RU2265000C1 - Method of storage of bacterial fertilizers - Google Patents

Abstract

FIELD: agriculture; namely to production and storage of bacterial fertilizers.

SUBSTANCE: the invention is pertaining to the field of agriculture, namely to production and storage of bacterial fertilizers and may be used at cultivation of leguminous grasses and other agricultural plants. The method of production and storage of bacterial fertilizers includes growing of the maternal crop, making of a working crop, after that it is necessary to conduct an inoculation of the prepared peat - a massive introduction of the liquid crop into the peat. The invention provides, that the organic carrier - transitional peat - is introduce with a water solution of sodium selenate so, that the final concentration of the sodium selenate makes 1·10^-4 g/kg of peat. The method allows to decrease significantly percentage of the loss of the bacterial cells on the carrier, to improve vitality and activity of the nitrogen fixers brought from a nutrient medium onto the organic carrier, and considerably to increase duration and a temperature range of storage of the bacterial preparation.

EFFECT: the invention ensures significantly decreased losses of the bacterial cells on the carrier, improved vitality and activity of the nitrogen fixers brought from a nutrient medium onto the organic carrier, increased duration and a temperature range of storage of the bacterial preparation.

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Description

The invention relates to agriculture and relates to the production and storage of bacterial fertilizers and can be used in the cultivation of legumes and other agricultural plants.

The unsolved problems of the production and practical use of bacterial preparations in the field are the relatively low reproduction rate of nodule bacteria and their low viability during storage and use. Phenolic substances contained in the main known carrier, peat, have an adverse effect on nodule bacteria, as a result of which the nitrogen fixation process is weakened.

It is known to use peat compost as an organic carrier, and the resulting mixture, inoculated with the bacterial component, is incubated with stirring for 3-5 days [RF patent No. 2084431 6 C 05 F 11/08, 1997. Yudkin L.Yu., Kovalev N. G., Khotanovich A.V., Temnova O.V. A method of obtaining a bacterial fertilizer]. The disadvantages of these methods are, accordingly, the narrowness of use and, in the latter case, the high complexity and energy costs.

A known method for producing biofertilizers, where dry granulated chicken droppings is used as an organic carrier [RF patent No. 2130005 C 05 F 11/08, 1999. Raymanov IT, Alimova F.K., Oziganova G.U., Khabibullin R. E., Krylova N.I., Fattakhova A.N. A method of obtaining biofertilizer]. The weak point of this method is the drying and immobilization of biomass on a dry organic carrier, which calls into question the shelf life of the fertilizer, and also, given the acid reaction of the carrier, requires pH adjustment, which implies additional manipulation. In addition, the indicated organic carrier in itself is a valuable organomineral fertilizer and is not always available for farms.

A known method of storing cultures of microorganisms, involving the use of an aqueous solution of sodium selenate [RF patent No. 2185435 C 12 N 1/04, 2002. Ilyin D.Yu., Blinohvatov AF, Ilyina GV, Ivanov AI A method of storing cultures of microorganisms]. However, this method involves the storage of crops on nutrient media enriched with minerals and sources of nitrogen and carbon, and in the case of storage of nitrogen-fixing media, transition peat is used only as a carrier, but not a nutrient medium. In addition, the volumes of the substrate for storing cultures of nitrogen-fixing bacteria are so significant that it is not always possible to use a refrigerator, which is recommended in the description of this method for storing small containers with medium inoculated with bacterial or fungal culture.

The closest in technological essence is the method of cultivation of nitrogen-fixing bacteria, involving the cultivation of the uterine culture of bacteria in an environment of the following composition: peas (decoction) 100.0 ml; glucose 10.0 g; (NH ₄ ) ₂ SO ₄ 1.0 g; KH ₂ PO ₄ · H ₂ O 0.5 g; K ₂ HPO ₄ 0.5 g; MgSO ₄ · 7H ₂ O 0.3 g; CaCO ₃ 1.0 g, then the manufacture of a working culture on a medium containing (g / l): glucose 20.0; corn extract 10.0; yeast extract 0.25; KH ₂ PO ₄ · H ₂ O 0.5; K ₂ HPO ₄ 0.5; MgSO ₄ · 7H ₂ O 0.3; CaCO ₃ 1,0, after which the implementation of the inoculation of prepared peat - a massive introduction of liquid culture into peat [Khotyanovich A.V. Cultivation methods of nitrogen-fixing bacteria, methods for producing and using preparations based on them (Methodical recommendations) Leningrad, 1991]. The disadvantages of this method are the risk of inhibition of culture when using salts of K, Mg, Ca; with violations in the aeration system and even with a slight change in the reaction of the environment, as well as a narrow range of storage temperatures of the crops.

The proposed method provides an increase in the viability and activity of the bacterial component of fertilizers on an organic carrier and an increase in the shelf life of the biological product while reducing material costs. This is achieved by the fact that during the cultivation of nitrogen-fixing bacteria in a way that includes growing the mother culture, then producing a working culture, after which the prepared peat is inoculated — the mass introduction of liquid culture into peat, according to the invention, an aqueous solution of sodium selenate is introduced into the organic carrier — transition peat. so that the final concentration of sodium selenate is 1 · 10 ^-4 g / kg of peat.

