RU2130264C1 - Preparation for plant growth stimulation and protection from sicknesses - Google Patents

Abstract

FIELD: microbiological industry, agriculture. SUBSTANCE: preparation has the following components, wt.-%: complex of strains of bacterium Pseudomonas species 7G, 7G2K, 17-2 taken at ratio (1:2:1)-(2:1:2) at concentration (2-5) x 10¹⁰ cells/ml, 4.5-5.5; humates, 5.0-10.0; trace elements, 0.02-0.05; and water, the balance. Preparation ensures to decrease vulnerability of plants by sicknesses caused by broad spectrum of phytopathogens. EFFECT: enhanced effectiveness of the preparation, increased yield of agriculture crops. 5 tbl, 7 ex

Description

 The invention relates to the microbiological industry and biotechnology, specifically to means for combating plant pathogens caused by ontopathogenic microorganisms, and can be used in agriculture to protect plants from diseases.

 To combat plant diseases, preparations based on various microorganisms are increasingly used, since the latter are harmless to beneficial fauna and do not pollute the environment, which is observed when agrochemicals are used [1, 2].

 Known, for example, is a preparation for combating potato scab containing a suspension of cells of the bacterial strains Pseudomonas fluorescence biobar V (strain MD-4f), Enterobacter agglomerans (strain 2-3B), Pseudomonas sp (strain F13-I), Acinetobactor sp (strain M24 -I) [3].

To combat potato scab, the latter is treated with a suspension of bacteria with a concentration of 1 • 10 ⁹ cells / ml.

 A disadvantage of the known preparation is a narrow spectrum of the fungicidal action of the latter, since it inhibits the reproduction of only fungi of the genus Streptomyces (bacteria that cause potato scab).

Known drug for stimulating plant growth, containing a suspension of cells of the bacterial strain Pseudomonas aureofaciens H16 in a concentration of (2 - 4) • 10 ⁹ cells / ml. The drug allows you to increase plant yield due to stimulating activity, but does not protect plants from various phytopathogens [4].

 Known drug for combating plant diseases, containing zeolite, molasses, wheat and rice bran, suspension of cells of the microorganisms Cellulomonas flavigene, Aspergillus oryzae, Nitrobacter gilis. The drug is effective in controlling plant diseases caused by phytopathogenic fungi of the genus Basidiomycetes, Rhizoctonia, and Helminthosporium [5].

 The disadvantages of the known drug are a narrow spectrum of antagonistic activity, multicomponent, the duration of receipt and weak stimulating activity on the growth and development of plants.

The closest to the claimed prototype is a drug to stimulate growth and protect plants from fungi of the genus Fusaruium and Erwinia bacteria [6], containing a suspension of the bacterial strain Pseudomonas putida at a concentration of 1 • 10 ⁹ cells / ml. Presowing treatment of plant seeds with this drug is carried out at the rate of 10 ⁶ - 10 ⁷ cells per 1 g of seeds. A disadvantage of the known drug is a narrow spectrum of its antagonistic effect (only on fungi of the genus Fusarium and bacteria Erwinia).

 An object of the invention is to obtain a new biological product with a wide spectrum of antagonistic activity against plant pathogens, as well as having a stimulating activity on plant growth and development.

The problem is achieved by the proposed drug to stimulate growth and protect plants from diseases, containing the following components, wt.%:
The complex of strains of Pseudomonasspecial 7G, 7G2K and 17-2 at a ratio of 1: 2: 1 - 2: 1: 2 with a concentration of (2 - 5) • 10 ¹⁰ cells / ml - 4.5 - 5.5
Humates - 5.0 - 10.0
Trace elements - 0.02 - 0.05
Water - Else
The strains Pseudomonas specias 7Г, Pseudomonas sp 7Г2К and Pseudomonas sp 17-2 were isolated from the rhizosphere population of microorganisms of wild cereal plants in the Horde forest-steppe of the Novosibirsk region. These strains were deposited at the Research Institute “Collection of microorganism cultures” SSC VB “Vector” under registration numbers V-694 (7G) B V-695 (7G2K) and V-696 (17-2). The strains of Pseudomonas sp 7G (NII KKM B-694), 7G2K (NII KKM B-695) and 17-2 (NII KKM B-696) used in the inventive preparation have the following cultural, morphological and physiological and biochemical characteristics.

