RU2800257C1 - Growth accelerator of the root system of plants (embodiments) - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: plant root system growth accelerator comprising culture liquid of microorganisms Bacillus licheniformis with a titre of at least 1.5*10⁷ CFU per cm³, Bacillus pumilus with a titre of at least 1.5*10⁷ CFU per cm³, Bacillus subtilis with a titre of at least 10⁷ CFU in cm³, Paenibacillus polymyxa with a titre of at least 2*10⁷ CFU in cm³, used as a 0.2% solution in deionized water for the treatment of young and adult plants during the growing season and flowering. A plant root system growth accelerator comprising culture liquid of microorganisms Bacillus mucilaginosus with a titre of at least 2.5*10⁷ CFU per cm³, Pseudomonas chlororaphis with a titre of at least 10⁷ CFU per cm³, Pseudomonas putida with a titre of at least 1.5* 10⁷ CFU in cm³, Trichoderma harzianum with a titre of at least 1.5 * 10⁷ CFU in cm³, Trichoderma viride with a titre of at least 2 * 10⁷ CFU in cm³, used as a 0.2% solution in deionized water, which increases the availability of mineral nutrition and increases protection against soil pathogens, for the treatment of young and adult plants during the growing season and flowering. A plant root system growth accelerator comprising culture liquid of microorganisms Arthrobacter globiformis with a titre of at least 6.6*10⁷ CFU per cm³, Bacillus coagulans with a titre of at least 1.2*10⁷ CFU per cm³, Bacillus licheniformis with a titre of at least 1*10⁷ CFU in cm³, Bacillus mucilaginosus with a titre of at least 1*10⁷ CFU in cm³, Bacillus pumilus with a titre of at least 3.3*10⁷ CFU in cm³, Bacillus subtilis with a titre of at least 1*10⁷ CFU in cm³, Paenibacillus polymyxa with a titre of at least 3.3 * 10⁷ CFU in cm³, used in the form of a 0.2% solution in deionized water, which is an accelerator of combined action for the treatment of young and adult plants during the growing and flowering season.

EFFECT: inventions make it possible to stimulate the growth of the plant's root system, to protect against soil pathogens, to accelerate the decomposition of dying roots, to increase the availability of mineral nutrition, to increase the abundance of flowering, crop yields and product quality.

3 cl, 2 tbl, 1 ex

Description

The field of technology to which the invention belongs.

SUBSTANCE: group of inventions relates to biotechnology and agricultural microbiology and concerns the production of preparations containing mixtures of bacterial and fungal strains based on culture fluids for accelerating growth and improving the microbial community of roots, accelerating the decomposition of organic residues, increasing the availability of mineral nutrition, increasing the yield of agricultural crops and the quality of the products obtained.

The basis of the invention is a mixture of culture liquids of microorganisms prepared in a certain proportion, in particular, the bacteria Bacillus licheniformis, Bacillus pumilus, Bacillus subtilis, Bacillus mucilaginosus, Bacillus coagulans, Paenibacillus polymyxa, Pseudomonas chlororaphis, Pseudomonas putida, Arthrobacter globiformis and fungi Trichoderma harzianum and Tricho derma viride, which have phytoprotective, cellulolytic and growth-promoting properties, can reduce the infectious background during the growing season and flowering of plants, ensure the improvement of the rhizosphere and increase the availability of mineral nutrition, which ensures the growth and development of plants.

State of the art

Known strains of microorganisms that directly stimulate growth and increase plant productivity, such as Pseudomonas putida VKPM B-4308 (RF Patent No. 2051586), Pseudomonas lemoignei VKM B-6615 (RF Patent No. 2121271), Streptomyces chrysomallus P-21 (RF Patent No. 2226214), Bacillus subtilis 8A (RF Patent No. 2495119,), Serratia ficaria VKPM V-11403 (RF Patent No. 2545398,), Bacillus pumilus A1.5 (RF Patent No. 2551968), Bacillus atrophaeus VKPM V-11474 (RF Patent No. 2570624), Bacillus cereus 875 TS (RF Patent No. 2624032). However, for none of the listed strains, the possible mechanisms of direct stimulation of plant growth are indicated, the effects exerted by these strains on the root system are not shown, and the complex effect of mineral nutrition mobilization and development of the rhizosphere is not achieved.

