WO2009072921A1

WO2009072921A1 - Method for producing combined microbiological fertilizer

Info

Publication number: WO2009072921A1
Application number: PCT/RU2007/000682
Authority: WO
Inventors: Vladimir Kuzmich Chebotar; Alexandru Efimovich Kazakov; Sergey Viktorovich Erofeev; Tatiana Nikolaevna Danilova; Tatiana Sergeevna Naumkina; Oxana Yurievna Shtark; Igor Anatolievich Tikhonovich; Alexey Yurievich Borisovich
Original assignee: Obshestvo S Ogranichennoy Otvetstvennoctyu 'bisol Bi-Inter'
Priority date: 2007-12-04
Filing date: 2007-12-04
Publication date: 2009-06-11
Also published as: DE112007003728T5; RS20100244A; RS54161B1

Abstract

The invention relates to agriculture, in particular to the processing of sugar production waste. The inventive method for producing combined microbiological fertilizer involves combining a microbiological component with a natural biocompatible carrier, wherein the inoculation material of arbuscular mycorrhizal fungi is used as a part of the microbiological component, a defecated juice in the form of a filtration washing precipitate of the sugar beet processing is used as a natural biocompatible carrier and the microbiological component is combined with the natural biocompatible carrier in the rhizosphere of mycorrhizal plants cultivated on the defecated juice. Afterwards, the remaining parts of the microbiological component, in particular rhizobium and rhizobacteria pre-cultivated on liquid nutritional media, are added to the thus produced substrate.

Description

METHOD FOR PRODUCING INTEGRATED MICROBIOLOGICAL FERTILIZER

Technical field

The invention relates to agriculture and the processing of food production waste, in particular, to obtain complex microbiological fertilizers to improve the nutrition of crops, protect plants from phytopathogenic microorganisms, reduce losses of agricultural products during storage, and also process sugar production wastes, and relates to a method for producing complex microbiological fertilizer.

State of the art

Currently, there are many microbiological preparations for agriculture for various purposes: growth-promoting, as well as inhibiting the development of phytopathogenic bacteria and fungi.

The known "Method of seed processing" described in the application N ° 2170987 of Great Britain with a priority of 02/14/85. MKI 4 A 01 C 1/06, published in ISM Ns 10 for 1987. Said method comprises treating the seeds with a composition consisting of microorganisms, a carrier, for example bran, and glue, such as gatti resin.

The most favorable results are obtained when processing wheat seeds. Microorganisms can stimulate their growth (Azospirilmm), bind nitrogen (Rhizobium), prevent plant diseases (Pseudomopas or Vasillus), or kill insects (Strotherms).

SUBSTITUTE SHEET (RULE 26) Also known is the "Method of protecting cabbage from bacterial diseases" according to the patent Na 1793878 with a priority of 10.12.90., Published 07.02.93g. in the Bulletin of inventions of the Russian Federation Ne 5.

According to the mentioned method, to protect cabbage from bacteriosis, cabbage seeds are treated with a suspension of the bacterial strain Pseudomonas sp. VKPM B-3481 at a concentration of 10 ^b - 10 ⁹ cells / ml at the rate of 20 ml per 1 kg of seeds after hydrothermal treatment. The use of the bacterial suspension "Pseudomonas sp. VKPM B-3481" reduces the development of vascular and mucous bacterioses and increases the yield of cabbage.

Also known is the "Method of obtaining a substrate for growing plants" according to the patent of the Russian Federation N ° 1829892 with a priority of 20.04.90. authors Vasyuchenko I.K., Perebyuk A.H., Pigulevsky V. S, Berezko M.N. and Puchko V.N.

The invention relates to methods for producing substrates for growing seedlings and vegetables using biomass of bacteria of the genus Pseudo-Monas - suppressors of phytopathogens.

According to this method, a substrate is obtained by applying calcareous, mineral, and bacterial fertilizers to peat, followed by mixing of the components, and the biomass of the bacterial strain Pseudomopas rutida fluoresceps CMPM B-3481 is used as bacterial fertilizer.

Also known is the "Method of obtaining bio-fertilizers" according to the patent of the Russian Federation Ns 2130005 with a priority of 04/06/1996. authors Raymanova I.T., Alimova F.K., Ozhiganova G.U., Khabibullina P.E., Krylova H.I., Fattakhova A.N. The method consists in the fact that a strain or community of microorganisms is sterile cultured in a nutrient medium until a bacterial mass titer of 10 ⁸ -10 ⁹ cells / ml is reached. The resulting biomass is separated from the medium, concentrated. Concentrated biomass is applied to the dried granulated chicken droppings. The biomass thus immobilized is dried. This method provides increased survival of microorganisms and increased biological activity of the soil.

