RU2616284C1 - Microbiological preparation for increasing crop yields - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: microbiological preparation for increasing crop yields consists of a mixture of a bacterial suspension concentrate of Micrococcus luteus PBT-1 bacteria strain and a suspension concentrate of Penicillium sp microscopic fungus strain. PBT-2, products of metabolism and a sterile nutrient medium with a final titre of viable cells in liquid Micrococcus luteus PBT-1 bacteria strain which is not less than 2.0×10⁶ cfu/ml and Penicillium sp microscopic fungus strain. PBT-2 is not less than 1.0×10³ cfu/ml.

EFFECT: invention enables to increase crop yields.

4 tbl, 2 ex

Description

The invention relates to agricultural biotechnology, in particular to the preparation "Mycobact brand RF", based on the bacterial strain Micrococcus luteus PBT-1 and a strain of the microscopic fungus Penicillium sp. PBT-2 for use in agricultural production and personal subsidiary plots as a microbiological fertilizer to increase the productivity of the agroecosystem and to accelerate the decomposition of organic residues of natural origin.

Known biological preparation Sternifag SP based on a fungus of the genus Trichoderma (Trichoderma), which has phyto-protective and growth-promoting properties and decomposes even high polymer components of plant residues, Sternifag technologistics.ru/products/...

Known Trichodermin - an environmentally friendly biological product created on the basis of fungi of the genus (trichoderma) Trichoderma lignorum, the antagonistic properties of which are manifested in two ways: firstly, when propagated, the fungus produces antibiotics that destroy plant pathogens, and secondly, the fungus using alien fungi as a nutrient medium, destroys pathogen fungi, enriching the soil with nutrients, stimulating the growth and development of plants, Trichoderma lignorum agrocounsel.ru/trihodermin ...

Known biological product for the restoration of disturbed environmental objects and restoration of soil fertility, including a porous carrier based on glassy metaphosphates and physiologically active microorganisms, including the genus Micrococcus, RU No. 2282525 C1, B09C 1/10, A01B 79/00, 20.03.2005.

Known strain of the fungus Penicillium vermiculatum Dang. VIZR-24 for preparation against phytopathogenic fungi, RU No. 2322490 C1, C12N 1/14, A01N 63/00, 04/20/2008.

Known strain Penicillium funiculosum KM MSU 433 - a product of glucose oxidase, RU No. 2170762 C2, C12N 1/14, C12N 9/04, 07.20.2000.

Known strain of the fungus Penicillium chrysogenum BKMF-3330D, used to obtain biomass that extracts radionuclides and heavy metals from liquids, RU No. 2024078 C1, C02F 3/34, 11/30/1994.

A known strain of Micrococcus luteus with catalase activity, transforming organic residues of natural origin and increasing the productivity of the agroecosystem, deposited in the Departmental Collection of Useful Microorganisms for Agricultural Purpose "Russian Agricultural Academy (RCAM) (GNU VNIISCHM) under registration number RCAM 0101630, 1288 No. 24 / 26, C02F 3/34, B09C 1/10, C12R 1/265, 07.27.2013.

Known strain Penicillium sp., With multifunctional properties, transforming organic residues of natural origin and increasing the productivity of the agroecosystem, deposited in the Departmental Collection of Useful Microorganisms of Agricultural Purpose of the Russian Agricultural Academy (RCAM) (GNU VNIISCHM) under registration number RCAM 01017, RU No. 24879, C1 No. 128483, RU No. 24879 / 26, C02F 3/34, B09C 1/10, C12R 1/38, 07.20.2013.

Known microbiological preparations and strains of the genus Micrococcus and the genus Penicillium are individual and the joint use of bacterial strains of Micrococcus luteus and the microscopic fungus Penicillium sp. not found.

Known biological product for soil cleaning, including a consortium of strains of microorganisms cultured on a nutrient medium to obtain suspensions, with their further processing and obtaining a liquid form of microbiological preparation, RU No. 2535978 C1, C12N 1/20, C02F 3/34, B09C 1/10, C12R 1/01, 12.20.2014.

This technical solution is made as the closest analogue of the present invention.

