RU2100932C1 - Plant growth biostimulating agent symbiont-universal and microorganisms consortium - Google Patents

Abstract

FIELD: microbiology, agriculture. SUBSTANCE: biostimulating agent is an alcoholic extract of biomass of microorganism consortium consisting of micromycetes Acremonium panaxeorum INMI F-160, yeast of two species Candida muscorum INMI Y-41 and Rhodotorula glutinis INMI Y-42 and bacterium of two species Azomonas macrocytogenes INMI B-28 and Xanthobacter sp. INMI B-29. Above indicated consortium is isolated from ginseng roots. Consortium is cultured at 20-25 C on medium containing, g/l: glucose 8-10; KH₂PO₄ 0.9; KH₂PO₄ 0.3; MgSO₄ 0.2; K₂SO₄ 0.1; asparagine 0.02; MnSO₄,Fe₂(SO₄)₃ and CuSO₄ - trace amounts. Preparation is isolated by adsorption with activated carbon and then by extraction with 70% ethanol from sorbent. EFFECT: improved method of preparing. 2 cl, 16 tbl

Description

 The invention relates to agriculture and for the products of microorganisms that stimulate the development and increase the yield of agricultural plants.

Known plant growth stimulants obtained from pure cultures of endophyte fungi (symbiont fungi) isolated from plant roots, such as, for example, sea buckthorn, coltsfoot, strawberries, etc. [1-4]
Known biostimulator of plant growth and development Symbiont-1, which is an alcoholic extract of natural biologically active compounds accumulated in the culture of the cylindrocarp fungus Cylindrocarpon radicicala Woll, isolated from micromycete biomass, which is obtained from ginseng roots [5]
The proposed biostimulator of plant growth and development Symbiont-Universal is an alcohol extract of natural biologically active substances that are produced by the microbial community, also isolated from ginseng roots, but consisting of five components belonging to the two kingdoms of microorganisms, eucaryotes and prokaryotes. The eucaryotic component is represented by three types of imperfect fungi: the mycelial organism Acremonium panaxeorum INMI F-160 and two yeast Candida muscorum INMI Y-41 and Rhodotorula glutinus INMI Y-42; prokaryotic component by two bacteria - Azomonas macrocytogenes INMI B-28 and Xantobakter sp. INMI V-29. A mycelial fungus dominates in the community, yeast and bacteria are complementary components. However, the role of these components is important, and all of them together form a stable microbial community that produces physiologically active substances called Symbiont-Universal. The component composition of this community is stable during prolonged cultivation. The ratio of components (fungus, bacteria and yeast) in a mature community to the time of extraction of physiologically active substances, which are the active principle of the Symbiont-Universal drug, is: (94-98.5) :( 4-1) :( 2-0.5). The ratio of different species among yeast and bacteria is ≈1: 1.

Isolated from the roots of ginseng natural mixed culture of microorganisms is cultivated at a temperature of 22-25 ^o C on an artificial nutrient medium of the following composition, g / l glucose 8-10, KH ₂ PO ₄ 0.9, K ₂ HPO ₄ 0.3, MgSO ₄ 0.2, K ₂ SO ₄ 0.1, asparagine 0.02, MnSO ₄ , Fe ₂ (SO ₄ ) ₃ , CuSO ₄ traces. pH of the medium is 6.5. Biologically active substances produced by the microbial community that make up the active principle of Symbiont-Universal are extracted from the medium in two stages: adsorption with sterile activated carbon and extraction with 70% ethanol from coal.