SUBSTITUTE SHEET (RULE 26) A known group of inventions "Method for presowing treatment of seeds of vegetable crops and a method for producing a preparation for presowing treatment of seeds of vegetable crops)) according to the patent of the Russian Federation JVs 2140138 with a priority of 11/13/98, authors Chebotar V. K., Bykova HB, Dark OB, Orlova N .BUT. and Khotanovich VV These methods are carried out using a biofungicidal preparation containing the bacterial strain Vasillis sibtilis 4-13 (deposited under the registration number VNIISM D-606 in the group of epiphytic microorganisms). A method of obtaining a biofungicidal preparation consists in mixing a culture fluid containing a strain of B.s. 4-13, which is preliminarily cultivated in a sterile liquid nutrient medium, with a PVA emulsion, an aqueous diluent and sterile chalk or dolomite in certain proportions. EFFECT: inventions make it possible to increase the efficiency of protecting vegetable crops from phytopathogenic fungi by pre-sowing seed treatment.

Known invention "Method for producing biofertilizers)) according to the patent of the Russian Federation

JVs 2241692 with a priority of 10/11/2002 authors Chebotar B.K., Kozakova A.E., Erofeeva CB. This method consists in combining granular mineral fertilizers with microbial biomass in the form of a bacterial preparation based on the Bacillus sibtilis 4-13 strain, which has antagonistic properties to phytopathogens and growth-promoting effect. The combination is made by applying a bacterial preparation in the form of a dry powder (dusting) on the granule surface of the granules of mineral fertilizers, either in the form of a finely divided liquid fraction (spraying), or in the form of a liquid fraction in the composition of moisture-proof mineral fertilizer compositions. The specified method provides an increase in the efficiency of plant protection against infectious diseases that penetrate along with mineral fertilizers.

It is also known "Bacterial fertilizer" HIKA "and a method for producing bacterial fertilizers)) according to the application JVs 99100664 with a priority of 01/21/99. authors Vinogradova E.Ya., Vinogradova A.E. (prototype).

SUBSTITUTE SHEET (RULE 26) Granular bacterial fertilizer contains a culture of the bacterial strain Vasillus mucilagiposus (VKMB 1451D), a sorbent and the remains of the culture fluid with metabolites, with a certain ratio of ingredients. Moreover, as a sorbent, this fertilizer may contain ground vegetable mass, peat, and vermiculite. In a particular case, the cultivation is carried out using an aqueous extract of bird droppings with the addition of molasses as a nutrient substrate.

A method for producing bacterial fertilizers based on Vasillus mucilagiposus includes cultivation of this strain on liquid nutrient media - hydrolysates of industrial and agricultural wastes, after which the resulting product is mixed with a sorbent and granulated.

Microbiological preparations have been known for over a hundred years, but often their effectiveness turned out to be insufficient or unstable, which is why they could not play a significant role in increasing the productivity of agricultural production.

Recently, the phenomenon of integration of the genetic systems of microorganisms and plants in the process of their interaction has been discovered. It was possible to show that both bacteria and plants possess sets of symbiotic genes that are silent in the absence of an appropriate partner, and in the presence of partners with certain genotypes, symbiosis is established. At the same time, the process of establishing symbiotic relationships is an environmental process that occurs in the soil. With the help of microorganisms, the plant provides its needs for nutrients (nitrogen, phosphorus and others), microorganisms are able to protect the plant from phytopathogens, and the most dangerous soil infections, for which there are no effective means to combat them. Such a system controls the production of supraorganism signs or adaptations that neither bacteria nor plants possessed before interaction. This aspect of microbial-plant interaction is now being actively studied.

SUBSTITUTE SHEET (RULE 26) Disclosure of the invention and options for its implementation

The objective of the invention is to increase the efficiency of protection of agricultural plants (crops) from phytopathogenic microorganisms and increase their productivity.

This problem is solved due to the fact that in the method of producing microbiological fertilizer, which consists in combining the microbiological component with a natural biocompatible carrier, inoculation material of arbuscular mycorrhiza fungi (AM fungi) is used as one of the parts of the microbiological component, and defecate is used as a natural biocompatible carrier, which is a filter-washing precipitate of sugar beet production, and their combination occurs in the rhizosphere of mycorrhizal plants growing these plants defe- kata, after which the resulting substrate is added remaining part of the microbiological component, namely the pre-cultured in liquid media nodule bacteria and rhizobacteria.

It is known that arbuscular mycorrhiza (AM) is a mutually beneficial plant-fungal symbiotic system (Labutova L.A., Provorov HA, Tikhodeev OH, Tikhonovich I.A. et al. Genetics of plant development. St. Petersburg, Science 2000, p. . 344-384) and that AM fungi can develop only in symbiosis with the plant. The genetic factors of plants that control the interaction are universal with respect to symbiotic fungi and bacteria and are present in most cultivated (mycorrhizal) plants. Symbiotic genes or some of them provide the possibility of entering into mycorrhizal symbiosis that optimizes mineral, including phosphorus plant nutrition. The ability to form mycorrhiza on the roots has 80-90% of all plants.

