The community of microorganisms for receiving a plant growth regulator, a method of obtaining a controller and a plant growth regulator

 

(57) Abstract:

The invention relates to biotechnology and the receipt of plant growth regulator on the basis of the microorganisms. From the roots of ginseng dedicated community of microorganisms consisting of micromycete Acremonium sp, yeast Rhodotorula glutinis bacteria Enterobacter agglomerans and Azotobacter beijerinckii. The community is deposited in the collection of microorganisms MSU under 452. It is intended for receiving a plant growth regulator. The controller is obtained by cultivating community in a nutrient medium containing sources of carbon, nitrogen, mineral salts within 1 month. From the culture fluid is separated mycelium. The mycelium is extracted with ethanol, incubated the mixture, remove the sediment. Get colorless transparent or slightly opalescent with a specific smell solution containing gibberellin, purine bases, amino acids. The drug increases the germination and vigor of seeds, crops and reduces the incidence of plants. 3 C. I. 1 C.p. f-crystals.

The invention relates to biotechnology and the receipt of funds, increasing the yield of agricultural crops from the community micro is the plant growth regulator, including mushrooms-endophyte isolated from the roots of a ginseng, sea buckthorn (1-2). The disadvantage of this community is uncertain composition of the microorganisms belonging to the community.

Known community of microorganisms for receiving a plant growth regulator, consisting of micromycete Acremonium panaxeorum UHMU F-160, yeast Candida muscorum UHMU U-411, Rhodotorula glutinis UHMU U-42 and bacteria Azomonas macrocytogenes UHMU B-28 and Xantobacter sp UHMU B-29 (3).

Known methods for producing plant growth regulators by culturing one of the above communities on nutrient medium containing a carbon source, mineral salts, amino acids, followed by separation of the biomass of biologically active substances that stimulate the growth of plants (1-4).

Known methods received drugs the Symbiont - 1(1), Symbiont-Estate (3). These drugs are alcohol extracts from biomass communities of microorganisms. The aim of the invention is the expansion of the sources of raw materials from new communities of microorganisms and increasing the efficiency and stability of drugs.

The objective is achieved by the fact that the roots of a ginseng allocated a new community of microorganisms consisting of micromycete Acremonium sp, yeast Rhodotorula glut the society is kept in the collection of the Moscow state University under room 452.

Included in the community of microorganisms have the following characteristics.

Cultural and morphological characteristics of bacteria Enterobacter agglomerans. When grown on nutrient and BSA form colonies white, slimy, smooth up to 3-4 mm in diameter. On the environment Saburo with a high content of glucose forms the exopolysaccharides. Culture on liquid medium is a single movable sticks sometimes in pairs. Cells growing on solid medium, a day later form short chains, pairs or represented by single forms of different lengths. The size of the sticks of 1.2 to 3 μm, a diameter of ~1 µm. Endospore do not form. Culture refers to a group of "Facultative anaerobic gram-negative rods". Optimum temperature for growth is 25-30oC, there was an increase at 37oC, and 15oC. Gelatino thins, hydrolyzes casein, starch is not hydrolyzes, pigments are not formed. Used as a nitrogen source ammonium salts, organic compounds and nitrate. Restores the nitrate to nitrite. Grows well on citrate, urease does not synthesize (growth medium with urea missing). The hydrogen sulfide from cysteine and indole from tryptophan does not form. Grows well on medium Ashby and liquid nitrogen-free environment, Fedora is ukazu, glycerin, D-mannose, D-xylose, maltose, melibiose, raffinose, arabinose, L-rhamnose, D-sorbitol and sucrose. Many of these carbohydrates forms a gas. Identification of bacterial species is made on the basis of the determinant Bergey''s Manual of Systematic Bacteriology (in four volumes), 1984, Baltimore /London.

Cultural and morphological characteristics of the yeast Rhodotorula glutinis.

When growing yeast on solid agar medium containing 10oB-wort, and glucose-peptone medium with addition of yeast extract formed small colonies of pink with a diameter of 2-3 mm Colonies rounded edges pink, the center of the colony is elevated, surface matte, with age appreciable annular grooves. Bar a bright pink color, smooth, shiny. Pseudomycelium culture does not form. Cells oval or oblong shape, size 1,52,0 to 3.0 μm. Multiply by budding yeast. Spores do not form. As a source of nitrogen for growth uses both organic and inorganic forms of nitrogen. As the sole source of carbon and energy uses glucose, sucrose, maltose, cellobiose, raffinose, mannitol, ethanol and succinate. Not assimilating lactose, melibiose, inulin, soluble starch, rhamnose, citrate, meta is th range growth 10-40oC. Optimum 15 - 28oC.