The trace element contained in the substance, selenium, is able to increase the activity of almost all parts of the antioxidant system of microorganisms, which limits vital activity, without negative impact on the enzymatic apparatus. This can significantly reduce the percentage of death of bacterial cells on the carrier, increase the viability and activity of nitrogen fixers transferred from the nutrient medium to an organic carrier, but also significantly increase the shelf life and temperature range of the bacterial preparation.

Another sign that distinguishes the proposed method from the known ones is that the use of ultra-low doses of selenium salt does not require significant economic costs, since the amount of the introduced substance (sodium selenate) used is very small.

Example.

For growing a uterine culture, a liquid nutrient medium of the following composition is prepared: peas (decoction) 100.0 ml; glucose 10.0 g; (NH ₄ ) ₂ SO ₄ 1.0 g; KH ₂ PO ₄ · H ₂ O 0.5 g; K ₂ HPO ₄ 0.5 g; MgSO ₄ · 7H ₂ O 0.3 g; CaCO ₃ 1.0 g, which is poured into 200 ml into 750 ml flasks and sterilized as usual. Then swabs from tubes with a strain of certain types of bacteria are placed in a liquid nutrient medium. Flasks are placed in a rocking chair for two days until they reach a certain titer of bacterial cells. The culture fluid from the flasks is then placed in working bottles with a capacity of 5 - 10 l, filled with nutrient medium, including glucose 20.0; corn extract 10.0; yeast extract 0.25; (NH ₄ ) ₂ SO ₄ 1.0; KH ₂ PO ₄ · H ₂ O 0.5; K ₂ HPO ₄ 0.5; MgSO ₄ · 7H ₂ O 0.3; CaCO ₃ 1,0, which are also sent for two days on a rocking chair. Thus, a concentrated suspension of bacterial cells is obtained, which is then introduced into an organic carrier. Sifted transition peat is used as a carrier substrate for finished fertilizers, the pH of which is brought to 7.2 units. To prepare the finished product for 60 kg of peat is taken: culture fluid (working culture) 8 l, molasses 3.6 kg. Humidity of the drug is 55-60%. The method of preparation is non-sterile.

To solve the problem at the time of preparation of the peat mixture, sodium selenate is added so that its final concentration is 1 · 10 ^-4 g / kg of substrate. Finished products are packed in plastic bags and bagging. The shelf life of the finished bacterial fertilizer according to the regulation is 4 months.

When the selenium compound was added to the organic carrier, peat, the control titer of the drug on the 10th day exceeded the control indices by 28.1-47.4% (Table 1), which indicates the doping of the protective systems of bacterial cells.

After the regulated shelf life of the bacterial preparations (4 months), the control samples showed a sharp decrease in the titer and, accordingly, the activity of the drug, however, in the experimental versions, on the enriched selenium organic carrier, the titer remained acceptable, even with significant fluctuations in storage temperatures (Table 2 )

The enrichment of the organic carrier for symbiotic nitrogen fixers with selenium salt stimulates the formation of nodules and the production of plant biomass, as shown in experiments on spring vetch and pea (Table 3.4).

Yields upon inoculation of pea seeds with rhizotorfin (control) increased by 0.28 t / ha, and rhizotorfin enriched with selenium increased by 0.40 t / ha (Table 5). Ultimately, the economic benefit of using bacterial fertilizers containing a low molecular weight compound of selenium in the organic carrier is reliable and amounts to 1331.01 rubles / ha of conditional net income.

On a selenium-enriched organic carrier, after a specified shelf life, the bacterium titer remains acceptable, even when storage temperature fluctuations are outside the range. The enrichment of the organic carrier for symbiotic nitrogen fixers with selenium salt stimulates the formation of nodules and the production of plant biomass.

The economic benefit of using bacterial fertilizers containing a low molecular weight compound of selenium as an organic carrier is reliable and amounts to 1331.01 rubles / ha of conditional net income.

Table 1 The titer of the drug on the 10th day after inoculation (billion cells in 1 g of the drug) A drug Transition peat Transitional peat + sodium selenate Agrofil 66.45 88.5 ± 3.4 Deviation from the control,% - 28.1-38.3 Risotorfin 6.85 9.8 ± 0.3 Deviation from the control,% - 35.7-47.4 table 2 The titer of preparations (billion cells in 1 g) after 4 months of storage at various temperatures on a carrier enriched with sodium selenate (10 ^-4 ) A drug Temperature +2 - + 5 ° С +20 - + 22 ° С Experience The control Experience The control Risotorfin 3.75 4.16 - 1.13 Rhizoagrin 3.48 3.75 - 1.24 Table 3 The number of nodules in legumes when applying bacterial fertilizers with enriched sodium selenate at a concentration of 10 ^-4 organic carrier, pcs. on 1 plant Type of plant, phase Experience Options Control (no processing) Risotorfin Risotorfin + Selenium Peas, flowering phase 90 130 170 bean phase 130 210 260 Spring vetch, flowering phase 120 170 230 bean phase 220 300 370

Claims (1)

A method of storing bacterial fertilizers, including growing a maternal culture, making a working culture, and then inoculating the prepared peat - massively introducing a liquid working culture into peat, characterized in that an aqueous solution of sodium selenate is introduced into the organic carrier - transition peat so that the final the concentration of sodium selenate was 1 · 10 ^-4 g / kg of peat.