JP2003274931A - Method and medium for cultivation of microorganism belonging to genus fusarium - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method supplying conidia of a microorganism belonging to nonpathogenic genus fusarium which can control seed infectious diseases of Oryza sativa, in a short time, stably and in an industrial scale. <P>SOLUTION: The method uses cultivating medium containing peptone as an essential ingredient that is not used ordinarily as the ingredient for fungi, and extract of yeast and glucose. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a method for culturing Fusarium spp. More specifically, it stabilizes conidia of a microorganism belonging to the genus Fusarium having an excellent control effect against plant diseases, particularly rice seed infectious diseases. The present invention relates to a culture method and a culture medium for collecting a large amount at low cost.

[0002]

2. Description of the Related Art As an attempt to control a rice seed infectious disease by using a microorganism having substantially no pathogenicity to rice, for example, JP-A-5-65209 and JP-A-1-65209.
Japanese Unexamined Patent Publication No. 1-89562 describes a microbial preparation belonging to the genus Fusarium that has no pathogenicity to rice.

As a method for culturing microorganisms belonging to the non-pathogenic Fusarium genus, JP-A-7-184637 describes a method using beer lees. As a method of culturing microorganisms, a method of culturing using waste materials such as bran, rapeseed meal and soybean shells is generally used. Microorganisms cultivated by these methods are not particularly problematic when applied as a solid, but when used as pesticides, many methods are used in which they are dissolved or suspended in water and sprayed with a sprayer. There is. Further, when fungi are used as pesticides, it is preferable to use conidia from the viewpoint of effect and preservation. In order to obtain a large amount of target conidia of fungi, not all media with good hyphal growth are suitable, and it is necessary to thoroughly examine the media composition and the like.

[0004]

DISCLOSURE OF THE INVENTION The present invention provides a technique for stably supplying a large amount of conidia of a microorganism belonging to the non-pathogenic Fusarium genus capable of controlling seed infectious diseases of rice in a short time. The task is to do.

[0005]

[Means for Solving the Problems] In order to solve the above problems, the present inventors have conducted diligent studies, and as a result, by culturing using a medium mainly composed of peptone, which is usually rarely used as a medium component of fungi, The inventors have found that a large amount of conidia of a microorganism belonging to the genus Fusarium can be stably collected, and have completed the present invention.

That is, the present invention is, firstly, a method for culturing a microorganism belonging to the genus Fusarium characterized by culturing using a medium containing peptone, yeast extract and glucose. The Fusarium microorganism is a microorganism having substantially no pathogenicity to plants, and Fusarium
Fusarium moniliforme, Fusarium oxysporum, Fusarium solani, Fusarium acuminatum, Fusarium avenaceum, Fusarium calmorum
sarium culmorum), Fusarium graminearum (Fus
arium graminearum) and Fusarium nivare (Fusar
ium nivale, also known as Micronectriella nivalis), which is at least one kind selected from the group consisting of Fusarium monilforme.
iforme) (FERM P-18726).

Further, the present invention is the culture medium for a Fusarium genus microorganism, which contains peptone, yeast extract and glucose.

[0008]

DETAILED DESCRIPTION OF THE INVENTION The present invention will be described in detail below.
In carrying out the culturing method of the present invention, mycelium and / or conidia of Fusarium spp., Which are purely cultivated in a medium such as potato dextrose agar medium, are inoculated into the above liquid medium. Then, the culture may be carried out by a commonly used culture method such as shaking culture, culture in an aeration and agitation culture tank, culture in a bubble column culture tank. The medium of the present invention comprises peptone,
It contains yeast extract and glucose, and the content of each component is preferably 0.5 to 1.5% of peptone, 0.1 to 0.2% of yeast extract, and 0 to 0.20% of glucose. For example, Fusarium m
The optimal medium for culturing oniliforme) (FERMP-18726) is 0.5% peptone, 0.125 yeast extract.
%, Glucose 0.025%.

The medium components peptone, yeast extract and glucose may be any commercially available ones and are not particularly limited.

The pH of the medium need not be adjusted,
It is usually 5.0 to 9.0, and more preferably 5.0 to 6.0 to prevent bacterial contamination. The culturing temperature may be any temperature at which the Fusarium genus can grow, but it is 15 to 35 ° C., preferably 2 for efficient production.
It is 5 to 30 ° C.

The culture method and culture medium of the present invention are known to be Fusarium moniliforme and Fusarium, which are known as microorganisms belonging to the genus Fusarium that have no substantial pathogenicity to plants, especially rice.
Fusarium oxysporum
Fusarium solani, Fusarium acuminatum, Fusarium avenaceum, Fusarium culmorum, Fusarium graminearum, and Fusarium graminearum. .

According to the culture method of the present invention, conidiospores of Fusarium spp. Of high concentration of about 1.0 × 10 ⁸ cfu / ml can be obtained usually in 3 to 4 days. The culture solution can be easily separated into hypha and spores by filtration or the like. The separated spore suspension can be made into spore pellets by centrifugation or the like, and can be formulated into a form in which ordinary pesticides can be taken, for example, powders, wettable powders, emulsions, flowables, granules and the like. . Such a formulation can be usually mixed with a carrier, a surfactant, a dispersant, an auxiliary agent and the like to give a plant disease controlling composition by a conventional method.

