RU2140138C1 - Method of presowing treatment of vegetable crop seeds and method for preparing vegetable crop seed treatment preparation - Google Patents

Abstract

FIELD: agriculture. SUBSTANCE: treatment is effected with bifungicidal preparation containing strain of spore bacterium-antagonist Bacillus subtilis. Optimum results are obtained with 30 to 130 g of preparation per 1 g of seeds depending on their size and roughness of their surface. Treated seeds are immediately dried by one of known techniques at 40-50 C to moisture content 12-14%. Bifungicidal preparation is prepared by mixing polyvinylacetate emulsion and biologically-active substances, in particular, mixture from culture broth containing Bacillus subtilis strain with concentration 10⁶-10⁷ clones in 1 ml, filler (sterile chalk or dolomite) and aqueous diluent (water or aqueous solution of substances nontoxic for seeds and bacteria). Bacillus subtilis strain is cultured in liquid sterile nutrient medium containing 4% of corn bran and 0.3% of chalk on shaker or fermenter at 30 ± 1 C for 24-36 hr. Preparation components are mixed in following amounts, wt %: polyvinylacetate emulsion 20 ± 2, culture broth 20 ± 2, aqueous diluent 20 ± 2, filler - the balance. As aqueous diluent, in particular, nontoxic dye is used. EFFECT: increased infection protection of seeds, accelerated their germination, and increased crop capacity. 5 cl, 2 tbl, 2 ex

Description

 The invention relates to the field of agriculture and can be applied in crop production, including vegetable growing for presowing treatment of seeds of vegetable crops.

 The main way to combat infectious plant diseases is traditionally the use of chemical pesticides. The widespread development of the chemical industry has led to a significant reduction in the cost of chemicals used to protect plants, and the prevalence of their use.

 A known method of presowing treatment of plant seeds, described in the application of the GDR N 268140, etc. 14.01.88, published in the bulletin "ISM" N 4 for 1990, and called the "Method of encrustation of seed material".

 According to this method, the seed material is pickled with a mixture containing insecticides and fungicides in a ratio of 0.5-10 l of seed dressing per deciton of seed material, depending on the type of seed.

 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 the collected 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. Possessing relatively high antimicrobial activity, they practically do not upset biological balance, and the quality of plant products compares favorably with those obtained with the help of mineral fertilizers and chemical protective agents.

 The well-known German chemical company Bayer came to the conclusion that a biomethod with high environmental potential in plant protection was promising (Kolb F. "Biologischer Pflanzenschutz als Bestandtiel des integrierten pflazenbaus in Europe", Pflanzenschutz, Nachricht Bayer, 1992, V 45, No. 1, p. 99-112).

 The literature describes numerous attempts to use antagonist microbes in crop production. The use of biological methods has been shown to be more cost-effective than traditional chemical methods (Linderman RS, Moore LW "Strategies for detecting and characterizing System for biological control of Soilborne plant pathogens" Plant Discase, 1983, V 67, N 10, p. 1058 -1064).

 Despite the obvious preference for biological measures, they have received limited use primarily due to the lack of active producer strains, production technologies for biological products, and reliable methods for their use in crop production. Available drugs are used mainly for field crops (cereals, technical) and to a small extent in vegetable growing. At the same time, the importance of obtaining environmentally friendly products in vegetable growing is higher than in the case of obtaining such products using field crops.

 In Russia in recent years, drugs based on bacteria from the Pseudomonas group have found application. The drug under the commercial name Rizoplan, the active principle of which is the living cells of the bacteria Pseudomonas sp., Selected for the ability to suppress phytopathogens - pathogens of root rot, is used for presowing treatment of seeds mainly of grain crops. The use of Rizoplan protects plants from defeat by the causative agents of late blight, downy mildew, and bacteriosis (collection of articles "Genetics of economically valuable traits of higher plants", editors-in-chief A. Shchapova, L. Kolosova, Institute of Cytology and Genetics, SB VOGiS N.I. Vavilova, Novosibirsk, 1990, p. 194-198).

