Strain mycelia sterilia-producer of biologically active substances with postregulatory properties

 

(57) Abstract:

The invention relates to the microbiological industry. The strain of the fungus Mycelia sterilia stored in the all-Russian collection of industrial microorganisms, collection number VKPM F-833 is a producer of biologically active substances with postregulatory properties. The invention allows to obtain a drug used as a bio-stimulator 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 corn, gooseberries, currants, potatoes, cucumbers, tomatoes, lawns, 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 (U.S. Pat. Of the Russian Federation No. 2100932). The disadvantage of this community is the difficulty and complexity of its cultivation, the instability of the drug received.

Known drugs is RNA wheat, oats, potatoes, wheat grass, mother and machihe. Each product consists of dissolved in 70% ethyl alcohol products of metabolism associated endophyte. Processing one of the listed drug of wheat seeds, cucumbers or potatoes before sowing increased the yield (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 ginseng root. The drug is effective only under good growing conditions (AV.St. The USSR № 370932).

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

Most near what wantom of plant growth regulator (patent of Ukraine No. 17558). The strain isolated from the root system of sea buckthorn. As the nutrient medium used, g/l: glucose, KN2RHO4TO2NRA4, MgS4TO2SO4, MnS4, FeS4, 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 presence of alcohol in the drug.

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 wild rosemary isolated a new strain of Mycelia sterilia. The strain is stored in the all-Russian collection of industrial microorganisms, collection number VKPM F-833. The strain is used for obtaining a biostimulant plant growth.

The strain isolated from roots of wild rosemary. Species the name of culture Mycelia sterilia var. Ledum., the name of the strain NZ-13. Pedigree strain: natural isolate. This type of the selected fungus similar to mushrooms “Heath mycorrhizal fungi”.

Cultural and morphological characteristics of strain: Aerobe. The fungus gives slow-growing appressed silky colony of black (growth coefficient on Booze, 1988 - equal to 10). At aneesa. When using the microscope revealed that the fungus is dark-colored, with a high content of drops of oil (the diameter of the hyphae does not exceed 3 μm). On hyphae formed a significant number of intercalary collected in chains of spherical chlamydospores.

The product synthesized by the strain, is a complex of biologically active substances.

When the cultivation of the strain in the course of 3 weeks in liquid medium geltzer, followed by separation of complex secondary metabolites obtained the drug gives the increase in the yield of agricultural crops by 20-30%, and the growth of the hypocotyl experimental lines mustard 30-60% of 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

TO2NRA41,8

MgS4 0,2

TO2SO40,1

MnS4Traces

FeS4Traces

L-asparagine 0,02

Agar-agar 15,0

Water Up to 1000 ml

The reproduction of culture in the implementation of the medium composition for fermentation: liquid nutrient medium geltzer culturing duration 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

KN2RHO40,6

TO2NRA41,8

MgS40,2

TO2SO4 0,1

MnS4Traces

FeS4Traces

L-asparagine 0,02

Water Up to 1000 ml

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

Drug-based strain NZ-13 received when grown in culture flasks. Conducted cultivation in flasks, then carried out the coarse and fine filtration. Then the obtained product is freeze-dried. When conducting biotests and chemical analyses used lyophilized culture filtrate liquid.

The content of phytohormones in the preparation: 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. Odnevall above.

Indelicacy acid (IAA) is poststimulation, but due to poor solubility 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.

The method of obtaining strain allows you to scale the production process that is a prerequisite for the commercialization of the project. The optimal concentration of strain in the product - 10 ppm.

Table 2 shows the elemental composition of lyophilized.

Elemental composition analysis showed that the product contains almost no heavy metals. In the drug compared with a nutrient medium znachitelnie free carbohydrates, mg/g dry. weight:

Sucrose 178

Glucose 376

Fructose 91

Mannitol 106

The drug contains the following amino acids: lysine - 0,0066, histidine - 0,0547, asparagine - 0,0221, serine - 0,0050, glutamine - 0,0121, glycine - 0,0218, alanine - 0,0410, valine - 0,0513. Amino acids were determined by the method ionoobmennoi chromatographie. When holding tight of hydrolysis (to determine corresponding amino acid) in the preparation is a fairly wide range of amino acids present in small quantities. The qualitative composition of free amino acids are less diverse, they are also present in small quantities.

The drug contains fatty acids, including arachidonic acid, which is an important component of the complex metabolites that stimulate the growth and development of plants. The content of fatty acids was determined by gas chromatography.

The drug also contains ascorbic acid 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 18,478

Amino acids 0,0660

Carbohydrates 751,000

Ash elements 27,737

Received the biostimulator sadisti microorganisms.

The invention is characterized by the following examples.

Example 1.

Strain li sterilia grows on rich organic medium (liquid wort) in conditions of aeration at 20-30°C for 48 hours produces 25 g permeabilites 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

TO2SO40,1

MnSO4Traces

FeSO4Traces

L-asparagine 0,02

Water Up to 1000 ml

As a source of carbon supply used glucose concentrations of 1.0% or 2.0 wt.%.

During cultivation in liquid nutrient medium declare strain requires additional growth factor - fermentolizat BVK in the amount of 0.2%. The 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 hours on a rocking chair (220 rpm). It was found the optimum pH value for growth of culture Mycelia sterilia 6,8-7, is its metabolites in five consecutive fermentation.

The liquid preparation on the basis of the proposed strain was carefully mixed and used for processing (soil) planting seedlings of spruce, gooseberry, currant (top dressing), potato (watering of seedlings), late cabbage (fertilizing seedlings before planting in the ground), 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 for individual crops are given 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 that are useful for plants. The drug, made on the basis of the proposed strain, will find wide application in modern agricultural biotechnology.

Permeability, the composition of which is given in table 1, synthesized by the claimed strain was also used in the processing of crops by seed treatment and spraying of plants on shoots. In addition, the positive R the m increased on average 32-34%.

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 - producer of biologically active substances with postregulatory properties, collection number VKPM F-833.

 

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