1. Cultural and morphological characteristics Gram-negative motile rods of size (0.5 - 0.7) • (1.2 - 3.8) microns, with polar flagella, do not form spores. The morphology of the colonies on nutrient media was determined after growth for 2-3 days at 28 - 30 ^o C in MPA. Strains form colonies: strain 7G - medium (6 - 8 mm (convex, transparent with a matte finish, smooth surface, mucous consistency, strain 7G2K - large (10 - 12 mm), whitish, highly convex, densely mucous, strain 17-2 - small (3 - 5 mm), slightly convex, transparent, smooth, round.

On King diagnostic environments:
King A - pigment is absent for all strains;
King B: strain 7G - pigment yellow-green, strain 7G2K - intensely green, strain 17-2 - pigment bright green.

 2. Physiological and biochemical properties.

All aerobic strains, optimum growth temperature 28 - 30 ^o C, grow to 41 ^o C, pH 5.5 - 7.5, the optimum 7.0 - 7.2. Glucose, glycerin, mannitol are used as carbon sources and do not absorb raffinose, galactose, maltose, sucrose, sorbitol, dulcite.

 The strains are resistant to antibiotics: rifampicin, ampicillin, novobiocin, chloramphenicol, kanamycin and are sensitive to tetracycline.

The strains are stored in 25% glycerol at (-20) - (70 ^o C) or in a freeze-dried state. Strains are non-pathogenic for warm-blooded animals and humans.

The drug is prepared as follows. In three flasks with a capacity of 750 ml, pour 250 ml of LB broth containing (g / l): tryptone or peptone - 10.0; yeast extract - 5.0; sodium chloride - 10.0; water is the rest. Then, individually 1-2 ml of a stationary culture of Pseudomonas sp. 7G, Pseudomonas sp. 7G2K and Pseudomonas 17-2. The cultivation of three cultures is carried out at 30 ^o C on a rocking chair (250 rpm) with aeration for 12 to 16 hours to the final titer (3 - 6 • 10 ¹⁰ cells / ml).

Then humates, microelements are added to a 1 dm ³ capacity container, 200-300 ml of water are added, the cell suspension of the complex of strains of Pseudomonas sp. Is added and added. 7G, 7G2K, 17-2, taken in a certain ratio, up to 1 liter. The concentration of bacterial cells in terms of dry weight in the preparation is 4.5 - 5.5 wt.%, And the titer of living cells is pseudomonassp. in the preparation (2 - 5) • 10 ¹⁰ cells / ml suspension.

 The resulting preparation is an aqueous suspension of yellowish-brown color, with a specific odor, readily soluble in water.

 The defining differences of the proposed drug in comparison with the prototype are: as a bacterial culture using a complex of strains of Pseudomonas sp. 7G, 7G2K, 17-2, taken in the optimal ratio, which allows to increase the fungicidal activity of the drug by expanding the spectrum of antagonistic action against phytopathogenic microorganisms. Each of the strains included in the preparation has high specificity with respect to a specific phytopathogen, and all together provide a high level of stimulating and fungicidal action. A decrease in the content of the complex of microorganism cell strains in the preparation below 4.5% leads to a decrease in the biological activity of the drug, and an increase above 5.5% practically does not lead to an increase in the biological effect, but can inhibit plants, therefore, it is economically and practically not practical, humates and microelements in experimentally selected optimal concentration are introduced as a useful additive, which allows to increase the stimulating activity of the drug on plant growth and development. Humates released from soils and ends are a mixture of high molecular weight humic and fulvic acids, as well as low molecular weight organic acids (amino acids, oxalic, citric, etc.) and their complexes with metals - iron, aluminum, manganese, zinc, copper. Under the influence of humates, metabolic processes are activated in the plant organism, respiration and the intake of mineral nutrition from the environment are enhanced.

 The addition of microelements to the preparation in an amount of 0.02-0.05% allows to increase the stimulating activity of the latter due to the balanced mineral nutrition of plants.

 The invention is illustrated by the following examples of specific performance.