The biological product of the closest analogue is used in conjunction with complex mineral fertilizer.

The biological product of the closest analogue contains a consortium of bacterial strains: Rhodococcus qingshengii BAK-PAH-1 VKPM AC-1946, Pusillimonas ginsegisoli BAK-PAH-2 VKPM B-11370, Shinella granuli BAK-PAH-3 VKPM B-11371.

A consortium of bacterial strains of the closest analogue includes bacterial strains of the genera Rhodococcus, Pusillimonas and Shinella, and is not otherwise provided for therein.

The present invention is based on the solution of the problem, which allows to obtain a new microbiological preparation "Mycobact brand of the Russian Federation", to reduce the time of its receipt, to increase and expand opportunities when using.

The technical result of the present invention consists in the joint cultivation of bacterial strains of Micrococcus luteus PBT-1 and the microscopic fungus Penicillium sp. PBT-2, in ensuring the optimal time for cultivation of the strains with the optimal use of the starting materials, in the transformation of organic residues and increasing the yield of the subsequent crop, in expanding the possibility of using it in both agricultural production and personal subsidiary plots.

According to the invention, this problem is solved due to the fact that the microbiological preparation is used as a microbiological fertilizer to accelerate the biological transformation of organic residues of natural origin and to increase crop yields. A microbiological preparation consists of a consortium of microorganism strains. A consortium of microorganism strains includes a bacterial strain. The microbiological preparation has a liquid form containing a bacterial suspension obtained by deep cultivation of bacterial strains of the consortium.

In the microbiological preparation, two strains were taken as a consortium of microorganism strains: the bacterial strain Micrococcus luteus PBT-1 and the strain of the microscopic fungus Penicillium sp. PBT-2.

The bacterial strain Micrococcus luteus PBT-1 consists of the biomass of bacteria Micrococcus luteus PBT-1 with a titer of 5.0 × 10 ⁶ CFU / ml - 7.0 × 10 ⁶ CFU / ml.

The strain of the microscopic fungus Penicillium sp. PBT-2 consists of the biomass of the microscopic fungus Penicillium sp. PBT-2 with a titer of 5.0 × 10 ³ CFU / ml - 7.0 × 10 ³ CFU / ml.

The liquid form of the microbiological preparation consists of a mixture of a bacterial suspension concentrate of the bacterial strain Micrococcus luteus PBT-1 and a suspension concentrate of the microscopic fungus of the strain of the microscopic fungus Penicillium sp. PBT-2, metabolic products and a sterile nutrient medium, when the titer of viable cells in liquid form of the bacterial strain Micrococcus luteus PBT-1 is not less than 20.0 × 10 ⁶ CFU / ml and the strain of the microscopic fungus Penicillium sp. PBT-2 is not less than 10.0 × 10 ³ CFU / ml.

Applicants have not identified sources containing information on technical solutions identical to the present invention, which allows us to conclude that it meets the criterion of "novelty."

Due to the implementation of the distinguishing features of the invention (in conjunction with the features indicated in the restrictive part of the formula), important new properties of the object are achieved.

Microbiological preparation “Mycobact brand KF” is a new microbiological preparation obtained by co-cultivation of two strains: the bacterial strain Micrococcus luteus PBT-1 and the strain of the microscopic fungus Penicillium sp. PBT-2.

The microbiological preparation “Mycobact brand KF” has a high concentration and was obtained during the maximum cultivation mode.

The microbiological preparation “Mycobact brand KF” has the ability to conduct and accelerate the transformation of organic residues and increase the yield of the subsequent culture.

The microbiological preparation “Mycobact brand KF” has the ability to be used in agricultural production.

The microbiological preparation "Mycobact brand KF" has the ability to be used in personal subsidiary plots.

Applicants are not aware of any publications that would contain information on the influence of the distinguishing features of the invention on the achieved technical result. In this regard, according to the applicant, it can be concluded that the claimed technical solution meets the criterion of "inventive step".

The present invention is carried out as follows.

In the microbiological preparation "Mycobact brand of the Russian Federation" as a consortium of strains of microorganisms taken bacteria strain Micrococcus luteus PBT-1 and a strain of the microscopic fungus Penicillium sp. PBT-2.