Known drug strain of bacteria Azospirillum zeae OPN-14, deposited in the All-Russian collection of industrial microorganisms under the number VKPM B-12542, as a biological agent with growth-stimulating activity in relation to plants (RF Patent No. 2678755). A known mechanism for stimulating the growth of Azospirillum zeae is due to the release of phytohormone-like compounds that directly affect plant growth. However, this effect does not make it possible to qualitatively improve the saturation of the plant with mineral salts, since it does not have a complex effect on the rhizosphere.

Known phosphate-mobilizing strain of soil bacteria Lelliottia nimipressuralis SSM 32-3, a producer of phytohormones, deposited in the All-Russian collection of industrial microorganisms of the Federal State Budgetary Institution GosNIIgenetika under registration number VKPM B-12783 (RF Patent No. 2676926). The mechanism of action of the microorganism on plant growth is due to an increase in the availability of phospho-containing compounds in the soil for root nutrition.

Known effective strain of soil phosphate-dissolving bacteria Pseudomonas chlororaphis ssp chlororaphis Vsk-26a3, which has the ability to suppress phytopathogenic fungi and bacteria. The Pseudomonas chlororaphis ssp chlororaphis Vsk-26a3 strain releases phosphates from insoluble minerals, has a broad spectrum of action against phytopathogenic fungi (inhibits 15 species) and bacteria, and also stimulates plant growth and increases their yield. The strain was deposited in the State Collection of Pathogenic Microorganisms and Cell Cultures "GKPM-Obolensk" under the number B-7427 (RF Patent No. 2603281).

A strain of the microscopic fungus Trichoderma asperellum is known, deposited in the All-Russian Collection of Industrial Microorganisms under the number F-1323, which has a complex effect to protect agricultural plants from diseases caused by phytopathogenic fungi, stimulate growth and increase plant yields, as well as biodegradation of crop residues and improve the phytosanitary condition of soils (RF Patent No. 2634415).

A consortium of lactic acid, nitrogen-fixing and phosphate-mobilizing bacteria is known, which enhances mineral nutrition and allows to increase productivity by 25-35%, plant survival by 20-40% and the quality of standard planting material (RF Patent No. 2127509). The consortium is used for pre-sowing treatment of seeds, for direct application to the soil before sowing or planting, for treatment of the vegetative part of plants or application to the soil during the growth period. However, there is no data on the ability of the consortium to reduce the number of pathogenic microorganisms, decompose plant residues in the soil and directly affect the growth of the root system.

Known consortium of microorganisms, consisting of four strains of soil bacteria antagonists Bacillus polymixa, Bacillus thuringiensis, Pantoea agglomerans and Pantoea chlororatis, with phytoprotective properties (RF Patent No. 2149552). The consortium is used to protect against gray rot and powdery mildew in tomato and cucumber, bacterial cancer in clover, vascular bacteriosis in cabbage, angular spot in cucumber, Fusarium wilt in tomato, helminthosporosis in barley, root rot in cotton. The protective effect exerted by the drug on the root system is shown. However, there is no data on the ability of the consortium to directly influence the growth and development of the root system.

The closest technical solution is a preparation for protecting plants from pathogens of agricultural crops, which has phytoprotective, cellulolytic and growth-promoting properties, including the cultural liquid of strains of microorganisms, characterized in that it contains culture liquids of bacterial strains Bacillus amyloliquefaciens VKPM B-11986, Bacillus amyloliquefaciens VKPM B-11987, Pseudomonas brassicacear as the cultural liquid of strains of microorganisms um VKPM V-11984, Rahnella aquatilis VKPM V-11985, Serratia plymuthica VKPM V-12008 in a ratio of 1:1:1:1:1 with a titer of each bacterial strain of at least 1 × 10 ⁹ CFU per 1 cm ³ , see RF patent No. 2595405.