 The drug Symbiont-Universal, as well as the well-known drug Symbiont-1, is effective when applied to cereals and potatoes in negligible doses (when processing seed material 1 ml / t, vegetative plants 1 ml / ha). At the same time, Symbiont-Universal stimulates the development of cereals (buckwheat, millet), legumes (peas), vegetables (cucumber, pepper, eggplant, cabbage, onions, carrots, radishes), fruit (apple, cherry), berry (strawberries, currants, Sea-buckthorn, gooseberry), flower (aster) crops. Moreover, due to improved growth and development of plants, their resistance to diseases and pests is increased. Symbiont-Universal is easily included in the technology of protecting crops from harmful organisms and is promising for use in the practice of cultivating crops, both open and protected soil. The drug can be used in compositions with trace elements, for example, copper sulfate or other salts, as well as, if necessary, simultaneously with pesticides. With the application of Symbiont-Universal on grain crops, the yield increase of winter wheat (processing by vegetation) reached 17 kg / ha, spring wheat (processing of seeds) 4.5-5.4 kg / ha, spring barley (processing of seeds) 3.6 kg / ha .

 Description and identification of microorganisms that make up the microbial community that produces the drug Symbiont-Universal.

 Research Methods.

 Wednesday. Acremonium panaxeorum mycelium was isolated from the community and maintained on a standard agarized Chapek medium with 2% sucrose and wort agar. Preliminary, the medium was acidified with lactic acid to pH 4-5.

For the cultivation of yeast used acidified to pH 4-5 agarized wort. When identifying them, a standard synthetic medium of the following composition was used, g / l: (NH ₄ ) ₂ S) ₄ 3.5; K ₂ HPO ₄ 0.15; KH ₂ PO ₄ 0.85; MgSO ₄ 0.5; NaCl 0.1; CaCl ₂ 0.1. Distilled water.

Bacteria were grown on diluted with distilled water 1: 5 MPB (MPA) or mineral medium recommended for growing the community, with the addition of a nitrogen source and adjusting the pH to neutral [4]
Microscopy of cultures of microorganisms was carried out using a phase contrast microscope, MBS-10 and a transmission electron microscope JEM 100 C. In the latter case, cell preparations were contrasted with a 1% solution of uranyl acetate.

The identification of pure cultures of microorganisms was carried out in accordance with standard methods used in these procedures [6-8,10,11] and using appropriate determinants [6,9,12,13-18]
The quantitative ratio of microorganisms in the community was determined by the amount of biomass of each component. The biomass of the fungus, yeast and bacteria was determined by standard methods [11]
Description and identification of micromycetes. A mycelial fungus isolated from the microbial community that produces the physiologically active substances that make up the Symbiont-Universal drug should be assigned to the class Deuteromycetes (Fungy imperfecti), order Hyphomycetales (Moniliales), family Moniliaceae, genus Acremonium (Link ex Fr.), Acremonium panoxeorum [9,12,17,18]
Acremonium panoxeorum micromycete has the following characteristics: a 12-day-old culture is represented by white open colonies, the growth rate is moderate. The mycelium is plentiful, cotton-like, poorly removed from agar. With age, the reverse side of the colony on Chapek's medium becomes brown, mycelium septic with many strands. Conidiophores form on the aerial mycelium. Conidiophores are solitary, sometimes in whorls. Micromycete does not form any aggregates from the conidiophores such as koremia, sporodochia or pionnot. Conidia are formed at the apex of the conidiophores, solitary and / or in false heads. Conidia are colorless, usually cylindrical with rounded ends. Sizes of conidia: length up to 17 microns, thickness up to 5 microns. Some conidiospores sometimes have a septum. Along with large cylindrical conidia, there are small conidia, from egg-shaped to spherical. When growing micromycetes in liquid culture, the formation of chlamydospores is observed. The preferred growth temperature is -20 ^o C.

 Currently, the genus Acremonium includes more than 70 species. Species of this genus are spaced in three sections; Simplex, Gleomastix, and Necrioidee [13] Based on the totality of the identified characters, Acremonium sp. Corresponds to the Simplex section. In the Simplex section, 38 species are described [18] Based on the source of isolation of this micromycete, the established characters do not correspond to any of the known species, and, therefore, the organism must be described as a new species. Due to the fact that the source of isolation of Acremonium sp was ginseng roots, Acremonium panaxeorum sp. nov. Acremonium ginseng. The culture is stored in the culture collection of the Laboratory of Soil Microbiology of the Institute of Microbiology of the Russian Academy of Sciences under the number F-160. (Acremonium panaxeorum INMI F-160).