Bacterial strains of Micrococcus luteus PBT-1 and the microscopic fungus Penicillium sp. PBT-2 is deposited in the Departmental Collection of Useful Microorganisms of Agricultural Purpose “Russian Agricultural Academy (RCAM) (GNU VNIISKHM).

The bacterial strain Micrococcus luteus PBT-1 is deposited under registration number RCAM 01016.

The strain of the microscopic fungus Penicillium sp. PBT-2 is deposited under registration number RCAM 01017.

Bacterial strains of Micrococcus luteus PBT-1 and the microscopic fungus Penicillium sp. PBT-2 are stored in the collection of Petersburg Biotechnology LLC.

Bacterial strains of Micrococcus luteus PBT-1 and the microscopic fungus Penicillium sp. PBT-2 consists of the biomass of bacteria Micrococcus luteus PBT-1 with a titer of 2.0 × 10 ⁶ CFU / ml - 4.0 × 10 ⁶ CFU / ml and the biomass of the microscopic fungus Penicillium sp. PBT-2 with a titer of 1.0 × 10 ³ CFU / ml - 3.0 × 10 ³ CFU / ml, respectively.

Bacterial strains of Micrococcus luteus PBT-1 and the microscopic fungus Penicillium sp. PBT-2 was jointly cultivated in a sterile nutrient medium to obtain a concentrate of bacterial suspension Micrococcus luteus PBT-1 and a suspension concentrate of the microscopic fungus Penicillium sp. PBT-2 with titers of 20.0 × 10 ⁶ CFU / ml - 40.0 × 10 ⁶ CFU / ml and 10.0 × 10 ³ CFU / ml - 30.0 × 10 ³ CFU / ml, respectively.

The microbiological preparation “Mycobact brand RF” has a liquid form, consisting of a mixture of a concentrate of bacterial suspension Micrococcus luteus PBT-1 and a suspension concentrate of the microscopic fungus Penicillium sp. PBT-2, metabolic products and a sterile culture medium, when the final titer of viable cells in liquid form of Micrococcus luteus PBT-1 is not less than 2.0 × 10 ⁶ CFU / ml and Penicillium sp. PBT-2 is not less than 1.0 × 10 ³ CFU / ml.

Obtaining the microbiological preparation "Mycobact brand RF" is as follows.

The composition of the microbiological preparation "Mycobact brand of the Russian Federation" includes:

- the biomass of bacteria Micrococcus luteus strain PBT-1, RCAM 01016 (RU No. 2488630 C1, C12N 1/26, C02F 3/34, B09C 1/10, C12R 1/265, 07.27.2013);

- biomass of the microscopic fungus Penicillium sp. strain PBT-2, RCAM 01017 (RU No. 2487933 C1, C12N 1/26, C02F 3/34, B09C 1/10, C12R 1/38, 07.20.2013);

- metabolites formed during cultivation;

- the remains of the nutrient medium.

According to organoleptic, physico-chemical and biological indicators, the microbiological preparation must comply with the requirements and standards specified in table 1.

To store the bacterial strain Micrococcus luteus PBT-1, meat-peptone agar with glycerin is used.

For a strain of microscopic fungi Penicillium sp. PBT-2 uses Chapek's medium.

Obtaining a bacterial strain Micrococcus luteus PBT-1 and a strain of microscopic fungi Penicillium sp. PBT-2 is as follows.

The bacterial strain Micrococcus luteus PBT-1 and the strain of microscopic fungi Penicillium sp. PBT-2 is obtained in flasks.

Prepare a liquid nutrient medium of the following composition, g / l of purified water (1 l): KH ₂ PO ₄ - 1.0; CaCO ₃ - 0.3; MgSO ₄ - 0.3; NaCl - 0.1; NaNO ₃ - 2.0; cellulose - 5.0. Before sterilization, the pH of the nutrient medium is adjusted to 6.0-6.5. Nutrient medium is sterilized at 1 atm for 30 minutes.