At the same time, it is also known from literary sources that sorghum plants, sorghum-Sudan hybrid, Sudan grass and millet are the most effective in obtaining inoculation material of arbuscular mycorrhizal fungi from fungi.

SUBSTITUTE SHEET (RULE 26) example, Labutova L.A., Provorov HA, Tikhodeev OH, Tikhonovich I.A. et al. Genetics of plant development. SPb. Science, 2000, pp. 344-384). Therefore, to obtain the inoculation material, one of the above plants is used (sorghum, sorghum-Sudan hybrid, Sudan grass, or millet). In order to create a symbiosis for the formation of AM, when planting seeds of mycorrhizal plants in the substrate (soil) on which they are grown, a pure culture or a mixture of cultures of AM fungi is introduced in the form of a soil-root mixture from under mycorrhized plants (I. Karatygin Co-evolution of fungi and plants // Transactions of the Botanical Institute of the Russian Academy of Sciences, 1993, Vyp. 9, pp. 1-118).

Moreover, it was experimentally established that good results for the formation of AM are achieved when defecate, a filter-washing cake of sugar production from sugar beet, is used as a substrate (soil) for growing mycorrhizal plants. Defecate - defecation mud, beet sugar production waste containing lime is formed in the process of cleaning beet juice. The yield of defecation mud is 8-12% of the mass of processed beets. In fresh defecate, about 40% of water. Dried up to a loose state (humidity 25-30%), the defect contains: 60-75% carbonate lime (mixed with caustic), 10-15% organic matter, 0.2-0.7% nitrogen, 0.2-0 , 9% phosphorus (P ₂ O ₅ ), 0.5-1% potassium (K ₂ O), a certain amount of magnesium, sulfur and trace elements. Defecate is a good calcareous fertilizer that is used for liming sod-podzolic and gray forest soils, podzolized and leached chernozems, mainly in beet-growing areas (Great Soviet Encyclopedia).

That is, the defecate, which is a waste of sugar beet production, is at the same time a good soil reclamant, improves its aggregation, structure (porosity, friability), stabilizes its acidity (PH), increases the exchange capacity of soil moisture absorption (moisture capacity). In addition, the use of defecate in the method of producing microbiological fertilizer allows the disposal of sugar production wastes.

SUBSTITUTE SHEET (RULE 26) In this case, the inoculation material of arbuscular mycorrhiza fungi is obtained by growing mycorrhizal agricultural plants, for example, sorghum, Sudan grass, sorghum-sudan hybrid, millet, using defecate as a filter and washing cake for sugar production from sugar beets. To do this, the seeds of sorghum or other mycorrhizal plants are planted in vessels, boxes or other containers with a defect; however, a pure culture or a mixture of cultures of AM fungi, for example, “Glomus intraradices”, i.e. the soil-root mixture from under mycorrhizal plants at a level of occurrence of mycorrhized roots (usually along the length) of at least 60% based on 3-5 g of such soil-root mixture per seed.

It has been experimentally established that the use of a soil-root mixture with a level of occurrence of mycorrhized roots less than 60% does not lead to a sufficient amount of AM in the inoculation material. It was also established empirically that the optimal ratio of the amount of soil-root mixture per seed (plant) is 3-5 g, because a smaller amount leads to a decrease in the effect, and a larger one does not increase it.

Planted plants (sorghum or others) are grown during the growing season 90-120 days. During this time, plants form a well-developed mycorrhiza. Typically, the percentage of mycorrhization of plants (eg, sorghum) at the end of this period is 90-100%.

Then the aerial part of the plants is cut and removed. The remaining roots together with the substrate (defecate) are dried at t ° 30-50 ⁰ C to a moisture content of 10-12% and ground to obtain a fraction of 0.05-0.2 mm.

From the obtained powder of inoculation material of AM fungi, a drazhirovanny mixture is prepared for subsequent treatment of seeds of mycorrhizal plants with it, mixing inoculation material with an adhesive in the ratio: 80 - 90% - inoculation material of AM fungi; the rest is adhesive.

SUBSTITUTE SHEET (RULE 26) It was experimentally established that the optimal result is obtained with the above ratio of ingredients.

Adhesive (adhesive) is used to attach the inoculum material of AM fungi to plant seeds. As an adhesive, any glue non-toxic to microorganisms and treated seeds can be used, for example, gati resin.

Then the seeds of sorghum (or other mycorrhizal plants) are treated with the resulting dragee mixture at the rate of 10-15r mixture per 100 g of seeds, dried at a temperature of 30-50 ⁰ C to a moisture content of 10-12%.

The seeds treated in this way are planted in a defecate, which is a filter-washing precipitate of sugar production from sugar beets, in special containers, greenhouses, or in places for storing defecate after filtration (for example, filtration fields) located next to sugar refineries, and grown using the recommended cultivation technology of the appropriate crop.

