KR100269417B1 - Microorganism of the genus ascochyta and use thereof - Google Patents

Abstract

The present invention uses a strain belonging to the genus Ascorkta, which shows pathogenicity to cooked bones, tadpoles, suwons, or Sylrpus lineolatus, without affecting crops such as rice. Provides means for selectively killing or inhibiting growth of highland weeds.

Description

Microorganisms belonging to the genus Ascokita and their uses

Chemical herbicides are mainly used as a method for controlling weeds in munon or fields and for weed crabs in Scirpus. Recently, the development of weed control agents that do not rely on chemical pesticides and their methods of use have been demanded due to the environmental pollution caused by the large use of chemical pesticides, and the expectation of microbial pesticides using phytopathogens is particularly high. To date, DeVine (United States, Weeds: Stranglervine), Collego (United States, Weeds: Northern jointvetch, Legumes), BioMal (Canada, Round-leaved mallow, Malvaceae) have been registered and commercialized. . However, no microbial herbicides have been developed for weeds and tadpoles. In addition, ascorbic pathogens in Munnon's major weeds (Scirpus juncocides var.ohwianus), tadpole gourd (Scirpus juncoides var, hotarui), Scirpus wallichii, or Scirpus lineolatus. Microorganisms belonging to the genus Kita are not known.

[Problems to Solve Invention]

An object of the present invention is to provide a means for controlling microbial weeds and tadpoles weeds.

[Start of invention]

As a result of intensive studies to solve the above problems, the present inventors have found that several strains belonging to the genus Ascorkita have a very excellent control against weeds of tadpoles and genus larvae and have completed the present invention.

That is, the present invention is a strain belonging to the genus Ascorkta which exhibits pathogenicity to any of cooked bone, tadpole high-necked roe, Suwon high-necked roe or Sylpus lineolatus.

In addition, the present invention is a strain belonging to the genus Ascorkta which exhibits pathogenicity in any of cooked bone, tadpole high-necked roe, Suwon high-necked roe or Sylpus lineolatus.

Moreover, this invention is a control method of weeds using the herbicide of the said description.

Hereinafter, the present invention will be described in detail.

First, the new strains of the present invention, askokita sp.JT-1006, askokita sp.JT-1007, askokita sp.JT-1008, askokita sp.JT-1009, and askokita sp.JT-1010 Explain the note. These strains collected leprosy and tadpoles weeds from all over Japan, isolates strains with pathogenicity to gilts and tadpoles weeds, and cultivates them purely to control the weeds in the genus. At the same time, it is newly created by a screening means that examines the non-pathogenicity of other crops, especially rice.

The main things of the mycological properties of the novel strains of the present invention are as follows.

The aerobic bacteria that can grow are in the range of 3 to 13, preferably in the range of 5 to 7. The growth temperature is in the range of 15 to 30 ° C, and the temperature suitable for growth is in the range of 25 to 30 ° C. Growth on potato dextrose agar medium flourishes, and colonies produce a large number of orange-brown granules in the form of gray-olive mycelium in a spiral form from radial rays. Is less. Conidia are colorless, similar to the genus Phoma, but oval divided into two chambers with one partition wall in the center. The size of conidia is about 50-10 μm × about 2-5 μm.

These fungal properties were described in the plant fungal description (Kobayashi et al., 1992, P20 ~ 52, 384. National Association for Rural Education) and Illustrated Genera of imperfect Fungi, Fourth Edition (Barnett, HL and Hunter, BB 1987. P6 39, 178. As a result of searching by Macmillan Publishing Company, the strain was identified as a microorganism belonging to the genus Ascorkita. Microorganisms of several species of Ascorkita are known as pathogens, such as cucumbers and tomatoes. Ascorkita codystar, which shows pathogenicity to Scirpus maritimis, is a microorganism of the genus Ascorkta, which shows pathogenicity in plants of the genus Panax and tadpoles. Two examples of Ascochyta lacustris, which show pathogenicity to Ascochyta kurdistanica and Scirpus lacustris, are known, but the cooked bone, tadpole, and suwon gourd which are the subject of the present invention are known. It is not known to exhibit pathogenicity to this or Sylpus lineorats.