Identification of the yeast culture was carried out on the basis of the determinant edited Babeva I. P., Golubev, C. I., "Methods for isolation and identification of yeast", M: Food industry, 1979, The Yeasts/ A taxonomic stady/ Sec cd/ by the Lodder, Amsterdam-London, 1970.

Cultural-morphological signs of micromycete Acremonium sp. When growing micromycete Acremonium sp on agar medium of čapek formed colonies white, after a few days of the colony have creamy color, and in old cultures dark brown up to black. Mycelium abundant vatoobraznye. Badly removed from the agar. With age, the reverse side of the colony becomes brown. And in old cultures black. Air mycelium formed conidiophores solitary, sometimes in whorls. Conidia are formed on the top of canadianese, alone and/or in false heads. Conidia are hyaline, mostly cylindrical with rounded ends. The length of the conidia is about 17 μm, a thickness of 5 μm. Meet conidiospore with a partition. Along with the large cylindrical conidia are fine from oval to spherical. In liquid medium, the mycelium of the fungus presents the septate hyphae with many strands, surrounded is by the formation of chlamydospores. Optimum growth temperature is 20oC. Pure culture of micromycete supported and stored on agar medium of čapek and wort-agar. Micronized is stored in the collection of microorganisms MSU under N 441.

Identification of micromycete carried out on the basis of the following sources: Litvinov, M. A. "the Determinant of microscopic soil fungi", Leningrad : Nauka, 1967, pidoplichko N. M. "Mushrooms-parasites of cultivated plants", Kiev, "Naukova Dumka", 1977, vol 2, Gilmal J. C. "A manial of soil fungi" Second ed. The Iowa State Univ. Press. Ames. Lowa, USA, 1984.

Cultural-morphological and physiological biohimicheskie signs of Azotobacter beijerinckii.

When culturing bacteria Azotobacter beijerinckii on meat-peptone agar (MPA) forms dense white and smooth, slightly convex colonies with aging purchasing beige shade. Colonies 1-2 mm in diameter, with age, acquiring a wavy edge. Colony soft and easily removed from the agar. Cells not less than 1.5 microns in diameter, coccoid, arranged in pairs, forming an eight. Cells daily culture on solid and liquid media stationary. When negative staining with India ink visible mucous capsule surrounding cells. During aging culture forms cisterian oxygen in the environment. Grows well on medium Ashby. Culture produces a lot of mucus. Fix molecular nitrogen in acid medium at a pH of 5.5 to 6.0. Grows well at 20-30oC, at 37oC no growth. Uses nitrate and restores it to nitrite, servoport of cysteine does not form. Gelatine does not dilute. Starch and casein not hydrolyses. As the sole source of carbon assimilates fructose, glucose, sucrose, ethanol, lactic and propionic acid. Rhamnose is not used.

Identification of this type of bacteria was carried out by Bergey's Manual of systematic Bacteriology", 1984.

The invention is illustrated by the following examples:

Example 1: receiving a plant growth regulator "Symbiont".

Microbial community consisting of micromycete Acremonium sp, yeast Rhodorula glutinis, bacteria Azotobacter beijerinckii and Enterobacter agglomerans, KM MSU 452, seeded in medium containing (%):

Glucose - 0,8

K2HPO4- 0,03

MgSO4- 0,02

K2SO4- 0,01

FeSO4and MnSO4- Traces

Asparagine - 0,001

Distilled water Up to 1 liter

Cultivated in stationary conditions when the initial pH of 6.5 and a temperature of 20oC during the month. At the end of the process of cultivation of biomass otdelyalos and get the drug "Symbiont", which is an aqueous-alcohol colorless, transparent or slightly opalescent with a specific smell solution metabolic products of the above-mentioned microorganisms. The drug contains gibberellin not less than 0.05 mg/ml, purine bases not less than 0.02 mg/ml and amino acids not less than 1 mg/ml, the content of heavy metals does not exceed 0.001%, arsenic and mercury are not available, the pH of 5.4 to 5.8.

The ethanol content 48-54%.

The dry residue is not less than 0.25%.