At the end of the growing season, after 90-120 days, the aerial part of sorghum plants (or other mycorrhizal plants) is cut (mowed) and removed. The upper 20 cm of defecate soil together with the roots of the plants is cut off and nodule bacteria previously grown on liquid nutrient media are added to it in the form of an aqueous suspension with a titer of 10 -10 CFU / ml in an amount of (0.5-5)%, and rhizobacteria in the form of an aqueous suspension with a titer of 10 ⁶ -10 ⁸ CFU / ml in an amount of (0.5-5)% of the weight of the soil.

Nodule bacteria are well known (for example: 1). Genetics of symbiotic nitrogen fixation with the basics of selection. Ed. I.A. Tikhonovich, N.A.

Provorova. St. Petersburg, Science, 1998; 2). Novikova N.I. Modern concepts of phylogeny and taxonomy of nodule bacteria. Microbiology, 1996, T 65,

SUBSTITUTE SHEET (RULE 26) Nodule bacteria contribute to the fixation of molecular nitrogen from the atmosphere and transfer it to the host plant, which increases the nitrogen nutrition of the plant and leads to increased yield. In addition, part of the nitrogen remains in the roots and in the soil, improving its agro-technical condition. It was experimentally established that the optimal result is obtained using an aqueous suspension of nodule bacteria with a titer of 10 ⁷ -10 CFU / ml in an amount of (0.5-5)%, because their lower concentration (titer) and amount (%) leads to a decrease in the effect, and a larger one does not increase it.

Nodule bacteria are pre-grown for 48-72 hours at a temperature of t ⁰ 28 ⁰ C and pH = 6.8 in a liquid mannitol-yeast medium of the following composition (g / l): 0.5 - K ₂ HPO ₄ ; 0.2 - MgSO ₄ x7H ₂ O; 0.1 - NaCl; 10.0 - mannitol, 0.5 - yeast extract; 15.0 - agar-agar, to obtain a titer of 10 -10 CFU / ml.

Associative rhizobacteria are also well known (e.g. Kameneva

SV., Muronets E.M. Genetic control of the processes of interaction of bacteria with plants in associations. Genetics, 1999, T.35, N ° l l, pp. 1480-1494).

In the case of associative symbiosis, no new visible structures are created on the roots of the plants, however, rhizospheric bacteria colonies are formed on the surface of the roots in strictly defined places, which can provide a number of useful functions for the plant, including, in particular:

- associative nitrogen fixation in the amount of up to 50 kg of nitrogen per hectare per year;

- the production of plant hormones that can accelerate the growth of the root system and thereby ensure the success of the plant in capturing the required nutritional area, as well as regulate plant development;

- optimization of the assimilation of phosphorus hard-to-reach compounds;

- induction of a systemic reaction to protect against phytopathogens;

- restriction (biocontrol) of the growth of phytopathogens on plant roots using mechanisms such as the isolation of antibiotic compounds, the dissolution of hyphae of pathogenic fungi, competition for places of population on the roots, interception of nutrients necessary for the development of a phytopathogen, etc.

SUBSTITUTE SHEET (RULE 26) It was experimentally established that the optimal result is obtained when using an aqueous suspension of rhizobacteria with a titer of 10 ⁶ -10 ⁸ CFU / ml in an amount of (0.5-5)%, because their lower concentration (titer) and amount (%) leads to a decrease in the effect, and a larger one does not increase it.

Rhizobacteria are pre-grown for 48-72 hours at a temperature of t ° 28 ⁰ C and pH = 6.8 in TSB liquid standard medium (tryptone soy broth), manufactured by SIGMA, to obtain a titer of 10 ^b -10 ⁸ CFU / ml.

The obtained defecate substrate with mycorrhizal plant roots mixed with a suspension of nodule bacteria and a suspension of rhizobacteria is used to prepare complex microbiological fertilizer (CMU) in powder form or in granular form.

To obtain microbiological fertilizer (CMU) in powder form, a mixture of a defecate substrate with plant roots mixed with suspensions of nodule bacteria and rhizobacteria obtained by the above method is dried at a temperature of t ° ⁽ 30-50) ° C to a moisture content of 10-12%, grind to a fraction of 0.05-0.2, and then pack. In this case, the inoculation material treated with a seed pelleting mixture, as well as the resulting complex microbiological fertilizer, is dried in any way at a temperature that does not have a heat tracing effect on the plant seeds and arbuscular mycorrhiza on the remaining plant roots in the fertilizer and bacteria, i.e. not more than 55 ⁰ C, and also to a state of humidity not lower than natural for seeds, AM and bacteria, i.e. to humidity not lower than 10%.