Inoculation of flasks is carried out in 10 ml suspensions of the bacterial strain Micrococcus luteus PBT-1 and the strain of microscopic fungi Penicillium sp. PBT-2, respectively.

Cultures are grown until micrococcus luteus PBT-1 bacteria biomass titers of up to 2.0 × 10 ⁶ CFU / ml - 4.0 × 10 ⁶ CFU / ml and microscopic fungus Penicillium sp. PBT-2 up to 1.0 × 10 ³ CFU / ml - 3.0 × 10 ³ CFU / ml, respectively.

The bacterial strain Micrococcus luteus PBT-1 and the strain of microscopic fungi Penicillium sp. PBT-2 is stored in the refrigerator for up to 3 months at a temperature of + 4 ° C and is used to obtain a concentrate of bacterial suspension Micrococcus luteus PBT-1 and a suspension concentrate of the microscopic fungus Penicillium sp. PBT-2.

Preparation of Micrococcus luteus PBT-1 bacterial suspension concentrate and Penicillium sp. Microscopic fungus suspension concentrate PBT-2 is as follows.

Micrococcus luteus PBT-1 bacterial suspension concentrate and Penicillium sp. PBT-2 microscopic fungus suspension concentrate are obtained in bottles or fermenters on similar media (a 2-fold dilution of the nutrient medium is allowed).

The optimal cultivation time is 7 days at a temperature of 28 ° C.

A concentrate of the bacterial suspension of Micrococcus luteus PBT-1 and a suspension concentrate of the microscopic fungus Penicillium sp. PBT-2 with the achievement of titers of 20.0 × 10 ⁶ CFU / ml - 40.0 × 10 ⁶ CFU / ml and 10.0 × 10 ³ CFU / ml - 30.0 × 10 ³ CFU / ml, which are used to obtain liquid form of the microbiological preparation "Mycobact brand of the Russian Federation."

Obtaining a liquid form of the microbiological preparation "Mycobact brand RF" is as follows.

Micrococcus luteus PBT-1 bacterial suspension concentrate and Penicillium sp. Microscopic fungus suspension concentrate PBT-2 is introduced into a sterile nutrient medium with cellulose.

The composition of the sterile nutrient medium (g / l): KH ₂ PO ₄ - 1.0; CaCO ₃ - 0.3; MgSO ₄ - 0.3; NaCl - 0.1; NaNO ₃ - 2.0; cellulose - 5.0. Before sterilization, the pH of the nutrient medium is adjusted to 6.0-6.5. Nutrient medium is sterilized at 1 atm for 30 minutes.

For mixing, stainless steel tanks with a volume of 200-400 l, previously sterilized with steam, are used. Mixing Micrococcus luteus PBT-1 bacterial suspension concentrate and Penicillium sp. Microscopic fungus suspension concentrate PBT-2 with a sterile nutrient medium is carried out at a ratio of 1:20, respectively, to achieve the final titer in liquid form of Micrococcus luteus PBT-1 not less than 2.0 × 10 ⁶ CFU / ml and Penicillium sp. PBT-2 is not less than 1.0 × 10 ³ CFU / ml.

After 10 days of storage of the microbiological preparation “Mycobact brand of the Russian Federation” packing units are selected and transferred to the laboratory for quality analysis. If the TU preparation is suitable, at least two additional packing units are taken from each batch and stored until the expiration of the warranty period.

A single sample for analysis is taken from each packaging unit after thorough mixing. The mass of a single sample should be at least 30 ml.

The appearance and color of the microbiological preparation “Mycobact brand of the Russian Federation” is determined visually for each packing unit at the time of sampling.

The assessment of physical and biological indicators is carried out on the basis of laboratory analysis of a single sample of each packing unit.

Determine the humidity of the studied microbiological preparation "Mycobact brand RF", placed in an oven, the ability to give moisture when heated to a temperature of 105 ° C.

Determination of the number of bacterial cells Micrococcus luteus PBT-1, microscopic fungus Penicillium sp. PBT-2 and extraneous microorganisms are carried out by known methods based on the preparation of a series of serial dilutions of the microbiological preparation “Mycobact brand RF” in sterile water and plating in Petri dishes, followed by the introduction of an appropriate agar nutrient medium and counting the grown bacteria or microscopic fungi.

