LT6142B - Process for treating seed-corn and seed - Google Patents

Abstract

The present invention relates to the field of agricultural biotechnology. It concerns the process for treating seed-corn and seed with preparations of lactic acid bacteria to protect seed against mould microorganisms. To produce antimicrobial bio-products the lactic acid bacteria with the best activity against pathogenic microorganisms were selected, namely, Lactobacillus sakei (MI806), Pediococcus acidilactici (MI810) and Pediococcus acidilactici (MI807) which had been deposited at the JSC "Biocentras" museum of microorganisms. The invention claims the use of lactic acid bacteria preparations for treatment of seed-corn and seed.

Description

The present invention relates to the field of agricultural biotechnology. It describes a method for treating seed grains and seeds by treatment with lactic acid bacteria to protect against mold microorganisms. The present invention describes a method of using lactic acid bacterial preparations for treating seed grains and seeds prior to sowing.

TECHNICAL LEVEL

Cereals used in the food industry are often contaminated with pathogenic microorganisms. Higher concentrations of pathogenic micro-organisms completely damage the grain and cannot be used for food production. When the infected grains are used as seed, they germinate poorly, significantly reducing grain yield. The most common seed grains are Fusarium spp., Cochliobolus sativus, Alternaria spp. or hard stumps in Tilletia caries spores. Cereals feed on cereals to release secondary metabolites, mycotoxins, which cause fusariosis rot. It is harmful not only to the plants themselves, but also to the grains themselves, which make them unsuitable for food and feed.

A number of chemical and biological agents are used to degrade or stop the growth of such pathogenic microorganisms. However, these preparations have various disadvantages:

1) lack of efficacy due to the narrow specificity of antimicrobial activity;

2) short-term effects as only grains or seeds are processed;

3) short life span of anticicrobial agents.

Various types of bacteriocin-synthesizing bacteria are described in the patent literature. The peptides isolated from them are widely used in various fields such as veterinary, agriculture, food industry, cosmetic industry, medicine and others.

US004980163 discloses broad-spectrum bacteriocin compositions. These compositions may be dissolved or suspended in a suitable solvent or matrix and exhibit a broader antimicrobial activity than each of the individual components of the composition. Dissolved or suspended bacteriocin formulations have broader practical applications. The compositions of the compositions looked like this:

lysostaphin and lanthionine-containing bacteriocin;

lysostaphin, a lanthionine-containing bacteriocin and chelating agent;

lysostaphin, a lanthionine-containing protein, chelating agent, and surfactant.

US005348881A describes N. Lactococcus microorganisms producing two or more different bacteriocins. Lactococcus lactis is used in the food industry for inhibiting Lactobacillus casei and other food contaminants. Also described are bacteriocin compositions, bacterial mixtures and methods for their use in the food industry.

US005683890A discloses Streptococcus thermophilus bacteriocins having the amino acid sequences of SEQ ID NO: 1 and SEQ ID NO: 2. Also described is a method for obtaining a supernatant extract containing at least one of the two bacteriocins. Finally, the use of these bacteriocins as agents against pathogens in the manufacture of foodstuffs, particularly cheese and sour milk products, and cosmetics is described.

US005691301A describes bacteriocin compositions comprising lanthionine-containing bacteriocins and non-bacteriocidal agents. When bacteriocin compositions are immobilized on suitable carriers, they become improved, fast-acting, broad-spectrum bacteriocins suitable for a wide range of applications.

US20120164119A1 describes a novel antimicrobial agent. It has a similar antimicrobial action to nisin. Bacteriocin is named lacticin 3147 with a molecular weight of 2.8. It inhibits the activity of lactococci, lactobacilli, enterococci, bacilli, leuconostocks, pediococci, clostridia, staphylococci and streptococci. It is also capable of inhibiting nisin-producing bacterial strains.

US006753024B2 describes the outer skins of tubers and the tubers cut into pieces. The pieces are crushed into a puree, which is dried, diluted with water and boiled. It is then inoculated with selected bacteriocin-producing bacteria and left to ferment until a nutrient preservative is obtained.