It was experimentally established that the optimal result is obtained by drying inoculation material, seeds, as well as complex microbiological fertilizer at t ° (30-50) ⁰ C to a moisture content of (10-12)% and its further grinding, for example, in a Nossep 825 mill ( VEB Maschinen Anlagenbau Nossep), to obtain a fraction of 0.05-0.2 mm.

SUBSTITUTE SHEET (RULE 26) To prepare a complex microbiological fertilizer (CMU) in granular form, a mixture of defecate with plant roots mixed with suspensions of nodule bacteria and rhizobacteria obtained by the above method is fed to a granulator (for example, OGM-0.8 or OGB-1.5) and depending on the required modification, granules of 1-10 mm are obtained, which are then packaged.

In the first particular case of the implementation of the inventive method for producing complex microbiological fertilizer (CMU), in the preparation of a coating mixture for treating the seeds of mycorrhizal plants (sorghum or others), not only AM inoculum material obtained at the first stage of the claimed method is used, but and nodule bacteria previously cultured in a liquid nutrient medium.

For this, nodule bacteria are pre-grown for 48-72 hours at t ° = 28 ⁰ C and pH = 6.8 in a liquid mannitol-yeast medium of the following composition (g / l): 0.5 - K ₂ HPO ₄ ; 0.2 - MgSO ₄ x7H ₂ O; 0.1 - NaCl; 10.0 - mannitol, 0.5 - yeast extract; 15.0 - agar-agar, to obtain ort. titer 10 ⁷ -10 ⁹ CFU / ml.

Thus, a dragee mixture for treating the seeds of mycorrhizal plants (sorghum or others) based on the inoculation material of AM fungi obtained as described above in the first stage of the claimed method, the following composition is prepared (in% by weight of the mixture): 70-75% - inoculation AM mushroom material;

10-15% - an aqueous suspension of nodule bacteria previously grown on a liquid nutrient medium with a titer of 10 ⁷ - 10 ⁹ CFU / ml; the rest is adhesive.

Then the seeds of sorghum or other mycorrhizal plants are treated with the resulting pelleting mixture, dried and grown from them in defecation

SUBSTITUTE SHEET (RULE 26) those plants until the end of the growing season, as described above, after which the aerial part of these plants is removed, and the upper 20 cm of soil-defect together with the roots of the plants are cut and used to prepare complex microbiological fertilizer (CMU) in powder form or in granular form adding to it, as described above, nodule bacteria and rhizobacteria previously grown on liquid nutrient media.

At the same time, it was experimentally established that during the period of plant growth from seeds and during the growing season, the concentration (titer) of nodule bacteria on the roots of sorghum or other mycorrhizal plants sharply decreases compared to their titer in a dragee mixture. Therefore, at the last stage of the method for producing CMC, not only rhizobacteria previously grown on liquid media, but also nodule bacteria are added to the defecate with plant roots to ensure their optimal titer in CMC.

Especially effective is the use of CMU obtained by this (first) special case of the implementation of the claimed method as fertilizer for legumes of mycorrhizal crops. Nodule bacteria contribute to the fixation of molecular nitrogen from the atmosphere in symbiosis with plants, especially legumes, in special symbiotic nodule organs and its transfer to the host plant, which increases the nitrogen nutrition of the plant and leads to an increase in its yield. In addition, part of the nitrogen remains in the roots and in the soil, improving its agro-technical condition.

In the second particular case of the implementation of the inventive method for producing complex microbiological fertilizer (CMU), when preparing a dragee mixture for treating the seeds of mycorrhizal plants (sorghum or others), not only AM fungi inoculation material obtained at the first stage of the claimed method is used, but also previously cultivated on a liquid nutrient medium rhizobacteria.

SUBSTITUTE SHEET (RULE 26) For this, rhizobacteria are pre-grown for 48-72 hours at t ° = 28 ° C and pH = 6.8 in TSB liquid standard medium (tryptone soy broth), manufactured by SIGMA, to obtain an ort titer of 10 -10 CFU / ml.

Thus, a dragee mixture for treating the seeds of mycorrhizal plants (sorghum or others) based on the inoculation material of AM fungi obtained as described above in the first stage of the claimed method, the following composition is prepared (in% by weight of the mixture):

70-75% - inoculation material of AM fungi; 10-15% - an aqueous suspension of rhizobacteria with a titer of 10 ⁶ - 10 ⁸ CFU / ml; the rest is adhesive.

Then, the seeds of sorghum or other mycorrhizal plants are treated with the resulting pelletizing mixture, dried and grown from them in the defect of the plant until the end of the growing season, as described above, after which the aerial part of these plants is removed, and the upper 20 cm of soil is defective together cut off with the roots of plants and used for the preparation of complex microbiological fertilizer (CMU) in powder form or granular form, adding to it, as described above, pre-grown on liquid pits media-negative nodule bacteria and rhizobacteria. At the same time, it was experimentally established that during the period of plant growth from seeds and during the growing season, the concentration (titer) of rhizobacteria on the roots of sorghum or other mycorrhizal plants sharply decreases compared to their titer in a coating mixture. Therefore, at the last stage of the method for producing complex microbiological fertilizer, not only nodule bacteria previously grown on liquid media are added to the defecate with plant roots, but also rhizobacteria to ensure their optimal titer in CMU. Colonies of rhizospheric bacteria provide plants with a number of useful functions, including associative nitrogen fixation, accelerate the growth of the root system, improve the absorption of phosphorus compounds, protect against phytopathogens, etc.