However, a preparation based on a mixture of these bacterial strains with complex phytoprotective, cellulolytic and growth-stimulating activity is intended to reduce the infectious background during the growing season of plants, as well as accelerate the decomposition of straw and crop residues, activate soil microbiocenosis and increase crop yields and product quality. The proposed drug is not intended for use in order to influence the accelerated development and protection of the plant root system from soil pathogens, the activation of mobilization processes and increase the availability of mineral nutrition.

Disclosure of the invention.

Based on this original observation, the present invention mainly aims to provide a plant root growth accelerator based on a mixture of microorganism strains with complex activity to stimulate the growth of the root system, the plant itself, including protection against soil pathogens, accelerated decomposition of dying roots, increase the availability of mineral nutrition, direct stimulation of root growth by secreted phytohormones and stabilization of the rhizosphere community of microorganisms, resulting in an increase in the abundance of flowering and crop yields and quality of products.

The problem is solved by using consortiums of microorganisms of the following composition:

1. A root system growth accelerator for treating young and adult plants during the growing season and flowering, containing a culture fluid of microorganisms Bacillus licheniformis with a titer of at least 1.5×10 ⁷ CFU per cm ³ , Bacillus pumilus with a titer of at least 1.5×10 ^{7 CFU per cm 3 , Bacillus subtilis with a titer of at least 10 7 CFU} per cm ³ , Paenibac illus polymyxa with a titer of at least 2×10 ⁷ CFU cm ³ used as a 0.2% solution in pure water ^.

1. A root system growth accelerator that increases the availability of mineral nutrition and increases protection against soil pathogens, for treating young and adult plants during the growing season and flowering, containing the culture fluid of Bacillus mucilaginosus microorganisms with a titer of at least 2.5 × 10 ⁷ CFU per cm ³ , Pseudomonas chlororaphis with a titer of at least 10 ⁷ CFU per cm ³ , Pseudomonas putida with a titer at least 1.5×10 ⁷ CFU per cm ³ , Trichoderma harzianum with a titer of at least 1.5×10 ⁷ CFU per cm ³ , Trichoderma viride with a titer of at least 2×10 ⁷ CFU per cm ³ , applied as a 0.2% solution in pure water.

2. Accelerator of growth of the root system of combined action for the treatment of young and adult plants during the growing season and flowering, containing the cultural liquid of microorganisms Arthrobacter globiformis with a titer of at least 6.6×10 ⁷ CFU per cm ³ , Bacillus coagulans with a titer of at least 1.2×10 ^{7 CFU per cm 3 cm 3 , Bacillus mucilaginosus with a titer of at least 1 × 10 7 CFU per} cm ³ , Bacillus pumilus with a titer of at least 3.3 × 10 ⁷ CFU per cm ³ , Bacillus subtilis with a titer of at least 1 × 10 ⁷ CFU per cm ³ , Paenibacillus polymyxa with a titer of at least 3.3 × ^{10 7 CFU} per ^{cm 3 applied} as a 0.2% solution in pure water.

The technical result obtained when using these preparations is to stimulate the growth of the root system, leading to an increase in the plant itself, an abundance of flowering, an increase in crop yields and the quality of the products obtained.

Cultural and morphological characteristics of bacterial strains included in the claimed composition

Cultural characteristics during surface cultivation of bacteria on ΜΠΑ:

Arthrobacter globiformis forms smooth, round, convex colonies with a smooth edge.

Bacillus licheniformis form round grayish colonies with an uneven edge, 3-5 mm in diameter, mucous, raised in a young culture and rough, flat in an aging one;

Bacillus pumilus form round colonies of light gray color, with a smooth edge, 3-5 mm in diameter, mucous, raised in a young culture and rough, flat in an aging one;

Bacillus subtilis form round colorless colonies, dry, finely wrinkled, velvety, the edge of the colony is wavy;

Bacillus mucilaginosus form convex, colorless, transparent, mucous colonies, with a smooth edge, viscous consistency, 3-5 mm in diameter;

Bacillus coagulans form small to medium or large colorless colonies with a slightly wavy margin. Differences in colony size on the same agar plate may give the impression that the culture is contaminated;

Paenibacillus polymyxa forms colorless, flat or convex, smooth and slimy colonies with a serrated edge;

Pseudomonas chlororaphis forms large, opaque, flat colonies with irregular margins, colorless to orange;

Pseudomonas putida form translucent, colorless, shiny colonies of a round-angular shape with smooth, solid edges.