Example 1. Getting the drug. Into a 1 L volumetric flask, add 50 g of humates, 0.3 g of trace elements and 200 to 300 ml of water, then a cell suspension (with a titer of 3 - 6 • 10 ¹⁰ cells / ml), consisting of a mixture of strains of Pseudomonas sp. 7G, 7G2K, 17-2, in a ratio of 1: 2: 1, bring the volume to 1 liter. The result is a preparation with the following content of components, wt.%:
The complex of strains of Pseudomonas sp. 7G. 7G2K, 17-2 with a ratio of 1: 2: 1 - 5.0
Humates - 5.0
Trace elements - 0.03
Water - 89.97
The resulting preparation is predominantly treated with seeds and vegetative cereal plants.

Before use, the drug is diluted with water in a ratio of 1: 200. The working concentration of the solution of the claimed drug is (1 - 2.5) • 10 ⁸ cells / ml.

Example 2. The drug is prepared analogously to example 1 and get the target product with the following content of components, wt.%:
The complex of strains of Pseudomonas sp. 7G. 7G2K, 17-2 with a ratio of 1: 1: 2 - 4.5
Humates - 5.5
Trace elements - 0.02
Water - 89.98
The drug with this ratio of components is mainly used for presowing treatment of seeds and vegetative plants of legumes and oilseeds.

Example 3. The drug is prepared analogously to example 1 and get the target product with the following ratio of components, wt.%:
The complex of strains of Pseudomonas sp. 7G. 7G2K, 17-2 with a ratio of 2: 1: 2 - 5.5
Humates - 10.0
Trace elements - 0.05
Water - 84.45
The drug with this ratio of components most effectively proved to be on potatoes and vegetables.

Example 4. Field tests of the drug obtained according to example 1, containing, wt.%:
The complex of strains of Pseudomonas sp. 7G. 7G2K, 17-2 in a ratio of 1: 2: 1 with a concentration of (2) • 10 ⁹ cells / ml - 5.0
Humates - 5.0
Trace elements - 0.03
Water - 89.97
conducted in the Kirzinsky CJSC Ordynsky district of the Novosibirsk Region on a field of 3.6 thousand hectares to stimulate growth and protect soft spring wheat Novosibirsk-89 from root rot damage. For this, pre-sowing seed treatment with the claimed preparation was carried out at the rate of 50 ml per 1 t. Before treatment, the preparation was diluted in 10 l of water, the working concentration of the preparation solution was 1 • 10 ⁸ cells / ml. Control seeds were treated with a chemical preparation - Vitavax 200 ff.

 The results are presented in table. 1.

 From the table. 1 shows that the treatment of seeds with the claimed preparation reduces the development of root rot on the primary and secondary roots, contributes to a better development of secondary roots and will increase productivity by 3 kg / ha (in conditions of cold spring-summer drought).

Example 5. The plot tests of the preparation obtained according to example 1 were carried out at the Institute of Agriculture of the Institute of Agriculture and Chemicalization of Agriculture of the RAAS. The size of the accounting plots was 30 m ² , the repetition was fourfold, the seeds were treated by the method of semi-dry etching.

To stimulate the growth and protection of spring wheat Lutescens-25 from damage by root rot, pre-sowing treatment of seeds with the claimed preparation was carried out at the rate of 50 ml per ton of seeds. Before treatment, the drug was diluted with water (10 l) in a ratio of 1: 200. Control seeds were treated with water. To compare the effectiveness of the claimed complex preparation, a similar seed treatment was carried out with the following preparations: Vitavax 200 ff (3 kg / t), a suspension of the complex of strains (SKS) Pseudomonas sp. 7g, 7G2K, 17-2, taken in a ratio of 1: 2: 1 at a concentration of 5 • 10 ¹⁰ cells / ml at a dose of 50 ml / t; aqueous solution of humates at a concentration of 0.025%.

 The results are presented in table. 2.

 From the table. 2 shows that the claimed drug is characterized by a higher stimulating and fungicidal effect than drugs containing a suspension of bacterial cells or humates separately (synergism).

 Data on the effect of pre-sowing treatment of spring wheat seeds Lutescens-25 on the development of root rot on primary and secondary roots in May-June 1997 are presented in table. 3.