Description and identification of yeast. Yeast cultures isolated from the microbial community that produces physiologically active substances that make up the Symbiont-Universal drug were classified as imperfect aspartogenic yeast of the genera Candida, Candida muscorum and Rhodotorula, Rhodotorula glutinis [6,7,13,15,16] The studied yeast has the following symptoms :
Candida muscorum. The colonies are whitish, taking a creamy tint with age. The size of the colonies is from 1-2 to several mm. The colonies are round, the edge of the colony is uneven, the center is convex, shifted to one side. The surface of the colony is matte, with age it becomes covered by radial grooves. Culture forms pseudomycelia, differentiating into pseudohypses and blastospores. In a monthly culture, the stroke is a yellowish-cream color with a matte surface and a wavy edge, pasty consistency. Candida muscorum cells are oval and oblong in size 2.2-4.5 x 4.0-8.0 microns. Propagated by budding. Organic and inorganic (nitrate) forms of nitrogen are used. When testing the body’s ability to assimilate certain substances as the sole source of carbon and energy, the following properties were revealed: glucose (+), sucrose (+), maltose (-), cellobiosis (±), lactose (-), melibiosis (-), raffinose (+), inulin (-), soluble starch (-), L-arabinose (±), D-xylose (-), L-ramnose (-), ethanol (+), glycerin (+), mannitol (- ), methanol (-), lactate (-), citrate (-), succinate (-).

 Notation: “+” substance used; "-" the substance is not used; "±" detected weak growth.

The body is able to grow on a vitamin-free environment. There is no growth on a 5% glucose-yeast autolysate. It does not form starch-like compounds. Urea does not hydrolyze. The temperature range of growth of 10-40 ^o C, the temperature optimum of growth of 15-28 ^o C.

 According to the totality of the identified characters, the isolate corresponds to the species Candida mucrocorum (di Menna) [6,7,13,15,16] Strain differences are the inability of the identified strain to assimilate lactose, maltose, xylose and lactate. The culture is stored in the culture collection of the Laboratory of Soil Microbiology RAS under the number INMI Y-41 (Candida mucrorum INMI Y-41).

 Rhodotorula glutinis. The colonies are pink, ranging in size from 1-2 to 5 mm in diameter. The colonies are rounded, the edge is even, the center of the colony is raised, can be shifted to one side. The surface of the colony is matte, with age with well-defined annular grooves. Pseudomycelia does not form. The stroke is bright pink, smooth, shiny. Rhodotorula glutinis cells are oval and oblong in size 1.5-2.3 x 2.0-4.0 microns. Propagated by budding. Organic and inorganic forms of nitrogen are used. When testing the body’s ability to assimilate certain substances as the sole source of carbon and energy, the following properties were revealed: glucose (+), sucrose (+), maltose (-), cellobiose (+), lactose (-), melibiosis (-), raffinose (+), inulin (-), soluble starch (-), L-arabinose (±), D-xylose (±), L-ramnose (-), ethanol (+), glycerin (±), mannitol (+ ), methanol (-), lactate (±), citrate (-), succinate (+).

 Designations: "+" substance is used, "-" substance is not used; "±" detected weak growth.

The body is able to grow on a vitamin-free environment. There is no growth on a 5% glucose-yeast autolysate. The temperature range of growth of 10-40 ^o C, the temperature optimum of growth of 15-28 ^o C.

 According to the totality of the identified characters, the isolate corresponds to the species Rhodotorula glutinus (Fres.) Harrison [6,7,13,15,16] The culture is stored in the culture collection of the Laboratory of Soil Microbiology RAS under the number INMI Y-42 (Rhodotorula glutinis INMI Y-42).

