Strain mycelia sterilia lh-1-producing complex of biologically active substances with postregulatory properties

 

(57) Abstract:

The invention relates to the microbiological industry. The strain of the fungus Mycelia sterilia HL-1 stored in the all-Russian collection of industrial microorganisms, collection number VKPM F-862, is a producer of biologically active substances with postregulatory properties. The invention allows to obtain a drug used as biostimulants of plant growth. 3 table.

The invention relates to the microbiological industry and relates to a new culture of microorganisms, producing biologically active substances with postregulatory properties that can be used in agriculture in the processing of such crops as potatoes, cucumbers, tomatoes, etc.

Known community of microorganisms for receiving plant growth regulators, consisting of micromycete Acremonium panaxeorum UHMU F-160, yeast Candida muscorum UHMU Y-411, Rhodotorula glutinis UHMU Y-42 and bacteria Azomonas macrocytogenes UHMU B-28 and Xantobacter sp UHMU B-29 /Pat. RF 2100932/.

The disadvantage of this community is the difficulty and complexity of its cultivation.

Known drugs that stimulate the development and growth of plants, obtained from the net which comprises dissolved in 70% ethyl alcohol products of metabolism associated endophyte. Handling one of these drugs wheat seeds, cucumbers or potatoes before sowing increased the harvest /geltzer F. Y. New producers stimulating substances to plants. Reports of agricultural Sciences, No. 5, 1975/.

Known drug Symbiot-2”, stimulating the productivity of plants, obtained from the products of metabolism of fungi endophyte (AV.St. The USSR №921488). As raw materials for its manufacturing use endophyte isolated from the roots of sea-buckthorn.

A disadvantage of the known stimulators of growth is the low activity of preparations derived from endophyte plants with the highest medicinal value.

Known drug Symbiont-1 carrier made of endophyte isolated from the roots of a ginseng. The drug is effective only under good growing conditions (AV.St. The USSR №370932).

A common disadvantage of known products is the inability to identify produced using known strains of products, the content of significant quantities of alcohol in the product.

The closest technical solution to the claimed invention is a known strain Cylinddrocarpon magntsianum, which is the producer of the plant growth regulator /patent UH, KH2PO4, K2HPO4, MgSO4TO2SO4, MnSO4, FeSO4, asparagine.

Its main drawback is the complexity of its receipt on an industrial scale, the strain has efficacy only in the root zone of a particular host plant.

The aim of the invention is the expansion of sources of raw materials of new strains and increasing the activity of growth regulator.

The objective is achieved by the fact that from the roots of Loch isolated a new strain of Mycelia sterilia LH-1. The strain is stored in the all-Russian collection of industrial microorganisms, collection number VKPM F-862. The strain is used for obtaining a biostimulant plant growth.

The strain isolated from the roots of the Loch. Species the name of culture Mycelia sterilia, the name of the strain HL-1. Pedigree strain: natural isolate.

Cultural and morphological characteristics of strain: Aerobe. The fungus gives slow-growing filamentous colonies are grey-brown in color, growing into the agar. The coloring of the environment under the influence of the fungus does not change. When using the microscope revealed that the fungus is dark-colored, with a high content of drops of oil. On hyphae formed a significant number of intercalary collected in chains erowid substances.

When the cultivation of the strain in the course of 3 weeks in liquid medium geltzer, followed by separation of complex secondary metabolites received the drug increases adsorbing surface cereals at 30-70%, and the growth of the hypocotyl experimental lines mustard 50-70% compared to control (water).

Biological activity was determined using the standard methods of biotests on the seeds of plants.

The strain was kept on agar nutrient medium at a temperature of 4-7°C With re-seeding once in 3 months.

Agar medium geltzer (HA), g/l, pH 6,2-6,6:

Glucose 8,0

KN2RHO40,6

TO2NRA4 1,8

MgSO40,2

TO2SO40,1

MnSO4Traces

FeSO4Traces

L-asparagine 0,02

Agar-agar 15,0

Water Up to 1000 ml

The reproduction of culture carried out by subculture on agar medium geltzer, the composition of which is presented above.

Conditions and the composition of the medium for fermentation: liquid nutrient medium geltzer culturing duration 20-30 days at a temperature of 261°C. the Composition of the medium: liquid medium geltzer, g/l, pH 6,0-6,4:

Glucose 8,0

KN2RHO40,6ptx2">FeSO4Traces

L-asparagine 0,02

Water Up to 1000 ml

The strain is not zoopathogenic, is not pathogenic, there is no danger to the environment.

Preparation on the basis of strain HL-1 was obtained when grown in culture flasks. Cultivation in flasks was carried out in two stages - after the first cycle was selected culture fluid was added to fresh medium in the amount of 30% of the original volume and was carried out by the second cultivation cycle. When conducting biotests and chemical analyses used lyophilized culture filtrate liquid.

