KR100791983B1 - Microorganisms for the control of the genus Atsurobotris and plant parasitic nematodes comprising the same - Google Patents

Abstract

A novel Arthrobotrys oligospora KAN-2 strain is provided to secure strong adaptability to domestic soil, have excellent nematicide effect and nutritive growth effect of a plant and supply large amount of strain time appropriately, thereby being usefully used for controlling plant parasitic nematodes such as Meloidogyne species. An Arthrobotrys oligospora KAN-2 having the plant-parasitic nematodes preventive effect is deposited as a deposition no. KACC93052P. A microbial agent for controlling nematoda comprises the Arthrobotrys oligospora KAN-2 as an effective ingredient. A method for mass-producing the microbial agent comprises the steps of: (a) culturing the Arthrobotrys oligospora KAN-2 in a solid or a liquid culture medium; and (b) inoculating the cultured microorganism hypha into cooked barley, cooked oat or a mixture thereof and then culturing it.

Description

Microorganisms for controlling plant parasitic nematodes, including the Astrobotris microorganism and the like {Microorganism of Arthrobotrys sp. and Microbial Agent for Preventing Plant-parasitic Nematodes Comprising the Same}

1 to 4 are conidia photographs (FIG. 1) and conidiophore photographs (FIG. 1) of the nematode predatory fungus Arthrobotrys oligospora KAN-20 strain of the present invention, respectively (FIG. 2). ), A chlamydospore picture (FIG. 3) and a three-dimensional sticky net picture (FIG. 4), a nematode capture organ.

5 is a photograph of nematodes attached to a three-dimensional sticky net.

Figure 6 is a photograph showing that the nematode of nematode predatory fungi grew in the nematode body after the nematode was attached to the nematode trapping organ.

Figure 7 shows the image of the nematode trapping organs and the appearance of nematodes attached to the three-dimensional sticky net.

Figure 8 is a photograph showing the results of culturing on boiled barley and boiled oats strain Atsurobotris oligospor KAN-20 strain of the nematode predatory fungus of the present invention.

Figure 9 is a photograph showing the results of the mass culture of the microbial preparation for controlling plant parasitic nematodes of the present invention using a mushroom spawn disease.

The present invention relates to a novel microorganism, and more particularly, to the genus Astrobotris microorganisms and their use present in the soil exhibiting nematicidal effect on plant parasitic nematodes damaging plants.

Most of farmland in Korea is farming in the form of cultivation or facility house cultivation, and various obstacles or damages occur accordingly. Among them, damage by soil nematodes, which inhabit the soil and cause growth disorders based on plant roots, is caused annually. It is spreading and suffering huge economic losses.

Plant parasitic nematodes are small organisms in the soil and belong to the nematode, and most of them are very small and difficult to identify with the naked eye. Plant parasitic nematodes are generally thin in the shape of threads or fusiform, bilaterally symmetrical, the length of which is very small, 0.25 to 0.45 mm, no body, but wrinkles on the body surface. The body wall is largely composed of epidermis, dermis, and muscle, and the nervous system is composed of the central nervous system and the nerve rings around the esophagus. Breeding is the female adult lays eggs, sometimes through hermaphrodite or sympathetic reproduction. Plant parasitic nematodes, which hatch from eggs, grow into adult larvae after 4 stages of larval stage and survive for about 1 month, with characteristic development of the digestive and reproductive organs. Females have approximately 50 to 600 eggs during their survival. Lays These plant parasitic nematodes have a cough around the mouth and use them to puncture the roots of plants, absorb nutrients from plant tissues or invade into root tissues.

Plant parasitic nematodes are parasitic in all parts of the plant, including seeds, leaves, stems, and roots, and depending on the species, external parasitics affecting the plant from the outside of the plant, internal parasitics and invading into the plant tissue It only has a variety of lifestyles, such as half-parasitic parasites, which are inserted into plants to ingest nutrients. In addition, internal parasitic nematodes are those that are mobile and impaired, and those that are immobile that do not migrate once added.

Representative plant parasitic nematodes are Root-knot nematodes (Meloidogyne sp.). Root-knot nematodes mainly found in Korea include carrot root-knot nematodes (M. hapla), sweet potato root-knot nematodes (M. incognita), Java root-knot nematodes (M. javanica) and peanut root-knot nematodes (M. arenaria). Through the plant cultivation in the melon, cucumbers, tomatoes, watermelon, strawberries, peppers, ginseng, etc. are causing great damage. The damage caused by plant parasitic nematodes is minor when the density of nematodes in the soil is low, but as the growing period increases as the crop grows, host plants become fatally damaged or die. Plants infected with plant parasitic nematodes have reduced resistance to pathogens to encourage or aggravate the onset of pathogen invasion, and some nematodes may mediate phytopathogenic viruses. Most of the damage caused by plant parasitic nematodes occurs at the roots in the soil, and since the damage progresses slowly, other symptoms on the ground are not clear and it is often difficult to make an accurate diagnosis. Therefore, damage is difficult to identify before the plant is rooted out, so the damage is likely to proceed very seriously.

Various methods for alleviating the damage caused by these plant parasitic nematodes and controlling nematodes have been attempted.

The cultivation methods include fallow, non-cultivation, crop rotation, cropping adjustment, planting of resistant varieties and planting attracted plants that are not cultivated for a certain period of time. However, fallow cultivation is not realistic due to the nature of cultivation in Korea, and the immersion treatment method is not widely accepted due to the hassle of using the field as a field and then using it as a field again. In addition, while crop rotation has traditionally been used as a method of reducing damage caused by crop production, it is very limited to change crops to crops that guarantee high income and to select crops suitable for regional characteristics. There is no choice but to. In addition, it is not realistic to adjust the planting time as the fallow method because of the climate characteristic of Korea, where the four seasons are clear and the growing season is limited, and the same is true for planting manned plants. This cultivation control is only a mitigating and mitigating damage, rather than a fundamental control of plant parasitic nematodes.

As a physical method, hot water disinfection method, electrothermal heating method, ultraviolet irradiation and ultrasonic method have been tried. However, hot water disinfection method is mainly performed only in the summer when the temperature rises and the sun is strong, and the parasitic nematode escapes. Given this, there is a lack of confidence in the effectiveness of hot water disinfection. Electrothermal heating is a method of controlling plant parasitic nematodes by installing heating wires in the ground and using them to warm arable land. The above methods can not only be applied to the intermittent period when the crop is not cultivated, there is a disadvantage that excessively expensive facilities and maintenance.