 From the table. 3 shows that the claimed drug has increased phytosanitary fungicidal activity than the standard drugs.

Example 6. The plot tests of the preparation obtained according to example 2 were carried out at the experimental site of the Institute of Agriculture and Chemicalization of Agriculture of the Russian Academy of Agricultural Sciences on wheat Novosibirsk 89, hybrid corn Moldavsky-23, soybean varieties SBNIIK-315. Before sowing, the seeds of wheat, corn and soybeans were treated with the drug at the rate of 50 ml per 1 ton of seeds, and before treatment the drug was diluted with water in a ratio of 1: 200 (50 ml of the drug per 10 l of water), and the seeds treated with water served as a control. The repetition of the experiment is 4 times. The area of each plot is 30 m ² .

 The test results are presented in table. 4.

 From the table. Figure 4 shows that the drug has a stimulating effect on the growth and development of plants; pre-sowing seed treatment 2 times accelerates the emergence of seedlings after sowing seeds.

 Example 7. Field trials of the preparation obtained according to example 3 were carried out at Prigorodny AOZT in the Novosibirsk Region to protect potato plants from diseases caused by phytopathogenic fungi of the genus Phytophora and Rizoctonia. For this, pre-sowing treatment of seed was carried out with the preparation obtained in Example 3 at the rate of 50 ml per 1 ton of tubers.

 Planting material weighing 3.6 tons was planted in a pilot plot of 0.4 ha. The comparison data of the crop from the experimental and control plot of the same area are presented in table. 5.

 From the table. Figure 5 shows that the proposed drug allows you to protect potato plants from phytopathogens and increase potato productivity by 25 - 30%.

 Thus, the use of the proposed drug allows, in comparison with the prototype drug, to provide a wider antagonistic effect on phytopathogens (not only on fungi of the genus Fusarium, but also on phytopathonens of the genus Alternaria, Bipolaris, Rizoctonia, Phytophora, etc.); provide stronger stimulation of plant growth and development due to the optimal set of basic starting components for plant nutrition (humates and trace elements) and bacteria that stimulate the development of the root system, especially secondary roots.

The combination of all components of the claimed drug is useful and justified by the fact that they exhibit noticeable synergism, and the drug as a whole is characterized by high rates of stimulating and fungicidal effects. In this case, the consumption rate of the drug can be reduced by 4-10 times compared with the prototype drug (the working concentration of the aqueous solution of the claimed drug is (1 - 2.5) • 10 ⁸ cells / ml, and the prototype - 1 • 10 ⁹ cells / ml

Sources of information
1. Aleshina OA, Biological methods of combating diseases and pests of agricultural plants. M., "Knowledge", 1974.

 2. Khizhnyak P.L. and other Chemical and biological plant protection, 1971, 171 - 172.

 3. Japanese application N 1-193203A, cl. A 01 N 63/00. Means for combating the scab of potatoes, publ. 08/03/89.

4. RF patent N 200195-C, cl. A 01 N 63/00 "The bacterial strain Pseudomonas aureofaciens - to obtain a plant growth stimulator; publ. 10/30/93. BI N 39-40 /
5. Japanese application N 2-91010A, CL A 01 N 63/04. The drug to combat plant diseases, publ. 03/30/90

 6. RF patent N 1805849A3, cl. A 01 N 63/00. The bacterial strain pseudomonas putida to obtain a drug used to stimulate plant growth and protect plants from fungi of the genus Fusarium and Erwinia bacteria; publ. 03/30/93, BI N 12.

Claims (1)

A preparation for stimulating the growth and protection of plants from diseases, including bacterial cells of a microorganism of the genus Pseudomonas and water, characterized in that the preparation contains a complex of bacterial strains of Pseudomonas species 7G, Pseudomonas sp. 7G2K, Pseudomonas sp. 17-2, taken in a ratio of 1: 2: 1-2: 1: 2 with a concentration of (2-5) • 10 ¹⁰ cells / ml and additionally contains humates with trace elements in the following ratio of components, wt.%:
The complex of strains of Pseudomonas sp. 7G, 7G2K, 17-2 - 4.5 - 5.5
Humates - 5.0 - 10.0
Trace elements - 0.02 - 0.05
Water - Else

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