Cultural characteristics during surface cultivation of mushrooms on wort agar:

Trichoderma harzianum quickly forms large zonal, hairy colonies with flaky aerial mycelium, the reverse side is unstained;

Trichoderma viride quickly forms large, non-pigmented, hairy colonies. Conidial sporulation in small blue-green pads

Morphological features

Arthrobacter globiformis in the process of culture aging, the cell morphology changes from the state of gram-negative rod-shaped cells to gram-positive coccoid cells.

Bacillus licheniformis cells are Gram-positive, spore-forming rods.

Bacillus pumilus cells are Gram-positive rods with endospores and peritrichous flagella.

Bacillus subtilis cells are Gram-positive spore-forming (spores located centrally) rods measuring 3 µm by 0.4 µm.

Cells of Bacillus mucilaginosus are large rods of the correct form with rounded ends, single, with a mucous membrane.

Bacillus coagulans cells are Gram-positive, spore-forming, motile rods measuring approximately 0.9 µm by 3.0–5.0 µm.

Paenibacillus polymyxa cells are gram-positive rods 0.8 µm by 2-5 µm in size, form a thermostable oval endospore that exceeds the size of the cell (lemon-shaped).

Pseudomonas chlororaphis cells are gram-negative rods that do not form spores with one or more polar flagella.

Pseudomonas putida cells are gram-negative rods that do not form spores and have one or more flagella.

The conidiophores of Trichoderma harzianum are pyramidal in outline, with opposite branches at the nodes, with whorls of 2-5 flask-shaped phialides 8 µm by 4 µm in size.

The conidiophores of Trichoderma viride are loosely branched, the branches are paired, the phialides are also paired, narrowly flaccid, 10 µm by 3 µm, conidia are green, spherical 4.5 µm by 3.5 µm, distinctly warty.

All of these strains are not zoopathogenic and phytopathogenic.

cultivation

Potato agar is used as media for propagation and maintenance of pure cultures. Cultivation is carried out at 25°C. The grown cultures are stored in bevelled tubes under a gauze stopper at 5°C.

For fermentation, potato medium is used as a nutrient medium. The cultivation time in an aerated 20 liter fermenter is 36-48 hours at a temperature of 25°C.

Fermentation is carried out jointly: culture liquids of microorganisms combined in the indicated ratios are inoculated into the medium in the form of a mixture. In the process of cultivation, the quantitative ratios of microorganisms in the consortium are preserved. After cultivation, the solution is packaged and stored at room temperature.

Industrial applicability.

The plant root system growth accelerator in all three variants (hereinafter referred to as preparation 1, preparation 2 and preparation 3) according to the test results was found to comply with the requirements of TU 20.20.13.120-001 - plant growth regulators.

Determination of the effect of the line of preparations on plant growth was carried out at the Department of Botany and Mycology of the Faculty of Medicine and Biology, Voronezh State University. In the course of research, the effectiveness of all formulations was demonstrated by examples.

Example 1. Evaluation of the effect of drugs on the growth of dry and fresh mass of shoots and on the abundance of flowering.

The cuttings of test cultures, which quickly took root, were transplanted into pots with a volume of 0.5 l. containing a substrate. Four plants were provided for each experiment.

Irrigation was designed in such a way that the solution flows evenly under the plant. In the daytime, one or two irrigations were carried out (using 0.07-0.08 liters of solution per plant for each irrigation). Plants were grown in a phytochamber at a temperature of 25-26°C during the daytime (18 hours during the first four weeks of the vegetative stage and 10 hours during the next nine weeks, in the so-called flowering phase) and at a temperature of 18-20°C at night. They were lit by lamps producing 600 watts per square metre.

The test plants were irrigated with a 0.2% solution of biopreparations in deionized water, corresponding to compositions 1, 2, 3 (mentioned above), control plants were irrigated with deionized water.

After cultivation, the wet and dry weight of the shoots was evaluated. As a result, an increase in the fresh weight of shoots by 20%, 14% and 5% was observed with the use of preparations 1, 2 and 3, respectively. A multiple increase in weight was also observed for dry shoots.

