The strain of the fungus trichoderma sp. mg-97 used to protect seedlings from coniferous fusariosis

 

(57) Abstract:

The invention relates to the field of Microbiology, relates to a new strain of Trichoderma sp. MG-97 and can be used to protect seedlings from coniferous fusariosis. Strain Trichoderma sp. MG-97 isolated from the soil of the nursery. Deposited in the collection of industrial microorganisms under F-765. The strain has a high antagonistic activity against phytopathogenic microorganisms of the genus Fusarium - pathogens lodging of coniferous seedlings. The strain of the fungus Trichoderma sp. MG-97 is used to protect seedlings from coniferous fusariosis. Feature of the strain is high competitiveness and growth rate. In addition, to isolate is characterized by a rapid accumulation of biomass and active sporulation. 6 Il.

The invention relates to the field of Microbiology, relates to a new strain of the fungus Trichoderma MG-97 and can be used to protect seedlings from coniferous fusariosis.

Fungi of the genus Trichoderma are used in the practice of plant protection more than 20 years. Available experimental data indicate a large set of strains used as byproduct in the production of trichodermin. However, the lack of a unified assessment of the pathogen complex of bpalogin [1, 2].

Institutions-developers: VNIIMS and Bellaisa recommended accordingly strains-producers of Trichoderma harzianum TF-17 and Trichoderma lignorum - 205-M-10, with a broad range of combat complex diseases of vegetable crops [3, 4].

Currently microbiological industry produced trichodermin used to protect vegetable crops in open and closed bottoms, and not production of trichodermin for the protection of coniferous seedlings against soil pathogens.

The technical result of the invention is a strain of the fungus Trichoderma sp. MG-97 VKPM F-765 with high antagonistic activity against phytopathogenic microorganisms of the genus Fusarium - pathogens lodging of coniferous seedlings.

The technical result is achieved by using a strain of the fungus Trichoderma sp. MG-97 VKPM F-765 for the protection of coniferous seedlings from fusariosis. Feature of the strain is high competitiveness and growth rate. In addition, to isolate is characterized by a rapid accumulation of biomass and active sporulation.

Offered the use of this strain as bioproducts for creation on its basis of environmentally friendly biological products.

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The strain is characterized by the following features:

Morphological characteristics: Forming a relatively slow-growing colonies that reach 2-3,5 cm in diameter over 4 days (at 20oC), the color of the aerial mycelium white to slightly greyish, at temperatures above 30oC the increase of substrate mycelium is slowing down, there is a sharp decline in the growth of the colonies and the fungus goes to the stage of conidiogenesis. Sporulation begins 5-7 days., abundant in 12-15 days (Fig. 1).

With the growth on potato-dextrose agar strain gives a dull yellow pigment (Fig. 2).

The isolate forms a transparent smooth hyphae up to 3-8 μm in diameter. Conidiophores are also transparent, thickened at the base, just branching. The size of 3-4 microns for the most part. At the base the size of the major axis to 6-8 microns in diameter. The primary processes are relatively equal in length, are placed in pairs at the same distance. The apex of the conidiophores have 2-4 process. Branches of conidiophores form whorls from 6-7 failed sometimes may be formed of a combination of failed and secondary processes (Fig. 3).

Conidia vary in shape from ovate to widely elliptic to slightly pointed grounds, the wifi of fialed are combined into a large slimy mass (Fig. 4).

Cultural characteristics:

Grows well in liquid synthetic medium of čapek; nutritionally dense environments: natural (wort-agar and potato-dextrose agar) and semi-synthetic malt-extract agar.

Chapek's medium (g/l): glucose - 30; NaNO3- 3; KH2PO4- 1; MgSO4to 0.5; KCl - 0,5; FeSO4- 0,01.