In addition, the strain was deposited in the Institute of Biotechnology Industrial Technology Research Institute (Higashi Nigugoku Ibaragi 껜 Tsugubashi 1-chome 1 half 3 Goo) under the following numbers.

Asokita sp.JT-1006 note: FERM BP-5692 (deposit date October 3, 1996)

Ascot Kita sp.JT-1007 note: FERM BP-5693 (deposit date October 3, 1996)

Asokita sp.JT-1008 note: FERM BP-5694 (deposit date October 3, 1996)

Ascot Kita sp.JT-1009 note: FERM BP-5695 (deposit date October 3, 1996)

Ascot Kita sp.JT-1010 note: FERM BP-5696 (deposit date October 3, 1996)

There is no need to use a special method for culturing this strain, and the same method as that of a known strain belonging to the genus Ascorkita can be used. As the medium, any medium that can be used can be used as long as it contains a magnetizable carbon source, nitrogen source, inorganic matter and necessary growth promoting substances. Examples of the specific medium include potato dextrose agar medium (PDA), zapek agar medium and the like. In culture, it is preferable to keep the temperature at 15 to 30 ° C, preferably at 25 to 30 ° C, and the pH at 3 to 13, preferably at 5 to 7. When cultured for 7 to 14 days under the above conditions, a sufficient amount of conidia are formed on the surface of the medium. This strain kills the mud or rapeseed weeds and tadpoles, and does not affect rice, wheat, barley, corn and other crops such as eggplant, beans, cabbage and cucumber. This strain can be cultured in large quantities and a large amount of spores can be easily obtained.

Next, the herbicide and control method of the present invention will be described.

As microorganisms used in the herbicide of the present invention, ascorkita sp.JT-1006 strain, ascorkita sp.JT-1007 strain, ascorkita sp.JT-1008 strain, ascorkita sp.JT-1009 strain, ascorkita sp.JT -1010 strains and the like, but may be used a mutant strain derived from these strains.

The herbicide of the present invention can be formulated by suspending spores obtained by culturing the strain in water, but the method of formulation is not limited thereto. In this case, the concentration of spores is preferably 10 ^{6-10 12} / ml, but is not limited thereto. Moreover, in suspending in water, you may add adjuvant, such as surfactant and an electrodeposition agent. The microorganism as a subject may be a fresh one immediately after the cultivation, or one that has been preserved and restored mainly from water. The preservation method is widely known as a microorganism preservation method. Cryogenic preservation (-80 ℃), vacuum freeze drying can be used.

The control method of this invention is performed by spreading the said herbicide in the field. Dispersion amount is preferably assuming according to rank the degree of weed, but spraying of the dog ¹⁵ so that the amount of the normal field 10 ahreudang spores 10 ^11-10.

Plants to which the herbicide and the control method of the present invention are subjected are not particularly limited, but are particularly effective for plants belonging to the genus Tadpole. Examples of tadpole high-rise plants here include Sciripus juncoides var.hotarui, cooked bone (Scirpus juncoides var.ohwianus), Scirpus wallichii, Scirpus lineolatus, Perth Nipponicus, Scirpus triqueter, Scirpus triangulatus, Scirpus yagara, Scirpus planiculmis, etc. no. Moreover, there is no restriction | limiting in particular also about the growth time of the plant made into control object, It can be made to control object widely from the plant immediately after germination to the plant which grew.

The present invention relates to a novel strain belonging to the genus Ascochyta and a method for controlling herbicides and weeds using the strain.

