JP2001149066A - Method for producing sporangium of bacillus popilliae - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an industrial method for producing a sporangium of Bacillus popilliae, capable of affording a large amount of sporangium of Bacillus popilliae containing a spore and a parasporal body by using an artificial medium not using insects of the family Scarabaeidae, to obtain an excellent controller for insects of the family Scarabaeidae, greatly damaging roots of lawn grass, agricultural and horticultural crops, trees, etc., comprising the sporangium as an active ingredient and to provide a method for controlling insects of the family Scarabaeidae. SOLUTION: This method for producing the sporangium of Bacillus popilliae containing a spore and a parasporal body is characterized in that a bacterium belonging to Bacillus popilliae is cultured in a medium containing 0.01 wt.% to 0.5 wt.% of activated carbon. This controller against insects of the family Scarabaeidae comprises the sporangium as an active ingredient. The method for controlling insects of the family Scarabaeidae uses the sporangium.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a method for producing a Bacillus popirie sporangia containing a spore and a parasporal body by culturing a bacterium belonging to the genus Bacillus popirie in an artificial medium, and using the sporangia as an active ingredient. And a method for controlling scarabidae insects.

[0002]

2. Description of the Related Art Larvae of scarab beetles feed on the roots of a wide range of plants such as turf, agricultural and horticultural crops and trees, and cause great damage to them. Since these larvae live in the ground, chemical pesticides sprayed from the ground are difficult to work and their habitats are difficult to identify, so in order to obtain a sufficient insecticidal effect, a large amount of insecticidal Pesticides must be sprayed and penetrated into the ground. As a result, environmental pollution is likely to be caused, so that its control and control have been practically difficult.

[0003] On the other hand, bacteria belonging to Bacillus popiliae are:
The parasites of these Scarabaeid insects parasitize and cause emulsification disease, eventually causing the Scarabaeidae insects to die.Therefore, an attempt is made to use bacteria belonging to Bacillus popirie for the control of Scarabaeidae insects, which are difficult to work with chemical pesticides. Attempts to do so have been made since ancient times.

[0004] As a method for cultivating bacteria belonging to Bacillus popiliae, a method for producing sporangia by growing bacteria using living larvae of the family Scarabaeidae, which is a parasitic host, has been conventionally known.

That is, the sporangia of Bacillus popirie is sprayed on a breeding medium in which larvae of Scarabaeid insects are present, and the sporangia of Bacillus popirie is orally ingested by the larvae of Scarabaeid insects. After injection, the larvae are bred for 3 to 4 weeks, Bacillus populiae is propagated in the larvae, and then the bodily fluid is collected by piercing or incising the larvae, and the obtained bodily fluid is centrifuged. This is a method of obtaining sporangia increased in the larva by separating or filtering.

However, according to this method, it is necessary to breed a large number of larvae of Scarabaeid insects, and as a result, it has been difficult to obtain a large amount of sporangia in a large amount in a short time and economically. For this reason, many studies have been conducted on a culture method using an artificial medium that does not use larvae of Scarabaeidae.

[0007] As a method of cultivating the artificial medium without using the larva of the insects of the order Scarabaeidae (hereinafter sometimes referred to as an insect extracorporeal culture method), there are solid culture and liquid culture. In solid culture, FOSTER et al. (JWFoster et al., 1950, J. Bacte
riol, 59, 463-470), STEINKRAUS (Keith H. Steinkra
US, 1957, J. Bacteriol, Vol. 74, pp. 625-632), but in liquid culture, HAYNES et al. (WCHaynes et al., Journey
nal of Bacteriology, June, 1966, 91, No6, 2270-2274
Page)). However, in any case, the insecticidal effect of the obtained spores or sporangia on the larvae of Scarabaeidae is neither specified nor led to commercial production.

