Strain mycelia radicis var. ginseng-producer of biologically active substances with postregulatory properties
The invention relates to the microbiological industry. The strain of the fungus Mycelia radicis var stored in the all-Russian collection of industrial microorganisms, collection number VKPM F-860 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 crops such 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 disadvantages of this community are the complexity and the complexity of its cultivation, the instability of the final product.
Known drugs that stimulate the development and growth rate-and-machihe. Each product consists of 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 for 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 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 tableevent of Ukraine No. 17558/. The strain isolated from the root system of sea buckthorn. As the nutrient medium used, g/l: glucose, KH2PO4, K2HPO4, MgSO4, K2SO4, 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 ginseng isolated a new strain of Mycelia radicis var. ginseng. The strain is stored in the all-Russian collection of industrial microorganisms, collection number VKPM F-860. The strain is used for obtaining a biostimulant plant growth.
The strain isolated from ginseng root. Species the name of culture Mycelia radicis var. ginseng, the name of the strain NTN-1. 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 grey-brown colour, with the lower side of the black color. The growth rate of 0.17 cm/day. The coloring of the environment under isim content drops of oil, very often glued in arimiopidardy patterns. On hyphae formed a significant number of intercalary collected in chains of spherical chlamydospores (from 3 to 5, rarely 7 pieces).
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 30-60%, 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:
Water ml 1000
The reproduction of culture carried out by subculture on agar medium geltzer, the composition of catagorically 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:
Water ml 1000
The strain is not zoopathogenic, is not pathogenic, there is no danger to the environment.
Drug-based strain NTN-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: endoxana acid (IAA), cytokinins (CC), gibberellins (GA) 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.
Endoxana acid (IAA) is poststimulation, but due to poor solubility in water and relatively high cost has a narrow range recommended for 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. Elemental composition analysis showed that the product does not contain heavy metals. The drug significantly compared with the culture medium increased the potassium content and molybdenum content of other elements omegaedit - 97.
The drug contains the following amino acids: histidine 0,0352, asparagine 0,0207, serine 0,0068, glutamine 0,0115, glycine 0,0204, alanine 0,0164, valine 0,0091.
Amino acids were determined by the method ionoobmennoi chromatography. 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. Free amino acids in the composition 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 is mg/g dry. weight:
Amino acids 0,0970
Ash elements 21,637
Received the biostimulator precipitated with ethanol, acetone, heat resistant up to 150°C, characterized biological stability, resistant to impact the Strain Mycelia radicis var. ginseng 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:
Water ml 1000
As a source of carbon supply used glucose in a concentration of 1.5 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 flasks with a volume of 750 mm3with 100 ml of medium of the composition within 48 hours 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 culo retains the ability to synthesize metabolites in five consecutive fermentation.
The metabolites, the composition of which is given in table 1, synthesized by the claimed strain, used in the processing of crops of potatoes, cucumbers, tomatoes.
The liquid preparation on the basis of the proposed strain was carefully mixed and used for processing (soil) planting seedlings of spruce (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 are shown in table 3. Control (100%) - performance products from areas not treated by the claimed 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.
The inventive strain when conducting patent and information search on the technical level is not detected, and therefore can make is the law of the Russian Federation.
The strain of the fungus Mycelia radicis var. ginseng - producer of biologically active substances with postregulatory properties, collection number VKPM F-860.