liquid nutrient medium, which contains the following: corn extract - 10 ± 0.01 ml, lactose - 20 ± 0.005 g, chemically deposited chalk - 0.5 ± 0.01 g, triple-substituted citric-acid sodium - 7.5 ± 0.05 g, 3-water acetous sodium - 2 ± 0.01 g, iron sulfate - 0.1± 0.001 g, 5-water manganese sulfate - 0.16 ± 0.002 g, hydrolysate of dry nonfat milk - up to l. Subsequent extraction of the target product in liquid condition or in the form of a dry powder is carried out with preliminary addition of a stabiliser to a cultural liquid or to a protective medium used when producing the probiotic in the form of the dry powder. The stabiliser is selected from: sodium metabisulfite, sodium hydrosulfite and ascorbic acid.

EFFECT: invention provides for stable production of a probiotic with high values of a titre of antimicrobial bodies not only in a cultural liquid, but also after its treatment required for production of different pharmaceutical forms of a preparation.

3 cl, 2 tbl, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to agriculture. Pesticide combination contains at least two active ingredient components together with one or more conventional additives. Component (I) is imidacloprid and component (II) is one or more growth activators selected from a group comprising harpin and acibenzolar-S-methyl. To control or prevent damage by pests and/or pathogenic damage to plant propagation material, plants and/or plant organs which grow at a later point in time, said pesticide combination is applied onto a plant, part of a plant or the area surrounding the plant.

EFFECT: invention enables to increase biological activity of the combination.

12 cl

FIELD: agriculture.

SUBSTANCE: method relates to agriculture, namely, to protection of crops. The method is implemented by obtaining a salt extract of biologically active substances extracted from a biomass of soil oligochaetes Nicodrilus caliginosus after their treatment with potassium chloride in crystalline form, and pre-sowing treatment of seeds or spraying plants during the growing season with optimum dose of the extract of biologically active substances in a proportion of 1:30 parts of the biomaterial.

EFFECT: method enables to reduce the dosages of biologically active substances and to increase crop yields.

3 tbl, 4 ex

FIELD: food industry.

SUBSTANCE: microbicidal or microbiostatic composition comprises bacteriocin and extract containing phenolic diterpenes and obtained or extracted from plants of family Labiatae. The extract contains phenolic diterpenes in a quantity greater than 1.0 wt % in conversion to the composition weight. The plant of family Labiatae is rosemary. This composition is applied in method of preventing and/or suppression and/or destruction of microorganism in food product. The method is carried out by contacting food with the composition. Also a set for obtaining the composition and food products containing the composition are offered.

EFFECT: invention has a capacity to inhibit growth of pathogenic microorganisms in food products.

39 cl, 11 dwg, 7 tbl, 6 ex

FIELD: agriculture.

SUBSTANCE: invention refers to agriculture, particularly to horticulture. The method consists in treatment of growing trees with a preparation corresponding to a mixture of suspensions of micro-organism strains Pseudomonas species 17-2 and Bacillus subtilis B-14 taken at ratio 1:1 with concentration (2-5)×107 of cells/ml. Also trees are treated in terms of 5-10 ml of suspension per one fruit tree. Vegetative fruit trees are treated starting from a phenological stage "end of blossoming" to a phenological phase "beginning of fruit growth" including.

EFFECT: increased efficiency of fruit trees protection from fungus diseases and reduced consumption rate of preparation.

2 cl, 3 tbl, 3 ex

FIELD: food industry.

SUBSTANCE: encapsulated antimicrobial material includes (i) a kernel containing an antimicrobial material and a shell (ii) out of an encapsulated material which is a hydrophobic shell with a melting temperature higher than 45°C. It is selected out of a group including animal oils and fats, completely hydrogenated vegetable or animal oils, partially hydrogenated vegetable or animal oils, unsaturated hydrogenated or completely hydrogenated fatty acids, unsaturated hydrogenated or completely hydrogenated monoglycerides and diglycerides of fatty acids, unsaturated hydrogenated or completely hydrogenated etherified monoglycerides and diglycerides of fatty acids, unsaturated partially hydrogenated or completely hydrogenated free fatty acids, other emulsifiers, animal waxes, vegetable waxes, mineral waxes, synthetic waxes, natural and synthetic resins as well as their mixtures. The hydrophobic shell is impermeable for the antimicrobial material and is selected to provide release of the antimicrobial material from the encapsulated antimicrobial material while in contact with food products and the antimicrobial material is bacteriocin selected from the lanthionine-containing bacteriocins, bacteriocins formed by Lactococcus, bacteriocins formed by Streptococcus, bacteriocins formed by Pediococcus, bacteriocins formed by Lactobacillus, bacteriocins formed by Camobacterium, bacteriocins formed by Leuconostoc, bacteriocins formed by Enterococcus and their mixtures. The composition contains the antimicrobial material and a carrier. The food product contains the product and the antibacterial material. The method is implemented by introducing the antimicrobial material into the food product.