Chemical control using pesticides is currently the most attempted, but agents used to control plant parasitic nematodes are usually organophosphate compounds that inhibit the activity of acetylcholine esterase. Insecticides work by increasing the amount of acetylcholine, a type of neurotransmitter. Most of these organophosphate insecticides are defined as hazardous substances. In addition, such pesticides indiscriminately kill not only plant parasitic nematodes but also crop symbiotic beneficial microorganisms present in the soil, causing salt accumulation, and eventually causing further damage to the soil. In addition, crops contaminated with residual pesticides pose a threat to the health of consumers, and at the same time, cause farmers to lose their crops and discard them.

As an alternative to solve this problem, a biological control method using natural microorganisms is proposed. Biological control methods for controlling plant parasitic nematodes include methods using predatory nematodes, predatory insects, mites, protozoa, bacteria, fungi, viruses, etc., but the most effective method is plant parasitic nematode predatory or nematode parasites. It's a fungus. The control method of the plant parasitic nematode using such a fungus is very easy to carry out, and has the advantage of long-term control effect through proper soil management. In addition, there is no problem of pesticides, growers' health, soil pollution and residual pesticides compared with chemical control methods, which is consistent with the reality of eco-friendly organic farming.

As part of this study, the overseas genus Atsurobotris (US Pat. Nos. 5,811,092, 4,666,714, 4,421,544; European Union Patent Nos. 0,623,284, SU 1,88,818, RU 2,106,092) and Tritothesi There were studies using microorganisms such as Um Sok (UK Patent No. 857,161) and Monacrossporium (French Patent No. 2,747,016). 0186866), the fungus of the genus Monacrossporium and the study of microbial agents using the same (Korean Patent Registration No. 10-0392852), and the like has been performed. However, since these microorganisms do not show an effective nematicidal effect in the soil to replace chemical pesticides, a new microorganism having a superior nematicidal effect is required.

Accordingly, the present inventors have newly isolated and identified nematode predatory microorganisms from domestic soil, and by showing that the microorganisms exhibit excellent control effects against plant parasitic nematodes, the present invention can be usefully used in the preparation of environmentally friendly microbial pesticides. Was completed.

It is an object of the present invention to provide a microorganism having a nematicidal effect on plant parasitic nematodes and a microbial preparation for controlling plant parasitic nematodes.

In order to achieve the above object, the present invention provides an Astrobotris microorganism having an nematode effect.

The present invention also provides a microbial preparation for nematode control containing the microorganism as an active ingredient.

Hereinafter, the present invention will be described in detail.

The present invention provides an Astrobotris microorganism having an nematode effect.

The microorganisms of the present invention were isolated from the melon and tomato cultivated soil into fungi that act as nematode predators. It was confirmed that. The microbial strain of the present invention exhibits superior growth and capture capacities than other strains of the genus Astrobotris.

In the Astrobotris nematode predatory fungal strain of the present invention, the development of hyphae and the formation of traps during the development cycle of the fungus, the secretion of attractant and the production of adhesive material, the generation of conidia and the dormant spores To form a mechanism of predatory behavior for soil nematodes. In the early stages of the predation process, the target organisms, such as nematodes, are attracted by the interaction between the attractant substance produced by the mycelium and the substance secreted by the nematode, and after the nematode is captured by the capture organ, the fungal mycelium penetrates the epidermis It destroys the plasma membrane of nematode cells and produces conidia and dormant spores with its nourishment. The viability of the nematode premature fungi in the soil and the production capacity of conidia and dormant spores enable the mass cultivation of the microbial strain of the present invention, and when the selected medium conditions are used, it shows more vigorous growth ability.

As a method for screening and culturing the microbial strain of the present invention, after separating the plant parasitic nematode inhabiting the soil using a suitable means such as a funnel, the soil sample is a known method (for example, FF Soprunov, 1958.Predacious Hyphomycetes and their application in the control of pathogenic nematodes.Academy of Science of the Turkmen SSR.Ashkabad (translated in 1966 by the Israel Program for Scientific translation, Jerusalem) .p. 1-365) Place and put the soil nematode isolated from the above plate and culture method can be used. The production of conidia was confirmed under a stereoscopic microscope, and pure strains were isolated by subcultured five or more times in an appropriate medium (see FIGS. 1 to 4).

Colonies formed by pure culture of isolated microbial strains are white or flesh-colored, and mycelia are colorless and transparent. The microbial strain of the present invention forms a large number of conidia spores, which is formed upright from the medium and forms a plurality of conidia at the end of the conidia. Conidia are egg-shaped and have a slightly recessed diaphragm. The conidia were 15.19 to 32.55 (24.56) × 8.68 to 15.19 (12.04) μm, and there was one diaphragm, located horizontally in one-third of the spores, dividing the conidia into two cells (Tables 1 through Table). 6). When the nematode is administered to the medium, the microbial strain of the present invention forms a three dimensional adhesive net in 1 to 2 days (see FIG. 5).

As a result of confirming the registration status of the strain bank by analyzing the ITS and 18S rDNA for the isolated strain, the microbial strain of the present invention was identified as a variety of microorganisms belonging to the genus Astrobotris. We isolated the isolated strains of A. sinensis KAN-2, Astrobotris Shinenssis KAN-4, Astrobotris Shinenssis KAN-11, A. oligospora (A. oligospora) ) KAN-9, A. musiformis KAN-12, Astrobotris Oligospora KAN-13, Astrobotris Oligospora KAN-20 and Astrobotris Oligospora KAN-21 Among them, KAN-2, KAN-9, KAN-13, KAN-12, and KAN-20 strains, which have excellent predatory ability against plant parasitic nematodes, were identified as the Korean Agricultural Microbiological Resources Center (KACC) on December 26, 2006. Korean Agricultural Culture Collection) (accession numbers; KACC93048P, KACC93049P, KACC93050P, KACC93051P, KACC93052P, respectively).

The present invention also provides a microbial preparation for nematode control containing at least one of the microorganisms as an active ingredient.

The microbial preparations of the present invention may be prepared by the use of surfactants and / or extenders which can be used as biochemical pesticides for the purpose of stable formulations suitable for use in actual packaging of hydrous, granular, liquid, emulsifying, water soluble, water soluble or encapsulating agents. It may be prepared in the form.

In the microbial preparation of the present invention, the microorganism may be supplied as contained in the nematode control formulation, or may be stored separately for long-term storage and mixed before use. When the microorganisms are stored separately for a long time and then supplied separately, the microorganisms may be stored in a glycerol storage solution at -70 ° C. or lower, or lyophilized at −20 to −80 ° C.