US00689370B2 describes novel bacteriocins and strains of novel bacteria produced. These antimicrobial peptides are used to reduce the growth of pathogenic bacteria in animals (especially poultry).

US007449311B2 relates to hitherto unknown bacteriocin-producing microorganisms. More particularly, the invention provides a novel bacteriocin with a lantibiotic. It is named macedocin because it is produced by Streptococcus macedonicus ACA-DC 198 microorganisms. Makedocin inhibits a broad spectrum of lactic acid producing bacteria as well as several food-borne pathogenic bacteria (including Clostridium tyrobutyricum). Makedocin is temperature stable, active and has a wide pH range. The patent also describes the use of non-pathogenic microorganisms for macedocine biosynthesis in food enzymes such as dairy products (butter, cheese), fermented meat and vegetables. The use of macedocin in non-fermentative processes such as raw meat, vacuum meat products, fast food and canned food is also described.

US20080248953A1 describes a method for improving plant vegetation and / or disease resistance. For this purpose, a purified polypeptide is used which has improved antimicrobial properties by protecting plants, plant seeds or the vegetation environment from pathogenic microorganisms.

US20110236359A1 describes a novel subtype of lactic acid producing strain Lactococcus lactis lactis MM19 (Cat. No. NML-080508-01) and Pediococcus acidilactici MM33 (Cat. No. NML-080508-02), isolated from the human gut. The L. lactis strains MM19 and P. acidilactici MM33 and the bacteriocins produced are useful for inhibiting microbial growth in foodstuffs and inhibiting microbial infection or colonization in mammals.

US20120028799A1 describes a method for controlling plant diseases caused by bacterial or fungal pathogens. It is based on the application of a protein or polypeptide isolated from Streptomyces scopuliridis strain RB72 containing an amino acid from SEQ ID NO: 1 for the protection of plants or plant seeds. It also describes the specific conditions under which effective treatment of plant diseases caused by bacterial fungal pathogens is achieved. In addition, a planting composition for plants and plant seeds treated with said antimicrobial agents is described.

The invention described in patent RU 2409661 C2 can be applied to the prophylaxis and treatment of infectious animals (especially birds). The strain Enterococcus feacium LVP1073 is isolated from the intestine of a healthy chicken. This producer synthesizes bacteriocin E1073, which is active against bacterial pathogens. This 34 amino acid-length bacteriocin is classified as a cationic peptide group (pi = 8.2) with a molecular weight of 3256 Da. The bacteriocin E1073 biosynthesis inducer Lactobacillus plantarum 1LVP was also identified. A signal peptide regulating bacteriocin biosynthesis has also been identified. The patent describes an invention effective against many gram-positive and gram-negative pathogenic microorganisms. Likewise, the treatment of the chickens with said bacteriocin prior to slaughter ensures that the resulting product is protected against pathogenic micro-organisms harmful to humans.

Patent LT 4209 B describes a method for treating plant grain using a bacterial preparation of lactic acid to improve grain quality. The present invention also describes a method of using this preparation for spraying grain ripening plants in fields. It also describes the treatment of malting beer with biopreparation.

However, there remains a need to develop a faster-acting, more efficient, next-generation bioproduct with a broad antimicrobial range. Safe antimicrobials based on lactic acid bacteria have narrow specific antimicrobial activity. These deficiencies eliminate and increase the germination and yield of grains and seeds, and a new way of treating seed grains and seeds has been developed.

THE SUBSTANCE OF THE INVENTION

SUMMARY OF THE INVENTION The object of the present invention is to reduce microbiological contamination of seed grains and seeds and to improve their germination by natural biopreparations by seeding seed grains and seeds in a plant substrate enriched with mineral substances and amino acids, a mixture of lactic acid bacteria. Some chemical antimicrobials (stains) have a long lifespan. For this reason, steamed grains can also enter the human body during food production.

SUMMARY OF THE INVENTION The present invention relates to the successful use of bio-preparations of lactic acid bacteria for protection of agricultural grains against pathogenic microbes.