[0008] A US patent (USP No.4824671) is an example in which bacteria belonging to Bacillus popirie are cultured by liquid culture using an artificial medium. However, although the obtained sporangia has spores but no parasporal body, the insecticidal effect on the larvae of Scarabaeidae is weaker than that of the sporangia formed in the larva. Fukuhara also reported that infection did not occur in sporangia obtained from artificial media (by Toshihiko Fukuhara)
Insect pathology p. 57).

As shown in the schematic diagram of FIG. 1, a sporangia is a sac containing spores and a body called a parasporal body (or parasporium), and is produced by a bacterium belonging to Bacillus populiae. In the conventional literature on a culture method using an artificial medium of Bacillus populiae, sporangia and spores are often used without distinction, and the term spore in the literature means only spores, It is often unclear whether it refers to a sporangium containing spores but not a parasporal body, or a sporangium containing spores and a parasporal body.

Therefore, it is unclear whether the spores of Bacillus populiae obtained by the method of culturing in an artificial medium described in the conventional literature are spores containing spores and parasporal bodies. Although the insecticidal effect of spores or spores on the larvae of Scarabaeidae is not clear, many of them are not spores containing spores and parasporal bodies, but only spores or only spores. It was presumed that the insecticidal effect on Scarabaeidae insects was weak.

[0011]

SUMMARY OF THE INVENTION An object of the present invention is to provide a large amount of spores of Bacillus populiae containing spores and parasporal bodies using an artificial medium without using larvae of Scarabaeidae. An industrial method for producing a Bacillus popirie sporangia to obtain the sporangia as an active ingredient,
It is an object of the present invention to provide an excellent control agent and an excellent control method for insects of the order Scarabaeidae which cause great damage to the roots of turf, agricultural and horticultural crops, trees, and the like.

[0012]

Means for Solving the Problems The present inventors have conducted intensive studies to solve the above-mentioned problems. As a result, effective elimination of insects of the order Scarabaeidae is not limited to spores of Bacillus populiae but also spores. It has been found that a sporangia containing a parasporal body is required. Furthermore, in order to grow Bacillus popirie outside of the larva of Scarabaeidae, and to form a sporangium containing spores and a parasporal body, the bacterium is excreted out of the cell with the growth of the bacterium. It has been found that it is necessary to remove metabolites that inhibit sporangium formation, and that it is not necessary to remove components required for sporangial formation when removing the inhibitory metabolites.

[0013] Even in the conventional reports, in the cultivation of Bacillus populiae in an artificial medium, it is necessary to remove a culture inhibitor for the growth of the bacterium, and an adsorbent such as activated carbon or soluble starch is added to the artificial medium. Was added. However, the present inventors have found that a sporangium containing spores and a parasporal body is not formed unless the culture is carried out with a limited amount of the activated carbon as a material for removing an inhibitory substance.

That is, the present inventors have found that if the amount of activated carbon added to the artificial medium of Bacillus popirie exceeds 0.5% by weight, even nutrients necessary for forming sporangia are removed. Alternatively, sporangia formation including spores and parasporal bodies was found to be sharply reduced. On the other hand, the amount of the conventionally added inhibitor adsorbent is 0.6% by weight or more with respect to the medium, and the degree of formation of spores or spores that exhibit the insecticidal effect of Scarabaeidae insects is reported. Had not been.

The inventors of the present invention have reported that culturing in an artificial medium containing 0.6% by weight or more of activated carbon, which has been reported so far, allows the growth of the bacterium (vegetative cell) itself to be good, but the spores and the parasporal body The production amount of sporangia containing spores is extremely small, and a sporangium containing sufficient spores and a parasporal body to exhibit an insecticidal effect of a scarabid insect is not obtained,
Furthermore, it was also found that when the artificial medium contains a certain amount of glucose or more, it is more difficult to obtain sporangia.

[0016] Therefore, the conventionally reported cultivation of Bacillus populiae in an artificial medium containing activated charcoal does not provide sufficient spores containing spores and parasporal bodies, and the larvae of the definite insects of the order Scarabaeidae cannot be obtained. It is highly probable that no insecticidal effect was reported because the insecticidal effect could not be confirmed.

