RU2760336C1 - Preparation for increasing the yield of spring barley - Google Patents

Abstract

FIELD: biotechnology; microbiology; agriculture.SUBSTANCE: claimed preparation contains culture liquids of bacterial strains Acinetobacter johnsonii A1 RNCIM B-13867, Pseudarthrobacter equi S2 RNCIM B-13868 and Pantoea agglomerans F19 RNCIM B-13869 in a ratio of 1:1:1 with a titer of each bacterial strain of at least 1х107CFU/cm3. The invention can be used in agriculture.EFFECT: invention allows improving the yield of spring barley.1 cl, 2 tbl, 4 ex

Description

The inventions relate to biotechnology, microbiology and agriculture and concerns the production of microbial preparations based on a mixture of bacterial strains to improve plant nutrition, improve soil biocenosis and increase the yield of grain crops with nutritional properties, phytoprotective and growth-stimulating activity.

The following bacterial fungicides based on bacteria of the genus Bacillus against diseases of various cultures were registered and included in the List of the State Chemical Commission of the Ministry of Agriculture of the Russian Federation: Integral preparation - liquid based on the Bacillus subtilis strain 24D, Fitosporin-M preparation - powder based on the Bacillus subtilis strain 24D (p. RF No. 2129375, IPC A01N 63/00, C12N 1/20, publ. 04/27/1999; 2129004, IPC A61K 35/74, C12N 1/20, publ. 04/20/1999), the preparation Bactofit wettable powder based on the Bacillus subtilis strain IPM 215 (p. RF No. 2019966, A01N 63/00, publ. 30.09.1994). The drug Bactophosphine (based on Bacillus mucilaginosus) is designed to enrich the soil, as well as restore its fertility, increase the germination and productivity of many types of crops in open and protected ground.

There are known strains of bacteria that not only show antagonism towards various phytopathogens and protect plants from diseases, but also significantly increase productivity. As examples, we can cite the strains Bacillus sp. 739 (p. RF No. 1743019, IPC A01N 63/00, C12N 1/20, publ. 05/30/1994), B. subtilis 4-13 - the basis of the drug "Extrasol" (p. RF No. 2259397, IPC C12N 1/20, A01C 1/06, publ. 27.08.2005), Bacillus mycoides var. V.A. VNIISHM D138 - the basis of the drug "Extragran" (p. RF No. 2284353, IPC C12N 1/20, A01N 63/00, A01C 1/00, publ. 09/27/2006), Bacillus spp. KR-083 (p. RF No. 2295562, IPC C12N 1/20, A01N 63/00, publ. 03/20/2007), Bacillus licheniformis VKPM B-10561, B-10562, B-10563, B-10564 (p. RF No. 2440413, IPC C12N 1/20, A01N 63/00, A61K 35/74, publ. 20.01.2012), B. subtilis 8A (p. RF No. 2495119, IPC C12N 1/20, C12R 1/125, A01N 63 / 00, publ. 10.10.2013), Bacillus amyloliquefaciens VKPM V-11475 (p. RF No. 2528058, IPC C12N 1/20, A01N 63/00, C12R 1/07, publ. 09/10/2014).

However, in the literature it was not possible to find information about their ability to improve nitrogen, phosphorus and potassium nutrition of plants.

A known biological product for the active enrichment of soil with nitrogen and phosphate, containing a nitrogen-fixing effective amount of Azospirillum sp. M32, an amount of Bacillus polymixa M7 sufficient to produce polymyxin, an antimicrobial peptide active against gram-negative bacteria that converts phosphorous materials fixed in the soil to which it is applied to phosphate in plant-accessible forms and a bacterial nutrient containing cellulose material containing cellulose as an integral component selected from the group consisting of straw, xylan, hemicellulose and lignin, as well as other organic carbon or nitrogen materials (US Pat. US No. 5147441 A, IPC C05G3 / 08, publ. 09/15/1992) ...

However, the properties of the consortium have been studied only in laboratory conditions; there is no data on the effectiveness of its action in the field.

It is known to use a biologically active drug based on a consortium of lactic acid bacteria 5842 VKPM B-5972, nitrogen-fixing and phosphate-mobilizing bacteria, followed by treatment of vegetative plants with a drug obtained on the basis of a consortium of lactic acid bacteria 3843 VKPM B-5974 and El-1. From nitrogen-fixing bacteria, the bacterial strain Azotobacter chroococcum 3064 VKPM B-3721 is used at a dose of 0.2-0.4 kg / ha, from phosphate-mobilizing Bacillus mucilaginosus VKPM B-5987 at a dose of 0.1-0.4 kg / ha (p. RF No. 2127509, IPC A01G 7/00, publ. 03/20/1999).