SUBSTITUTE SHEET (RULE 26) The most effective is the use of CMU obtained by this (second) special case of the implementation of the claimed method, as fertilizer for non-legume mycorrhizal crops.

In the third particular case of the implementation of the inventive method for producing complex microbiological fertilizer, when preparing a coating mixture for treating seeds of mycorrhizal plants (sorghum or others), not only AM fungi inoculation material obtained in the first stage of the claimed method is used, but also nodule bacteria and rhizobacteria pre-cultured in liquid culture media under the conditions described above.

Thus, a dragee mixture for treating the seeds of mycorrhizal plants (sorghum or others) based on the inoculation material of AM fungi, obtained as described above in the first stage of the claimed method, the following composition is prepared (in% by weight of the mixture):

60 - 65% - inoculation material of AM fungi;

10 - 12.5% - aqueous suspension of nodule bacteria with a titer

10 ⁷ -10 ⁹ KOE / ml; 10 - 12.5% - an aqueous suspension of rhizobacteria with a titer of 10 ⁶ -10 ⁸ CFU / ml; the rest is adhesive.

Then, the seeds of sorghum or other mycorrhizal plants are treated with the resulting dragee mixture, dried and grown from them in the defect of the plant until the end of the growing season, as described above, after which the aerial part of these plants is removed, and the upper 20 cm of soil is defect together cut off with the roots of plants and used to prepare complex microbiological fertilizer (CMU) in powder form or granular form, adding to it, as described above, pre-cultivated on a liquid x nutrient media nodule bacteria and rhizobacteria.

SUBSTITUTE SHEET (RULE 26) Nodule bacteria contribute to the fixation of molecular nitrogen from the atmosphere and its transfer to the host plant, which increases the nitrogen nutrition of the plant and leads to an increase in its yield. In addition, part of the nitrogen remains in the roots and in the soil, improving its agro-technical condition.

Rhizospheric bacteria provide plants with a number of useful functions, including associative nitrogen fixation, accelerate the growth of the root system, improve the absorption of phosphorus compounds, and protect against phytopathogens.

At the same time, it was experimentally established that during the period of plant growth from seeds and during the growing season, the concentration (titer) of nodule bacteria and rhizobacteria on the roots of sorghum or other mycorrhizal plants sharply decreases compared to their titer in a pelleting mixture. Therefore, at the last stage of the method for producing CMC, nodule bacteria and rhizobacteria previously grown on liquid nutrient media are added to the substrate from the defecate with plant roots to ensure their optimal titer in CMC.

In the particular case of implementation, a 15% aqueous solution of polyvinyl acetate emulsion (ITB A) can be used as an adhesive. Moreover, unlike gati resin, PVA glue has a low cost.

The complex microbiological fertilizer (CMU) obtained by the described method allows using it to simultaneously improve the nitrogen and phosphorus nutrition of plants, increase their protection against biotic and abiotic phytopathogens, and also solve the problem of sugar production waste utilization.

Industrial applicability

SUBSTITUTE SHEET (RULE 26) Example 1

First, inoculation material of arbuscular mycorrhiza mushrooms is obtained, for which sorghum seeds (10-15 seeds per vessel) are planted in vessels with a defect capacity of 4 l and a depth of 25 cm; at the same time, a soil-root mixture from under mycorrhizal plants with a level of occurrence of mycorrhized roots along the length of at least 60% is calculated in the defecate at the rate of 5 g of such soil-root mixture per seed. Planted sorghum plants are grown for 90-120 days. Then the aerial part of the plants is cut and removed. The remaining roots along with the defect are dried at t ⁰ (30-5O) ⁰ C to a moisture content of 10-12% and ground in a Nossep 825 mill (VEB Maschinen Anlagenbau Nossep) to obtain a fraction of 0.1-0.15 mm.

Then, a coating mixture is prepared by mixing 85% of the obtained inoculum material of AM fungi with 15% PVA glue.

Then the sorghum seeds are treated with this prepared pelleting mixture at the rate of (10-15) g of the mixture per 100 g of seeds, dried at a temperature of (30-5O) ⁰ C to a moisture content of 10-12%.