[0017] From these, the present inventors restricted the amount of activated carbon and glucose in the artificial medium to thereby produce a large number of sporangia containing insecticidal spores and parasporal bodies against insects belonging to the family Scarabaeidae. Bacillus with
A method for cultivating poppyri and a control agent for Scarabaeidae comprising a sporangia obtained by the culturing method as an active ingredient have been completed.

That is, the present invention provides (1) spores and a parasporal body, wherein a bacterium belonging to Bacillus popiliae is cultured in a medium containing 0.01% to 0.5% by weight of activated carbon. A method for producing a Bacillus popirie sporangia comprising:

(2) The method according to (1), wherein the ratio of the number of sporangia to the total number of bacteria is 0.1% or more, which is represented by the sporangialization rate of the formula 1,

[0020]

[Formula 1] Sporulation rate (%) = (number of sporangia) / (total number of bacteria) × 100

(3) The production method according to (1) or (2), wherein the concentration of glucose contained in the medium is 0.01% by weight or less, and (4) spores and a parasporal body. The method according to any one of (1) to (3), wherein the sporangia comprising: has insecticidal activity against insects of the order Scarabaeidae; and

(5) The production method according to (4), wherein the insect of the family Scarabaeidae is Scutellariae, and (6) the production method according to any one of (1) to (5), wherein the culture is a solid culture. When,

(7) The production method according to (6), wherein the medium contains agar, polypeptone, yeast extract and phosphate, and (8) the bacterium belonging to Bacillus popiliae is・ Semadara (FERM P-1
6818), the method according to any one of (1) to (7),

(9) The sporangium of Bacillus popirie containing spores and parasporal body, which is produced by the production method according to any one of the above (1) to (8), is used as an active ingredient. An insecticide for scarabidae,

(10) Any one of the above (1) to (8)
And a method for controlling Scarabaeidae insects in which a spore bacillus of Bacillus popirie containing a spore and a parasporal body produced by the production method described in (1) is applied to Scarabaeidae.

[0026]

BEST MODE FOR CARRYING OUT THE INVENTION The bacterium belonging to the genus Bacillus populiae used in the present invention is Bacillus sp.
us popilliae). The bacteriological properties of the Bacillus spp. Species are described in the Bargeys Manual of Deterministic Bacteriology (Bargeys Ma
According to the Natural of Determinative Baceriology), the morphological properties are 1.3-5.2 μm in length and 0.5-0.5 μm in width.
8 μm gram-negative bacilli, growth temperature 20-35 ° C., with a parasporal body in the sporangia.

Bacillus popilli
The bacteria belonging to ae) include Bacillus popilliae semadara (Bacillus popilliae semadara) (May 2, 1998).
The Bacillus popirie dutoki strain (deposited with the Accession No. FERM P-16818 at the Institute of Biotechnology and Industrial Technology, Ministry of International Trade and Industry)
popilliae dutky) (American Type Culture Collecti
on No. 14706, hereafter referred to as ATCC 14706), Bacillus popirie meloronsa (Bacilluspopi)
lliae melolontha) and the like.

Hereinafter, a method for producing a Bacillus populiae sporangia according to the present invention will be described. As a method for culturing the microorganism, ordinary solid culture can be used. Polysaccharides such as agar, gellan gum, curdlan, and xanthan gum can be used as a substrate for solid culture, but culture using agar is particularly preferred.

In general, a carbon source such as a saccharide is added to the medium to promote good growth of microorganisms. Bacteria belonging to the genus Bacillus popillier used in the present invention are also promoted by the addition of a sugar source such as glucose. However, in order to form a sporangia containing spores and parasporal body, the addition of glucose is not preferable, and the concentration of glucose contained in the medium is preferably 0.01% by weight or less.