Hydration, which is one form of the microbial preparation of the present invention, may be prepared by drying and grinding a solid medium inoculated with microorganisms, followed by mixing by adding a surfactant and an extender / nutrient. In addition, the granules of the present invention may be prepared by drying and grinding a solid medium inoculated with microorganisms and then adding a surfactant, an extender, and a disintegrant.

In the above, as the surfactant, alkyl (C ₈ -C ₁₂ ) arylsulfonate, dialkyl (C ₃ -C ₆ ) arylsulfonate, dialkyl (C ₈ -C ₁₂ ) sulfosuccinate, lignin sulfonate, Sodium salts of sulfonate compounds such as naphthalenesulfonate condensates, naphthalenesulfonate formalin condensates, alkyl (C ₈ -C ₁₂ ) naphthalenesulfonate formalin condensates, and polyoxyethylene alkyl (C ₈ -C ₁₂ ) phenylsulfonates Or calcium salt, alkyl (C ₈ -C ₁₂ ) sulfate, alkyl (C ₈ -C ₁₂ ) aryl sulfate, polyoxyethylene alkyl (C ₈ -C ₁₂ ) sulfate, polyoxyethylene alkyl (C ₈ -C ₁₂ ) phenyl Sodium salts or calcium salts of sulfate compounds such as sulfates, anionic surfactants such as sodium salts or calcium salts of succinate compounds such as polyoxyalkylene succinates, sodium benzoates, alkylcarboxylates, and polyoxyethylene alkyls ( C _12- C ₁₈ ) ether, poly With nonionic surfactants such as oxyethylene alkyl (C ₈ -C ₁₂ ) phenyl ethers, polyoxyethylene alkyl (C ₈ -C ₁₂ ) phenyl polymers, ethylene oxide propylene oxide copolymers, polycarboxylates, tritons 100 and tween 80 One or two or more selected from the group consisting of may be used in combination, and other surfactants may be used. It will be readily understood by those skilled in the art. Extenders include bentonite, talc, clay, kaolin, calcium carbonate, silica sand, pumice, diatomaceous earth, acidic clay, zeolite, pearlite, white carbon, ammonium sulfate, urea, glucose, dextrin, soy flour, rice, wheat It may be easily understood by those skilled in the art that ocher, glucose and starch, water, and the like may be used alone or in combination of two or more thereof, and other extenders may be used. will be. As the disintegrant, one or more selected from the group consisting of bentonite, talc, dialite, kaolin and calcium carbonate can be used.

In addition, the granules of the present invention are one or two selected from the group consisting of surface active agents, inert carriers, preservatives, wetting agents, feed accelerators, attractants, encapsulating agents, binders, emulsifiers, dyes, UV protectors, buffers and flow agents for the microorganism. It may be prepared by further adding the above.

In applying the microbial preparation of the present invention, a method of intensively spraying the microbial preparation on the leaves, stems or roots of the plant or the soil adjacent thereto may be used, wherein the amount of the microorganism is 1 of the soil volume per cultivated land area (m 2). It is desirable to spray the% drug. On the other hand, 1 g per 1.0 × 10 ⁶ to 1.0 × 10 1.0 × 10 per ⁷ and a solution diluted with a biomass concentration plant or in a direct spray in the adjacent soil or, 1 g of ⁶ to 1.0 × 10 ⁷ microorganisms It is also possible to immerse plants for 1-2 hours in dilute solutions.

The microbial preparations of the present invention can be used for controlling plant parasitic nematodes. The plant parasitic nematodes include Root-knot nematodes (Meloidogyne species), Root-lesion nematodes (Pratylenchus Species), Cyst Nematodes (Heterodera species), and the like. Carrot root knot nematode, sweet potato root knot nematode, Java root knot nematode, peanut root knot nematode.

In addition, the present invention comprises the steps of culturing the Astrobotris genus in a solid or liquid medium; And inoculating the cultured microbial mycelium on boiled barley, boiled oats, or a mixture thereof to provide a method for mass-producing a microbial agent for controlling nematodes, or the nematode, which contains the same.

CMA II medium (corn meal agar; cornmeal 30 g / l, glucose 10 g / l, peptone 5 g / l, NaCl 5 g / l, CaCO ₃ 0.5 g / l, agar 15 g / l) As the liquid medium, CMII medium (cornmeal 30 g / l, glucose 10 g / l, peptone 5 g / l, NaCl 5 g / l, CaCO ₃ 0.5 g / l) is preferably used, but is not limited thereto. It is not. When the microorganism of the present invention and a microbial preparation including the same, which are mass-cultured and manufactured by the above process, are sprayed on the soil, the Astrobotris microorganism of the present invention is optimally grown using barley and boiled oats in the soil as nutrients. In addition, by effectively feeding the plant parasitic nematodes in the soil has the effect of controlling the plant from the plant parasitic nematodes.

EMBODIMENT OF THE INVENTION Hereinafter, this invention is demonstrated in detail to an Example.

However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited by the following examples.

Example  From soil eelworm  Isolation of Predatory Fungi

<1-1> eelworm  Collection of Soil Samples for Predatory Fungal Separation

Soil samples were collected from the natural habitats of fields and fungi in Okcheon, Chungcheongnam-do, Nonsan, and Daejeon Metropolitan City.

<1-2> from soil Nematode  detach

Free-living nematodes in the soil were separated by sugar gradient centrifugation and beer-man funnel separation (Pramer, D. and E.L. Schmidt. 1964. Experimental soil microbiology. Burgess. Minneapolis. P. 20-21). The isolated nematode was placed in sterile water and refrigerated at 4 ° C. In addition, after harvesting the soil and roots from the field affected by root-knot nematode damage, the root-knot nematode 2 nymphs were separated from the soil using a hollow but funnel method, and the root-knot nematode females were separated from the roots using an anatomical microscope. The isolated nematodes were used to isolate the nematode predatory fungi and to capture the nematode predatory fungi.

<1-3> eelworm  Isolation of Predatory Fungi

Nematode prey fungi were isolated using a soil sample filtered to 2 mm. The filtered soil samples were placed on 2% Water Agar medium and incubated at 25 ° C. for 3 days. When the nematode predatory fungi were not detected while using a dissecting microscope, the nematodes isolated in Example <1-2> were further inoculated, and observed by dissecting microscope for 4 to 14 days while incubating at 25 ° C. Nematodes were fed and grown conidia of fungi were transplanted into sterile CMA medium using sterile needles and incubated at 25 ° C. As a result, 8 nematode predatory fungi strains were isolated purely and named as KAN-2, KAN-4, KAN-11, KAN-9, KAN-13 KAN-20, KAN-21 and KAN-12 strains, respectively. (See FIGS. 1-4).

