RU2284353C1 - Strain of microorganism bacillus mycoides var. b. a. for preparing biopreparation used in stimulation of plant growth and protection against diseases - Google Patents

Abstract

FIELD: agricultural biotechnology, microbiology.

SUBSTANCE: invention relates, in particular, to the strain of microorganism for preparing a biopreparation used in stimulation of growth and protection of plants against diseases. The strain of microorganism Bacillus mycoides var. B. A. is isolated in Volgogradskaya district from common millet rhizosphere zone. The strain is deposited in the collection of microorganism of All-Union scientific-research institute of agricultural microbiology (VNIISCHM) at № D138. The isolated strain shows expanded protective properties against phytopathogenic microflora and promotes to stimulation of plants growth.

EFFECT: valuable properties and enhanced effectiveness of strain.

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Description

The invention relates to agricultural biotechnology, namely to the production and use of biological agents to combat pathogens of plants and to stimulate their growth.

A known strain of Pseudomonas fluorescence biovar V MD-4f [1], which inhibits the propagation of fungi of the genus Streptomyces, which are the causative agents of potato scab. Seed potatoes are treated with a suspension of cells of this strain with a concentration of 10 ⁹ cells / ml.

The disadvantage of this strain is the limited scope (it acts only on phytopathogenic fungi of the genus Streptomyces) and requires a large consumption of suspension for seed treatment.

Also known is the strain Pseudomonas cepacia AMMA [2], which inhibits the growth of phytopathogenic fungi of the species Aphanomyces.

However, there is no information about its antagonistic activity against other phytopathogens.

The closest analogue to the claimed strain is a strain of Pseudomonas aureofaciens [3], which inhibits the growth of virulent agrobacteria that cause bacterial cancer of grapes and other plants. Mentioned strain deposited in VKPM under the number B-4117. Cells of this strain are grown for 24 hours at 30 ° C on Hottinger broth or potato agar to a cell titer of 10 ⁸ -10 ⁹ cells / ml.

The disadvantage of this strain is a narrow area of its antagonistic action, as well as the fact that it does not have properties to stimulate plant growth.

The aim of the invention is the selection of a natural strain of bacteria with an extended area of antagonistic action on pathogenic microflora with the possibility of stimulating plant growth.

Isolated strain of Bacillus mycoides var. B.A. possesses enhanced protective properties against phytopathogenic microflora, and also helps to stimulate plant growth in the composition of the biological preparation "Extragran".

The strain is deposited in the collection of microorganisms of the All-Russian Research Institute of Agricultural Microbiology (VNIISKHM) under the number D 138.

Strain Bacillus mycoides var. B.A. isolated in the Volgograd region from the rhizosphere zone of common millet (Panicum miliaceum L).

Cultural and morphological characters.

Morphologically, Bacillus mycoides var. B.A. identical to Bacillus cereus.

The form of colonies on agar is grayish, spreading, rhizoid.

On oblique agar - whitish, friable, brilliant, rhizoid with a plaque thickening in time.

Represents gram-positive sticks (0.8-1.0 × 2.0-4.0) microns, connected in long chains. On glucose broth, the sticks are much longer. They move slowly.

By the nature of the location of flagella - peritrichi.

The dispute is centrally located. Germination is polar.

Physico-biochemical properties.

Strain Bacillus mycoides var. B.A. - aerob.

The optimum growth and spore formation temperature is 28-30 ° С.

It assimilates sucrose, starch, nitrates.

It shows antagonism towards harmful objects Fusarium verticilium, Gelminto-sporium - forms zones on agar cultures.

The main difference is the size of the zones of suppression of growth of Fusarium on potato-sucrose agar.

The sizes of the zones against Fusarium reach 9-10 cm in Petri dishes.

The strain is phage resistant.

Attitude to antibiotics - resistant to chloramphenicol, penicillin, novobicin and sensitive to tetracycline and streptomycin.

The strain is not phytotoxic and harmless to humans and warm-blooded animals.

Storage: on dry perlite with chalk at a temperature of 5-6 ° C.