As the nitrogen source used for the medium, organic nitrogen such as polypeptone and dried yeast extract can be used. These concentrations range from 0.1% to 3.0% by weight, preferably 0.25% to 1.5% by weight. These nitrogen sources often contain glucose, but it is preferable to select and use a nitrogen source so that the glucose concentration in the medium does not exceed 0.01% by weight.

As the phosphate used in the medium, phosphates such as potassium phosphate and dipotassium hydrogen phosphate or inorganic salts such as sodium salts thereof can be used. These concentrations are desirably used in the range of 0.05% by weight to 1.0% by weight. The concentration of polysaccharides such as agar and gellan gum added to make a solid medium is 0.5% by weight to 5.0% by weight, preferably 1.0% by weight to 3.0% by weight.

Activated carbon is added to the medium, and the type of activated carbon to be added may be any type of activated carbon such as powder, granule, sheet and the like. There is no particular limitation, but powdered activated carbon is preferred. The activated carbon concentration in the medium is one of the important requirements of the present invention, and is 0.01% to 0.5% by weight.
Needs to be When the concentration of activated carbon in the medium is 0.01% by weight or less, the effect of removing substances that inhibit the formation of sporangia including spores and parasporal body is not sufficient. And the formation of sporangia including parasporal body is sharply reduced.

The activated carbon may be added to the medium before sterilization or may be added to the medium after sterilization. However, it is important that the activated carbon is sufficiently sterilized and is not contaminated by other microorganisms. is necessary. The culturing temperature is 25 ° C to 35 ° C, preferably 27 ° C to 30 ° C. The culturing time varies depending on the culturing temperature, but is usually 10 to 1 at 27 ° C to 30 ° C.
5 days.

After completion of the culture, a method for obtaining sporangia from the culture is as follows. Since the cells containing the sporangium are on the surface of the solid culture medium, a buffer solution such as water or a phosphate buffer or Tris-HCl is used. The cells and the sporangia are suspended and washed off by using, and then the cells and the sporangia are obtained by a general separation method such as centrifugation or filtration.

In cultivation in a conventional artificial medium containing at least 0.6% by weight of activated carbon, almost no spores of Bacillus popirie containing spores and parasporal body are obtained, and sporangia of Formula 1 is obtained. The ratio of the number of sporangia to the total number of bacteria, expressed as a percentage, is usually less than 0.05% by weight.

[0036]

[Formula 1] Sporulation rate (%) = (number of sporangia) / (total number of bacteria) × 100

On the other hand, the number of sporangia obtained by the method for producing sporangia of Bacillus populiae of the present invention is much larger than the number of sporangia obtained by the conventional culture method.

The obtained bacterial cells and sporangia may be used as a control agent for insects of the order Scarabaeidae in the form of a buffer in which they are suspended, or may be dried and used as a powder. After drying, it may be used as a suspension of water or a buffer solution. Further, these sporangia may be used in admixture with components such as known and commonly used excipients, carriers and nutrients used for agricultural chemicals. It is also possible to use the spores of the present invention in combination with other microbial preparations.

The insecticide of the present invention, which comprises the sporangia of Bacillus popiliae as an active ingredient, is an insect of the order Scarabaeidae, which is a well-known insect pest of lawns and agricultural crops.
itopertha orientalis, Bean beetle (Popillia japo)
nica), phyllopertha diversa,
Both exhibit excellent insecticidal and control effects on Chinese locust bean (Adoretus tenuimaculatus) and Japanese corn beetle (Anomala rufocuprea), and are particularly excellent against the larvae of relatively large beetles, Douganebuibui and Semarakogane. Show the effect.

[0040]

The present invention will be described more specifically with reference to the following examples and test examples, but the scope of the present invention is not limited to these examples.

(Example 1) (Preparation of culture medium) Two 300 ml Erlenmeyer flasks each containing distilled water 1
00 ml, tryptone 0.5% by weight (DIFCO
), 1.5% by weight of yeast extract (manufactured by Oxoid), 0.3% by weight of dipotassium hydrogen phosphate (special grade manufactured by Wako Pure Chemical Industries), 2.0% by weight of agar (special grade manufactured by Wako Pure Chemical Industries) ) Was added and mixed. (Hereinafter, a medium containing these medium components is referred to as medium A.)