Such biopreparations are suitable for use in the prevention of phytopategens and secondary metabolites formed during plant cultivation and storage of grain.

SUMMARY OF THE INVENTION The present invention provides a novel method of protecting grains and seeds that completely or at least partially solves existing problems. The invention differs from known antimicrobial treatment of cereals in that complexes are treated with antimicrobial agents derived from different lactic acid bacteria to treat the contaminated pathogenic microorganisms.

It is also different that different lactic acid bacteria are selected so that the atimicrobial biopreparations obtained from them have a broad antimicrobial effect on different pathogenic microorganisms, depending on the grain contamination.

The third difference is that antimicrobial agents can be used either in purified form or in combination with lactic bacteria.

The fourth difference is that the biopreparation can be used for complex treatment of seed grains or seeds before or after sowing.

Microorganisms in seed grains and seeds can be beneficial or harmful. The seed microflora of seed grains and seeds includes mold microorganisms such as Fusarium spp., Cochliobolus sativus, Alternaria spp., Root rot and hard and dusty stumps. This microflora reduces grain yield and disadvantages the storage of seed grains and seeds.

Mold fungi are an undesirable impurity in the seed grain used in the food and feed industry. Therefore, it is necessary to control mold contamination of seed grains in their storage areas, as factors such as temperature and humidity can initiate mold growth. Lactic acid bacteria are used to stop the growth of the mold microflora.

PREFERRED EMBODIMENTS

Lactobacillus sakei (MI806), Pediococcus acidilactici (MI810) and Pediococcus acidilactici (MI807), the most pathogenic microorganisms of which have been isolated from the rye cultures of the rye bread "Musro", have been selected for the development of bio-products with antimicrobial properties.

Lactobacillus sakei (MI806): Gram-positive, anaerobic, non-spore-forming, rod-shaped bacteria belonging to the Firmicutes type, Lactobacillales, family Lactobacillaceae, genus Lactobacillus. The culture is grown under anaerobic conditions; on agarized MRSA medium, grows in 48 hours at 37 ° C. May grow at temperatures between 35-50 ° C. Grows in the pH range 3.5-7.0 -8.5.

L. sakei has an enzymatic metabolism and the end product is lactate. Other possible products: acetate salts, ethanol, CO2, succinate.

The taxonomic dependence data for Lactobacillus sakei (MI806) are as follows (SEQ ID NO: 1):

NAGTCGAGCGGACAGAAGGGAGCTTGCTCCCGGATGTTAGCGGCGGA

CGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAA

ACCGGAGCTAATACCGGATAGTTCCTTGAACCGCATGGTTCAAGGATGAAAGAC

GGTTTCGGCTGTCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAG

GTAACGGCTCACCAAGGCGACGATGCGTAGCCGACCTGAGAGGGTGATCGGC

CACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGA

ATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAG

GTTTTCGGATCGTAAAGCTCTGTTGTTAGGGAAGAACAAGTGCAAGAGTAACTG

CTTGCACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCA

GCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGG

GCTCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGA

GGGTCATTGGAAACTGGGAAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCAC

GTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGAC

TCTCTGGTCTGTAACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGAT

TAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGGGGGTT

TCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACG

GTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAG

CATGTGGTTTAATTCGAAGCAACGCGAAGAAcCTTACCaGGTCTTGACATCCTCT

GANNNCCCTAGAGATAGGGCTTTCCCNTCGGGGNNN.

Pediococcus acidilactici (MI810) is assigned to the Firmicutes type, line

Lactobacillales, of the family Lactobacillaceae, genus Pediococcus. Bacteria are grown on MRSA or M-17 media. When grown at 37 ° C under anaerobic conditions, it grows within 48 -72 hours.

P. acidilactici is a coca-like, gram-positive, immobile, spore-free lactobacterium. Coke tetrad forms. Bacteria are classified as facultative anaerobes. Performs fermentative type metabolism. Fermentes sugars. The end product of metabolism is lactic acid. CO2 is not emitted. P. acidilactici grows well in acidic environments but can grow well over a fairly wide pH range of 4.5 to 8.0. They are also acid resistant.