The content of phytohormones in the preparation: indelicato acid (IAA), cytokinins (CC), gibberellin (HA) - was determined by the chromatographic method.

Cytokinins are the most active natural compounds involved in the hormonal regulation of plant ontogenesis and characterized by the ability to induce the division of plant cells.

Gibberellovaja acid HA3the most widely used in agriculture as a plant growth regulator. However, HA7represents the greatest interest, because it has a wider range of actions and fiziologicheskii in water and relatively high cost has a narrow range recommended for the cultivation of crops.

All these compounds are related to well-known plant growth regulators, are described in detail in the literature /plant growth Regulators. Edited by G. C. Muromtseva, M., 1979/.

The inventive strain allowed us to obtain simultaneously all these compounds, which significantly increased the biological activity of the drug and at the same time increased the economic efficiency of its application.

The biological activity of the dry preparation on the basis of the proposed strain on the bluegrass, grade Denim, (control - water) and the content of phytohormones in this preparation are listed in table 1.

Table 2 shows the elemental composition of lyophilized.

The content of free carbohydrates, mg/g dry. weight: sucrose - 431, glucose - 181, fructose - 134, mannitol - 145.

The drug contains the following amino acids:

lysine - 0,055, histidine - 0,024, asparagine - 0,005, serine - 0,096, glutamine is 0.025, glycine - 0,031, alanine - 0,040, valine - 0,061. Amino acids were determined by the method ionoobmennoi chromatographie.

The drug contains fatty acids, including arachidonic acid, which is vanim component of the complex of metabolites stimulating the growth and development of plants. The content of fatty to Isleta and citrine (vitamin P). Was determined by the method of color reactions.

Data on the chemical composition of the drug (mg/g dry. weight:

Phytohormones 28,450

Amino acids 0,166

Carbohydrates 834,000

Ash elements 14,500

Received the biostimulator precipitated with ethanol, acetone, heat resistant up to 150°C, characterized biological stability, resistant to microorganisms.

The invention is characterized by the following examples.

Example 1.

Strain Mycelia sterilia HL-1 grows on rich organic medium (liquid wort) in conditions of aeration at 20-30°C for 48 h produces 25 g formatable showing biostimulating activity. Cultivation of strain operate on liquid nutrient medium geltzer, duration of cultivation 20-30 days at a temperature of 26±1°C. the Composition of the medium: liquid medium geltzer, g/l, pH 6,0-6,4:

Glucose 8,0

KN2RHO4 0,6

TO2NRA41,8

MgSO40,2

K2SO40,1

MnSO4Traces

FeSO4Traces

L-asparagine 0,02

Water Up to 1000 ml

As a source of carbon supply used glucose in a concentration of 1.5%. During cultivation in liquid Cultivation is carried out at 28-30°C, under conditions of aeration and mixing in the flasks with a volume of 750 mm3with 100 ml of medium of the composition within 48 h on a rocking chair (220 rpm). When this occurs, the acidification of the medium to a pH of 4.7 by releasing organic acids and in the culture fluid. It was found the optimum pH value for growth of culture Mycelia sterilia of 6.8-7.0 and for the synthesis of metabolites - 6,8-7,2.

Investigated the synthesis of metabolites declare strain in the temperature range of 25-35°C. To achieve the desired pH values resulted in Podarok 10% NaOH solution.

The process conditions: temperature 28-30 ° C. pH of 6.8 to 7.2, the air flow of 0.5 l/l environment 1 min, stirring speed 300 rpm, time of cultivation 18-20 hours

The process is stable, the Association stably retains the ability to synthesize metabolite in five consecutive fermentation.

Metabolite, the composition of which is given in table 1, synthesized by the claimed strain used in the processing of crops of potatoes, cucumbers, tomatoes, processing seedlings of red currants and gooseberries.

The liquid preparation on the basis of the proposed strain was carefully mixed and used for processing (soil) planting seedlings soil), tomatoes (foliar application), cucumbers (watering of seedlings). Led morpho-physiological and phenological observations, take into account the yield of the products was determined by some parameters characterizing the quality of the products. The results are shown in table 3. Control (100%) - performance products with a plot that is not processed by the inventive strain.

The inventive strain carries a positive effect on plants through direct and stimulate plant growth through the synthesis of various metabolites, useful for plants. The drug, made on the basis of the proposed strain, will find wide application in modern agricultural biotechnology.

The inventive strain when conducting patent and information search on the technical level is not detected, and therefore it is possible to draw a conclusion on the compliance of the claimed invention for patentability under the Patent law of the Russian Federation.

The strain of the fungus Mycelia sterilia LH-1 - producing complex of biologically active substances with postregulatory properties, collection number VKPM F-862.

 

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