 Description and identification of bacteria. Bacterial cultures isolated from the microbial community producing the biologically active substances that make up the Symbiont Universal drug were assigned to the genera Azomonas, Azomonas macrocytogenes Xantobacter, Xantobacter sp. [14] Bacteria have the following symptoms.

 Azomonas macrocytogenes after three days of growth on MPA is represented by small colonies of 1-2 mm in diameter. The colonies are round, convex, mucous, shiny. The edge of the colonies is even, the color is transparently whitish. The cells of the three-day culture are mainly motile, short and thick sticks. The movement of the cells is somersaulting. During electron microscopic studies of cells, it was found that flagellation corresponds to a subpolar monotrich; the culture is rather pleomorphic. There are pear-like cells, cells with lateral outgrowths, etc. Cell sizes: length 1.5-1.8, thickness 0.57-0.85 μm.

The ability to form any resting forms has not been identified. Gram-negative cells. They are strict aerobes. Oxidase and catalase positive. Temperature limits of growth: 11-36 ^o C with optimum growth at 20-28 ^o C. The culture retains well viability on MPA. The culture is capable of nitrogen fixation (by acetylene reductase reaction).

 When a pure culture with glucose is grown on the medium recommended for culturing the microbial community [4], the bacterium forms exopolysaccharide in significant quantities. When checking the ability of bacteria to assimilate certain substances as the sole source of carbon and energy, the following properties were revealed: glucose (+), mannitil (+), molonate (-), malate (+), glutamate (+), galactose (+), sorbitol (+).

 Legend: "+" substance is used, "-" substance is not used.

 According to the ninth edition of Bergi's determinant of bacteria, the genus Azomonas (family Azotobacteraceac) is represented by three species: A.agilis, A.insignis, and A.macrocytogenes [14] For the first two species, the main natural habitat is water. A.macrocytogenes is a typical soil inhabitant.

 According to the totality of the identified characteristics, the strain Azomonas sp. can be classified as Azomonas macrocytogenes (ex. Baillie, Hodgkiss and Norris) [14] Differences from the type species are that the identified strain is able to assimilate glutamate and nitrate nitrogen.

 The culture is stored in the culture collection of the Laboratory of Soil Microbiology of the Russian Academy of Sciences under the number INMI V-28 (Azomonas macrocytogenes INMI V-28).

 Xantobacter sp. after three days of growth, MPA is represented by colonies up to 2 mm in diameter. The colonies are round, convex, strongly mucous (viscous), shiny. The edge of the colonies is even, the color in the young culture is pale yellow, bright yellow with age and with abundant growth. Ambient agar does not pigment.

The cells of the three-day culture Xantobacter sp. represented by chopsticks, from moderate to very long. The latter, apparently, are chains of unseparated cells. The most characteristic cell sizes: 1.7-3.7 x 0.5 microns. Solid-grown cells are immobile. However, in some cases, microscopy of the culture from liquid media showed the ability to move. Electron microscopic studies of the cells revealed flagella, and their attachment corresponds to the monotrichal type. Many cells contain slightly fluorescent inclusions resembling the glow of polyoxybutyrate granules. The ability to form any resting forms has not been identified. Gram-negative cells. The bacterium is a strict aerobic, oxidase-negative, catalase-positive. The temperature limits of growth are 11-37 ^o C. The optimal temperature range of growth is 22-28 ^o C. At extreme temperatures, growth is very weak. The culture is capable of nitrogen fixation (by acetylene reductase reaction). The growth was studied on: glucose (+), propionate (±), molonate (-), asparagine (±), methanol (±), cellobiose (+), arabinose (+), ramnose (+) and galactose (+).

 Notation: “+” substance used; "-" the substance is not used; "±" reaction is mild.

