RU2802346C1 - Bioinsecticide composition for protection of crops from soil pests - Google Patents

Abstract

FIELD: agriculture and biotechnology.

SUBSTANCE: bioinsecticide composition comprises a solid carrier material and a biological component. As a solid carrier material, a mixture of vermiculite with brewer's grains or brewer's grains is used, and as a biological component, immobilized biomass of a bacterial-fungal complex is used, which includes the biomass of micromycete Beauveria bassiana F-1357 in combination with bacteria Bacillus thuringiensis B-6335 or actinobacteria Pseudonocardia carboxydivorans AC-2046, taken at a ratio of 1:1-3 by volume from the biomass of fungi and biomass of bacteria for subsequent immobilization on the carrier material and content with a concentration of at least 1×10⁷ CFU in 1 g of the composition, in the following ratio of components, wt. %: carrier material 80-90; biomass of the bacterial-fungal complex 1-5; moisture is the rest. A mixture of vermiculite and brewer's grains is taken at a volume ratio of 1:3.

EFFECT: reduction of populations of harmful soil-living insects in the stage of adults or larvae and increased plant productivity.

2 cl, 3 tbl, 3 ex

Description

The invention is intended for use in agriculture, namely in the schemes of biological protection of plants from soil-dwelling insect pests in potato growing and soil structuring.

Harmful soil-dwelling insects, primarily the larvae of click beetles - wireworms are dangerous, widespread pests of seeds and seedlings of crops, especially in Solanaceae crops, causing economic damage to the crop.

A known method of protecting potatoes from wireworms, which consists in making a repellent drug, which is used as a decoction obtained after the release of tar from birch bark. Otgar is applied to the soil during planting along with a potato tuber in the amount of 1-2 tablespoons.

A known drug for combating soil-dwelling harmful insects 12% dust hexachloran (HCCH), which was widely used in the 50s of the last century by the method of continuous application with a drug consumption of 120-130 kg/ha (Lysine concentrate is an environmentally friendly drug for protecting tilled crops). crops from harmful soil insects // Recommendations M .: TsNTIPR, 1989. P. 4).

The disadvantage is that the use of preparations leads to the destruction of soil organisms and leads to extreme contamination of the soil with insecticide residues for a long time. In addition, the presence of horizontal and vertical migrations of insects and, above all, wireworms, reduced the effectiveness of chemical treatments with insecticide.

A known drug for the protection of row crops and industrial crops, including entomopathogenic nematodes and an attractant introduced when planting crops (Dobrovolsky B.V. et al. Environmentally safe protection of row crops from wireworm // Production of environmentally safe crop production-Pushchino: VNIIBZR, 1995 pp.308-310). The disadvantage of this drug is the low efficiency in the application and short shelf life. In addition, the use of nematodes is technologically difficult, since they are multicellular animals and quickly lose their viability.

A known composition for combating soil pests containing a solid carrier in the form of granular superphosphate - 50 kg and a feed concentrate of lysine - 2.5 kg applied per hectare (Zhdankin F.A. The effectiveness of insecticides with the addition of lysine concentrate in the fight against wireworms on vegetables - gourds in the conditions of the Moldavian SSR // The use of lysine feed concentrate in crop production. Publishing House of Rostov University, 1987. P. 64-65). However, this concentrate uses chemical carriers for the attractant, which is not always justified both from an economic and economic point of view. When dissolving superphosphate, there is a local accumulation of mineral salts in the zone of attractant activity, which can lead to a decrease in the attractiveness of the composition. In addition, mineral fertilizers are expensive to use over large areas. The use as an attractant is not economically justified due to its high cost.

Known microbiological composition for combating the larvae of click beetles (RU 2311779; EN 2311029; EN 2311774), containing a solid carrier and a microbiological source of lysine. In this case, wheat grain is used as a solid carrier, and a protein-vitamin concentrate is used as a source of lysine, and this composition additionally contains a mother culture of the entomopathogenic fungus Beauveria bassiana F-65 and succinic acid.

A known method for producing a biopesticidal preparation containing a solid carrier - brewer's grains and a microbial component - a strain of entomopathogenic fungi Beauveria bassiana F-145 (RU 2651487).