Medium A contains a special-grade powdered activated carbon manufactured by Wako Pure Chemical Industries, Ltd.
It was added so as to be 0.1% by weight and 1.0% by weight. Thereafter, the mixture was sterilized in an autoclave at 121 ° C. for 20 minutes and sufficiently stirred before the agar was solidified, and the diameter was 9 cm.
Was added in a plastic petri dish to prepare a plate medium. Using those plate media having different activated carbon contents, Bacillus popirie semadala strain and Bacillus
Popilier dutoki (ATCC 14706) strain was cultured.

Semadara strain (FERM P-16818)
Used sporangia in bodily fluids collected from larvae infected with scarab beetles infected with emulsified disease. The number of sporangia was measured by a direct detection method using a spore microscope, adjusted to 1 × 10 ⁴ cells / ml, taken in an eppen tube 0.5 ml, and heated at 70 ° C. for 20 minutes in a heat block. Was applied to the plate medium prepared above.

The Dutoki strain was prepared by lyophilizing and storing cells (excluding spores and sporangia) by 0.5% by weight of tryptone manufactured by DIFCO, 1.5% by weight of yeast extract manufactured by Oxoid, and Wako Pure Chemical Industries, Ltd. 0.3% by weight of special grade dipotassium hydrogen phosphate manufactured by Wako Pure Chemical Industries, Ltd.
Using a liquid medium that had been sterilized at 121 ° C. for 20 minutes with the addition of 20% by weight, and in a 30 ° C. incubator for 120 r.
The cells were cultured for one week under the stirring conditions of pm. 0.1 ml of the culture was aseptically applied to an agar medium. The plate medium coated with the above cells was placed in a 30 ° C. incubator for 1 hour.
Cultured for 4 days.

After completion of the culture, distilled water 2 is aseptically added to a petri dish.
After the resulting colonies were suspended by adding ml, bacterial cells and sporangia were obtained. The number of sporangia per unit volume in the obtained solution and the total number of bacteria were measured by direct microscopy with a microscope, and the sporangialization rate was calculated by Equation 1. Table 1 shows the test results in the case where the Semadara strain was used. Only when the activated carbon addition concentration was 0.1% by weight, sporangia was formed.

[0046]

[Formula 1] Sporulation rate (%) = (number of sporangia) / (total number of bacteria) × 100

FIG. 2 shows a micrograph of the sporangium of the semaphora strain obtained when the activated carbon addition concentration was 0.1% by weight.
The micrograph is of ECLIPSE E6, an optical microscope manufactured by Nikon Corporation.
00 to Sony Color Video Camera Module XC
-003 was connected and photographed (magnification: × 3800). It can be seen that the sporangium contains one spore and one parasporal body.

Table 2 shows the test results when the Dutoki strain was used. Similarly, sporangia was formed only when the activated carbon concentration was 0.1% by weight. Table 1 shows the formation of sporangia by the Semadara strain, and Table 2 shows the formation status of sporangia by the Dutoki strain.

[0049]

[Table 1]

[0050]

[Table 2]

(Example 2) The amount of activated carbon in the medium was examined. The components of the medium A were added to each of seven 300-ml Erlenmeyer flasks, and no special powdered activated carbon manufactured by Wako Pure Chemical Industries, Ltd. was added thereto, and 0.05% by weight, 0.1% by weight, and 0.2% by weight.
5 wt%, 0.5 wt%, 1.0 wt%, 2.0 wt%
It was prepared to contain. Thereafter, autoclave sterilization was performed at 121 ° C. for 20 minutes to prepare a plate medium in the same manner as in Example 1.