For example, Fusarium moniliforme (Fu
The plant disease control agent composition containing sarium moniliforme) (FERMP-18726) is a method for dressing rice seeds, immersing the seeds, or treating the soil, especially seedling-growing soil at the time of sowing or germination. It is possible to effectively control seed-borne diseases.

[0014]

EXAMPLES Next, the present invention will be described in more detail by way of examples, but the present invention is not limited to the following examples, and is non-pathogenic within the scope not departing from the gist of the present invention. The type of Fusarium, the composition ratio of the medium, etc. can be freely changed. In the following examples, Fusarium moniliforme (Fu
sarium moniliforme) FERM p-18726 was used.

Example 1 Polypeptone (manufactured by Nippon Pharmaceutical Co., Ltd.) 0.5% adjusted to pH 6.8, yeast extract (manufactured by Difco Laboratories)
0.125%, glucose (manufactured by Wako Pure Chemical Industries, Ltd.)
150 ml of a liquid medium containing 0.025% was poured into a 300 ml Erlenmeyer flask and sterilized under high pressure. To the sterilized medium, 1 platinum loop of Fusarium moniliforme precultured on potato dextrose agar was inoculated. Then 25
The cells were cultivated at 3 ° C with shaking for 3 days.

(Investigation method) After completion of the culture, the culture solution was filtered through double gauze to separate into hypha and conidia. The number of conidia in the obtained conidia suspension was investigated by virtue of the number of viable cells by the dilution plate method using potato dextrose agar medium.
The survey results are shown in Table 1.

(Comparative Example) Usually, a liquid medium of potato dextrose (Difco Labo), which is often used for culturing fungi.
ratories) (potato extract 0.4%, glucose 2)
%, PH 6.0) and Czapek Dox Broth (SIGMA) (glucose 3.6%, sodium nitrate 0.2%, potassium monohydrogen phosphate 0.1%, magnesium sulfate 0.05%, potassium chloride 0.05%) , Ferrous sulfate 0.001%, pH
6.8), Nutrient Broth widely used for bacteria
Fusarium moniliforme was inoculated into each (manufactured by Difco Laboratories) (meat extract 0.5%, peptone 1%, sodium chloride 0.5%, pH 7.0) in the same manner as in Examples, and cultured for 3 days. The investigation was conducted in the same manner as in the example.

Table 1 shows the conidia concentration when Fusarium moniliforme was cultured for 3 days under the medium composition and culture conditions of Examples and Comparative Examples. As shown in Table 1, Fusarium moniliforme used in Example 1 was cultured in a liquid medium containing 0.5% peptone, 0.125% yeast extract, and 0.025% glucose. Potato dextrose liquid medium and Czapek DoxBroth that are frequently used for use, and like the medium composition used in the Examples, Nutrient that is frequently used for bacteria
Compared with Broth, a large number of conidia were produced.

[0019]

[Table 1]

Examples 2 to 7 In the medium composition, the amount of glucose added was 0 (Example 2),
(Example 3) 0.025%, (Example 4) 0.05%,
(Example 5) 0.1%, (Example 6) 0.2%, (Example 7) 0.5%, (Example 8) 1.0% except that the same as Example 1. Cultured. Table 2 shows the conidia concentration obtained in each example.

[Table 2]

Industrial Applicability As described above, according to the culture method of the present invention, which uses a medium mainly composed of peptone, which is rarely used for culturing fungi, it belongs to the genus Fusarium which has an excellent effect of controlling plant diseases. A large amount of conidia of microorganisms can be collected.

Claims (8)

[Claims] 1. A method for culturing a microorganism belonging to the genus Fusarium, which comprises culturing using a medium containing peptone, yeast extract and glucose. 2. The method for culturing according to claim 1, wherein the microorganism belonging to the genus Fusarium is a microorganism having substantially no pathogenicity to plants. [Claim 3] Fusarium microorganisms are Fusarium moniliforme, Fusarium oxysporum, Fusarium solani, Fusarium acuminatum, Fusarium acumenum, Fusarium acumium.
sarium culmorum), Fusarium graminearum (Fus
arium graminearum) and Fusarium nivare (Fusar
ium nivale, also known as Micronectriella nivalis), at least one selected from the group consisting of:
The culturing method described. 4. A microorganism belonging to the genus Fusarium is Fusarium moniliforme (FER).
MP-18726), The culture method according to any one of claims 1 to 3. 5. A culture medium for a Fusarium genus microorganism, which comprises peptone, yeast extract and glucose. 6. The culture medium for Fusarium spp. According to claim 5, which contains 0.5 to 1.5% of peptone, 0.1 to 0.2% of yeast extract, and 0 to 0.20% of glucose. 7. The microorganism belonging to the genus Fusarium is Fusarium moniliforme, Fusarium oxysporum, Fusarium solani, Fusarium acuminatum, Fusarium acuminatum, Fusarium avenaum. Fusarium culmorum), Fusarium graminearum and Fusarium nivale, also known as Micronectriella nivali
The culture medium according to claim 5 or 6, which is at least one selected from the group consisting of s). 8. A microorganism belonging to the genus Fusarium is Fusarium moniliforme (FER).
MP 18726), The culture medium according to any one of claims 5 to 7.