 The well-known "Method of seed treatment" 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 N 10 for 1987. This method involves treating the seeds with a composition consisting of microorganisms, a carrier for example bran, and glue such as gatti resin.

 Seeds are first recommended to be covered with glue, and then with a binder and appropriate microorganisms. The most favorable results are obtained when processing wheat seeds. Microorganisms can stimulate their growth (Azospirillum), bind nitrogen (Rhizobium), prevent plant diseases (Pseudomonas or Bacillus), or kill insects (Streptomyces).

 In this method, along with bacilli, bacteria from the genera Pseudomonas, Azospirillum are used, which die when dried, which virtually eliminates the possibility of early seed treatment (dressing) and their subsequent long-term storage. In addition, an expensive and inaccessible adhesive material is used in it as an adhesive - gatti resin obtained from tropical plants.

 A significant drawback of this method is also the fact that in the case of using Bacillus bacteria, applying to the seeds such a nutrient substrate as bran causes an intensive development of mold on the seeds, which can lead to seed death.

 Also known and closest in technical essence to the claimed method "Method of protecting cabbage from bacterial diseases" according to patent N 1793878 with priority dated 12/10/90, published on 02/07/93 in the Bulletin of inventions of the Russian Federation N 5 (prototype for the method).

According to the named method, to protect cabbage from bacteriosis, cabbage seeds after hydrothermal treatment are treated with a suspension of the bacterial strain "Pseudomonas sp. BKPM B-3481" at a concentration of 10 ⁶ - 10 ⁹ cells / ml at the rate of 20 ml per 1 kg of seeds. Before planting in the field, the root system of seedlings of cabbage is treated again with a bacterial suspension, and then plants are sprayed with it in the phase of head formation.

 The use of the bacterial suspension "Pseudomonas sp.BKPM B-3481" reduces the development of vascular and mucous bacterioses and increases the yield of cabbage.

However, Rizoplan has the following disadvantages:
- firstly, the cultivation of bacteria "Pseudomonas sp. BKPM B-3481" is carried out on rich nutrient media (glycerin, peptone (King B medium), which are scarce and expensive;
- and, secondly, due to the fact that the bacteria Pseudomonas are non-spore bacteria, after processing (inoculation) of the seeds with Rizoplan, the 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.

 To the claimed method of producing a preparation for pre-sowing seed treatment the closest in technical essence is the composition of the disinfectant described in the above application GDR N 268140 (prototype for the preparation).

 In accordance with it, the disinfectant is a mixture of the following components, taken in ratios: 5-20 g of water glass per 100 ml of a dispersion of polyvinyl acetate (PVA) with fungicides and insecticides, the concentration of which is 3-16%. The adhesion ability of fungicides and insecticides of composition with seeds is on average 99.5%.

 The disadvantages of the use of fungicides and insecticides are indicated above.

 The task of the claimed group of inventions is to increase the efficiency of protecting vegetables from infectious diseases by pre-sowing treatment of their seeds, accelerate germination and increase yield.

 This problem is solved due to the fact that in the method of presowing treatment of seeds, they are treated with a biofungicidal preparation containing a strain of an antagonist bacterium, and the strain of spore bacterium "Bacillus subtilis Ch-13" is used as a strain of an antagonist bacterium.

 The strain "Bacillus subtilis Ch-13" was deposited under the registration number VNIISHM D-606 dated September 10, 1998, in the group of epiphytic microorganisms.

 It was experimentally established that the strain of the bacterium "Bacillus subtilis Ch-13" has a pronounced antibiotic effect with respect to all the main phytopathogenic fungi: Fusarium, Rhizoctonia, verticilla living in the soil or on the surface of the seeds. At the same time, bacteria form spores, which allows you to save the drug itself, prepared on their basis, 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 vegetable crops is carried out with a preparation containing the bacterial strain "Bacillus subtilis Ch-13", in relation to the weight of the treated seeds, which depends on the size of the seeds and the degree of roughness of their surface. It has been experimentally established that the following ratios are required for different types of seeds:
- for small and rough seeds, such as seeds of carrots, beets, tomatoes, lettuce, parsley, celery - 110-130 g of the drug per 1 kg of seeds;
- for medium smooth seeds, for example, such as seeds of cabbage, radish, cucumbers, zucchini, turnips, dill - 70-90 g of the preparation per 1 kg of seeds;
- for large seeds with a smooth surface, such as seeds of corn, peas, beans - 30-50 g of the drug per 1 kg of seeds.