1) Production process of novel strain of the present invention

After cultivating in munone or field, and showing the signs of disease, weeds and tadpoles of weed or tadpoles were harvested, cut off the leprosy and soaked the leaves in 30% to 60 seconds in 70% ethanol solution. A 1% sodium hypochloride solution was added to an equal amount of 0.1% Tween 80 solution, suspended in a solution for 1 to 3 minutes, and then surface sterilized. The leaves were washed once with 0.1% Tween 80 solution and twice with sterile water, and then placed on the PDA medium containing antibiotics, incubated for 2 to 5 days in a thermostat at 25 ° C, and expanded to the new PDA medium. And cultured to form conidia. In addition, the microbial cells were purified and separated by culturing conidia in a new PDA medium to obtain a single colony.

For each isolated strain, the pathogenicity of the weeds of Geumdongsa and tadpoles was reviewed again and the effects on rice, wheat, corn, tomatoes, and cucumbers were examined. New strains that do not affect rice, wheat, corn, tomatoes, and cucumbers. Ascokita sp.JT-1006, Ascokita sp.JT-1007, Ascokita sp.JT-1008, Ascokita sp.JT -1009 strains, Ascorkita sp. JT-1010 strains were isolated.

2) Identification of the prepared strain

Identification of the strain of the present invention was performed mainly by observing the form of the conidia. As a result, as described above, the strain was identified as a microorganism belonging to the genus Ascorkita.

3) Bulk culture and preparation method

Sterilized water was added to microorganisms belonging to the genus Ascorkta grown on PDA medium to form a high concentration of spore suspension, and about 100 μl was added dropwise onto a V-8 juice agar medium and diffused into a sterile L-shaped glass rod at once. Implantation of microorganisms in a large amount of petri dishes (diameter 9 cm) was enabled. As a result, the amount of spores produced was about 6 × 10 ⁷ spores per petri dish.

As described above, the inanimate organism of the present invention can obtain spores easily and in large quantities by culturing on a plate medium.

The spores thus obtained are suspended in 10% skim milk and dried in vacuo to form a hydrate. This hydrating agent can be suspended in water and applied by adding a surfactant such as Tween 80.

4) Pathogenicity test for cook bone

Cooked bone seeds were suspended in a solution in which an equal amount of 0.1% Tween 80 solution was added to sodium hypochlorite solution, soaked for 1-3 minutes and surface sterilized. The surface sterilized seeds were seeded in a previously sterilized in vitro medium for plant growth, and grown up to 1-2 leaves as a test material.

The microorganism of the present invention was inoculated in vitro by punching two disks with a cork borer on the mycelium obtained by culturing on potato agar medium. After 10 days in a constant temperature room at 25 ℃ the control rate of cooked bone was examined. The results are shown in Table-1. The mortality rate of cooked bone by this bacterium was 100% and control effect was 100%.

[Table 1] Control effect on cooked bone of each strain

* Mortality rate: mortality rate / inoculation number of times * 100

* Control rate: Decrease rate of whole plants

* JTKA-644: Ascochyta sp.

* JTH-102: Exserohilum monoceras from the field

* JTSS-001: Fusarium sp.

As apparent from Table 1, the microorganism of the present invention showed an excellent control effect on cooked bone (Scirpus juncoides var. Ohwianus).

5) Pathogenicity test on weeds in tadpoles

The test for the weeding of the tadpoles was carried out in the same manner as the pathogenicity test for the cooked bone.

The tadpoles, Suwons, Silfers lineorats and Silfers nipponicus were used as published plants. The test results are shown in Table-2.

[Table 2] Control Effects on Weeds of Tadpoles in JT-1008

+: Examination or growth inhibition

-:No influence

As apparent from Table 2, the microorganism of the present invention showed an excellent control effect against weeds in the genus and tadpoles. The same result was found in other strains.

6) Impact on crops

As for the crop test, the spore concentration of the inoculum used in the same manner as the pathology test for the cooked bone was 10 ⁷ / ml. Rice, wheat, corn, eggplant, soybeans, cabbage and cucumbers were used as reference plants. The test results are shown in Table-3.