<1-4> purely separated eelworm  Cultivation and Preservation of Predatory Fungi

Purely isolated nematode prey fungi were inoculated in CMA medium (cornmeal 30 g / L, agar 15 g / L) and CMAII medium and cultured for one week at 25 ° C., followed by subculture. Pure fungus was incubated for one week using CMA slope medium and CMAⅡ slope medium, and then paraffin oil was refrigerated and stored refrigerated for long-term storage.

Example  2. eelworm  Morphological features of predatory fungi

The colonies grown well in CMA medium in sterile 2% Water Agar medium cut to 10 mm in width and placed in the center of the plate and incubated for one week at 25 ° C, followed by conidiophore formed by administering nematodes to the plate. Conidia and the like were observed using a microscope.

As a result, colonies of KAN-2, KAN-4 and KAN-11 strains were white or flesh-colored, and mycelia were colorless or transparent. Conidia were formed upright from the medium, branched, branched, up to nine conidia. In addition, circular dormant spores were formed on the medium. Conidia had a rugby ball or fusiform shape and formed three septums. The conidia were 28.21 to 45.57 (36.48) x 17.36 to 21.70 (19.64) 탆, and the diaphragm consisted of two at the base and one at the terminal. The conidia were 141.12-388.08 μm in size, 4.34-66.5 μm in basal length and 2.17-44.3 μm in distal end, and did not form branches and no conidia node. The size of mycelia was 4.34-10.85 micrometers, and the size of dormant spores was 13.02-21.7 micrometers (Table 1).

Morphological Features of KAN-2, KAN-4, and KAN-11 spawn  diameter  4.34 to 10.85 μm  A vein  Fleece  Color  White Conidia  shape  Branched, nonconidia  Length  141.12-388.08 μm  diameter  Base  4.34∼6.51 μm  Distal end  2.17 to 4.34 μm  Distal shape  Blunt Conidia  shape  Rugby ball or fusiform, spindle  size  28.21 to 45.57 (36.48) x 17.36 to 21.70 (19.64) μm  Diaphragm  Mainly three  Diaphragm position  2 bases (5%, 12%), 1 distal end (85%)  Count  1 to 9 Dormant spores  size  13.02 to 21.7 μm Predator  Three dimensional sticky net

Colonies of KAN-9, KAN-13, KAN-20, KAN-21 strains were white or flesh-colored, and mycelia were colorless or transparent. The conidia were erected from the medium, non-branched, conidia were formed, and many conidia were formed. The conidia had the shape of honey egg or pear, and formed one diaphragm. The conidia were 15.19-34.72 (24.87) x 8.68-15.19 (12.37) 탆, and the diaphragm was formed at the 1/3 position. The conidia had a size of 185.22 to 493.92 µm, a base of 5.425 to 10.85 µm and a terminal of 2.17 to 6.51 µm. Mycelial size ranged from 2.17 to 10.85 μm, and no dormant spores were observed on the medium (Table 2).

Morphological Features of KAN-9, KAN-13, KAN-20 and KAN-21 spawn  diameter  2.17 to 10.85 μm  A vein  Fleece  Color  White Conidia  shape  Unbranched, large numbers of conidia  Length  185.22 to 493.92 μm  diameter  Base  5.425-10.85 μm  Distal end  2.17 to 6.51 μm  Distal shape  Swollen Conidia  shape  Honey Egg, Pear Shape  size  15.19 to 34.72 (24.87) x 8.68 to 15.19 (12.37) μm  Diaphragm  One  Diaphragm position  1/3  Count  5-30 Predator  Three dimensional sticky net

Colonies of KAN-12 strains were white or fleshy, and mycelia were colorless or transparent. Conidia were erected from the medium, branched, and conidia did not form. The conidia had an elongate-ellipsoid and formed one septum. The size of the conidia was 28.21 to 41.23 (32.77) x 8.68 to 13.02 (9.55) 탆, and the diaphragm was located at the center to divide the conidia into two cells. The conidia had a size of 335.16 to 643.86 µm, a base of 6.51 to 10.85 µm and a terminal of 2.17 to 4.34 µm. The mycelia were 2.17-8.68 mu m in size, and formed circular dormant spores on the medium, and the size was 13.02-21.7 mu m (Table 3).

Morphological features of KAN-12 spawn  diameter  2.17 to 8.68 μm  A vein  Fleece  Color  White Conidia  shape  Branched, nonconidia  Length  335.16 ~ 643.86 ㎛  diameter  Base  6.51 to 10.85 μm  Distal end  2.17 to 4.34 μm  Distal shape  Coral Conidia  shape  Elongate-ellipsoid  size  28.21 to 41.23 (32.77) x 8.68 to 13.02 (9.55) 탆  Diaphragm  One  Diaphragm position  Centrally located  Count  10 things Dormant spores  size  13.02 to 21.7 μm Predator  Two dimensional sticky net

The KAN-2, KAN-4, KAN-11, KAN-12, KAN-9, KAN-13, KAN-20 and KAN-21 strains were grown in white mycelium like wool in CMA medium, and many congenital Spores formed. The eight strains formed a three-dimensional sticky net or a two-dimensional sticky net as an organ for trapping nematodes. KAN-2, KAN-4, KAN-11 and KAN-12 did not form conidia nodes, while KAN-9, KAN-13, KAN-20 and KAN-21 formed a large number of conidia nodes. All 8 strains were conidia formed upright from the medium. KAN-2, KAN-4, KAN-11 and KAN-12 strains were observed to form conidia spores. The tops of conidia spores of KAN-2, KAN-4 and KAN-11 strains were blunt, and the tops of conidia spores of KAN-12 were coral-shaped. The conidia of the conidia were swelled in shape (Tables 1 to 3).