The closest is a fungal composition for combating click beetle larvae (RU2311031), containing a solid carrier and a microbiological source of lysine, while wheat grain is used as a solid carrier, and a protein-vitamin concentrate is used as a source of lysine, and this composition additionally contains an equal volume mixture of mother cultures of entomopathogenic fungi Beauveria bassiana F-65 and Metarrhizium anisopliae var. anisopliae F-596.

The known composition does not allow effective control of click beetle larvae using both chemical and biological components, the complexity of the preparation of the composition and the lack of information on the effect of the components on improving the structure and quality of the soil, as well as the crop yield.

The technical solution is to create a composition to ensure effective control of soil-dwelling insect pests of agricultural crops - click beetle larvae using a bio-insecticide composition based on a solid carrier material with an immobilized microbiological component, including a bacterial-fungal complex, the interaction of microorganisms of which provides a synergistic effect .

When introducing a biocomposition by bringing soil or parts of plants into contact with the claimed biocomposition, consisting of components that provide broad functionality or multifunctional properties, it helps to reduce populations of harmful soil-living insects in the adult or larval stage, and also helps to increase plant productivity.

The technical result is achieved by the fact that the bioinsecticide composition for combating soil-dwelling insect pests contains a solid biodegradable carrier material and a microbial biomass immobilized on it, namely, brewer's grain or a mixture of vermiculite and brewer's grain is used as a carrier material, and as a biological component, the biomass of the bacterial-fungal complex is used, which includes the biomass of bacteria and fungi, namely the entomopathogenic fungus Beauveria bassiana F-1357 in combination with the biomass of the bacterial strain: entomopathogenic bacteria Bacillus thuringiensis B-6335 or actinobacteria Pseudonocardia carboxydivorans AC-2046, taken at the following ratio components, wt. %:

carrier material 80-90 Biomass of the bacterial-fungal complex 1-5 Moisture Rest

A mixture of vermiculite and brewer's grains is mixed in an effective amount at a volume ratio of 1:3.

As a biological component for the biocomposition, a concentrated biomass of the bacterial-fungal complex obtained by growing the biomass of monocultures and then compiled into a bacterial-fungal complex at a ratio of bacterial biomass to fungal biomass of 1–3: 1 is used for subsequent immobilization by drying on a carrier material with a concentration of not less than 1x10 ⁷ CFU (colony-forming units) in 1 g of the biocomposition, respectively.

The claimed bioinsecticide composition for pest control of agricultural crops, including the larvae of click beetles, differs from the prototype in the nature of the carrier material, the sources of the biocomponent - the microbial bacterial-fungal complex, which creates a synergistic effect, represent a complex biological preparation for plant protection, including entomopathogenic microorganisms, capable of infecting an insect, causing disease, and then the death of the latter, or microorganisms that promote growth and have ligno-cellulolytic activity; carrier material that provides structuring of the composition and additional attractiveness for the pest, as well as provides nutrition to the biomass of fungi and bacteria immobilized on it and ensures their viability for a long time, which contributes to the intensification of sporulation of the fungus and accelerated germination of spores, causing increased infectivity of the microbial biomass of the preparation and, as a result, the high efficiency of the biocomposition as a whole.

The use of the proposed biocomposition provides high efficiency, since brewer's grains or a mixture of grains with vermiculite are used as a biodegradable solid carrier, which provide ease and uniformity of the application of the composition, being at the same time a biodegradable carrier of the composition components, and also ensure high preservation of spores, mycelium and cells of an entomopathogenic fungus B. bassiana F-1357 and bacteria Pseudonocardia carboxydivorans AC-2046 or B. bassiana F-1357 and Bacillus thuringiensis B-6335. Bacteria and fungi included in the composition have a different mechanism of action on insect pests, have the ability to interact in the microbial complex, the ability to adapt and exist in the soil of the endorhizosphere zone of plants. An entomopathogenic fungus, getting on the surface of insects, germinates on them, destroying the surface integument of the insect due to enzymes (protease, lipase, chitinase), ensures its death. As well as bacteria that produce specific entomotoxins (exotoxins) and enzymes that contribute to the effective infection, invasion and poisoning of various stages of development of insect pests. In this case, a dead insect is a source of entomopathogenic agents (spores and areas of mycelium of the fungus, as well as reproduction of bacterial cells), and can independently lead to infection of other individuals of the population.