The use of this drug allows you to increase the yield, survival of plants while improving the quality of the standard planting material, its yield, as well as to increase the winter hardiness of crops, especially fruit crops.

However, the use of this drug is quite laborious. In addition, it does not contain potassium-solubilizing bacteria, which provide plants with an important nutrient.

The closest to the claimed technical solution in terms of the number of essential features is a preparation for increasing the yield of grain crops, which has phytoprotective and growth-stimulating properties, containing culture liquids of bacterial strains Bacillus amyloliquefaciens VKPM B-11986, Bacillus amyloliquefaciens VKPM B-11987, Pseudomonas VKPM 11984, Rahnella aquatilis VKPM B-11985, Serratia plymuthica VKPM B-12008 in a ratio of 1: 1: 1: 1: 1 with a titer of each bacterial strain of at least 1 × 109 CFU in 1 cm ³ (p. RF No. 2595405, IPC C12N 1/20, A01N 63/00, publ. 08/27/2016).

Despite the fact that a microbial preparation based on a mixture of bacterial strains provides a healthier biocenosis, accelerates the decomposition of lignocellulose, increases crop yields and the quality of the resulting product, it has drawbacks. So, the drug is multicomponent, and this requires additional time and labor to obtain each component that is part of a known drug. In addition, it does not contain bacterial strains exhibiting nitrogen-fixing, phosphate-solubilizing and potassium-solubilizing activities.

The need for systems to improve plant growth conditions, in particular by increasing the levels of available nitrogen, phosphorus and potassium in soil systems, is still ongoing.

The technical problem posed to the invention is the expansion of the range of preparations for restoring soil biocenosis and increasing the yield of spring barley, by creating a new biological product of complex action (stimulating the growth and development of plants, increasing plant productivity and the level of soil fertility) based on the biomass of natural strains of bacteria, possessing nitrogen-fixing, phosphate-solubilizing, potassium-solubilizing, phytoprotective and growth-stimulating activities.

The technical problem posed is solved by the fact that in the known preparation for increasing the yield of grain crops, having phytoprotective and growth-stimulating activity, improving the microbiological activity of the soil, including the culture liquid of strains of microorganisms, according to the invention, spring barley is taken as the grain culture, and as the culture liquid of the strains bacteria it contains culture liquids of bacterial strains Acinetobacter johnsonii A1 VKPM B-13867, Pseudarthrobacter equi S2 VKPM B-13868, Pantoea agglomerans F19 VKPM B-13869 in a ratio of 1: 1: 1 with a titer of each bacterial strain of at least 1x10 ⁷ CFU in 1 cm ³ .

The introduction into the preparation of a new bacterial strain Acinetobacter johnsonii A1 VKPM B-13867 deposited in the All-Russian collection of industrial microorganisms (VKPM) under registration number VKPM B-13867 imparts nitrogen-fixing activity to the preparation, i.e. promotes better nitrogen nutrition of spring barley.

The introduction into the preparation of a new bacterial strain Pseudarthrobacter equi S2, deposited in the All-Russian Collection of Industrial Microorganisms (VKPM) under registration number VKPM B-13868, imparts potassium-solubilizing activity to the preparation, thereby contributing to better potassium nutrition of spring barley.

The introduction into the preparation of a new bacterial strain Pantoea agglomerans F19, deposited in the All-Russian Collection of Industrial Microorganisms (VKPM) under registration number VKPM B-13869, imparts phosphate-solubilizing activity to the preparation, thereby contributing to better phosphorus nutrition of spring barley.

Preparation of the culture liquid of the above bacterial strains in a ratio of 1: 1: 1 with a titer of each bacterial strain of at least 1x10 ⁷ CFU in 1 cm ³ provides the preparation with a complex effect, which helps to increase the yield of spring barley.

The invention is illustrated by the following tables:

Table 1. shows the average values of the salinity coefficients, in table. 2.- yield of spring barley.

Example 1. Preparation of the preparation.

Cultivation of each bacterial strain Acinetobacter johnsonii A1 VKPM B-13867, Pseudarthrobacter equi S2 B-13868, Pantoea agglomerans F19 B-13869 is carried out separately in GRM broth (composition g / l: pancreatic hydrolyzate of fish meal - 8.0; enzymatic peptone - 8.0; sodium chloride - 6.0). The duration of cultivation is 72-96 hours at a temperature of 24-26 ° C and constant stirring on an orbital-type shaker with a rotation speed of (180 ± 5) rpm.