 Of the characteristics of the organism, it should be noted its "capriciousness", in particular, a fairly rapid loss of life during storage on MPA at room temperature. However, at a low temperature (in the refrigerator), it can remain for a long time.

 The results of the studies show that the bacterium has many features characteristic of representatives of the genus Xantobacter. A key similarity with representatives of the genus Xantobacter is the ability of bacteria to nitrogen fixation, as well as the ability to form reserve substances in the cell, cell size, the spectrum of key consumed substrates, etc. The strain is prototrophen and does not need biotin.

 In the genus Xantobacter, two species are currently legalized: X. autotrophicus and X. flavus [14] An isolated and described organism does not correspond to any of these species and, apparently, claims to be a new species. However, due to the fact that at present the characteristics of the genus are being refined and it is actively replenished with new species (Reding HK Croes GLM Dijkhuizen L. Wiegel J. Emendant of Xantobacter flavus as a monile species. Personal message.), It is advisable to keep the described organism name Xantobacter sp.

 The culture is stored in the culture collection of the Laboratory of Soil Microbiology of the Russian Academy of Sciences under the number NIMI V-29 (Xantobacter sp. INMI V-29).

 Determination of the ratio of microorganisms in the community. Strict quantification of the various microorganisms that make up the community that produces the physiologically active substances of the Symbiont-Universal drug is difficult, because it contains both unicellular organisms, bacteria and yeast, and multicellular morphologically heterogeneous mycelial fungus. In this regard, the ratio of microorganisms was determined by an indirect criterion of the area of the projective coverage of the visual field under a microscope [11]. The ratio of fungus, bacteria and yeast in a mature community (by the time of extraction of physiologically active substances that are part of the Symbiont-Universal drug is: (94- 98.5) :( 4-1) :( 2-0.5). The ratio of different species among yeast is accepted ≈1: 1.

Example 1. Obtaining a biostimulator Symbiont-Universal. 1 g of ginseng roots are washed from the soil and sterilized for 2-3 minutes in alcohol, washed repeatedly with sterile water, ground in a sterile agate mortar, transferred to flasks with a sterile nutrient medium of the following composition, g / l: glucose 8-10; K ₂ HPO ₄ -0.3; KH ₂ PO ₄ - 0.9; MgSO ₄ 0.2; K ₂ SO ₄ - 0.1; asparagine 0.01-0.02, FeSO ₄ - 7 drops of a 2% solution / l; CuSO ₄ - 7 drops of a 2% solution / l; gibberellin (standard alcohol solution) -1 ml, MnSO ₄ - traces and leave to germinate for 5-12 days. The grown microbial community is subcultured on a nutrient medium of the same composition, but without the addition of gibberellin, and cultured for 30 days at a temperature of 22-25 ^o C. At the end, sterile activated carbon is added to the flasks at the rate of 0.5% by weight of the liquid. Stand 2-4 hours and filter the precipitate on a sterile filter. The filter residue is placed in a sterile flask, 100 ml of 70% ethanol are added, the mixture is kept for 8-10 hours and filtered. The filtrate is poured into sterile containers. From 300 ml of culture, 90 ml of an alcoholic solution of a biostimulant is obtained. An integral indicator of the activity of a biostimulator is its effect on the growth and development of test plants (wheat, watercress).

 Example 2. The effect of different concentrations of the drug Symbiont-Universal on the germination of seeds, the growth of roots and stems of wheat.

35 seeds of wheat. laid out in a Petri dish on sterile filter paper and moistened with a drug in a dilution of 10 ^-1 , 10 ^-3 , 10 ^-4 . Reference seeds treated with the culture medium used to grow the community in the same dilutions. Control the seeds treated with water defended for a day. Petri dishes are closed with lids and incubated in a thermostat for 5 days at 25 ^o C. After this time, determine the percentage of seed germination, the length of the roots and seedlings. The results are shown in table 1.

 Given in the table. 1 results show a steady positive effect of low concentrations of Symbiont-Universal on the growth of roots and stems of wheat.