 Thus, the optimal result of pre-sowing treatment is achieved depending on the type of seeds with a ratio of 30-130 g of the drug per 1 kg of seeds.

In the particular case of the implementation of the method of pre-sowing treatment of seeds of vegetable crops consists of processing them with a preparation containing the bacterial strain "Bacillus subtilis Ch-13", and then drying them using one of the known methods at a temperature of 40-50 ^o C to seed moisture 12-14%.

 The operation of drying the seeds after their treatment with a liquid antibacterial drug allows you to sow these seeds not only immediately after their processing, but also after a fairly long period of time - up to six months after treatment, which allows you to process the seeds for future use with their subsequent storage until sowing.

In this case, drying is carried out in any way at a temperature that does not have a traumatic effect on the seeds of plants, that is, not exceeding 55 ^o C. It has been experimentally established that the optimal result is obtained if the seeds are dried using one of the known methods at a temperature of 40-50 ^o C.

 In addition, the drying of seeds is carried out in any way to the state of their moisture, not lower than natural, that is, to a moisture content of at least 12%. It has been experimentally established that the optimal result is obtained by drying the seeds to a moisture content of 12-14%.

The implementation of the proposed method is carried out using a preparation for presowing treatment of seeds of vegetable crops, the method of preparation of which consists in mixing an emulsion of polyvinyl acetate (PVA) and biologically active substances, which use a mixture of culture fluid containing the bacterial strain "Bacillus subtilis Ch-13" in concentrations of 10 ⁶ - 10 ⁷ cells / ml, filler (sterile chalk or dolomite) and an aqueous diluent (water or an aqueous solution of substances non-toxic to seeds and bacteria), and the bacterial strain "Bacillus subtilis Ch-13" ku Litrovat in a sterile liquid nutrient medium containing 4% wheat bran and 0.3% chalk, on a rocking chair or in a fermenter at 30 ± 1 ^o C for 24-36 hours, and the components to obtain the drug are mixed in the following proportions, expressed in percent from the total weight of the drug:
PVA emulsion - 20 ± 2%,
culture fluid - 20 ± 2%
aqueous diluent - 20 ± 2%
filler (sterile chalk or dolomite) - the rest
The strain of the bacterium "Bacillus subtilis Ch-13" is grown on an accessible nutrient medium consisting of bran and chalk, which is a significant advantage compared, for example, with the cultivation method of pseudomonas, which use scarce and expensive nutrient media.

 With this method of cultivation, bran nutrients are almost completely utilized by bacteria, which prevents the development of molds on seeds after they are treated with a biofungicidal preparation.

The concentration (number of viable cells and spores) of the strain of the bacterium "Bacillus subtilis Ch-13" should be 10 ⁶ - 10 ⁷ cells per 1 ml of culture fluid. Using a concentration of less than 10 ⁶ cells / ml leads to a decrease in the effect, and an increase in concentration above 10 ⁷ cells / ml does not increase the effectiveness of the protective effect.

 Natural filler (building) or dolomite is used as a filler in the preparation because of the ability of this bacterium to actively colonize these materials. At the same time, chalk is first subject to mandatory sterilization (thermal or radiation) in order to remove microflora, including commonplace bacteria.

 Attachment to the seeds of bacteria and chalk is carried out using an adhesive (adhesive). A 15% aqueous solution of polyvinyl acetate emulsion (PVA) is used as an adhesive due to its non-toxicity to the proposed bacteria and processed seeds, as well as its low cost.

 As an aqueous diluent, water or aqueous solutions of harmless substances for plant seeds and the bacteria "Bacillus subtilis Ch-13" are used.

 The percentage of components for biofungicidal preparation is determined empirically. The use of components in a smaller amount than indicated leads to a decrease in the effect, and their use in a larger amount does not increase the effectiveness of the protective effect of the drug.