Table 1. The effect of drugs on the wet and dry mass of shoots.

Raw mass of shoots (grams) Wet mass of shoots (%) Dry weight of shoots (grams) Dry mass of shoots (%) Control 73.67 100 % 23.85 100 % Preparation 1 88.70 120.4% 28.26 118.5% Preparation 2 84.41 114.6% 27.3 114.5% Preparation 3 77.27 104.9% 26.5 111.1%

The area of green and red leaves of each plant was also estimated. Estimation of the area of red leaves made it possible to estimate the intensity of anthocyanin formation, which indirectly made it possible to evaluate the contribution of the studied preparations to the flowering efficiency. As a result, an increase in the proportion of red leaves was observed both within the series and relative to the control. So the proportion of red leaves increased by 118%, 68% and 12% with the use of preparations 1, 2 and 3, respectively.

Table 2. The effect of drugs on the increase in the proportion of red leaves.

Total area of green leaves (cm^2) Total area of red leaves (cm^2) The proportion of red leaves of the plant (%) Increase in the proportion of red leaves relative to control (%) Control 2203.13 543.61 19.8% 100 % Preparation 1 1792.4 1186.49 39.8% 218.3% Preparation 2 1916.95 914.33 32.3% 168.2% Preparation 3 1755.15 609.32 25.8% 112.1%

Thus, the technical result of the claimed invention is to stimulate growth and increase the productivity of agricultural plants. It is achieved by the fact that consortiums of microorganisms are used for processing plants, which have a pronounced growth-stimulating activity due to a phytohormone-like effect on plants, improve the rhizosphere community, improve their mineral nutrition, due to the mobilization of phosphorus nutrition, while being able to maintain viability and biological activity for a long time when stored in the form of a liquid culture.

Claims (25)

1. Accelerator of the growth of the root system of plants containing the culture liquid of microorganisms: Bacillus licheniformis with a titer of at least 1.5 * 10 ⁷ CFU in cm ³ , Bacillus pumilus with a titer of at least 1.5 * 10 ⁷ CFU in cm ³ , Bacillus subtilis with a titer of at least 10 ⁷ CFU in cm ³ , Paenibacillus polymyxa with a titer of at least 2 * 10 ⁷ CFU in cm ³ , applied as a 0.2% solution in deionized water, used for the treatment of young and adult plants during the growing season and flowering. 2. Accelerator of growth of the root system of plants containing the culture liquid of microorganisms: Bacillus mucilaginosus with a titer of at least 2.5 * 10 ⁷ CFU in cm ³ , Pseudomonas chlororaphis with a titer of at least 10 ⁷ CFU in cm ³ , Pseudomonas putida with a titer of at least 1.5 * 10 ⁷ CFU in cm ³ , Trichoderma harzianum with a titer of at least 1.5 * 10 ⁷ CFU in cm ³ , Trichoderma viride with a titer of at least 2 * 10 ⁷ CFU in cm ³ , applied as a 0.2% solution in deionized water, increasing the availability of mineral nutrition and increasing protection against soil pathogens, for the treatment of young and adult plants during the growing season and flowering. 3. Accelerator of growth of the root system of plants containing the culture liquid of microorganisms: Arthrobacter globiformis with a titer of at least 6.6 * 10 ⁷ CFU in cm ³ , Bacillus coagulans with a titer of at least 1.2 * 10 ⁷ CFU in cm ³ , Bacillus licheniformis with a titer of at least 1 * 10 ⁷ CFU in cm ³ , Bacillus mucilaginosus with a titer of at least 1 * 10 ⁷ CFU in cm ³ , Bacillus pumilus with a titer of at least 3.3 * 10 ⁷ CFU in cm ³ , Bacillus subtilis with a titer of at least 1 * 10 ⁷ CFU in cm ³ , Paenibacillus polymyxa with a titer of at least 3.3 * 10 ⁷ CFU in cm ³ , applied as a 0.2% solution in deionized water, which is an accelerator of combined action for the treatment of young and adult plants during the growing season and flowering.