The seeds treated in this way are planted in a defecate placed in boxes in a greenhouse with a layer of 20-25 cm, and plants are grown from them using the usual technology of cultivating sorghum. At the end of the growing season, after 90-120 days, the aerial part of the sorghum plants is cut off and removed, and the resulting substrate from the defecate and sorghum roots is shaken out of the boxes and used to prepare CMU, adding 5% aqueous suspension previously grown on liquid mannitol-yeast to it nodule bacteria with a titer of 10 ⁷ -10 ⁹ CFU / ml and a 5% aqueous suspension of rhizobacteria with a titer of 10 ^b -10 ⁸ CFU / ml, previously cultured in TSB standard liquid (tryptone soy broth).

CMU is prepared in powder form, for which it is dried at a temperature of (30-5O) ⁰ C to a moisture content of (10-12)%, grind in a Nossep 825 mill to a fraction of (0.05-0.1) mm, and then packaged in paper bags of 0.5 kg. CMU obtained in this way can be stored for up to 2 years.

SUBSTITUTE SHEET (RULE 26) Example 2

First, inoculation material of arbuscular mycorrhiza fungi is obtained, as described in Example 1.

Then a panning mixture is prepared by mixing 65% of the obtained inoculation material of AM fungi, 12% aqueous suspension of nodule bacteria (with a titer of 10 ⁷ -10 ⁹ CFU / ml), 12% aqueous suspension of rhizobacteria (with a titer of 10 ^b -10 ⁸ CFU / ml) and 11% PVA glue.

Then, the sorghum seeds are treated with this obtained dragee mixture based on (10-15) g of the mixture per 10Or of seeds and dried at a temperature of (30-5O) ⁰ C to a moisture content of (10-12)%.

The seeds treated in this way are planted in defecate, which is a filter and washing cake of sugar production from sugar beets, in the defect storage fields after filtration (filter fields) located next to sugar refineries, and grown using the recommended cultivation technology of sorghum culture.

At the end of the growing season, after 90-120 days, the aerial part of the sorghum plants are mowed and removed. The upper 20 cm of defecate soil, together with sorghum roots, is cut off and used to prepare CMU, adding 2% aqueous suspension of nodule bacteria previously grown on liquid mannitol-yeast medium with a titer of 10 ⁷ -10 ⁹ CFU / ml and 2% aqueous suspension previously cultured in a standard TSB liquid medium (tryptone soy broth) of rhizobacteria with a titer of 10 ^b -10 ⁸ CFU / ml.

CMU is prepared in granular form, for which it is fed to an OGB-1.5 brand granulator and 1.5 mm granules are obtained, which are then packed in 2 kg plastic bags.

The CMU obtained in this way can be stored for up to 2 years.

Experimental evidence of the effectiveness of the use of complex microbiological fertilizer (CMU) obtained by the claimed method are shown in the tables below. Moreover, to compare the effectiveness of the CMU, the microbiological fertilizer Extrasol, made on the basis of the above RF patent Ns 2140138, was taken as a reference.

SUBSTITUTE SHEET (RULE 26) Table 1. Economic and valuable properties of complex microbiological fertilizer

As shown in table 1, a complex microbiological fertilizer (CMU), obtained according to claim 1 of the proposed method, has a number of economically valuable properties, such as plant growth stimulation and growth inhibition of phytopathogenic fungi. At the same time, the activity of CMU is higher than that of Extrasol, taken for comparison as a reference.

Table 2. The effect of pre-sowing treatment of pumpkin seeds of the variety "Stofytovaya" with complex microbiological fertilizer on the yield of pumpkin (St. Petersburg)

SUBSTITUTE SHEET (RULE 26) From table 2 it is seen that when pre-sowing treatment of pumpkin seeds of the variety "Compound" complex microbiological fertilizer obtained according to the item of the inventive method, increased the yield of pumpkin by 3.35 times compared with the control. At the same time, CMU is more effective than Extrasol taken as a reference.

Table 3. The effect of pre-sowing seed treatment of peas of the sowing variety “Tatyana” with complex microbiological fertilizer on economically valuable plant traits (Oryol Oblast)

As can be seen from table 3, during the pre-sowing treatment of pea seeds of the sowing variety "Tatyana" and half (50% NPK) and full (100% NPK) dose of the complex microbiological fertilizer obtained according to claim 2 of the proposed method, a significant positive effect on pea plants. So, the mass of plants increased by 31.5-43.0%, and the mass of seeds from one plant by 21.4-37.8%. In all respects, CMU is more effective than Extrasol taken as a reference.

Table 4. The effect of pre-sowing treatment of spring wheat seeds of the variety "Kpestyanka" with complex microbiological fertilizer on economically valuable plant traits (Oryol Oblast)

SUBSTITUTE SHEET (RULE 26) Table 4

As can be seen from table 4, during the pre-sowing treatment of spring wheat seeds of the Krestyanka variety and half (50% NPK) and full (100% NPK) doses of the complex microbiological fertilizer obtained according to item 3 of the proposed method, a significant positive effect on the economic valuable traits of plants. So, with a full dose of fertilizers, the mass of plants increased by 81.6%, the tillering energy increased by 1.7 times, and the number of seeds on one plant increased by 91%. In all respects, CMU is more effective than Extrasol taken as a reference.