 Example 3. The stimulating effect of Symbiont-Universal on the growth and development of buckwheat.

 The experiment was carried out in a production environment. The experimental design included two options: 1 seed treatment by Symbiont-Universal at the rate of 1 ml / t on a PS-10 machine (26.6.94 g); 2 control without processing. During the growing season, twice in the budding phase (11.07.94 g.) And in the flowering phase (1.08.94 g.), The length of the plant was measured and the mass of the aerial part of buckwheat was determined. It was established that the use of Symbiont-Universal had a stimulating effect on the development of buckwheat plants. The results of the experiment are given in table.2.

 To analyze the structure of the crop, five test sheaves of 100 plants each were selected from each test variant. These crop patterns showed that when processing seeds with Symbiont-Universal there is an increase in plant productivity. The results are shown in table.3.

 Accounting for economic efficiency showed that buckwheat yield in the variant with presowing cultivation by Symbiont-Universal is 1.2-2.2 kg / ha higher compared to the untreated control.

 Example 4. The action of Symbiont-Universal on the development and yield of millet.

The experiment was carried out in the field on plots of 13.5 m ² .

 Five repetition. Seeds were treated with Symbiont-Universal 3 hours before sowing in doses of 1.0; 1.5; 2 ml / t. The seeding rate is 5.5 million germinating seeds per 1 ha. Standard seeds treated with water. Control untreated seeds. The results are presented in table. 4.

 The results of the experiment indicate that under the influence of Symbiont-Universal, growth processes are intensified, leading to an increase in the yield of millet.

 Example 5. Tests of Symbiont-Universal on peas.

 The seeds of food peas of the Orlovchanin variety were treated with Symbiont-Universal at the rate of 1 ml per 1 ton of seeds. The area of accounting plots is 0.5 ha. The duplication is twofold. Control untreated seeds. During the growing season, the height of the plants was measured twice, the mass of the ground part of the peas was determined, the number of nitrogen-fixing nodules was calculated, and the damage of the peas by nodule weevils was taken into account. The results are presented in table.5.

 Thus, the use of Symbiont-Universal had a stimulating effect on the growth and development of peas, and also increased the tolerance of peas to nodule weevils.

 Example 6. Tests of Symbiont-Universal on a set-onion.

 The field experiment on the onion set of the Myachkovsky variety was laid against a natural background. The experimental design included two options: 1 soaking the bulbs in Symbionte Universal (5-7 drops per 0.5 l of water) for 30 minutes, 2 - soaking the bulbs in Symbionte Universal (5-7 drops per 0.5 l of water) for 30 min + spraying of vegetative plants at a rate of 1 ml / ha. In the control variant, the bulbs were soaked in water. The results of the experiment are presented in tables 6 and 7.

 The results of the experiment indicate that the use of Symbiont-Universal contributes to a better development of plants, reduction of damage by onion moth, affection of peronosprosis and increase yield.

 Example 7. Tests of Symbiont-Universal on commodity crops of cucumber.

 Field experience was laid on cucumbers of the Electron variety. The experimental design included two options: 1 soaking seeds in Symbionte Universal (1 ml per 1 ton of seeds) for 30 minutes, 2 soaking seeds in Symbionte Universal (1 ml per 1 ton of seeds) for 30 minutes + spraying vegetative plants at the rate of consumption of the drug is 1 ml / ha in the phase 1-2 and 4-5 of these leaves. The flow rate of the working fluid is 100 l / ha. In the control embodiment, the seeds were soaked in water. The results of the experiment are presented in table.8.

 Example 8. Tests of Symbiont-Universal on cabbage.