The taxonomic dependence data for Pediococcus acidilactici (MI8010) are shown below in SEQ ID NO. 2):

NAANTGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCAGAA

GCAGGGGATAACACCTGGAAACAGATGCTAATACCGTATAACAGAGAAAACCGC

CTGGTTTTCTTTTAAAAGATGGCTCTGCTATCACTTCTGGATGGACCCGCGGCG

CATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGATGATGCGTAGCCGAC

CTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGG

GAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGC

CGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAGCTCTGTTGTTAAAGAAGAACG

TGGGTGAGAGTAACTGTTCACCCAGTGACGGTATTTAACCAGAAAGCCACGGCT

AACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATT

TATTGGGCGTAAAGCGAGCGCAGGCGGTCTTTTAAGTCTAATGTGAAAGCCTTC

GGCTCAACCGAAGAAGTGCATTGGAAACTGGGAGACTTGAGTGCAGAAGAGGA

CAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAG

TGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGG

TAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGATTACTAA

GTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGTAATCCGC

CTGGGGAGTN.

Pediococcus acidilactici (MI807) belongs to the Firmicutes type, genus Lactobacillales, family Lactobacillaceae, genus Pediococcus. The bacteria are grown on MRSA or M-17 medium. Growing at 37 ° C, takes 48 to 72 hours to grow anaerobically.

P. acidilactici is a coca-like, gram-positive, immobile, spore-free lactobacterium. Coke tetrad forms. Bacteria are classified as facultative anaerobes. Performs fermentative type metabolism. Fermentes sugars. The end product of metabolism is lactic acid. CO2 is not emitted. P. acidilactici grows well in acidic environments but can grow at fairly wide pH ranges of 3.5-8.0. Acid resistant.

The following are the taxonomic dependency data for Pediococcus acidilactici (MI807) (SEQ ID NO: 3):

NCCTTTAGTGCTGGACGTGTAAAGTAATCACGTTAGCTTGTTGGTAAAG

TTCTTCTAAAGTATCTACATACATGCCTTCCTTTTCTAATGCAGGATTACGGAAAA

CATCGTACGCAATTACCTTTGCGCCAAATCCTTTAAAAATGTCAATTGCAGCACG

GCCAATCCGTCCGGTACCAATTACCCCAACGGTCATTTGATTAAGTTCCAAACC

GATGTCTGGTTCCCAACGATAATCGCCATGAGCCATTTTGTATTCAAATTCAGGA

ATCTTACGGAGTAATGCTAACAGTTGGGTGACTGACAATTCAGCAATTGCTCTTG

GAGAATATGCTGGTACGTTCGAAATTTTGATATCATTTTTCTTGAGTGCATCTGC

GGGTNCNNNTCAAGTCCN.

The isolated microorganisms exhibit broad antimicrobial activity (Table 1).

table. Inhibitory effect of PRBs and anti-fungal preparations on spore formation and growth

PRB supernatants supernatants (BPMs) in Hybride spectrum of ants (S) and neutralized to pH 6.5 KTU05-06 KTU05-07 KTU05-08 KTU05-09 KTU05-10 s BPM S BPM s BPM S BPM S BPM Mushrooms R. stolonifer 2-KA P. expansum 23L + + + + + + + + - - A. nigerN \ 0-029 + + + + + + + + + + A. versicolor MI-130 + + - + + + + - - + A. fumigatus KO-5 - - - - + + - - + + P. chrysogenum 48-L - - + + + + + + + + A. altemata 1-4U P. cyclopium 21 AL A. tereus S-1

F. culmorum L-2 ++ + +++ ++ + +++ ++ + +++ ++ + +++ ++ + +++ F. solani 4-SA Yeast P. brutonii P.3 S. capsularis S.1 - - - - - - - - - - Z. ba // zz'Z.1 (T) K. marxianus K.7.1 (T) S. cerevisiae Sa.1.5 (T) A. parapsilosis C.7.2 ++ ++ - - ++ ++ ++ ++ - - Y. lipolytica Y.C.6.1 hansenii Deb.4 (T) + - - - - - - - + -

