RU2216173C2 - Method for obtaining preparation for presowing treatment of seeds and vegetating plants of agricultural varieties, fruit trees and berry bushes - Google Patents

Abstract

FIELD: agriculture, plant growing, potato growing, vegetable growing, horticulture, viniculture. SUBSTANCE: the suggested method deals with presowing treatment of seeds and spraying of vegetating plants due to mixing of biologically active substances as the mixture consisting of culture liquid containing bacterial strain of Bacillus subtilis 24 D at 5×10⁹ cells/ml concentration, humates , microelements. The abovementioned bacterial strain is cultivated in liquid nutritive medium upon fermenters in the following ratios expressed in percentage against final weight of preparation: culture liquid 70-75%, humates 20%, microelements 5-10%. Bacteria of mentioned strain are active towards a wide spectrum of phytopathogenic bacteria of fungi and develop spores that considerably increases safety of preparation on treated seeds. Storage period of preparation is 6 mo since the moment of its obtaining. Optimal dosage of preparation to treat seeds of grain varieties and potato corresponds to 1.5 l/t, seeds of vegetable varieties - 20 ml/kg, spraying of vegetating plants - 2.0 l/ha, fruit trees and berry bushes - 50 ml/2-3 trees or bushes. The present innovation enables to increase efficiency in protecting grain, vegetable cultures, potato, fruit trees and berry bushes against infectious diseases due to presowing treatment of their seeds and spraying of vegetating plants to stimulate and increase immunity to unfavorable conditions of growing. EFFECT: higher efficiency. 5 ex, 3 tbl

Description

 The invention relates to the field of agriculture and may be applicable in crop production, including potato and vegetable growing, horticulture, viticulture for presowing seed treatment and spraying of vegetative plants.

 The main way to combat infectious plant diseases is traditionally the use of chemicals. The use of pesticides in large volumes gave rise to the problem of resistance of harmful objects and the economic problem of rising costs for protective measures and the environmental safety of environmental pollution.

 The disadvantages of using fungicides, insecticides and similar artificial preparations are the deterioration of the environmental situation, the violation of the natural balance, the increased content of harmful substances in agricultural products.

 In this regard, in recent years, intensive research is being conducted on alternative methods of protecting plants from infectious diseases.

 One of these methods is the use of biological agents. With a relatively high antimicrobial activity, they practically do not upset the biological balance, and the quality of plant products improves significantly.

 The literature describes numerous attempts to use antagonist microbes in crop production. It is shown that the use of biological methods is economically more effective than traditional chemical methods (Plant Protection and Quarantine. 2 - 1997. - P. 13. Second All-Russian; Scientific and Theoretical Conference "Progressive environmentally friendly technologies for storage and integrated processing of agricultural products to create products nutrition of increased nutritional and biological value ", Part II, October 1-4, 1996).

 Despite the obvious preference for biological measures, they have received limited use, primarily due to the lack of active producer strains, technologies for the production of biological products and reliable methods of their use in crop production.

 In Russia in recent years, drugs based on bacteria from the Pseudomonas group have begun to appear. The drug, under the commercial name Rizoplan, whose active principle is live cells of the bacteria Pseudomonas sp., Selected for their ability to suppress phytopathogens - root rot pathogens, are used for presowing seed treatment. Rizoplan protects plants from damage by pathogens of late blight, downy mildew, bacteriosis (Protection and quarantine of plants. - 8, 1996 - S. 14-15). However, the drug has the following disadvantages: firstly, the cultivation of bacteria "Pseudomonas sp. VKPM B-3481" is carried out on rich nutrient media (glycerin, peptone, King B medium), which are scarce and expensive, and secondly, due to Since the bacteria Pseudomonas are non-spore bacteria, after seed treatment with Rizoplan, subsequent drying of these treated seeds leads to almost complete death of the bacteria within 2-3 days. This determines the impossibility of early treatment of seeds with Rizoplan, which creates great inconvenience when working with this drug.

 The biological preparation Pseudobacterin-2 has a protective and stimulating effect based on bacterial cells of the genus Pseudomonas aureofaciens. The active principle of the drug is live bacterial cells that colonize the roots, stems and leaves of plants, while inhibiting the growth and development of pathogens. The drug almost completely eliminates root rot from the plant; the increase in the yield of cucumber and tomatoes in greenhouses when using Pseudobacterin-2 was 18-20% (Plant Protection and Quarantine. - 7, - 1999. - P. 14-15). The disadvantage of this drug, as in the first case, is that the lifespan of bacterial cells is limited, which is highly undesirable for an agricultural producer.

 The effectiveness and profitability of biological products can be improved if they are used in combination with plant growth regulators. This technique directedly affects the growth and development of plants from seedlings to harvest, activates their immunity. Growth regulators, as a rule, have a multifunctional effect on plants - they increase their height, weight of the aerial parts and roots, productive bushiness, spike grains, contribute to the formation of larger grains, increase plant resistance to adverse environmental factors and diseases. Recently, biological preparations of complex action have been created that combine both fungicidal and stimulating symptoms. These include Fitosporin, Agat-25K, Albit.