 The experiment was laid on Cabbage Seeds of the Transfer variety in open ground conditions. The technology of growing cabbage is generally accepted in the conditions of the Moscow region. At the first stage, the seeds were sown in a heated greenhouse. The experiment scheme in the greenhouse included three options: 1 soaking seeds in Symbiont-Universal based on 1 ml / t of seeds (5-7 drops per 0.5 l of water) for 30 minutes; 2 - seed soaking in Symbionte-Universal at the rate of 1 ml / t of seeds (5-7 drops per 0.5 l of water) for 30 min + spraying of vegetative plants at a rate of 1 ml / ha and a working fluid flow of 100 l / ha; 3 - spraying of vegetative plants at a rate of 1 ml / ha and a flow rate of 100 l / ha. Control without processing. The results of the experiment are presented in tables 9 and 10.

 From table 9 it is seen that the treatment with Symbiont-Universal slightly affected the seedlings yield, however, the quality of seedlings was much better, and the affection with the “black leg” decreased by more than 2 times.

 The second stage of planting seedlings in the field. Three weeks after transplanting in option 3, additional spraying of plants was carried out at a flow rate of 1 ml / ha and a flow rate of 500 l / ha. In all variants with the use of Symbiont-Universal, a decrease in damage to cabbage by leaf-eating pests was noted, as well as earlier (by 7 days) ripening of the crop.

 Example 9. Tests of Symbiont-Universal for carrots.

 The experiment was laid in the field on carrots of the Nantes-4 variety. Sowing carrots was carried out on ridges in three rows. The plot length is 4 m. The experimental design included three options: 1 soaking seeds in Symbiont-Universal based on 1 ml / t of seeds (5-7 drops per 0.5 l of water) for 30 minutes; 2 seed soaking in Symbionte-Universal at the rate of 1 ml / t of seeds (5-7 drops per 0.5 l of water) for 30 min + spraying of vegetating plants at a rate of 1 ml / ha and a working fluid flow of 100 l / ha; 3 spraying of vegetating plants at a rate of 1 ml / ha and a flow rate of 100 l / ha. Control without processing. The results of the experiment are presented in table.11.

 The results of the experiment showed that in all variants with the use of Symbiont-Universal, the yield is higher than in the control.

 Example 10. Tests of Symbiont-Universal on the fruiting currant.

 The tests were carried out on the fruiting blackcurrant of the Vologda variety on a plantation in 1987. Currant bushes were sprayed with 0.001% solution of Symbiont-Universal with the addition of 0.0002% copper sulfate. Processing was carried out twice: at the beginning of bud blooming and during flowering. Kotrol - without processing. The flow rate of the working fluid is 0.125 liters per bush. The repetition of the experiment is fourfold. In each repetition, 10 bushes were processed. The yield of berries from each treated bush averages 1.72 kg, in the control 1.17 kg.

 Example 11. Tests of Symbiont-Universal on green buckthorn cuttings.

 100 green cuttings from uterine bushes of the Garden Gift variety were selected for the experiment. The treatment was carried out on June 28, 1994. The cuttings were immersed for 30 min in a 0.01% solution of Symbiont-Universal with the addition of 0.002% copper sulphate and immediately planted in the soil of a frame film greenhouse with a fogging plant. Planted cuttings were watered with a 0.001% solution of Symbiont-Universal with the addition of 0.0002% vitriol at the rate of 1 liter per plant. Control plants were soaked in water. In the second decade of September, eight plants were evaluated in five replicates in each case, the rooting and growth of plants. The results are presented in table.12.

 Example 12. Tests of Symbiont-Universal on green cherry cuttings.

 The processing and cultivation of the cuttings of the cherry variety "Vladimirskaya" was carried out in the same way as in the experiment with sea buckthorn. The experiment was repeated four times with 10 plants each. The control was plants soaked for 12 h in a solution of indolylbutyric acid (IMA) 50 mg / L. The Symbiont-Universal treatment significantly affected the number of roots formed (3.5 times more than with standard processing), the maximum root length and the yield of marketable seedlings. The results are shown in table.13.

 Example 13. Tests of Symbiont-Universal on green gooseberry cuttings.