Antimicrobial activity was determined by diffusion to agar using indicator microorganisms. For this, a thin layer of medium is applied to the Petri dishes, thus forming the base as a source of additional nutrients. Drying the lower layer to 10 mL of molten up to 45 ° C, the cold medium is added a suspension of pathogens ~ 10 ⁴ cfu cell lawn. The medium is poured onto prepared plates with the prepared bottom agar layer. Once the top layer of agar has solidified, a sterile cylinder is cut into 8 mm diameter cavities and the agar is removed. The wells are dosed with 50 μΙ of biopreparation. Petri dishes are incubated with the crop in a thermostat. The incubation time and temperature are selected with regard to the pathogenic micro-organism. The positive reaction - inhibition of growth of the microorganisms to be tested - is assessed by measuring the diameter of the clear area around the well by measuring the diameter of the clear area around the well.

The technological scheme of the fermentation of antimicrobial biopreparations is shown below, step -1 (Figure 1).

Fig. Technological scheme of fermentation of antimicrobial biopreparations (Phase I)

Lactobacillus sakei (MI806), Pediococcus acidilactici (MI810) and Pediococcus acidilactici (MI807) are used as bacteriocin-synthesizing lactic acid bacteria to produce a complex biopreparation.

Lactobacillus sakei (MI806) bacteria were grown in MRS broth without stirring for 18 hours at 35 ° C. Thereafter, 2% of the resulting inoculum is plated in a sterilized fermentation medium consisting of potato heavy, 5% rice extrudate, 0.1 g / L manganese sulfate (MnSO4), 0.2 g / L magnesium sulfate (MgSCU), 2 g / L cysteine hydrochloride ([HSCH2-CHNH3 ⁺ -CO2H] CI ·). The bacterial culture is grown for 6 hours at 35 ° C.

Pediococcus acidilactici (MI807) was grown without stirring in MRS broth for 18 hours at 35 ° C. 2% of the resulting inoculum is then seeded in a sterilized fermentation medium containing potato, 5% rice extrudate, 0.1 g / L manganese sulfate (MnSO-i), 0.2 g / L magnesium sulfate (MgSO4), 2 g / L of cysteine hydrochloride ([HS-CH2-CHNH3 ⁺ -CO2H] Clj.) The bacterial culture was grown for 18 hours at 35 ° C.

Pediococcus acidilactici (MI810) bacteria were grown without stirring in MRS broth for 18 hours at 35 ° C. Subsequently, 2% of the resulting inoculum is plated in a sterilized fermentation medium consisting of potato heavy, 5% rice extrudate, 0.1 g / L manganese sulfate (MnSO4), 0.2 g / L magnesium sulfate (MgSO4), 2 g / L cysteine hydrochloride ([HS-CH 2 -CHNH 3 ⁺ -CO 2 H] Cl). The bacterial culture is grown for 6 hours at 35 ° C.

The following are the main characteristics of the culture fluid (Table 2). Table 2. Main characteristics of the biopreparation

Indicator name Results P. acidilactici KTU05-10 P. acidilactici KTU05-07 L. sakei KTU0506 Appearance Thick consistency fluid Thick consistency fluid Thick consistency fluid Color Beige Beige Beige The smell Specific potato heavy odor Specific potato heavy odor Specific potato heavy odor Dry matter,% 10-12 10-12 10-12 CFU / ml. > 1O ^{9 '10} > 1O ^{9 '10} > 1O ^{9 '10} Diameter of indicator culture growth inhibition zones (without well diameter), mm Fusarium culmorum Not less than 8 Not less than 8 Not less than 8 Bipolar sorokiniana Not less than 20 Not less than 22 Not less than 24 Bacillus subtilis Not less than 20 Not less than 20 Not less than 23

The following is a schematic of the antimicrobial purification (step II) (Figure 2).

