The strain of the fungus entomophthora thaxteriana n19-11 (vizr) to obtain biological mihaylin t

 

(57) Abstract:

The invention relates to the field of agricultural Microbiology, and can be used to obtain drug mihaylin T to protect plants from aphids, spider mites and thrips. Strain Entomophthora thaxteriana 19-11 (VIZR) are obtained by stepwise selection of monocanalicular sieving natural isolate, isolated from peach aphid. The selection is carried out on the basis of coccinobaphi isolates of this fungus under cultivation in deep conditions. Strain Entomophthora thaxteriana deposited in the collection of pure cultures of their control under 19-11. The output of minoufiya T in strain is 2.5-2.6 g with 1 liter of nutrient medium consisting of 4% glucose, 1% soybean flour and water (the rest). The content of minoufiya T in biomass of 19.4-19.5 per cent. The use of a biological product can reduce the concentration of the working emulsion of the drug to 0.5%. Insecticidal activity 90-100% loss of pests of agricultural crops in the open and closed ground. table 2.

The invention relates to the field of agricultural Microbiology, and can be used to obtain drug mihaylin T to protect plants from aphids, spider mites and thrips.

As the closest analogue of the definition of substances basis of biologics, however, in the process of saving and multiple transfers this strain has lost its toxinogenesis properties and decreased its biological efficacy against insects.

The objective of the invention is to obtain a new strain of the fungus, with a higher (in comparison with the known) level of education of toxic substances and insecticidal activity against sucking pests.

Strain Entomophthora thaxteriana N 19-11 (VIZR) obtained by stepwise selection of monocanalicular sieving natural isolate, isolated from peach aphid. The selection carried out on the basis of coccinobaphi isolates of this fungus under cultivation in deep conditions. Strain Entomophthora thaxteriana deposited in the collection of pure cultures of their control under N 19-11. The output of minoufiya T in strain is 2.5 - 2.6 g with 1 liter of nutrient medium consisting of 4% glucose, 1% soybean flour and water (the rest). The content of minoufiya T in biomass of 19.4 - 19.5 per cent.

Morphological features

Vegetative mycelium of strain deceptively, colorless, disintegrating after 24 hours, short, straight or curved gehennae body. The formation of conidiophores with conidia occurs on the substrate m>Conidia are spherical with a papilla at the base contain a single large or several small fat inclusions.

The size of conidia 25 - 39 x 21 - 32 µm, on average, 32 x 26 μm. Conidia are single-layered cell wall, smooth surface.

The formation of resting spores begins to 66 hours of growth. Resting spores rounded with a double shell and large fat droplet in the center. The size of dormant spores 20 - 25 microns, an average of 22 μm. The surface of spores smooth, three-layered cell wall.

Cultural morphological characteristics of the strain studied in the wort (50% nikolenka beer wort), environments Saburo (1% peptone and 4% glucose) and soybean (1% soy flour and 4% glucose). In all these environments, the strain forms a flat, smooth and colourless colonies, which at the 24-hour lighting are covered with conidia, which gives the white colonies. The pigment in the environment is not allocated.

The nature of the surface of the smooth growth. The growth rate of the colony under standard conditions (pH 6.5 - 7.0 and temperature of cultivation of the 22oC) to the wort and the environment Saburo 3.0 mm/day, soy environment - 2.8 mm/day.

The output of minoufiya T for the strain of E. thaxteriana N 18 is of 4.99% or 0.35 g/l nutrient medium, respectively.

The effectiveness of drugs on larvae of the tobacco thrips were determined by the rapid method, similar to aphids. Treatment of spider mites is carried out by immersion of bean leaves with mites in 1% aqueous emulsion preparations. The sheets are then placed in a Petri dish filled with water to prevent rouzbehani ticks, the death of the account within 24 hours.

In tables 1, 2 shows insecticidal activity and release formulations of strains of the fungus E. thaxteriana N 18 and N 19-11.

Thus, the strain Entomophthora thaxteriana N 19-11 has a high level of coccinobaphi and biological efficiency, thereby reducing the concentration of the working emulsion preparation, obtained on its basis, up to 0.5% to achieve 90 - 100% loss of pests of agricultural crops in the open ground.

Color

On agarici eleesa. Well sorularda colony is white due to abundant white bloom of conidia.

Physiological signs

The pigment is not formed in the environment is not allocated. Of the carbon sources well metabolizes glucose, maltose, starch, fructose (which marked the best growth and sporulation). In the medium with amino acids can grow without carbon through the carbon part of the amino acids, however, weak growth and sporulation absent. Poorly growing and sporulate on media containing sucrose, lactose, galactose, arabinose, xylose and alcohols.

From sources of nitrogen assimilates a mixture of amino acids, peptides, proteins, can grow on media with asparagine. Nitrate nitrogen is not absorbed, on media with ammonium nitrogen - growth weak.

Temperature

Growth was observed in the temperature range + 3 - 36oC. the Temperature optimum for growth 27oC, for conidiophore 20 - 22oC.

Microaerophilic growth

The strain is a strict aerobe.

Check the viability of the strain is carried out by re-sowing of crops in the fresh wort agar, followed by cultivation in an incubator at 22oC.

Way to save

Duration of cultivation on KOs is activate your product

The strain can be stored on the shoals wort-agar at +4oC for 1 to 1.5 years without replanting. The state collection of microorganisms ibfm Russian Academy of Sciences, the strain is stored in liquid nitrogen for 3 years without loss of viability.

The presence of marker signs

The strain produces two types of endotoxins and has a selective toxic effect. Productivity E. thaxteriana N 19-11 on minoufiya T is 2.5 - 2.6 g/l environment, the insecticidal activity of 98 to 100% in different variants with the test insects. The level of coccinobaphi strain is a stable symptom: after 16 generations toxigenicity strain and efficiency of samples of drugs remains at the initial level.

Example. Strains of E. thaxteriana N 18 and N 19-11 grown on a medium containing 1% soy flour and 4% glucose, water - the rest (pH 6,5 - 7,0) at the 27oC for 3 days. Biomass is filtered off, dried at a temperature of 37oC (air-dry) and grind.

The selection of minoufiya T 1 g biomass spend ethanol for 60 minutes the resulting solution is separated by filtration from biomass and the alcohol is evaporated until its removal.

The output of minoufiya T for the strain of E. thaxteriana N 19-11 amounts to 19.4% or 2,53 g/l PA Microhydrin T against sucking insect pests and mites.

 

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FIELD: biotechnology.

SUBSTANCE: the present innovation deals with immunization in plants, particularly, compositions and methods for inducing plant resistance to phytopathogenic organisms, such as phytopathogenic fungi. An agent indicated for inducing plant resistance to phytopathogenic microorganisms is being an extract out of biomass of non-phytopathogenic microorganisms. The method to obtain the above-mentioned agent includes the following stages: a) resuspending against 50-200 g (dry weight) biomass of non-phytopathogenic microorganisms in 1 l either inorganic or organic solvent, b) mixing at room temperature for 1-12 h, c) incubating, d) resuspending, e) cooling up to room temperature at maturing and f) filtrating, not obligatory. The innovation enables to induce plant resistance to phytopathogenic microorganisms.

EFFECT: higher efficiency of immunization.

12 cl, 6 ex, 5 tbl

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