The conidia of KAN-2 had a rugby ball shape or spindle shape and usually had three diaphragms. The conidia were 28.21-45.57 (36.48) x 17.36-21.70 (19.64) [mu] m, with two of the three septa located at the base of the conidia and one at the end. KAN-4 conidia were rugby ball-shaped or fusiform, usually three septa. Conidia were 32.55-47.74 (40.58) x 15.19-21.70 (19.49) [mu] m, with two of the three septa located at the base of the conidia and one at the end. The conidia of KAN-11 were rugby or fusiform, usually three diaphragms. The conidia were 32.55-45.57 (39.17) x 17.36-23.87 (20.85) [mu] m, with two of the three septa located at the base of the conidia and one at the end. The conidia of KAN-12 had one diaphragm in the form of a banana. The diaphragm was located in the central part of the conidia, dividing the conidia into two cells. The conidia were 28.21 to 41.23 (32.77) x 8.68 to 13.02 (9.55) 탆. The conidia of KAN-9 had one diaphragm in the shape of an inverted egg or pear. The diaphragm was located about one third of the conidia, dividing the conidia into two cells. The conidia were 21.70 to 34.72 (24.87) x 8.68 to 13.02 (11.98) 탆. The conidia of KAN-13 had one diaphragm in the shape of an inverted oval or pear. The diaphragm was located about one third of the conidia, dividing the conidia into two cells. The conidia were 17.36-34.72 (23.22) x 8.68-15.19 (12.37) 탆, slightly longer than KAN-9. The conidia of KAN-20 had one diaphragm in an inverted egg or pear shape. The diaphragm was located about one third of the conidia, dividing the conidia into two cells. The conidia were 15.19-32.55 (24.56) x 8.68-15.19 (12.04) 탆, similar to those of KAN-9. The conidia of KAN-21 had one diaphragm in inverted egg-shaped or pear-shaped. The diaphragm was located about one third of the conidia, dividing the conidia into two cells. The conidia were 17.36 to 28.21 (22.50) x 8.68 to 13.02 (11.44) μm, similar to those of KAN-9 and KAN-13 (Tables 4 to 6).

KAN-2, KAN-4, KAN-11 and KAN-12 formed circular dormant spores in V8Agar medium (V8 vegetable juice 100 ml / l, Campbell Soup Company). The size of dormant spores was 13.02-21.7 micrometers.

The length of conidia, the diameter of the basal and distal ends, the diameter of the hyphae, and the form of conidia Strain Conidia Base Distal end spawn shape KAN-2 167.58-388.08 4.34-6.51 2.17 to 3.255 4.34-10.85 Branched. No spores KAN-4 141.12-352.8 4.34-6.51 2.17 to 3.255 4.34∼9.77 Branched. No spores KAN-11 229.32-343.98 4.34-6.51 2.17 to 4.34 4.34-10.85 Branched. No spores KAN-12 335.16-643.86 6.51-10.85 2.17 to 4.34 2.17-8.68 Branched. No spores KAN-9 185.22-444 6.51 to 9.765 3.255 ~ 4.34 2.17-10.85 Not branched. With spores KAN-13 238.14-444 6.51-10.85 2.17 to 6.51 2.17-10.85 Not branched. With spores KAN-20 211.68-493.92 5.425-10.85 3.255 to 6.51 2.17-10.85 Not branched. With spores KAN-21 264.6 ~ 441 6.51-10.85 3.255 to 6.51 2.17-10.85 Not branched. With spores

Conidia size and morphology, number of conidia formed and number of diaphragms in μm Strain size Diaphragm shape Number KAN-2 28.21 to 45.57 (36.48) x 17.36 to 21.70 (19.64) 3 Fusiform 1 to 9 KAN-4 32.55 to 47.74 (40.58) x 15.19 to 21.70 (19.49) 3 Fusiform 1 to 9 KAN-11 32.55 to 45.57 (39.17) x 17.36 to 23.87 (20.85) 3 Fusiform 1 to 9 KAN-12 28.21 to 41.23 (32.77) x 8.68 to 13.02 (9.55) One Elongated oval 10 KAN-9 21.70 to 34.72 (24.87) x 8.68 to 13.02 (11.98) One Honey egg or Western type 5-30 KAN-13 17.36 to 34.72 (23.22) x 8.68 to 15.19 (12.37) One Honey egg or Western type 5 to 29 KAN-20 15.19 to 32.55 (24.56) x 8.68 to 15.19 (12.04) One Honey egg or Western type 5-30 KAN-21 17.36 to 28.21 (22.50) x 8.68 to 13.02 (11.44) One Honey egg or Western type 5 to 27

Strain Each size of mold structure (㎛) Conidia Conidia spawn KAN-2 36.48 × 19.64 167.58-388.08 4.34-10.85 KAN-4 40.58 × 19.49 141.12-352.80 4.34∼9.77 KAN-11 39.17 × 20.85 229.32-343.98 4.34-10.85 KAN-12 32.77 × 9.55 335.16-643.86 2.17-8.68 KAN-9 24.87 × 11.98 185.22-441.00 2.17-10.85 KAN-13 23.22 × 12.37 238.14-441.00 2.17-10.85 KAN-20 24.56 × 12.04 211.68-493.92 2.17-10.85 KAN-21 22.50 × 11.44 264.60-441.00 2.17-10.85

Example  3. eelworm  Sequencing and Identification of Predatory Fungi

<3-1> ITS analysis

<3-1-1> DNA Isolation and PCR Amplification

ITS sequences between 18S-5.8S-26S were analyzed. Nematode predatory fungi were inoculated in PDA slant and incubated at 25 ° C. for 3 days, followed by inoculation with PD (potato dextrose) medium, followed by shaking culture at 130 ° C. for 2 days at 25 ° C. to obtain mycelia. The mycelium was lyophilized, put into 1.5 ml tubes, crushed, suspended in 400 μl of extraction buffer (200 mM Tris-HCl, pH 8.0), 200 mM NaCl, 30 mM EDTA, 0.5% SDS, and proteinase ( 50 μg of proteinase) K was added and reacted at 37 ° C. for 1 hour. 400 μl of 2 × CTAB solution (2% CTAB (w / v), 100 mM Tris-HCl (pH 8.0), 20 mM EDTA (pH 8.0), 1.4M NaCl, 1% polyvinylpyrrolidone (PVP) Mr 40,000) After adding the mixture, the mixture was extracted with chloroform: isoamylalcohol (24: 1), and 0.7 vol. Of isopropanol was added to the supernatant obtained by centrifugation to obtain DNA. DNA was dissolved in TE buffer (Tris-HCl (pH 8.0) 100 mM, EDTA (pH 8.0) 1 mM), RNase was added, quantified and stored at 4 ° C.

The ITS region of rDNA was amplified using the ITS1 primer set forth in SEQ ID NO: 1 and the ITS4 primer set forth in SEQ ID NO: 2. Amplification was performed with 15 ng of genomic DNA, 0.5 pmol of primer each, 200 μM dNTP, 2 units of Taq DNA polymerase, 10 mM Tris-HCl, 50 mM KCl, 1.5 mM MgCl ₂ was added by adding H ₂ O to a final volume of 50 μl. DNA amplification was performed for 1 minute at initial denaturation at 94 ° C., denaturation at 94 ° C. for 40 seconds, annealing at 60 ° C. for 1 minute, and extension at 72 ° C. for 1 minute. The process was repeated and finally extended at 72 ° C. for 5 minutes.

