RU2539025C1 - Agent for microbiological protection of plants and method of microbiological protection of plants using this agent - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: agent for microbiological protection of plants comprises a mixture of culture liquids Trichoderma viride, Azotobacter chroococcum, Bacillus megaterium, Beauveria bassiana, Metarhizium anisopliae with the necessary amount of water. The ratio of cultures in the mixture is 1:1:1:1:1, with a titre of each culture is not less than 1×107 CFU/ml. Using the said agent the method of microbiological protection of plants is implemented, comprising 2-3-fold spraying of plants during the growing season at a dosage of 12.5 l of the agent per 1 ha of crops. Spraying is carried out when the average temperature of air is not less than 15°C with an interval of 2-3 weeks.

EFFECT: stimulation of plant growth, increase in plant yield, improvement of soil fertility.

2 cl, 5 tbl, 4 ex

Description

The invention relates to agriculture and can be used in the technology of biological protection of plants from diseases and insect pests during the growing season.

Increasing the resistance of plants to pests and diseases is the main task both in crop production and in agriculture as a whole.

Currently, from preparations and mixtures containing various cultures of microorganisms, “Extrasol” is used to protect plants from pests and stress factors, containing mixtures of cultures of microorganisms Arthrobacter mysore, Flavobacterium sp: L-30, Agrobacterium radiobacter 204, Agrobacterium radiobacter 10, Azomonasas agilis 12, Bacillus subtilis 4-13, Pseudomonas fluorescens 2137, etc. It is used for seed treatment and at the stage of plant vegetation [7, 8].

The disadvantage of this drug is that in it all microorganisms are in conditions of the same pH value, although each microorganism has a certain optimum pH. In addition, the drug does not have a protective effect against insect pests.

Known drug for protecting plants from diseases (RU, Patent No. 2170511), containing, wt.%: The biomass of the strain of the fungus Trichoderma viride PersexS.F. Gray 16 CCM F-59M - 80-90; microcrystalline cellulose - 5-10; magnesium salts - 5-10. The drug has antagonistic activity against a wide range of pathogens of agricultural crops, can be used in conjunction with chemical pesticides.

However, in this preparation there are no cultures of microorganisms that accelerate plant growth and improve soil fertility.

For processing on the vegetative mass of nightshade crops during the budding period of plants before flowering, liquid fertilizer “Riverm” is used in a tank mix with Fitodoctor, Trichodermin, Planriz, Mikosan [6]. Those. phosphorobacter is in symbiosis with Bacillus subtilis, one of the species Trichoderma, depending on the source strain used, as well as with Pseudomonas fluorescens.

The disadvantage of this tank tool is the narrow recommended use for nightshade crops, as well as the lack of microorganism cultures that protect against insect pests.

Known microbial composition for controlling the larvae of nutcracker beetles (Patent RU No. 2311768), comprising an equal volume mixture of uterine cultures of entomopathogenic fungi Beauveria bassiana F-65, Metarhizium anisopliae var. Anisopliae F-596.

Also known are insecticides based on entomopathogenic microorganisms, such as Beauveria bassiana and Metarhizium anisopliae fungi and Bacillus turhingiensis bacteria, which include usnic acid as a synergist of insecticides, and the caterpillar beetles No. 4 and larvae 4 .

These insecticidal preparations based on the entomopathogenic microorganisms Beauveria bassiana and Metarhizium anisopliae have an insecticidal effect, but do not exhibit a curative effect from bacterial, fungal, and viral diseases of plants.

The closest in technical essence is a method of protecting perennial crops using a mixture of three to six biofungicides. (Patent RU No. 2458503). We take it for a prototype. Biofungicides of various antifungal and antibacterial nature are used, at a dosage of 50-100% per hectare. A mixture of six different cultures of microorganisms is used. Bactofit (Bacillus subtilis strain IPM-215 and the produced antibiotic) - 2.5 l / ha, Planriz (Pseudomonas fluorescens strain AP-33) - 1.5 l / ha, Trichoderma (Trichoderma viride) - 1.5 l / ha, Trichocin (Trichoderma harzianum) - 100 g / ha, Phytolavin (Streptomyces griseus or Streptomyces lavendulae strain 696) - 1 l / ha, Vermiculen (Penicillium vermiculatum) - 2.5 l / ha). Or use a composition of three cultures of microorganisms (Alirin-B (Bacillus subtilis strain B-10 VIZR) - 100 g / ha, Trichodermin (Trichoderma viride - 1.5 g / ha, Funiculosum (Penicillium funiculosum) - 2.5 l / ha ). Each is separately dissolved in a small amount of water (10 L). Then all solutions are mixed with water in the tank immediately before spraying and the prepared tank mixture is sprayed with the plantings available on the farm. The consumption rate per 1 ha is 800-1200 liters. Spraying is carried out through 5-15 days after harvesting. th spraying after 2-4 weeks.