 In the particular case of the implementation of the method for producing a preparation for presowing treatment of seeds of vegetable crops, an aqueous solution of a dye non-toxic to plant seeds and the bacterium "Bacillus subtilis Ch-13" is used as an aqueous diluent.

 The dye is used to mark treated seeds, which makes it easy to distinguish treated seeds from unprocessed ones and gives them a good marketable appearance. As a dye, almost any water-soluble paint non-toxic to bacteria and plants can be used. At the same time, preference is given to food colors, for example, such as “green strong” (E143), “erythrosine” and others.

 Example 1

The bacterial culture "Bacillus subtilis Ch-13" is grown in a sterile liquid nutrient medium containing 4% wheat bran and 0.3% chalk, on a rocking chair or in a fermenter at 30 ± 1 ^o C for 24-36 hours. 10 ⁷ cells / ml are accumulated in the culture fluid. Then get biofungicidal preparation by mixing:
50 ml of culture fluid
107 g - sterile chalk (fine powder),
50 ml - PVA emulsions (15% aqueous solution),
50 ml of water.

 10 kg of cabbage seeds are placed in a rotating drum (30 rpm), with a diameter of 700 mm and 800 g of biofungicidal preparation are added in portions; the drum is rotated for 2-3 minutes. Treated seeds are sown.

 Example 2

The bacterial culture "Bacillus subtilis Ch-13" is grown as described in Example 1, and a culture fluid with a titer of 10 ⁶ cells / ml is obtained.

Then get biofungicidal preparation by mixing:
50 ml of culture fluid
110 g - dolomite (fine powder),
50 ml - PVA emulsions (15% aqueous solution),
50 ml - dye green strong E-143 (1% aqueous solution).

The resulting preparation is treated with 8 kg of carrot or beet seeds (feed, table), as described in example 1, taking 960 g of the preparation to the specified weight of the seeds. 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 to the initial moisture content (12-13%). Store seeds before sowing for 6 months (warranty period).

 Table 1 presents some results characterizing the degree of effectiveness of biological protection of vegetables by presowing treatment of their seeds with Rizoplan and the proposed method in relation to the main phytopathogenic fungi.

 Table 2 shows comparative data on the germination rate of some types of vegetables, depending on whether their seeds were treated with a biofungicidal preparation or not, as well as on which biological product they were processed.

 From the above tables it can be seen that the effectiveness of the proposed method for the treatment of vegetable seeds using a preparation containing "Bacillus subtilis Ch-13" obtained by the proposed method is higher than when processed with the Rizoplan preparation.

Claims (4)

 1. The method of presowing treatment of vegetable seeds with a preparation containing a strain of an antagonist bacterium, characterized in that a strain of the spore bacterium "Bacillus subtilis Ch-13" is used as a strain of an antagonist bacterium. 2. The method according to claim 1, characterized in that the seeds of vegetable crops are treated with the drug in a ratio of 30 ^o C130 g of the drug per 1 kg of seeds, depending on the type of seeds. 3. The method according to claim 1, characterized in that the seeds after treatment with the drug are dried at a temperature of not more than 55 ^o C to a seed moisture of at least 12%. 4. A method of obtaining a preparation for presowing treatment of seeds of vegetable crops, which consists in mixing biologically active substances and an emulsion of polyvinyl acetate (PVA), characterized in that as a biologically active substance using a mixture of culture fluid containing the bacterial strain "Bacillus subtilis Ch-13" at a concentration of 10 ⁶ - 10 ⁷ cells / ml, sterile chalk or dolomite and an aqueous diluent, and the strain of the bacterium "Bacillus subtilis Ch-13" is cultivated in a sterile liquid nutrient medium containing 4% wheat bran and 0.3% chalk, rocking chair or in a fermenter at 30 ± 1 ^o C for 24 - 36 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:
PVA Emulsion - 20 ± 2
Culture fluid - 20 ± 2
Water diluent - 20 ± 2
Sterile Chalk or Dolomite - Else
5. The method according to claim 4, characterized in that an aqueous solution of a dye non-toxic to plant seeds and microorganisms is used as an aqueous diluent.

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