<3-1-2> Sequence Analysis and Systematic Analysis

PCR amplified ITS region is Wizard  Purified with PCR Preps DNA Purification System (Promega), sequencing was performed using BigDye ^™ terminator cycle sequencing ready reaction kit (Perkin-Elmer Co.), 40 ng of template, 3.2 pmol primer, terminator ready reaction mixture ) 8 μl of H ₂ O was added to adjust the volume to 20 μl, followed by repeated PCR reactions 25 times at 96 ° C. for 10 seconds, 50 ° C. for 5 seconds, and 60 ° C. for 4 minutes. At this time, the same primers used for the PCR reaction were used for the reaction. Then, in gel development and sequencing, the nucleotide sequence was determined using an ABI Prism 310 Genetic Analyzer (PE Applied Biosystem).

As a result, the KAN-2, KAN-4, KAN-11, KAN-9, KAN-13 KAN-20, KAN-21 and KAN-12 strains each have an ITS sequence as shown in SEQ ID NO: 3 to SEQ ID NO: 10 It was confirmed that there is.

<3-2> 18S rDNA analysis

The base sequence was determined in the same manner as in the ITS sequencing of Example <3-1>, except that the primer of NS1 of SEQ ID NO: 11 and the NS2 primer of SEQ ID NO: 12 were used as the primer.

As a result, the KAN-2, KAN-4, KAN-11, KAN-9, KAN-13 KAN-20, KAN-21, and KAN-12 strains each contained an 18S rDNA sequence set forth in SEQ ID NO: 13 to SEQ ID NO: 20. It confirmed that it has.

<3-3> Identification of Isolated Strains

Strains were identified based on the results of ITS and 18S rDNA analysis and morphological characteristics of each strain.

As a result, the KAN-2, KAN-4 and KAN-11 strains were Atsurobotris senensis, KAN-9, KAN-13, KAN-20 and KAN-21 strains were Astrobotris oligospora and KAN-12 strains. Was identified as belonging to the Astrobotris city. Accordingly, the present inventors disregard each strain of Astrobotris Shinensis KAN-2, Astrobotris Shinensis KAN-4, Astrobotris Shinensis KAN-11, Astrobotris Oligospora KAN-9, Astrobotris Formics KAN-12, Atsubotris oligospora KAN-13, Atsubotris oligospora KAN-20, and Atsubotris oligospora KAN-21, among them excellent predators for plant parasitic nematodes. KAN-2, KAN-9, KAN-13, KAN-12, and KAN-20 strains were deposited with the Korea Agricultural Microbiological Resource Center on December 26, 2006 (Accession Number; ).

Example  4. eelworm  Predatory fungus eelworm  Predatory capacity comparison

<4-1> Nematode Prediction Test Indoors

Free life nematodes, root-knot nematodes and insect pathogenic nematodes were used to compare the nematode preservation ability of the microorganisms of the present invention. Nematode premature fungus plate cultured in 2% Water Agar medium for 1 week and free life nematode isolated from Example <1-2> Pathogenic nematodes were administered to each of 200 animals in one plate. Observed by anatomical microscope, the number of live nematodes was examined to select strains with excellent nematode preservation ability.

In order to test the nematode attraction and predation effect of nematode predatory fungi, root-knot nematode suspension, entomopathogenic nematode suspension and free-living nematode suspension were respectively administered to a 5 cm diameter plate, and the colonies grown on CMA medium were cut into 3 mm diameter circles. Placed in the center of the plate. After 3 hours and 6 hours, the attracting effect was confirmed according to the number of nematodes collected in the colonies, and the predation effect was evaluated according to the number of nematodes (% from control) which died after one or two days.

As a result, all of the microbial strains of the present invention had excellent nematode attraction and capture, and among them, the Astrobotris oligospora KAN-20 strain was selected as the best strain (Table 7).

Strain Root-knot nematodes Insect Pathogenic Nematodes Free life nematodes Attraction effect Capture Effect (%) Attraction effect Capture Effect (%) Attraction effect Capture Effect (%) KAN-9 +++ 90 + 50 ++++ 90 KAN-12 ++ 70 + 40 +++ 80 KAN-13 +++ 90 + 50 ++++ 90 KAN -20 +++ 90 + 50 ++++ 90 KAN-21 +++ 90 + 50 ++++ 90 KAN-2 ++ 80 + 50 +++ 80 KAN-4 ++ 80 + 50 +++ 80 KAN-11 +++ 80 + 50 +++ 80

<4-2> Nematode Prediction Test at the Port

Seedlings were grown from the small pot containing the tomato to the two to three leaf stages (20 cm). 800 g of sterilized soil was placed in a pot of 12 cm × 9 cm × 11.5 cm, and tomato-controlled control 1, tomato and sweet potato root-knot nematode control 2 and tomato and 4,000 eggs of sweet potato root-knot nematodes and 1% (8 g) of nematode-causing fungal drugs were divided into experimental groups. While the pot was incubated for one month in a greenhouse maintained at 25 to 30 ° C., the growth size (length) of the tomatoes was measured, and the number of leaves, flower differentiation, and fruit number were examined. After one month, the stems were cut and the roots were thoroughly washed, and the roots were stained with phloxine B (Sigma Chemical Company; P2759) to investigate the number of root knots and the number of ovules.

As a result, all of the strains of the present invention showed superior nematode preservation ability, and among them, the Astrobotris oligospora KAN-20 strain was selected as the most excellent strain by reducing the number of root nodules and egg sacs most. (Table 8).

Strain Root nods Reduction Number of ovum Reduction Growth length Increase Number % ^* Number % ^* Mm % ^* KAN-9 3.6 72.0 28.0 3.0 125.0 0.0 652.0 172.5 72.5 KAN12 6.0 120.0 0.0 4.2 175.0 0.0 618.0 163.5 63.5 KAN-13 2.8 56.0 44.0 2.8 116.7 0.0 637.0 168.5 68.5 KAN -20 2.1 42.0 58.0 0.7 29.2 70.8 633.0 167.5 67.5 KAN-21 3.2 64.0 36.0 2.0 83.3 16.7 656.0 173.5 73.5 KAN-2 6.6 132.0 0.0 5.0 208.3 0.0 564.0 149.2 49.2 KAN-4 5.8 116.0 0.0 5.2 216.7 0.0 662.0 175.1 75.1 KAN-11 2.8 56.0 44.0 1.8 75.0 25 616.0 163.0 63.0 Control group 1 0.0 0.0 0.0 0.0 0.0 0.0 405.0 107.1 7.1 Control 2 5.0 100.0 0.0 2.4 100.0 0.0 378.0 100.0 0.0

*: Ratio to control 2

<4-3> Nematode Prediction Test in the Field

Microbial preparations for plant parasitic nematode control were sprayed on tomato fields prepared by inoculating mycelium obtained by incubating at 25 ℃ for one week in CMII liquid medium in boiled barley and boiled oats. The spraying amount was sprayed with 1% and 2% of the soil volume, and the experiment was divided into control groups not administered the microbial agent. One and two months after the application of the microbial agent, soil was collected and the nematode number per 1 g of soil was examined to test the nematode predation ability of nematode predatory fungi.