The strains of used bacteria, filamentous fungi are deposited in the All-Russian Collection of Industrial Microorganisms VKPM [BRC VKPM National Research Center "Kurchatov Institute" - GosNIIgenetika]. Bacterial and fungal strains have entomopathogenic and/or lignocellulolytic activity (Pseudonocardia carboxydivorans AC-2046 [RU 2685858 C1], Bacillus thuringiensis B-6335, micromycete Beauveria bassiana BKPM F-145) and are characterized as environmentally non-toxic. In the classification of microorganisms by pathogenicity groups of the Sanitary and Epidemiological Rules SP 1.3.2322-08 dated May 1, 2008 "Safety of working with microorganisms of III-IV pathogenicity (danger) groups and pathogens of parasitic diseases", this species (genus) is not listed.

Example 1. The preparation of the biocomposition is carried out in several stages. The biomass of the proposed microbial complex is obtained by preparing the inoculum of the strains: for the Beauveria bassiana fungi on agarized Czapek and Sabouraud nutrient media, as well as for the bacterial strains of Bacillus thuringiensis and actinobacteria Pseudonocardia carboxydivorans - on the MPA medium and the Kuster medium, respectively. Liquid-phase cultivation of fungi and bacteria is carried out on the following media: Sabouraud or Chapek broth, meat-peptone broth, Kuster broth, respectively, at a temperature of T=25±3°C for 2-5 days.

The carrier material - spent grains is treated with alkali to neutral pH. A mixture of vermiculite and brewer's grains as a carrier material is obtained by mixing in an effective amount at a volume ratio of 1:3.

On the carrier material (brewer's grains or a mixture of vermiculite and brewer's grains with a fractional composition of not more than 0.9 cm) make a biological component - the biomass of strains of bacteria and fungi. Monocultures or a composition of strains are preliminarily grown on standard nutrient media, and the culture liquid is mixed with the biomass of bacteria and with the biomass of fungi in a ratio of 1-3: 1 by volume and a titer of 1x10 ^7-9 CFU / ml with an initial content of dry biomass of monocultures of at least 5 g/l for bacteria and at least 10 g/l for a fungal strain. Mixing with the carrier material of the biomass of the microbial complex and subsequent drying to immobilize the biological component on the carrier is carried out at T=60±5 ⁰ C. The characteristics of the bioinsecticide composition are presented in table 1.

Table 1

The name of indicators Results of determination, n=3 Titer (content of immobilized biomass on the carrier material: a mixture of pellets and vermiculite, pellets), CFU / g Not less than (1.5±0.5) x10 ⁷ Moisture content,% 14±5 Appearance, form Air-dry bulk mass or pressed - granular form Fractions (particle size), cm 0.01-0.9 Bulk density, kg / m ³ 130-200

The production efficiency of the proposed composition for the control of click beetle larvae is illustrated by examples.

Field experiments to identify the effectiveness of this composition against the wireworm were carried out for several years. The studies were carried out on an experimental field (near the city of Syktyvkar, Republic of Komi), according to the Methodology for conducting variety trials in field experiments. Ecological tests of prototypes were carried out during the growing season on the experimental field according to the scheme of planting three varieties of potatoes on accounting plots-variants, where the area of each variant is 28 m2 in 4-fold repetition, with up to 40 potato bushes in a row. The depth of the arable layer is 22-25 cm. Potato varieties of different maturation groups of mass reproduction: early-ripening - Snegir, medium-early - Charodey (Originator of the Leningrad Research Institute of Agriculture "Belogorka") and mid-ripening - Cerata (Originator - STET HOLLAND B.V.) Weeds were dominated by: sow thistle, creeping wheatgrass. The soil of the area under the experiment is soddy-weakly podzolic, light loamy, heterogeneous; pH=4.47; humus - 1.69%; K2O-105 and P2O5-183 mg/kg. Planting, observation and accounting was carried out in accordance with the methods of potato variety testing. The cultivation of 3 varieties of potatoes of mass reproduction was carried out with the processing of prototypes, where composition No. 1 includes Beauveria bassiana F-1357 and Bacillus thuringiensis B-6335, and composition No. 2 - the composition of Beauveria bassiana F-1357 and Pseudonocardia carboxydivorans AC-2046, taken in effective quality at various ratios.