Ready microbiological preparation "Mycobact brand of the Russian Federation" is poured into containers. Storage of the microbiological preparation “Mycobact brand of the Russian Federation” is provided in rooms specially designed for these purposes.

Each packaging unit in consumer packaging must be marked in the form of a packaging label, which should contain recommendations for transportation, use and storage.

The microbiological preparation “Mycobact brand of the Russian Federation” contains a consortium of strains of microorganisms, programmed to perform specific tasks on the transformation of plant residues.

As a result, the joint organized activity of this consortium of microorganism strains in the form of bacteria and microscopic fungi allows the efficient transformation of plant residues into humic forms.

The microbiological preparation “Mycobact brand of the Russian Federation” allows to carry out and accelerate the biological transformation of organic residues after harvesting or before sowing (planting) of the next crop no later than one day before sowing (planting), and no earlier than one day after sowing (planting) . Microbiological preparation “Mycobact brand of the Russian Federation” increases the yield of the subsequent culture.

The microbiological preparation “Mycobact brand of the Russian Federation” is used both in agricultural production and in personal subsidiary plots to increase soil fertility.

The regulations for the use of the microbiological preparation “Mycobact brand of the Russian Federation” are presented in table 2.

Experimental studies on the use of the microbiological preparation "Mycobact brand RF" confirm examples 1-2.

Example 1

In the laboratory of field crop cultivation technologies and agro-ecological land valuation of the Kursk Research Institute of Agro-Industrial Production of the Russian Agricultural Academy, a model-field experience was studied to study the effect of the “Mykobact brand of the Russian Federation” preparation on the degree of decomposition of wheat straw and the yield of the subsequent crop (sugar beet). The studies were conducted under conditions of 2013 (decomposition of wheat straw) and 2014 (effect on sugar beet yield) in crop rotation during crop rotation, such as pure steam, winter wheat, sugar beet, spring barley.

The soil of the experimental site was represented by chernozem typical thick heavy loamy soils.

Soil cultivation with the microbiological preparation “Mykobact brand of the Russian Federation” was carried out immediately after harvesting winter wheat and chopping straw (July 20, 2013) with a backpack sprayer. The application rate of the microbiological preparation “Mycobact brand of the Russian Federation” was 3 l / ha, the consumption rate of the working solution was 250 l / ha.

When determining the application rate of the microbiological preparation “Mycobact of the RF brand”, it was assumed that 7.5 t / ha of straw was delivered to the soil (The yield of winter wheat in this field was 50 c / ha, with a grain: straw ratio of 1: 1, 5, the amount of straw was 7.5 t / ha). The dose of the microbiological preparation “Mycobact brand of the Russian Federation” with a quantity of straw of grain crops of 3-4 t / ha was 2 l / ha, with a quantity of straw more than 4 t / ha - 3 l / ha. After applying the microbiological preparation “Mycobact brand of the Russian Federation” plant residues were planted into the soil to a depth of 10-12 cm.

The decomposition of straw was observed 90 days after its processing on October 29.

The influence of the microbiological preparation "Mycobact brand RF" on the degree of decomposition of wheat straw is presented in table 3.

From table 3 it can be seen that under the conditions of 2013 in the control variant for 90 days of exposure, wheat straw decomposed by 44.5%.

The treatment of straw with the microbiological preparation “Mycobact brand of the Russian Federation” at a dose of 2 l / ha increased the degree of decomposition of wheat straw to 61.4% or 16.9% higher in comparison with the control. With an increase in the dose of the application of the microbiological preparation “Mykobact of the RF brand” to 3 l / ha, the degree of decomposition of wheat straw increased to 83.4%.

The effectiveness of the influence of wheat straw treated with the microbiological preparation “Mykobact of the RF brand” on the yield and quality of sugar beet root crops (the next crop rotation) was determined in 2014.

After the onset of physical ripeness of the soil and its loosening, sowing of sugar beets (hybrid seeds of Shanon) was carried out.