The disadvantage of these tank mixtures is the introduction of a significant amount (2.5 l / ha) of spore microorganisms in relation to the introduced volume of fungal cultures and the absence of mushroom cultures in the mixture that would have a detrimental effect on insects. In addition, the presence in the mixture of Fitolavin, which is a complex of streptotricin antibiotics, inhibits the soil microflora.

To date, biological plant protection products containing microorganism cultures: Trichoderma viride, Beauveria bassiana, Metarhizium anisopliae, Azotobacter chroococcum, Bacillus megaterium, which simultaneously increase resistance to bacterial, fungal and viral diseases and stress factors that protect against pests and improve soil fertility plant nutrition - no. Moreover, the inventive mixture is recommended for processing during the growing season.

Thus, the claimed tool meets the criteria of patentability - novelty.

The technical result of the claimed group of inventions is to obtain an environmentally safe, non-pathogenic for humans and animals biological means of combating plant diseases and insect pests, increasing the effectiveness of the tool by using a complex of biofungicides and bioinsecticides, providing the ability to stimulate plant growth and improve soil fertility, as well as the use of the drug during the growing season is the most favorable complex for reducing the number of pathogens sa diseases and insect pests.

The technical result is achieved by the fact that the agent for microbiological protection of plants includes a mixture of culture fluids: Trichoderma viride: Azotobacler chroococcum: Bacillus megaterium: Beauveria bassiana: Metarhizium anisopliae with the required amount of water at a crop ratio of 1: 1: 1: 1: 1 at a titer each culture is not less than 1 × 10 ⁷ CFU / ml.

The technical result is also achieved by the fact that the method of microbiological protection of plants using the agent according to claim 1 includes 2-3-fold spraying of plants during the growing season at a dosage of 12.5 liters of agent per 1 ha of crops, when the average air temperature reaches at least 15 ° C, with an interval of two to three weeks.

Timely treatment of plants during the growing season at a consumption rate of 12.5 liters of tank mixture per 1 ha will provide the plant with nutrition and protection from diseases and insects. At a lower consumption of funds, mainly soil phytopathogens and insect pests develop, as a result of which yield decreases. The use of funds in an amount of more than 12.5 l / ha leads to inappropriate cost overruns.

The microbiological protection agent is a mixture of culture fluids of five microorganism cultures: Trichoderma viride, Beauveria bassiana, Metarhizium anisopliae, Azotobacter chroococcum, Bacillus megaterium. It allows you to use a mixture of biofungicides and bioinsecticides at the same time and is a systemic tool that improves soil fertility and the process of plant nutrition, leading to a population of non-harmful levels and the gradual extinction of pests, suppressing pathogens and increasing resistance to them, as well as stress factors.

Due to the cellulolytic and lignolytic activity of Trichoderma viride, degradation and reduction of the molecular weight of polymer nutrient components in the soil occurs, which makes them available for nutrition of other microorganisms introduced into the tank mixture. In addition, due to the release of Trichoderma viride antibiotic substances, the soil is decontaminated, which protects plants from rot, root infections. Trichoderma viride inhibits the propagation of phytopathogens: Fusarium, Phytophthora, Alternaria, Pythium, Botrytis, Phoma. Trichoderma viride also inhibits the development of rust, powdery mildew pathogens. The fungus stimulates the root nutrition of plants and improves soil fertility. A synergistic effect on insects is achieved by the simultaneous introduction of Beauveria bassiana and Metarhizium anisopliae. The short development cycle and high reproductive ability of Beauveria bassiana and Metarhizium anisopliae increases the possibility of the parasite getting into insects: Colorado potato beetle, potato bug, meadow and corn moth, harmful tortoise, various types of moths, acacia moth, cotton scoop, pea weevil, beetroot weevil , wireworm, bear, larvae, relic lumberjack, termites, etc. The migration of infected insects with the initial stages of muscardinosis contributes to the spread of the disease. At the same time, the spectrum of endangered pests, including thrips, cereal flies, ground beetles, is expanding significantly. The addition of Azotobacter chroococcum, Bacillus megaterium to the tank mixture allows the plants to be provided with nutrient-friendly forms of nitrogen and phosphorus. Microorganisms also participate in the mineralization process. In addition, they are antagonists of root rot and bacterial diseases of plants.