Growth conditions and composition:

When the bacteria multiply the strain of Bacillus mycoides var. B.A. used meat and peptone broth. 4 meat broth cubes were dissolved in 1 liter of hot water. The broth was cooled, fat was removed from the surface, and filtered off from mechanical particles through a cotton-gauze filter. A sample of peptone and sodium chloride was added to the prepared meat water, dissolved by heating, poured into a 1000 ml flask, the volume was adjusted to distillation with distilled water, a pH of 8.1-8.2 was adjusted with a 10% sodium hydroxide solution, and adjusted to a boil, which ensured precipitation. Then it was acidified with hydrochloric acid (10% solution) to a pH of 7.1-7.5, again boiled and filtered through a paper filter. The resulting meat and peptone broth was poured into sterile dishes and sterilized at 1 atm for 30 minutes. 0.5% glucose and 0.5% sucrose were added to the prepared meat and peptone broth. A solution of alkali was adjusted to pH 7.6-7.8, 2% agar was added, the agar was melted in a water bath, it was poured into sterile tubes and flasks and sterilized at a pressure of 0.8 atm for 30 minutes.

After sterilization, the pH of the medium should be 6.8-7.2.

Cultivation is carried out on a rocking chair or vibroreactor at a temperature of 28-30 ° C during the day.

On beet molasses Bacillus mycoides var. B.A. grown for 48 hours at a temperature of 28-30 ° C with the addition of chalk.

Studies have shown that the drug is based on a strain of Bacillus mycoides var. B.A. allows you to fight against diseases of cultivated plants such as fusarium, helminthosporiasis, powdery mildew, various bacterioses, gray rot, late blight, scab, root rot, etc.

The action of the drug based on the strain of Bacillus mycoides var. B.A. consists in expanding the scope, since it exhibits an antifugal effect against a larger list of pathogenic bacteria than the prototype.

In addition, it provides a stimulating effect on plant growth, since when using the drug, the absorption activity of plant roots increases and the synthesis of physiologically active substances and compounds that suppress the harmful effect of phytopathogenic microorganisms, in particular Erwinia Rhizoctonia, is enhanced.

Due to this, crop yields are increasing.

The use of such a drug is more economical than the use of mineral fertilizers.

Example 1. The study of the activity of the strain against pathogenic microorganisms.

Fusariuma suspension was added to potato-sucrose agar.

Agar was poured into Petri dishes. Wells were cut out in agar and a culture of Bacillus mycoides var. B.A. at different stages of development. After 48 hours, zones of growth inhibition of pathogenic bacteria Fusariuma appeared.

Example 2. Stimulation of plant growth.

The seeds of wheat, corn and other crops were kept in the culture fluid of Bacillus mycoides var. B.A. for 30 minutes, as well as in a control cup. Then the seeds were laid out on wet filter paper in Petri dishes and incubated for 48 hours at a temperature of 28-30 ° C. After 48 hours, the length of the sprouts was measured and compared with a control plate. The sprout length of the treated seeds exceeded the sprout length in the control cup by several percent.

The strain in the composition of the drug was used for pre-sowing treatment of seed and planting material, while in some cases high efficiency was revealed when spraying seedlings of crops.

Field tests showed that the strain, which is part of the biological product, improves the nutritional conditions of plants, synthesizes physiologically active substances, and also suppresses phytopathogenic bacterial and fungal microflora.

Example 3. Increasing the yield of sunflower seeds whose seeds were processed before sowing.

Culture - sunflower, variety - hybrid "Opera".

Place of testing - Volgograd region.

The yield on the plot treated with the drug with the strain is 17.6 c / ha. Productivity in the control plot is 16 c / ha. The difference in yield in the treated and control plots amounted to +1.6 c / ha.

Example 4. Increasing the yield of wheat, the seeds of which were processed before sowing.

Culture - winter wheat, grade "Zernogradka-9".

Place of testing - Volgograd region.

The yield on the plot treated with the drug with the strain is 38.2 c / ha.

Productivity at the control site is 34 c / ha. The difference in yield in the treated and control plots was +4.2 c / ha.

The strain also showed good properties during storage of vegetable products.

Potato tubers treated with the preparation under the same storage conditions as the control ones were stored longer and were not susceptible to diseases.

The claimed strain of Bacillus mycoides var. B.A. (VNIISKHM No. D 138) is new and meets the criterion of inventive step.

The above examples confirm the achievement of the technical result and the compliance of the claimed strain with the condition of industrial applicability.

Information sources

1. Japanese application No. 1-19320, M. Kl. A 01 T 63/00, published. 1989 year

2. US Patent No. 5,244,658, M. Cl. C 12 N 1/20, published. 1993 year

3. RF patent No. 1482188, M. Cl. C 12 N 1/20, published. 1994

Claims (1)

The microorganism strain Bacillus mycoides var. B.A. VNIISKHM No. D138 to obtain a biological product used to stimulate growth and protect plants from diseases.