Table 5. The effect of presowing treatment of tubers of potato cultivar "Elizabeth" microbiological fertilizer on the yield and commercial quality of potatoes (St. Petersburg)

SUBSTITUTE SHEET (RULE 26) As can be seen from table 5, during the pre-sowing treatment of tubers of potato cultivar "Elizabeth" complex microbiological fertilizer obtained according to claim 4 of the proposed method, there was also a significant positive effect on the yield and commercial quality of potatoes. So, the use of the indicated KMU fertilizer increased the tuber yield by 35.5 c / ha, which is an increase of 30.7%. At the same time, the number of healthy tubers increased by 11%, and the number of marketable potato fractions (I + P) increased by 18.5% compared to the control.

SUBSTITUTE .LIST PTRAVITTO 26Ϊ SUBSTITUTE SHEET (RULE 26)

Claims

CLAIM

1. A method of obtaining a complex microbiological fertilizer, which consists in combining the microbiological component with a natural biocompatible carrier, and the like, which use the inoculation material of fungi as one of the parts of the microbiological component arbuscular mycorrhiza (AM fungi), a defecate is used as a natural biocompatible carrier, which is a filter-washing precipitate for the production of sugar beets, and their combination occurs in the rhizosphere of mycorrhizal plants cultivation of these plants on a defect, and the inoculum material of AM fungi is obtained by growing mycorrhizable agricultural plants on a defecate, for which their seeds are planted in a container with a defect, a pure culture or a mixture of cultures of AM fungi in the form of a soil-root mixture from under mycorrhizal plants, with the occurrence rate of mycorrhizal roots not less than 60%, at the rate of 3-5 g of such soil-root mixture per seed, planted plants are grown for 90-120 days before the end of the growing season, then the earthly part of the plants is cut and removed, and the remaining roots, together with the defecate, are dried at a temperature of 30-50 ⁰ C to a moisture content of 10-12% and crushed to obtain a fraction of 0.05-0.2 mm, after which a coating mixture is prepared, mixed vaya obtained inoculation material AM fungi with adhesive in the ratio (by weight):

80 - 90% - inoculation material of AM fungi; the rest is adhesive; then the seeds of sorghum or another mycorrhizal plant are treated with the resulting dragee mixture at the rate of 10-15 g of the mixture per 100 g of seeds, dried at a temperature of 30-50 ⁰ C to a moisture content of 10-12%, planted in defecate and plants are grown for 90 -120 days before the end of the growing season, then the aerial part of the plants is cut off and removed, and the upper 20 cm of the defect along with the roots of the plants are cut off, after which the remaining parts of the microbiological component are added to the obtained substrate — previously cultured on liquid nutrient In environments, nodule bacteria in the substitute sheet (RULE 26) de aqueous suspension with a titer of 10 ⁷ -10 ⁹ CFU / ml in an amount of (0.5-5)%, as well as rhizobacteria in the form of an aqueous suspension with a titer of 10 ^b -10 ⁸ CFU / ml in an amount of (0.5-5) % by weight of the substrate.

2. A method of producing a complex microbiological fertilizer according to claim 1, characterized in that the taunting mixture for treating the seeds of mycorrhizal plants based on the inoculation material of AM fungi is prepared in the following composition (in% by weight of the mixture):

70-75% - inoculation material of AM fungi; 10-15% - an aqueous suspension of nodule bacteria with a titer of 10 ⁷ - 10 ⁹ CFU / ml; the rest is adhesive.

3. A method of producing a complex microbiological fertilizer according to claim 1, characterized in that the taunting mixture for treating seeds of mycorrhizal plants based on the inoculation material of AM fungi is prepared in the following composition (in% of the weight of the mixture): 70-75% - inoculation material of AM fungi; 10-15% - an aqueous suspension of rhizobacteria with a titer of 10 ^b - 10 ⁸ CFU / ml; the rest is adhesive.

4. A method of producing a complex microbiological fertilizer according to claim 1, characterized in that the taunting mixture for treating the seeds of mycorrhizal plants based on the inoculation material of AM fungi is prepared in the following composition (in% of the weight of the mixture): 60-65% - inoculation material of AM fungi;

10-12.5% - an aqueous suspension of nodule bacteria with a titer of 10 ⁷ - 10 ⁹ CFU / ml;

10-12.5% - an aqueous suspension of rhizobacteria with a titer of 10 -10 CFU / ml; the rest is adhesive.

5. The method for producing the complex microbiological fertilizer according to claim 1, characterized in that PVA glue is used as an adhesive for preparing the coating mixture.

SUBSTITUTE SHEET (RULE 26)