As a prototype, the invention 2099947 dated 12/12/97 (Russian patents and applications for inventions, 1994-2001) was chosen as the “Biological product Phytosporin for protecting plants from diseases”. The preparation contains a strain of the spore bacterium antagonist "Bacillus subtilis VNIISKhM 128" with a cell concentration of 1 • 10 ⁹ -1 • 10 ¹⁰ cells in 1 ml of physiological saline in an amount of 92-98 vol. % and filler in an amount of 2-8 vol.%. The above strain is characterized by high antagonistic activity against a wide range of phytopathogenic bacteria and fungi and can be used to protect various types of plants, in particular agricultural, ornamental, woody. Fitosporin is an environmentally friendly drug and harmless to both plants and warm-blooded.

 The disadvantage of the Fitosporin preparation is that only spore-forming bacteria are present, while microelements and stimulants are additionally desirable for enhancing immunity, enhanced growth and development of plants.

 The task of the claimed group of inventions is to increase the efficiency of protecting cereals, vegetables, potatoes, fruit trees and berry bushes from infectious diseases by presowing treatment of their seeds, spraying vegetative plants.

 This problem is solved due to the fact that their seeds are treated with a biofungicidal preparation containing a strain of the antagonist bacterium, and the strain of the spore bacterium "Bacillus subtilis 24D (VNIISM 129)", humates and trace elements (the biological product Integral) are used as a strain of the bacterium antagonist the composition is sprayed with vegetative plants in order to stimulate and protect the emerged plants from pathogenic microflora.

 This method of plant protection and stimulation was tested in field experiments in various soil and climatic conditions and in various crops with high efficiency. The bacteria of this strain are active against a wide range of phytopathogenic bacteria from the genera Erwinia, Pectobacterium, Pseudomonas, Xanthomonas, as well as fungi of the fungus class and imperfect fungi. The bacteria "Bacillus subtilis 24D (VNIISCHM 129)" form spores, which significantly increases the safety of the drug and the seeds treated with this drug without loss of bacterial viability for a long time. The guaranteed shelf life of the composition is 6 months from the date of its receipt.

 In the particular case of the implementation of the proposed method, pre-sowing treatment of seeds of agricultural crops is carried out with a preparation containing the bacterial strain "Bacillus subtilis 24D (VNIISKHM 129)", humates and trace elements. It was experimentally established that for the processing of crops and potatoes the optimal dose of the drug was 1.5 liters per ton of seeds.

 In the particular case of the implementation of the method of presowing treatment of seeds of vegetable crops consists of processing them with a preparation containing the bacterial strain "Bacillus subtilis 24D (VNIISCHM 129)", humates, trace elements in a dose of 20 ml per 1 kg of seeds.

 In the particular case of the implementation of the method of presowing treatment of seeds and spraying vegetative plants consists of processing crops of grain, vegetables, potatoes with a preparation containing the bacterial strain "Bacillus subtilis 24D (VNIISHM 129)", humates, trace elements at a rate of 2.0 l / ha.

 In the particular case of the implementation of the method of presowing treatment of seeds and spraying vegetative plants consists of treating fruit trees and berry bushes with a preparation containing the bacterial strain "Bacillus subtilis 24D (VNIISKHM 129)", humates, trace elements at the rate of 50 ml per 2-3 trees / shrubs.

Implementation of the proposed method is carried out using a preparation for presowing treatment of seeds and vegetative plants of agricultural crops, fruit trees and berry bushes, the method of preparation of which consists in mixing biologically active substances, which are used as a mixture of culture fluid containing the bacterial strain "Bacillus subtilis 24D ( ARRIAM 129) "at a concentration of 5 • ¹⁰ September Cl. / mL, humates, trace elements, wherein the bacteria strain" Bacillus subtilis 24D (ARRIAM 129) "was cultured in a liquid nutrient medium at fe Menter for 48 hours, and the components are mixed to obtain a formulation in the following proportions, expressed as a percentage of the final weight of the formulation: culture medium - 70-75, humates - 20 trace - 5-10.

 The bacterial strain "Bacillus subtilis 24D (VNIISCHM 129)" is grown on an accessible nutrient medium with molasses, which is a significant advantage compared to cultivation methods of pseudomonas, which use scarce and expensive nutrient media.

 The humates that make up the drug fulfill a double role: firstly, for bacilli they are a preservative, which prevents the cell titer from falling for a long time - 6 months, and secondly, an excellent growth and development stimulator for plants at the first stage of development. Subsequently, this function is performed by microorganisms.