Fig. Technological scheme of antimicrobial biopreparation purification (Phase I):

The resulting culture fluid was centrifuged at 17,000 rpm in a continuous centrifuge. This separates the biomass. The supernatant obtained below is concentrated using an ultrafiltration device with a membrane pore diameter of 1000 to 3000 Da. The resulting concentrate is precipitated with acetone, ethanol or other organic solvent.

The following are schematic diagrams of the biopreparations of the pretreatment and secondary treatments (Fig. 3).

a) Fig. b). Schematic diagrams of biopreparations produced by primary (a) and secondary (b) treatment

Experimental field studies with spring wheat seeds treated with the bioproduct have shown significant influence of the preparation on crop yield.

Some examples of application of the invention are described below.

example

Preparation of bio-preparations of lactic acid.

Biopreparations of 3 different bacterial cultures: L. sakei KTU05-06, P. acidiiactici KTU05-07, P. acidilactici KTU05-10 are produced for the treatment of seed grains, seeds and crops. The crop and the amount of the biopreparation it produces depends on the degree of contamination of the seed grain and / or seeds with the undesirable microorganisms. The possible formulations of biopreparations and the amounts of components are given in Tables 3 and 4.

table. Compositions of Rounded Antimicrobial Biopreparations

Biopreparation No. A bacterial culture is used L. said KTU05-06 P. acidilactici KTU05-07 P. acidilactici KTU05-10 1a 0-100% 0-100% 2a 0-100% 0-100% 3a 0-100% 0-100% 4a 0-100% 0-100% 0-100%

table. Compositions of Concentrated Antimicrobial Biopreparations

Biopreparation No. A bacterial culture is used L. said KTU05-06 P. acidilactici KTU05-07 P. acidilactici KTU05-10 1b 0-100% 0-100% 2b 0-100% 0-100% 3b 0-100% 0-100% 4b 0-100% 0-100% 0-100%

example

Influence of lactic acid biopreparations on yield of wheat Tris

Before sowing, wheat grains were treated with biopreparations composed of 3 different bacterial cultures: L. sakei KTU05-06, P. acidilactici KTU05-07, P. acidilactici KTU0510. Biopreparation formulations with the greatest influence on spring wheat yield were selected for research (Table 5).

table. Influence of lactic acid biopreparations on yield of wheat Tris

Study variant Grain yield, t / ha Grain harvest additive, t / ha Yield in relative numbers Control 4.73 0 100 KTU05-6 5.15 0.42 108.8 KTU05-7 5.23 0.50 110.5 KTU05-10 5.08 0.35 107.4 KTU05-7 + KTU0510 5.06 0.33 106.9 KTU05-6 + KTU05-7 + KTU05-10 5.06 0.33 107.0

example

Effectiveness of lactic acid biopreparations against durum wheat in spring wheat Triso

Before sowing, wheat grains were treated with biopreparations composed of 3 different bacterial cultures: L. sakei KTU05-06, P. acidilactici KTU05-07, P. acidilactici KTU0510. In the research, biopreparation formulations were selected that are most effective against durum wheat in spring wheat (Table 6).

table. Influence of lactic acid biopreparations on durum wheat "Triso"

Study variant 1.5 m ^{2 of} infected bells % Decrease in violations Control 306 0 KTU05-6 18th 94.3 KTU05-7 4 98.8 KTU05-10 23rd 92.7 KTU05-7 + KTU05-10 4 98.8 KTU05-6 + KTU05-7 + KTU05-10 10th 96.7

example

Practical application of lactic acid biopreparations in field conditions

PRB was used in the same scheme with three plants (spring wheat "Triso" spring barley "Class" and "Simba").