The action of the claimed funds is based on the symbiotic relationship of cultures among themselves. Trichoderma viride metabolism products are used by Beauveria bassiana and Metarhizium anisopliae. At the same time, Beauveria bassiana and Metarhizium anisopliae have a different time peak of development. Also, the accumulation of organic matter is very important for the development of active aerobic cultures of Azotobacter chroococcum. In addition, the bacteria Bacillus megaterium translate phosphorus into a soluble state available to plants and its mineral forms necessary for plant nutrition and Azotobacter chroococcum.

Thus, included in the composition of the claimed means of culture of microorganisms ensure the achievement of a technical result. This determines the compliance of the claimed solution with the patentability condition - inventive step.

Preparing a culture medium for growing microorganisms is carried out as follows: 25 kg of sugar are poured into 1000 liters of water poured into a fermentation tank, then 30 kg of sifted wheat or corn bran are added, and potassium phosphate disubstituted 100 g / 1000 l of medium is added. Then set the required pH of the nutrient medium. For this, a 10% phosphoric acid solution (to acidify the medium) and a 10% potassium hydroxide solution (to alkalize the medium) are introduced in such an amount as to set the initial pH of the medium for Azotobacter chroococcum and Bacillus megaterium equal to 7.0 ± 0.5, and for Trichoderma viride, Beauveria bassiana, Metarhizium anisopliae -6.5 ± 0.5 (the number of added solutions of phosphoric acid and potassium hydroxide depends on the initial pH of the water). Mix thoroughly, after which bran swells for 2 hours. Next, 1 kg of each of the declared cultures of microorganisms in a dry form (freeze-dried cultures) is introduced into the nutrient medium. After that, the process of fermentation and propagation of microorganisms is carried out at a temperature of 15-20 ° C with periodic stirring and aeration of the mixture until a titer of at least 1 × 10 ⁷ CFU / ml is achieved for 12-14 days. Each culture of the claimed microorganisms Trichoderma viride, Beauveria bassiana, Metarhizium anisopliae, Azotobacter chroococcum, Bacillus megaterium is grown in a separate fermenter. Then the culture fluid is filtered. At the same time, certain portions of the culture fluid are taken from each container, placed in a separate container in a ratio of 1: 1: 1: 1: 1, the required amount of water is added and their uniform mixing is carried out. The resulting tank mixture of the inventive cultures of microorganisms is used to treat vegetative plants on the day of preparation.

The method of microbiological protection of plants using the tool according to claim 1 includes 2-3-fold spraying of plants with a tank mixture during the growing season at a dosage of 12.5 l of the mixture per 1 ha of crops, when the average air temperature reaches at least 15 ° C, with an interval of two -three weeks.

To prove the conformity of the proposed solution to the condition of industrial applicability, we give examples of a specific implementation.

Example 1

Each culture of the claimed microorganisms Trichoderma viride, Beauveria bassiana, Metarhizium anisopliae, Azotobacter chroococcum, Bacillus megaterium is grown in separate fermentation tanks. Next, certain portions of culture liquids with a titer of at least 1 × 10 ⁷ CFU / ml are selected from each container, placed in a separate container in a ratio of 1: 1: 1: 1: 1, the required amount of water is added and they are mixed uniformly. The resulting tank mixture is sprayed with plants during the growing season. The consumption of each culture of the microorganism in a dosage of 2.5 l / ha or 12.5 l / ha of the mixture. The plants are treated when the average air temperature reaches at least 15 ° C, with an interval of two to three weeks.

The effectiveness of using the product according to claim 1 against insect pests on the example of use on winter wheat is presented in table 1.

The increase in yield when using the inventive microbiological remedies in comparison with an insecticide occurs mainly due to the fact that the defeat of insect larvae by the fungi Beauveria bassiana and Metarhizium anisopliae occurs at all stages of their development. Also, infection of insects with these fungi affects not only the viability of the infected generation, but also affects the fertility of subsequent generations, reducing it. This is achieved as a result of the spread of mycelium of fungal cultures with infected migrating insects, as well as the ability of fungi to further asexual reproduction carried out by conidia. Those. their action is prolonged. While the insecticide, having a detrimental effect on insects, affects the plant.

The effectiveness of using the product according to claim 1 from diseases using winter wheat as an example is presented in table 2.

The effectiveness of using the product according to claim 1 from diseases using the example of use on sunflower is presented in table 3.