As a result, it was confirmed that the nematode prey fungi of the present invention all effectively reduce the number of nematodes (Table 9).

Nematode Control Rate of Nematode Predatory Fungi Spread Average number of nematodes isolated from soil (horses / g) % Reduction of nematodes to control group 1 month later 2 months later One% 70.5 48 40 2% 39.5 32 60 Control 86.5 80 0

<4-4> Comparative experiment with microbial nematode

An experiment was conducted to compare the nematode control effects with nematicids currently available in the market. Microbial solution for nematode control (B agent, comparative group) compared to house sprayed with 2% of the plant parasitic nematode control agent (A agent) prepared on the tomato field based on the nematode predatory fungus of the present invention (A agent) to the soil volume. -Nematodes; Korea Bio Chemicals) was compared to the house of the irrigation and the house to which nothing was administered. After the tomato harvest was finished, the soil of the house was taken to check the nematode count.

As a result, 86.2% of the soil treated with agent A showed a nematode reduction effect of 86.2% than the control group and 81.9% of the control group treated with B (Table 10). This means that the microbial preparations of the present invention are very effective in the field and superior in nematode control even when compared to conventional comparative agents.

Comparison of nematode control with nematode control microorganisms type Average nematode number (mari / g) % Nematodes to control group % Nematodes for control group % Nematode Reduction Reduction Rate for Control Reduction Rate for Comparative Group A agent 2% (experimental group) 24.2 13.8 18.1 86.2 81.9 B system liquid irrigation (Comparative group) 133.4 75.8 100 24.2 0 Control 175.8 100 131.8 0 -

Example  5. Nutritional Growth Effect of Plant

An experiment was conducted to investigate the nutritional effects on plants of the microbial preparation for controlling plant parasitic nematodes prepared on the basis of nematode prey fungi. The experiment was carried out in the greenhouse cucumber house, and the microbial preparation was divided into the experimental group administered 1%, 2% and 3% to the soil volume and the control group not administered the microbial agent.

As a result, the microbial strains of the present invention showed a nutritionally good effect on both the growth and yield of cucumber (Table 11 and Table 12).

type Spread Growth length (cm) Average yield (g) Root damage caused by root-knot nematodes 1 month later 2 months later 3 months later Experimental group One% 25 92 100 419 5.1 2% 23 99 119 217 4.4 3% 16 101 111 209 1.7 Control - 22 85 91 192 5.6

type Height (cm) Number of leaves Stem weight (g) Root Weight (g) One% 12.8 ± 1.9 6 7.7 ± 1.0 3.2 2% 10.4 ± 1.7 5 4.9 ± 0.9 3.1 3% 10.8 ± 1.35 5 4.7 ± 0.4 2.4 Control 7.14 ± 1.4 4 2.2 ± 0.4 2.1

Example  6. eelworm  Predatory fungus On carbon ratio (C / N)  Growth experiment

Eight strains of purely isolated microorganisms were grown by varying the amount of nitrogen. Growth changes were confirmed by measuring the diameter of the colonies of each strain on the plate, and the degree of sporulation was observed using a microscope.

As a result, KAN-2, KAN-4 and KAN-11 strains grew rapidly in 15: 1 medium and formed a large number of conidia. KAN-12 strains grew rapidly in 10: 1 medium, but the growth was large in 2: 1 medium and conidia were formed in 15: 1 medium. KAN-9, KAN-13, KAN-20 and KAN-21 strains showed rapid growth and excellent sporulation in 10: 1 medium (Table 13).

Nematode Capture Mold Growth Results in Efficient Culture Medium Strain Diameter of colonies after 6 days of culture (mm) Mm / day 15: 1 10: 1 5: 1 2: 1 CM SA KAN-9 84.0 84.0 84.0 84.0 73.6 59.0 15.6 KAN-12 78.6 81.3 78.6 75.3 72.0 64.0 10.1 KAN-13 84.0 84.0 84.0 84.0 77.0 55.0 14.0 KAN-20 84.0 84.0 84.0 84.0 78.3 60.0 13.7 KAN-21 84.0 84.0 84.0 84.0 73.8 57.0 14.1 KAN-2 84.0 84.0 84.0 77.6 68.3 59.6 14.7 KAN-4 82.6 79.3 80.6 76.0 67.6 64.6 12.6 KAN-11 84.0 84.0 80.6 75.0 72.6 62.0 14.3

In the table, 15: 1 is 20 g of dextrose and 1.32 g of peptone, 10: 1 is 20 g of dextrose and 2 g of peptone, and 5: 1 is 20 g of dextrose and peptone 4 g, 2: 1 represents 20 g of dextrose and 10 g of peptone, CM (cornmeal agar, 2: 1) represents 10 g of dextrose, 5 g of peptone, 30 g of corn flour, 5 g of NaCl and CaCO ₃ 0.5 g, SA (sabouraud agar, 4: 1) represents 40 g of dextrose, 10 g of peptone, 15 g of agar.

In addition, for mass production of mold, GYA (glucose 40 g / L, yeast extract 20 g / L, agar 15 g / L) medium, CMA II medium, cornmeal 30 g / L, glucose 10 g / L, yeast Growth experiments were performed in the same manner as above using extract 5 g / L, NaCl 5 g / L, CaCO ₃ 0.5 g / L, agar 15 g / L).

As a result, the strains of the present invention, even in the above three kinds of media, all grew vigorously in mycelia and formed a large number of conidia (Table 14).

Growth of Nematode Capture Fungi in Other Media (mm) Strain Badge GYA Badge Ⅱ Badge CMAⅡ 3 days 6 days 3 days 6 days 3 days 6 days KAN-9 31.5 57.0 69.0 84.0 37.6 73.6 KAN-13 31.5 58.0 73.5 84.0 41.6 77 KAN-20 32.5 60.5 73.5 84.0 43 78.3 KAN-21 30.5 56.5 75.0 84.0 39.4 73.8 KAN-2 26.0 45.5 47.0 80.0 38 68.3 KAN-4 27.5 47.0 49.0 76.5 39 67.6 KAN-11 37.0 59.5 47.5 78.5 40.6 72.6

Example  7. eelworm  Plant Parasitics Using Predatory Fungi eelworm  Formulation Activity and Growth Tests of Control Agents

Nematode premature fungi were inoculated into CMII liquid medium, CMA solid medium and CMAII solid medium and cultured for one week at 25 ° C. The mycelia obtained in this way were inoculated into sterilized spawn bottles containing different microbial delivery media. After the inoculation, the spawn was incubated for 3 weeks in a 25 ° C incubation chamber. After incubation, 1 g was taken and suspended in sterile sterile water to measure bacterial growth and cell number.