(a) for use in biopreparations in the form of round:

Control (water 20 ml / 1 kg seed)

Biopreparation no. 1a (20 ml / kg of seed)

Biopreparation no. 2a (20 ml / kg of seed)

Biopreparation no. 3a (20 ml / kg of seed)

Biopreparation no. 4a (20 ml / 1 kg seed)

(b) Concentrated biopreparations used:

Control (water 20 ml / 1 kg seed)

Biopreparation no. 1b (20 ml / kg of seed)

Biopreparation no. 2b (20 ml / kg of seed)

Biopreparation no. 3b (20 ml / kg of seed)

Biopreparation no. 4b (20 ml / kg of seed)

The agronomic efficiency of PRB was investigated using field test method. Each field experiment is equipped with 4 replicates, the fields are randomized.

The seed was treated with the Hege 11 seed dresser, which ensures even distribution of the product on the seed. The seeds were dried before sowing. The seed treated with the rounded biopreparations was dried in a field protected from the sun, then the required amount of seed for each variant was weighed for each field. Under field conditions, the fields were sown with a precision seeder Hege 80.

After sowing, the crops were treated with concentrated aqueous solutions of biopreparates. For this purpose, the following methods of application of the bio-formulation to the crop were used:

spraying, spraying and fogging. Spraying working pressure from 30 to 50 bar, spraying rate of working biopreparation solution from 2000 to 4000 L / ha. Working spraying pressure 50-60 bar, spraying rate of working biopreparation 200-300 l / ha, air volume 8000-9000 m ³ / h. Spraying working pressure 2-3 bar. Before spreading the biopreparation, the crop field was divided into 0.1 ha (12m x 83.3m) working areas. The crop field was treated with biopreparation at a wind speed not exceeding 4 m / s and at high atmospheric pressure.

In order to illustrate and describe the present invention, descriptions of preferred embodiments are set forth above. It is not a complete or limiting invention that seeks to determine the exact form or embodiment. The above description should be viewed as an illustration rather than a limitation. Obviously, many modifications and variations may be apparent to those skilled in the art. Embodiments are selected and described so that those skilled in the art will best understand the principles of the present invention and their best practical application for different embodiments with different modifications suitable for a particular application or embodiment, since the quantitative application rates of this embodiment may vary. The scope of the invention is intended to be defined by the appended definition and its equivalents, in which all of the foregoing terms have the widest meaning unless otherwise indicated. It will be appreciated that the embodiments described by those skilled in the art may make changes within the scope of the present invention as defined in the following definition.

1. A method for treating seed cereals and seeds, comprising treating the seed cereals and seeds with a secreted lactic acid bacterium selected from the group consisting of Lactobacillus sakei MI806, Pediococcus acidilactici MI810 and

Pediococcus acidilactici MI807.

DEFINITION OF INVENTION

Claims (13)

DEFINITION OF INVENTION A method for treating seed cereals and seeds according to claim 1, characterized in that said cereals and seeds are treated with at least two of said lactic acid bacteria. A method for treating seed cereals and seeds according to claim 1 or 2, characterized in that said cereals and seeds are treated with a composition comprising Lactobacillus sakei MI806, Pediococcus acidilactici MI810 and Pediococcus acidilactici MI807. A method for treating seed cereals and seeds according to any one of claims 1 to 3, characterized in that said cereals and seeds are treated with a composition containing at least two of said lactic acid bacteria. A method for treating seed cereals and seeds according to any one of claims 1 to 4, characterized in that said cereals and seeds are treated with a biopreparation comprising a composition according to any one of claims 3 to 4. A process for the treatment of seed cereals and seeds according to any one of claims 1 to 5, characterized in that the biopreparation used is rounded or concentrated. The process for treating seed cereals and seeds according to claim 6, characterized in that said biopreparation is rounded. A method for treating seed grains and seeds according to claim 7, characterized in that the dressed preparation is used before sowing. A method for treating seed cereals and seeds according to claim 6, characterized in that said biopreparation is concentrated. A method for treating seed cereals and seeds according to claim 9, characterized in that the concentrated biopreparation is used after sowing. 11. Use of a biopreparation for treating seed grains and seeds. 12. The use according to claim 11, wherein the biopreparation is used to treat the seed and seed before or after sowing. 13. Seed grains and seeds, characterized in that they are treated according to any one of claims 1 to 10.