 Trace elements - boron, molybdenum, magnesium, copper, zinc, cobalt, etc. are necessary for seeds to fully form the future crop. They are directly involved in all photosynthesis reactions. It is impossible to replace their deficit in soil with any other agrotechnical method; crop shortage in their absence can reach 50%.

 Example 1

The bacterial culture "Bacillus subtilis 24D (VNIISCHM 129)" is grown in a sterile liquid nutrient medium in a fermenter for 48 hours. In the culture fluid, cells accumulate at a concentration of 5 • 10 ⁹ . By mixing humates, trace elements, we obtain the drug. We prepare a working solution for seed treatment: dilute 1.5 liters of the resulting preparation in 10 liters of water. The solution is seasoned in the PS-10 pickling machine and we carry out semi-dry treatment of wheat or barley seeds. We process potato tubers in a similar way. Seed is sown in the field.

 Example 2

The bacterial culture "Bacillus subtilis 24D (VNIISCHM 129)" is grown in a sterile liquid nutrient medium in a fermenter for 48 hours. In the culture fluid, cells accumulate at a concentration of 5 • 10 ⁹ . By mixing humates, trace elements, we obtain the drug. We prepare a working solution for seed treatment: 200 ml of the drug per 10 kg of vegetable seeds (carrots, beets). Seeds are placed in a rotating drum (30 rpm) with a diameter of 700 mm and the preparation is introduced in portions, the drum is rotated for 2-3 minutes. Then the seeds are poured from the drum onto the pallet and dried in a stream of air with a temperature of 40-50 ^o C to a moisture content of 12%. Seeds processed in this way are stored until sowing.

 Example 3. Two liters of the drug, prepared according to the above scheme, is diluted in 200-300 liters of water. With a boom sprayer, this amount of solution is sprayed onto vegetative plants of crops, vegetables, and potatoes. It is preferable to carry out the first spraying at the early stages of plant development (in cereals - the tillering phase) in order to prevent the development of pathogenic microorganisms and to enhance plant growth. The second spraying of vegetative plants is carried out in order to increase the yield and its quality. The drug is 80% composed of living microorganisms, therefore, the most favorable time for the treatment is evening or morning hours (quiet, calm weather, lack of direct sunlight).

Example 4. A preparation prepared according to the above scheme is used on berry shrubs, fruit trees. 50 ml of the drug is taken and diluted in two liters of water. This solution is watered with 2-3 trees / shrubs or 100 m ^{2 of} strawberry plantings.

 Table 1 presents the results of a scientific experiment on the effect of biological products by processing the seed of barley on the growth and development of plants, the spread of root rot.

 Table 2 presents the results of production experience on the impact of the proposed drug on the reduction of diseases of potato plants by diseases.

 Table 3 presents the results of experiments on the effect of presowing treatment of seeds with a biological product on the yield of vegetable crops.

 4. An example of a method for producing the drug.

 Getting a uterine culture.

In a sterile nutrient medium, which contains (in g / l water): sodium nitrate 3, potassium phosphate monosubstituted 1, magnesium sulfate 0.3, chalk 1, molasses 20, and the pH of which is brought up to 7.0-7.2 , bring bacteria Basillus subtilis 24D in an amount of 2-3% (by volume) and grow for 44-48 hours at 32 ± 1 ^o C.

 Production cultivation of the strain in a fermenter.

Prepare a nutrient medium of the specified composition, sterilized. Cool to 28-30 ^o C and contribute 3-4 vol.% Seed, obtained by the above method. Cultivation is carried out at 32 ± 1 ^o C for 44-48 hours with constant aeration (0.6 atm) and stirring.

 The resulting culture fluid is cooled. Take 70% of the culture fluid, add humates in an amount of 20% and trace elements in an amount of 10%; or 75% of the culture fluid, add humates in an amount of 20% and trace elements in an amount of 5%. A biological preparation consisting of culture fluid, humates and trace elements (70 + 20 + 10)% and (75 + 20 + 5)%, is one and the same preparation, has equivalent fungicidal properties on plants.

The preparation is a light brown liquid with a sediment of a nutrient medium and a possible thin bacterial film on the surface and contains Basillus subtilis 24D cells at a concentration of 5 • 10 ⁹ , humates and trace elements.

Claims (1)

A method of obtaining a preparation for presowing treatment of seeds and vegetative plants of crops, fruit trees and berry bushes, which consists in mixing biologically active substances, characterized in that as a biologically active substance using a mixture of culture fluid containing a bacterial strain Bacillus subtilis 24D (VNIISM 129 ) at a concentration of 5 • September ¹⁰ cells / ml, humates, trace elements, wherein the Bacillus subtilis bacterial strain 24D (ARRIAM 129) cultured in a liquid nutrient medium in fermentors for 48 hours, Formulation components were mixed in the following ratios,% by weight of the final preparation:
Culture fluid - 70-75
Humates - 20
Trace elements - 5-10

Images