As a result, it was confirmed that the growth was the best when using boiled barley as the delivery medium (Table 15).

Growth degree of nematode prey fungi type Months media Growth Cell count Pollution degree barley Boiled barley ++++ 8.1 × 10 ⁷ - Boiled Barley + CaCO ₃ ++++ 7.8 × 10 ⁷ - Boiled Barley + 4% Glucose ++++ 8.0 × 10 ⁷ - Boiled Barley + Rice Bran + Vermiculite +++ 7.5 × 10 ⁷ - wheat Boiled wheat ++ 3.4 × 10 ⁵ - Boiled Wheat + CaCO ₃ ++ 3.1 × 10 ⁵ - Boiled Wheat + Rice Bran ++ 2.8 × 10 ⁵ - Boiled Wheat + Rice Bran + Chaff ++ 2.5 × 10 ⁵ - Boiled Wheat + Rice Bran + Chaff + Vermiculite ++ 2.1 × 10 ⁵ - oat Boiled oats ++++ 7.5 × 10 ⁷ - Boiled Oats + CaCO ₃ ++++ 7.8 × 10 ⁷ - Boiled oats, rice bran, rice bran, vermiculite, granulated powder +++ 6.8 × 10 ⁷ - rice plant Boiled rice +++ 4.3 × 10 ⁶ - Boiled rice + CaCO ₃ +++ 5.1 × 10 ⁶ - Etc Rice bran + chaff + vermiculite + granulated powder + 1.2 × 10 ⁵ - Rice bran + rice bran + vermiculite + granulated granules + 1.5 × 10 ⁵ - K-1 (rice husk + vermiculite + oat flour + granule powder) + 1.1 × 10 ³ + K-2 (rice bran + powder) + 1.4 × 10 ² - K-3 (rice bran + oat flour) + 3.6 × 10 ³ ++ K-4 (rice bran + granulated powder) + 2.2 × 10 ² -

< Formulation example  1> Preparation of Microbial Agents

<1-1> Preparation of Hydrating Agent

For the purpose of stable formulation using the microbial strain of the present invention was prepared a hydrated hydrate when diluted in water as powder phase. Specifically, the hydrating agent was prepared by heating and drying the solid medium, followed by grinding by adding a surfactant and an extender. That is, the raw powder was dried and pulverized for about 2 hours in a dryer at 70 ° C., and then coarsely ground, surfactant and extender were evenly mixed and mixed. The mixture was ground through a Jet-O-mill (manufactured by Aljet) and the ground powder was hydrated before preparing a microbial formulation in the form of a hydrate.

<1-2> Granular  Produce

For the purpose of stable formulation using the microbial strain of the present invention was prepared granules that can be used as it is granular. Specifically, the raw powder was pulverized by drying for about 2 hours in a dryer at 70 ° C., the pulverized powder was pulverized by passing through a jet-O-mill, and then the microbial pulverized powder, surfactant, extender / nutrient and disintegrant were separated from the strain. 50% spore, 3% polycarboxylate, 3% sodium lignosulfonate, 1% sodium dialkyl sulfosuccinate, 10% starch, 20% bentonite, 13% talc About 35% of the amount was hydrolyzed and kneaded. After the dough was finished, it was granulated using a granulator (diameter 1 mm, length 1-5 mm) and dried in a dryer at 70 ° C. The dried product was passed through a 16-30 mesh sieve to remove the powder to prepare a granule.

<1-3> Preparation of Encapsulated Formulations

Encapsulated formulations were prepared for the purpose of stable formulation with the microbial strain of the present invention. Specifically, 11% soybean powder, 6% green beans, 4% rice flour, 4% potato starch, 4% rye flour, 3% ocher soil, and autoclaved at 121 ° C. for 20 minutes, and then mixed again before cooking to cool in cold water. I was. After thoroughly mixing for 10 minutes or more in a stirrer (d = 5 cm, 200 rpm or more), the additive was mixed with the microbial strain of the present invention while stirring was continued to prepare a capsule formulation.

As described above, the microbial strains of the present invention are very adaptable to domestic soil, excellent insecticidal effect and nutrient growth effect of the plant, and can supply a large amount of strains in a timely manner, plant parasitic nematodes, In particular, it can be usefully used for the control of root-knot nematodes.

Attach sequence list electronic file  

Claims (9)

Atsurobotris oligosporane KAN-20 (Accession No. KACC93052P) showing nematode effect on nematodes. A microbial preparation for nematode control, comprising Asthrobellis oligospora KAN-20 (Accession No .: KACC93052P) as an active ingredient. The method of claim 2, The nematode is a microbial agent, characterized in that the plant parasitic nematodes. The method of claim 3, The plant parasitic nematode is selected from the group consisting of root-knot nematodes, root rot nematodes and cyst nematodes. The method of claim 4, wherein The root-knot nematode is a microbial agent, characterized in that selected from the group consisting of carrot root-knot nematode, sweet potato root-knot nematode, Java root-knot nematode and peanut root-knot nematode. The method according to any one of claims 2 to 5, The microbial agent is a microbial agent, characterized in that the formulation is selected from the group consisting of a hydrating agent, granular hydrating, liquid hydrating, liquid, water-soluble, water-soluble granules and encapsulating agent. Culturing the Astrobotris oligosporane KAN-20 (Accession No. KACC93052P) of claim 1 in a solid or liquid medium; And A method of mass-producing a microorganism preparation for controlling the genus Astrobotris of claim 1 or the nematode of claim 2, comprising inoculating the cultured microbial mycelium on boiled barley, boiled oats or a mixture thereof. The method of claim 7, wherein The solid medium is CMA II medium (corn meal agar; cornmeal 30 g / l, glucose 10 g / l, peptone 5 g / l, NaCl 5 g / l, CaCO ₃ 0.5 g / l, agar 15 g / l) How to feature. The method of claim 7, wherein The liquid medium is CMII medium (cornmeal 30 g / l, glucose 10 g / l, peptone 5 g / l, NaCl 5 g / l, CaCO ₃ 0.5 g / l).

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