[Table 3] Impact on Crops

+: Examination or growth inhibition

-:No influence

As is clear from Table-3, the microorganism of the present invention did not affect the growth of rice, wheat, corn, eggplant, cucumber, cabbage and soybean.

[Step 1] (Liquid)

Conidia (10 ¹⁰ ) and Tween 80 (1 g) of Ascorkita sp. JT-1008 strain were added to 1 L of water and mixed to form a liquid.

[Formulation 2] (Hydrating)

10 ⁸ conidia (JT-1007 strain) were suspended per 1 ml of a mixture of 9% maltose, 1% clay and 90% water. After drying, the dried product was mixed and ground to form a hydrating agent.

[Formulation 3] (hydrating agent)

10 ⁸ conidia (JT-1007 strain) were suspended per 1 ml of a mixture of 10% skim milk and 90% water. After drying, the dried product was mixed and ground to form a hydrating agent.

Preparation Example 4 (Powder)

10 ⁸ conidia (JT-1009 strain) per 1 g of a mixture of 14% hydroxypropyl-β-cyclodextrin, 12% white carbon, 74% clay were mixed. It was dried and then ground uniformly to form a powder.

[5] (former)

Assembly as β- cyclodextrin 15%, 2% starch, 18% bentonite, minutes per 1g of a mixture of 36% calcium carbonate, 29% water living person (JT-1010 Note) 10 ⁸ won after granulator (造粒機) was added to Granulation was carried out by drying.

Preparation Example 6 (Emulsion)

10 ⁸ conidia (JT-1008) per 1 g of a mixture of 18% polyoxyethylene nonylphenyl ether ammonium phosphate, 6% polyoxyethylene nonylphenyl ether, triethyl phosphate, and 47% tributyl phosphate Suspended to form an emulsion.

Preparation Example 7 (Emulsion)

An emulsion was prepared by suspending 10 ⁸ conidia (JT-1008 strain) in 1 ml of a mixture of 95% spindle oil, 4% castor oil, and 1% silicone oil.

Preparation Example 8 (Dry Flowing Agent)

Condenser (JT-1006 strain) 10 ⁸ was suspended in 1 ml of a composition of 12% of alkylbenzenesulfonate and 88% of polyethylene glycol ether to form a dry fluid.

Preparation Example 9 (Made in Capsule)

In ⁸ ml of a mixture of 0.7% sodium arginate, 5% kaolin, 15% glycerin, and 79.3% water, 10 ⁸ conidia (JT-1007 strain) were suspended and added dropwise to a 0.2 mol calcium acetate solution to obtain a capsule product. This was chopped, sieved and dried to form a capsule.

[Effects of the Invention]

The herbicide of the present invention can be controlled without affecting crops such as rice by selectively killing or inhibiting growth of the weeds of the genus Bronze and Tadpole. It also does not pollute or destroy the environment like chemical pesticides.

Claims (5)

A strain belonging to the genus Ascorkta which exhibits pathogenicity to cooked bone, tadpole high, suwon high, or silpus lineorats. According to claim 1, the strain belonging to the genus Ascorkita sp.JT-1006 strain, Ascorkita sp.JT-1007 strain, Ascorkita sp.JT-1008 strain, Ascorkita sp.JT-1009 strain, Ascorkita sp.JT-1010 strain characterized in that. A herbicide comprising as a active ingredient a strain belonging to the genus Ascorkta which exhibits pathogenicity to cooked bones, tadpoles, suwons, and silpus lineorats. The strain belonging to the genus Ascorkta is ascorkta sp.JT-1006 strain, Ascorkita sp.JT-1007 strain, Ascorkita sp.JT-1008 strain, Ascorkita sp.JT-1009 strain, or Asco Kita sp.JT-1010 The herbicides characterized by the above-mentioned. A method for controlling weeds using the herbicides according to claim 3 or 4.