Example 2. Field experience on environmental testing of prototype biocompositions based on brewer's grains

The biopreparation form immobilized on the carrier, the bioinsecticidal composition, was introduced when potatoes were planted in accordance with the experimental scheme. The study of the effect of bioinsecticide composition in the field conducted observations during the growing season for 91 days. Testing of prototypes using brewer's grains as a carrier with immobilized biomass of the microbial complex at a ratio of 3:1 of bacterial biomass and fungal biomass. The treatment was carried out once when planting potatoes by introducing the biocomposition into the soil at the rate of 60 g/m ² . Accounting for damage was carried out during the period of tuber formation, as well as accounting for damage to tubers after harvesting in accordance with the method of determination. Indicators of wireworm damage and the yield of marketable potatoes by options are presented in table 2.

table 2

Potato variety Accounting for indicators of damage by wireworm (%);
Control (no processing) No. 1. B. bassiana + B. thuringiensis No. 2. B. bassiana + P. carboxydivorans Wireworm damage,% Average yield, % /
Commercial yield, % Wireworm damage,% Average yield,% /
Commercial yield, % Wireworm damage,% Average yield,% /
Commercial yield, % Early ripe (Bullfinch) 0-5 1 - 3.3 0 - 2 Mid Early (Wizard) 0.5-7 0 - 6.7 18 Mid-season (Cerata) 9–11 5.5 - 11 1 - 11 NSR _0.5 33.5

Example 3. Field experience on environmental testing of prototypes of biocomposition based on a mixture of brewer's grains and vermiculite

As a carrier material, a mixture of vermiculite and spent grains was used at a volume ratio of 1:3. A test using a mixture of vermiculite with brewer's grains as a carrier for biomass was carried out using a microbial complex at a biomass ratio of bacteria and fungal culture of 1:1.

The study of the effect of bioinsecticide preparations in the experiments was carried out by observations in one field by planting potatoes in plots according to the options for a period of 101 days. The treatment was carried out once when planting potatoes by introducing the biocomposition into the soil at the rate of 30 g/m ² . Accounting for damage was carried out during the period of tuber formation, as well as accounting for damage to tubers after harvesting in accordance with the method of determination. Indicators of wireworm damage and the yield of marketable potatoes according to the options are presented in table 3.

Table 3

Potato variety Accounting for indicators of damage by wireworm (%);
Control (no processing) No. 1. B.bassiana + B.thuringiensis No. 2. B.bassiana + P.carboxydivorans Wireworm damage, % Average yield, % /
Commercial yield,% Wireworm damage, % Average yield, % /
Commercial yield,% Wireworm damage, % Average yield, % /
Commercial yield, % Early ripe (Bullfinch) 1.5 - 4.1 1 - 2.7 2-4 Mid Early (Wizard) 2-4 0 - 2 1 - 2.5 Mid-season (Cerata) 1.5 - 2.8 1 - 1.8 1 - 1.6 NSR _0.5 13.5

The bioinsecticide composition with both ratios and composition had a positive effect on increasing the yield of marketable products - potatoes, as well as reducing damage to potato tubers by wireworms (larvae of the nut beetle) under various weather conditions of the growing season. At the same time, the most promising is the use of biopesticides, namely, bioinsecticide preparative forms - biocompositions for growing mid-ripening and medium-early varieties of potatoes, which is important for the soil and climatic conditions of the Komi Republic - a zone of risky farming, especially for potato cultivation.

Thus, a technical result has been achieved in creating a bioinsecticide composition for protecting crops from soil pests, namely from the wireworm - the larvae of the click beetle.

Claims (3)

1. Bioinsecticide composition for combating soil pests, larvae of click beetles, containing a solid carrier material and a biological component, characterized in that a mixture of vermiculite with brewer's grains or brewer's grains is used as a solid carrier material, and immobilized as a biological component biomass of the bacterial-fungal complex, which includes the biomass of the micromycete Beauveria bassiana F-1357 in combination with the bacteria Bacillus thuringiensis B-6335 or the actinobacteria Pseudonocardia carboxydivorans AC-2046, taken at a ratio of 1: 1-3 by volume from the biomass of fungi and biomass of bacteria for subsequent immobilization on the carrier material and content with a concentration of at least 1×10 ⁷ CFU in 1 g of the composition, in the following ratio, wt. %: carrier material 80-90 Biomass of the bacterial-fungal complex 1-5 Moisture Rest 2. Bioinsecticide composition according to claim 1, characterized in that the mixture of vermiculite and spent grains is mixed at a volume ratio of 1:3.