EFFECT: invention ensures a long antimicrobial effect.

43 cl, 16 ex, 4 tbl

FIELD: food industry.

SUBSTANCE: invention relates to biotechnology namely to preparation for stimulation of growing and developing of plants based on strain Halobacterium salinarum All-Russian collection of industrial microorganisms B-9025. To obtain the preparation strain Halobacterium salinarum All-Russian collection of industrial microorganisms B-9025 is grown during 6 days. After that biomass is separated by centrifugal process at 7000 g and incubated in distilled water. After that deposit is thrown away and obtained supernatant is centrifugally processed at 50000 g, supernatant is thrown away and distilled water of the same volume is added to the obtained deposit. Deposit is dissolved. Such suspending process is repeatedly performed till ratio of optical density of solution at wave-length 280 nm to solution optical density at wave-length 570 nm (D280\D570) less than 2,5, after that supernatant is standartised till bacteriorhodopsin concentration makes from 10-9 M to 10-6 M.

EFFECT: preparation allows to reduce general fertiliser consumption while growing plants which leads to production of more ecologically clean products with ample quantity of nutrient substances and green mass.

5 ex, 3 tbl

FIELD: agriculture.

SUBSTANCE: compound acceptable for agriculture, application of the compound and method for stimulating early blossom and budding and/or fruitification for non-legumes and legumes crops are described. The compound stimulates early blossom and budding for non-legumes and horticultural croppers, ornamental plants, crop or bedding plants and contains one or several lipochitooligosaccharides (LCOs) in the efficient quantity.

EFFECT: increased harvest connected with stimulating early blossom, quantity increase of buds and flowers, earlier fetal development for non-legumes and legumes crops comparing with the condition without using LCOs.

14 cl, 14 tbl, 22 dwg

FIELD: agriculture.

SUBSTANCE: invention can be used during establishment of industrial plantations of valuable nut crop - pistache (Pistacia vera L) in the North Caucasus. Seeds of pistache are soaked in aqueous solution of kinetin having temperature of 35-40°C and in concentration of 500 mg/l during 10-12 days. The soaked seeds are planted in pits covered with substrate, that is manure, straw and mixture of sand with humus placed layer by layer.

EFFECT: survival rate is increased without preparatory cold stratification.

1 tbl, 2 ex

FIELD: agriculture.

SUBSTANCE: when plants are growing under coverage, low molecular chitosan dissolved in a 0.025% aqueous solution of amber acid is applied under the roots at 25 mg per plant. Complex mineral fertiliser is also put at the same time. Treatment is repeated twice a month for the whole vegetation period of the plants.

EFFECT: protecting cucumber plants from gallic eelworm, Meloidogyne incognita.

6 tbl, 5 ex

FIELD: chemistry.

SUBSTANCE: invention is used in agriculture. Plant growth and development regulator is based on products, separated from placental tissues of human or agriculture animals. It contains in its composition (in wt %): polypeptides 2.5-7.0; amino acids 40.0-60.0; amino saccharides 4.0-6.0; hexuronic acids 8.0-10.0; mucopolysaccharides 25.0-40.0; inorganic salts -remaining part. Placental tissue is milled, processed with chlorous acid solution, reaction mixture is neutralised with alkali solution and is converted in work form in form of suspension by fine-dispersed milling.

EFFECT: regulate growth and development of plants.

4 cl, 4 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: agent for preplant treatment of seeds contains a composition of trace elements in doses less than usually recommended for trace elements, wt %: copper 1.00±0.20, zinc 0.90±0.20, magnesium 1.50±0.25, cobalt - 0.04±0.005, iron 0.20±0.03, manganese 0.20±0.05, molybdenum 0.15±0.03, boron 0.15±0.03, selenium 0.005±0.001, water -the balance.

EFFECT: invention increases safety of application when used with pesticides and fungicides, and increases resistance of crops to different diseases.

2 tbl

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