The experiment was carried out in two versions on cuttings of gooseberry cultivar "Sadko". In the first embodiment, the cuttings were treated with a solution of Symbiont-Universal under the same conditions as when testing the drug on sea buckthorn. Four repetitions, 15 cuttings each. In the second embodiment, untreated cuttings were planted in a substrate saturated with 0.01% solution of Symbiont-Universal + 0.002% vitriol at the rate of 10 l of solution per 1 m ^{2 of} substrate. In the control embodiment, the substrate was watered. Evaluation of the results was carried out in the second decade of September. The results are presented in table.14.

 From the table. 14 shows that the influence of Symbiont-Universal on the survival rate of gooseberry cuttings depends on the method of its application. When the soil is impregnated with the preparation, this indicator significantly increases by 1.5 times.

 Example 14. Tests of Symbiont-Universal on eggplant sheltered soil.

 The experiment was laid in a film unheated greenhouse on eggplant varieties Universal-6. The experimental design included two options; 1 soaking seeds in a 0.01% solution of Symbiont-Universal with the addition of copper sulfate (0.2 g per 10 l of water) for 1 h; 2 soaking seeds in Symbiont-Universal under the same conditions + spraying vegetative plants with a 0.001% solution of Symbiont-Universal with the addition of copper sulfate (0.2 g per 100 l of water) at a rate of 1 ml / ha of the preparation in the flowering phase. The flow rate of the working fluid is 100 l / ha. In the control embodiment, the seeds were soaked in water. Prepared according to the experimental scheme, eggplant seeds were sown in peat pots. Seedlings were planted in a film greenhouse according to the scheme: (80 + 60) / 20 x 30 cm.

 The results of the experiment are given in table.15.

 From table 15 it is seen that in the variants with the use of Symbiont-Universal, the yield is higher than in the control.

 Example 15. Tests of Symbiont-Universal on flower crops.

 The experiment was laid in open ground on asters of the Yubileynaya Belaya variety. The experimental design included two options: 1 seeds were soaked in Symbiont-Universal in a 0.01% solution (5-7 drops per 0.5 L of water) for 30 min, dried to a granular state and sown in boxes placed at the light installation (April 15, 1994 ) In a phase of 3-4 leaves, seedlings were planted in open ground and healthy plants and the number of plants affected by the "black leg" were counted; 2 soaking seeds in a 0.001% solution of Symbionta-Universal (5-7 drops per 0.5 l of water) for 30 minutes + spraying vegetative plants with a 0.01% solution at a flow rate of 500 l / ha. Control without processing. The results of the experiment showed that soaking the seeds in a 0.01% solution of the drug increases the germination energy and germination by 10-12% compared to the control. The yield of healthy seedlings was higher than in the control, and the percentage of affected plants with a “black leg” was 2.5 times less. The results are presented in table.16.

 Assessment of the general condition of plants according to the experimental variants was carried out according to a five-point system during budding and flowering. Good condition was rated higher. The best condition of the plants was “4.65 points” in the 2nd variant, in the control “4.5 points”. At the same time, the inflorescences on the main stem blossomed a whole week earlier.

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Claims (2)

 1. Biostimulator of plant growth based on microorganisms, characterized in that it contains an alcoholic extract of the biomass of a community of microorganisms, consisting of micromycete Acremonium panaxeorum INMI F-160, two types of yeast Candida muscorum INMI U-41 and Rhodotorula glutinis INMI U-42 and two species bacteria Azomonas macrocytogenes INMI B-28 and Xantobacter sp. INMI V-29.  2. A community of microorganisms consisting of the micromycete Acremonium panaxeorum INMI F-160, two types of yeast Candida muscorum INMI U-41 and Rhodotorula glutinis INMI U-42 and two types of bacteria Azomonas macrocytogenes INMI B-28 and Xantobactep sp. INMI B-29 to obtain a biostimulator of plant growth.

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