Wort agar: beer wort (9oB) -1 l; agar - 15 g

Malt - extract agar (g/l): malt extract Difco - 15; agar - 15 g

According to the experts of the Krasnoyarsk regional veterinary laboratory, this strain of avirulent, non-toxic, not toxicogenic in relation to warm-blooded organisms and does not cause infectious diseases.

In Fig. 1-6 presents the following picture:

Fig. 1 - sporulation of the fungus Trichoderma sp. strain MG-97 on malt-agar;

Fig. 2 - formation of diffuse pigment of the fungus Trichoderma sp. strain MG - 97;

Fig. 3 - structure conditoner forms of the fungus Trichoderma sp.strain MG-97;

Fig. 4 - the structure of the conidia of the fungus Trichoderma sp. strain MG-97;

Fig. 5 - the cultivation of the strain MG-97 on the environment Zakharchenko;

Fig. 6 - the antibiotic activity of the fungus Trichoderma sp. strain MG-97.

Implementation using Trichoderma sp. MG-97 as producer of bioprep cultivation involves several steps:

Inoculation of the fungus produce spore culture of conidia at the rate of 1 x 1071 ml of culture medium. As inoculum used spore culture of the strain grown on the shoals with wort-agar. From tubes do flush in flasks with sterile water and then determine the concentration of conidia in the suspension in the chamber Goryaeva or method brida; and the method of conversion for the entire volume of the nutrient medium using the required amount of suspension to be planted. Sowing produced in the Erlenmeyer flask with a volume of 250 ml per 100 ml of culture medium. For maximum sporulation growing produce on optimized medium Zakharchenko of the following composition (g/l): sucrose - 25,44; corn extract to 31.8; KH2PO4-3,14; MgSO4- to 0.645; cultivated strain for 15 days at 28oC (Fig.5).

After culturing filamentous film is separated from the nutrient medium, dried to a constant weight and ground in a homogenizer; in this form of biological product is missing nutrient medium on which the fungus was grown.

The resulting biological product is a dry powder dark green color, with a title dispute 2 x 1011spores/g

Example 2. Slice 5 and 10 days of cultivation take samples of the culture fluid, which is filtered through a membrane filter with a pore diameter of N 8. Then the culture fluid moistened with sterile paper disks with a diameter of 5 mm, Dried disks are laid on a nutrient medium in Petri dishes, pre-seeded "lawn" of test organisms: Fusarium avenaceum, Fusarium sporotrichiella v. poae, Fusarium momliforme v. lactis; Fusarium nivale, Fusarium gibbosum. Cup incubated in a thermostat at 26-28oC for 5-7 days. Control are paper discs moistened with sterile water.

The lack of growth of the test organism around the paper disk indicates the presence of the antibiotic properties of the fungus Trichoderma sp. MG-97. The ordinate axis lay a zone of no growth of the test organism in mm. So, all of the investigated pathogens Fusarium lodging of coniferous seedlings had medium and high sensitivity to the action of metabolites of strain (Fig. 6).

Thus, the strain Trichoderma sp. MG-97 can be used as a producer of a biological product of trichodermin for the biological control of pathogens Fusarium lodging of coniferous seedlings.

Sources of information

1. Tretyakov, A. P., Shcherbakov, I., Shirmanova N. And. Strain micromycete Trichoderma lignorum for the production of drug against phytopathogenic Gribov harzianum SK-55 fungus, fungicide containin it, and method of manufacture of the same and its use: Patent 5422107 USA, M. CI.6A 01 N 63/00, A 01 N 1/00. Hokkaido Green Kosan, Inc. N 172273; 23.12.1993; N 4-359484.

3. Grin'ko N. N. The use of trichodermin in the vegetable greenhouse. - Minsk: Bellamente. 1992. - 60 S.

4. Strains recommended for the production of drugs in biological laboratories / / / plant Protection. - 1989. -N 12. - S. 56-57.

The strain of the fungus Trichoderma sp. MG-97 VKPM F 765 (all-Russian collection of industrial microorganisms) used to protect seedlings from coniferous fusariosis.

 

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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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