To assess the effectiveness of the microbiological preparation “Mycobact brand of the Russian Federation” throughout the entire period of vegetation of sugar beet, we observed the growth and development of plants, the dynamics of root crops, yield and quality of root crops.

The influence of the microbiological preparation "Mycobact brand RF" on the yield of sugar beets are presented in table 4.

From table 4 it is seen that treatment of wheat straw with the microbiological preparation “Mykobact of the RF brand” at a dose of 2 l / ha increased the yield of sugar beet root crops by 23 c / ha or 5.5%, and with the yield in the control variant - 415 c / ha. With an increase in the dose of the application of the microbiological preparation “Mykobact of the RF brand” to 3 l / ha, the increase in sugar beet yield increased and amounted to 36 c / ha or 8.7%. The use of the microbiological preparation “Mycobact brand of the Russian Federation” for the processing of wheat straw paid off with sugar beet yield.

Example 2

A test on the use of the microbiological preparation “Mycobact of the brand of the Russian Federation” was conducted on the experimental field of the Department of Plant Production of St. Petersburg State Agrarian University.

In order to conduct research, a seed experiment was laid on studying the influence of the microbiological preparation “Mykobact of the RF brand” on the degree of decomposition of rye straw and the yield of the subsequent crop - buckwheat of the Kazan large-fruited variety. In this culture, the duration of the growing season is from 64 to 82 days.

In the month of August, after winter rye was harvested, an aqueous solution of the microbiological preparation Mikobakt Mark of the Russian Federation was applied to rye straw at a rate of 4 l / ha with a straw weight of at least 3.0-4.0 t / ha. The consumption rate of the working solution was 200.0 l / ha.

After three months (October), the degree of decomposition of winter rye straw was determined. It was found that in the control version without the microbiological preparation “Mykobact of the RF brand” (straw treatment with water) for 79 days, rye straw decomposed by 40.2%, and when the microbiological preparation “Mykobact of the RF brand” was added, this indicator amounted to 86.4%.

The effectiveness of the influence of rye straw treated with the microbiological preparation “Mykobact of the RF brand” on the yield, growth and development of buckwheat of the Kazan large-fruited variety was determined the following year.

The growth and development of buckwheat of the Kazan large-fruited variety was more intense, the fruit yield was 1.29 t / ha, which is 0.77 t / ha higher than in the control without treatment with the microbiological preparation “Mycobact RF brand” (straw treatment with water).

A dose of 4 L / ha of the microbiological preparation “Mycobact of the RF brand” is preferable to accelerate the degree of decomposition of winter rye straw before laying it in the soil.

Examples 1-2 confirm the feasibility of using the microbiological preparation "Mycobact brand RF" for the biological transformation of organic residues of natural origin and to increase the yield of subsequent crops (sugar beets and buckwheat, respectively).

The proposed microbiological preparation "Mycobact brand RF" refers to the microbiological industry, includes the known bacteria strains Micrococcus luteus PBT-1 and the microscopic fungus Penicillium sp. PBT-2, methods for their cultivation, production technology and its implementation, confirmed by experimental work and research of Petersburg Biotechnologies LLC, which determines, according to the applicants, its criterion of "industrial applicability".

Microbiological preparation "Mycobact brand of the Russian Federation" allows you to:

- carry out joint cultivation of bacterial strains of Micrococcus luteus PBT-1 and the microscopic fungus Penicillium sp. PBT-2;

- provide optimal time for the cultivation of strains;

- reduce the time to obtain a microbiological preparation;

- transform organic residues of natural origin;

- increase the yield of the subsequent crop;

- use it both in agricultural production and in personal subsidiary plots.

Claims (1)

Microbiological preparation for increasing crop yields in liquid form, consisting of a mixture of a bacterial suspension concentrate of the bacterial strain Micrococcus luteus PBT-1 and a suspension concentrate of the strain of the microscopic fungus Penicillium sp. PBT-2, metabolic products and a sterile culture medium with a final titer of viable cells in liquid form of the Micrococcus luteus strain PBT-1 at least 2.0 × 10 ⁶ CFU / ml and a strain of the microscopic fungus Penicillium sp. PBT-2 is not less than 1.0 × 10 ³ CFU / ml.