Cultural liquids of bacteria containing living bacterial cells, their metabolites, obtained by separate cultivation of producer strains, are mixed in equal proportions (1: 1: 1). The finished preparation is a liquid from light yellow to light brown in color with a loose sediment formed during storage, pH - 7-7.2, the total number of viable bacterial cells in 1 cm ³ is not less than 1 * 10 ⁷ . The shelf life of the drug at a temperature of 4-10 ⁰ С is 3 months, at a temperature of 20-25 ⁰ С - no more than 8 days.

Example 2. The effect of the drug on the microbiological activity of the soil.

The use of the preparation for treatment of seeds and soil by seedlings stimulates the vital activity of soil microorganisms, increasing soil biogenicity by 3 times (780.56 million CFU / g) compared to the control (261.52 million CFU / g). The method of application includes 2 stages of treatment: seed treatment and spraying of plants at seedlings. At the first stage, the seeds were treated by a single spraying at the rate of 10 liters of working fluid per 1 ton of seeds, the norm of the preparation was 300 ml. for 100 liters. water. At the second stage, a single spraying of plants was carried out in the germination phase at the rate of 300 l / ha of working fluid, the rate of the preparation was 1 l / ha. The control was the treatment of seeds and vegetative plants with water.

One of the indicators of the intensification of mineralization processes in the soil, which characterizes the initial stage of the transformation of organic matter, may be the ratio of bacteria assimilating organic and mineral nitrogen (Mishustin's mineralization coefficient). In soils with a more vigorous process of mineralization, microorganisms that assimilate mineral nitrogen usually outnumber the microflora that develops at the expense of organic nitrogen. We found that the mineralization coefficient when using the proposed drug is 4.5 times higher than the control indicators, the standard mineralization coefficient is 3 times higher than the control (see Table 1).

Thus, the claimed preparation, when processing seeds and soil by seedlings, has a positive effect on an increase in the total microbial number of the soil and leads to an intensification of mineralization processes.

Example 3. Growth-stimulating effect of drugs in the treatment of seeds of spring barley.

The growth-stimulating effect was studied by identifying the potential synergistic effect of the studied bacteria when acting on the germination of seeds of cereal plants. For this, working suspensions of microorganisms were combined in a 1: 1: 1 ratio. Plant seeds, in the amount of 100 pieces, were soaked in the claimed preparation, the exposure time was 30 minutes. Then all the seeds were laid out in 10 pieces on the surface of filter paper moistened with sterile water in sterile Petri dishes for germination. The seeds were germinated at 23 ° C. On the 3rd day, the germination energy was determined, on the 7th - laboratory germination, and the shoot length and root length were assessed. As a result of the effect of the drug, an increase in the germination energy of wheat seeds was observed on average for barley seeds - by an average of 34.5% (from 15 to 54%) (p ≤ 0.05) compared to the untreated control; an increase in laboratory germination of spring barley seeds by 30%; an increase in the length of the aboveground part of spring barley by 36.7%; an increase in the length of the spring barley root by 46.7.

Example 4. Increasing the yield of spring barley.

Field studies were carried out on the experimental fields of the Federal Research Center of Agrobiotechnology of the Far East named after V.I. A.K. Seagulls (Primorsky Territory, Ussuriysk, settlement Timiryazevsky. The object of research was a zoned variety of spring barley Hordeum vulgare L., variety Vostochny of the Federal Research Center of Agrobiotechnology of the Far East named after A.K. Chaika. The plot area is 15 m2, the location is randomized, threefold repetition. Method of application includes 2 stages of treatment: seed treatment and spraying of plants on seedlings. At the first stage, seeds were treated by a single spraying at the rate of 10 liters of working fluid per 1 ton of seeds, the norm of the preparation is 300 ml per 100 liters of water. the second stage - a single spraying of plants in the germination phase was carried out at the rate of 300 l / ha of working fluid, the rate of the preparation was 1 l / ha. The control was the treatment of seeds and vegetative plants with water.

The yield by options is presented in table 2. As follows from the presented results, the use of the proposed drug contributes to an increase in the yield of spring barley 0.7 t / ha, which exceeds the control indicators by 25.9%.

Thus, the use of the proposed drug, which includes new natural strains of bacteria with nitrogen-fixing, phosphate-solubilizing, potassium-solubilizing, phytoprotective and growth-stimulating activities, expands the range of drugs for restoring soil biocenosis and increasing the yield of spring barley.

Claims (1)

A preparation for increasing the yield of a grain crop, including a culture liquid of strains of microorganisms, characterized in that spring barley is taken as a grain culture, and as a culture liquid of microorganism strains it contains culture liquids of bacterial strains Acinetobacter johnsonii A1 VKPM B-13867, Pseudarthrobacter equi S2 VKP B-13868 and Pantoea agglomerans F19 VKPM B-13869 in a 1: 1: 1 ratio with a titer of each bacterial strain of at least 1x10 ⁷ CFU / cm ³ .

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