From the controller includes amino acids glutamine, glycine, alanine, valine, histidine, tsitizin, guanine.

The drug stimulates the development of mycorrhizal fungi, which leads to improved plant nutrition. Received mycorrhizal fungi ahead in its development of pathogenic, occupying the same ecological niche, which helps reduce the degree of destruction of fungal phytopathogens.

The Foundation protects the plant during the growing season. The speed of impact after 8-10 days after the growing season.

The controller is compatible with all crop protection products: disinfectants, insecticides, fungicides, herbicides and plant growth regulators. The drug was tested on different plants.

ISS is Ontario with a consumption rate of the product 2 ml/1 kg over 30 minutes and subsequent spraying of plants 3 weeks after transplanting into the soil at the rate of consumption of the drug 1 ml/ha provided a true increase yield of cabbage in 11-30%. The use of the drug has increased the germination power of seeds, their germination and improve the quality of seedlings. The incidence of cabbage seedlings "black leg" decreased by 15%, cabbage various bacterial diseases by 15-20%, downy mildew of Lucerne by 5-12%.

Studies of biological effects of Symbiont on cucumber (variety not female). The most effective was the soaking of seeds in the plant growth regulator with the consumption rate of the product of 1 ml/kg, rate of flow of the working solution of 2 l/kg, a soak time of 30 minutes, 2x spraying: 1st phase of education 1-2 leaves, the 2nd phase of education 4-5 leaves. The consumption rate of the drug 1 ml/ha, the rate of flow of the working solution of 300 l/ha Increase in yield of cucumber was 16.7% or 1,53 kg/m2. In field trials the yield of cucumber increased by 33%.

In the 2-year trials of plant growth regulator on the pepper cultivar Vigor determine the most effective regulation of growth regulator application: soak seeds for 30 min, the flow rate of the drug 2 ml/kg, rate of the working solution of 2 l/kg and spraying flowering 1 ml/ha, at a rate of flow of the working solution of 300 l/ha of Studies have shown that the use of the "Symbiont" fruit.

Tests conducted on the eggplant varieties Wagon 3 showed that the most effective treatment of the eggplant is soaking for 30 minutes in a solution of "Symbiont" and spraying of plants flowering at the above rates. The yield of eggplant increased 13.8-16.8% and amounted to 1.13 - 1,12 kg/m2. Received more developed and healthy seedlings of eggplant, increased weight 1 of the fetus and the average rate on 1 plant.

The drug increases the yield of buckwheat in the range of 9-11%, this has increased the germination and vigor of seeds, grain weight average 1 plants. Buckwheat yield increased by 9-11%. The drug also increases the yield of sugar beet by 10-20%, a sugar content of roots by 0.5-0.8%. The drug is tested when growing asters. The application of the regulator increased the height of plants asters, increasing the number of inflorescences and their diameter, % blooming flowers, reducing the infestation of seedlings asters "black leg".

Treatment of seeds of watermelon varieties of Light and Kherson early growth regulator at a dose of 4 ml/kg, at a flow rate of the working solution of 2 l/kg, increased the yield of watermelon in the 36-40%, the yield increase was 42-lucency the drug helps to increase crop yields and plant resistance to pathogens.

Sources of information:

1. Author's certificate N 370932, 22.11.73, A 01 N 65/00.

2. Author's certificate N 921488, 23.04.82, A 01 N 65/00.

3. Patent N 2100932, 10.01.98, A 01 N 63/00.

4. Author's certificate N 1824148, 30.06.93, A 01 N 65/00.3

1. The community of microorganisms consisting of micromycete Acremonium sp, yeast Rhodotortula glutinis, bacteria Enterobacter agglomerans and Azotobacter beijerinckii KM MSU 452 for receiving a plant growth regulator.

2. A method of producing a plant growth regulator, providing for the cultivation of microorganisms with subsequent extraction of biologically active substances obtained alcohol from biomass, characterized in that the microbial community use community under item 1.

3. The plant growth regulator containing alcohol extract of the biomass of the microbial community, characterized in that it contains alcohol extract of the biomass of the microbial community under item 1.

4. The plant growth regulator on p. 3, characterized in that it contains gibberellin not less than 0.05 mg/ml, purine bases not less than 0.02 mg/ml, amino acids not less than 1 mg/ml, alcohol 48 - 54%, the dry residue of trace elements is not less than 0.25%, while arsenic and mercury are missing.

 

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