A comparative assessment of the use of the claimed protective agent with a standard for plant diseases shows that, with equal results from the influence of the standard and the claimed microbiological agent on plants, the crop is 2 kg / ha higher when processed by the claimed agent. This is due to the fact that there is a gradual acquisition of resistance of pathogens to chemicals. And the simultaneous use of several cultures of microorganisms, as producers of antibiotic substances and enzymes to combat pathogens, allows you to extend the action of the product during the period of plant development.

Example 2

Evaluation of the effectiveness of the method of application of the proposed tool is carried out during field trials.

Field tests were carried out at Aksayskaya Zemlya LLC, Rostov Region, where they evaluated the effectiveness of the content of humus in the soil and wheat yield.

Prepare a tank mixture of the declared culture fluids of the microorganisms Trichoderma viride, Beauveria bassiana, Metarhizium anisopliae, Azotobacter chroococcum, Bacillus megaterium with a titer of at least 1 × 10 ⁷ CFU / ml, taken in a ratio of 1: 1: 1: 1: 1. Then, the prepared wheat crop is sprayed with winter wheat crops during the growing season, with the average air temperature not lower than 15 ° C. After 2-3 weeks, repeat the treatment of crops. The use of funds by the claimed method allowed to increase the content of humus by 0.16% for 5 years. At the same time, wheat productivity increased from 11 kg / ha to 35-40 (up to 50 in some years) kg / ha. This is because the decomposition of organic matter of the soil by the claimed microorganisms contributes to the accumulation of humus. In addition, the dead cells of microorganisms themselves are a reserve for their decomposition by other microorganisms, and this helps to increase productivity. Similar tests were carried out on wheat at the Aleksandrovsky Production and Industrial Complex, Rostov Region, Temizhbeksky LLC, Stavropol Territory.

The use of a mixture of culture fluids of microorganisms reduces the number of pathogens of a complex of diseases and insect pests through the simultaneous use of a complex of biofungicides and bioinsecticides; stimulating plant growth and improving soil fertility. Spraying with a mixture of plant culture liquids at the stage of growth and development is due to the fact that this is the most favorable period, because throughout the development of plants, a decrease in the number of pathogens of the disease complex and a gradual extinction and expansion of the spectrum of populations of insect pests are achieved.

Example 3

Confirmation of the possibility of using the tank mixture in the required (any) amount of water is given in table 4. For this purpose, wheat crops were sprayed with the declared agent in an amount of 12.5 l / ha, applied in 100, 300, 500, 800 and 1000 liters of water (excluding soil moisture and precipitation), followed by% damage by insects and% root rot. The results are presented in table 4. A comparative assessment confirms that in this case, a prerequisite for achieving the desired result is the introduction of exactly 12.5 l / ha of a mixture of culture fluids containing cells, spores, conidiomycelial mass of microorganisms and their metabolic products.

Example 4

The possibility of using different strains is given in table 5. The species affiliation of microorganisms characterizes their narrowly targeted enzymatic activity and the direction of action against pathogens of diseases and insects. Despite the varying degrees of secretion of biologically active substances, antibiotics and fungicides of various strains, when using the claimed cultures in the tank mixture, there was no significant difference in% expression in the suppression of diseases and the entomopathogenic effect during the treatments. This confirms the possibility of using different strains within the same type of the claimed microorganisms.

Information sources:

1. Kotlyarov V.V. Physiology of Plant Immunity: A Training Manual. / V.V. Kotlyarov - Krasnodar, 2006 .-- 102 s.

2. Mosichev S. M., Skladnev A. A. Kotov V.B. General technology of microbiological production. - M .: Light and food industry, 1982. - 264 p.

3. Biological plant protection / M.V. Sternšys, F.S.-U. Jalilov. I.V. Andreeva, O.G. Tomilova; under the editorship of M.V. Sternsys. - M.: KolosS, 2004 .-- 264 p.

4.http: //ru-patent.info

5. www.ovoshevodstvo.com/joumal/brovse/2010 10 /

6.http: //colhoz.com

7.http: //beltorg.org/product_info.php

8.http: //www.bisolbi.ru

Claims (2)

1. Means for microbiological protection of plants, including a mixture of culture fluids Trichoderma viride, Azotobacter chroococcum, Bacillus megaterium, Beauveria bassiana, Metarhizium anisopliae with the required amount of water in the ratio of cultures in the mixture 1: 1: 1: 1: 1 with a titer of each culture not less than 1 × 10 ⁷ CFU / ml. 2. The method of microbiological protection of plants using the tool according to claim 1, including 2-3-fold spraying of plants during the growing season at a dosage of 12.5 l of the product per 1 ha of crops, when the average air temperature reaches at least 15 ° C, with an interval two to three weeks.