The sedara strain was prepared by adding activated carbon in Example 1.
Cells containing sporangia obtained by culturing at a concentration of 0.1% by weight
It was used. The obtained solution is aseptically diluted with distilled water and 1 ×
10 ^FiveIt was prepared so that the number of cells / ml. Eppe the solution
0.5 ml into a heat tube and 70 with a heat block.
After heating at ℃ for 20 minutes, take 50μl and prepare as above
The activated carbon was applied to plate media with different amounts of activated carbon.
The plate medium coated with the cells was incubated at 30 ° C.
The cells were cultured for 14 days in a container.

After completion of the culture, distilled water 2 is aseptically added to a petri dish.
After the colonies generated were suspended by adding ml, sporangia and bacterial cells were obtained. The number of sporangia per unit volume and the total number of bacteria in the obtained solution were measured by direct microscopy with a microscope, and the sporangialization rate was calculated by Equation 1. FIG. 3 shows the relationship between the activated carbon concentration in the medium and the sporangialization rate. The vertical axis shows the sporangialization rate (%), and the horizontal axis shows the activated carbon concentration (% by weight) in the medium.
Represents When the activated carbon concentration is 0.5% by weight or less, good sporangia formation is observed.

Example 3 The influence of the concentration of glucose in the medium on sporangia formation was examined. A plate medium was prepared so that the activated carbon concentration of the medium A was 0.1% by weight, and a plate medium prepared such that the glucose concentration was 0.2% by weight and the activated carbon concentration was 0.1% by weight in the medium A. (Hereinafter, referred to as plate medium P and plate medium Q, respectively).

A plate was prepared by adjusting the concentration of activated carbon to 0.1% by weight in a medium in which the concentrations of tryptone and yeast extract in the components of the medium A were each doubled (hereinafter referred to as a plate medium R). ).

Further, the medium components were 1.0% by weight of tryptone manufactured by DIFCO, 0.6% by weight of yeast extract manufactured by Oxoid, 0.3% by weight of special grade dipotassium hydrogen phosphate manufactured by Wako Pure Chemical, and Wako Pure Chemical. 1.5% by weight of special grade agar manufactured, 1.0% by weight of special grade starch made by Wako Pure Chemical Co., 0.1% by weight of special grade glucose made by Wako Pure Chemical, 0.1% by weight of special grade fructose made by Wako Pure Chemical, Wako Pure Chemical Special grade sucrose 0.
1 wt%, Wako Pure Chemical Special Grade Maltose 0.1 wt%, Wako Pure Chemical Special Grade Mannose 0.1 wt%, Wako Pure Chemical Special Grade Salicin 0.1 wt% Plate medium with 0.6% by weight (referred to as plate medium S)
It was created.

The glucose concentration in the medium component A was measured to compare the glucose concentration in the medium. A solution was prepared by removing agar from the medium A component, and the glucose concentration was measured by an enzymatic method using an F kit from Boehringer Mannheim. The resulting glucose concentration was 38 mg / l (0.0038% by weight).

As described above, the glucose concentration in each plate medium was 0.0038% by weight for the plate medium P, 0.2% by weight for the plate medium Q, 0.0076% by weight for the plate medium R, and 0.1% for the plate medium S. 1% by weight. The measured value of the plate P was doubled because the concentration of the tryptone / yeast extract was doubled in the plate medium R. Since glucose is added to each of the plate media Q and R, the concentration of the existing glucose is equal to the added glucose concentration.

The Bacillus popirie semadara strain was cultured using the above four types of plate punishment. The culture method used is the same as in Example 1. The sporangialization rate was calculated in the same manner as in Example 1. Table 3 shows the glucose concentration in the medium and the sporangialization rate. It can be seen that the sporangialization rate is good when the glucose concentration is 0.01% by weight or less.

[0060]

[Table 3]

Example 4 An insecticidal test was conducted on a beetle insect using the sporangia obtained by the production method of the present invention. Humic soil 2 in 10 plastic cups 6cm in diameter
0 g was added at a time. Five of them had 1 sporangia.
The sporangia was sprayed so as to give × 10 ⁸ / cup. For the sporangial spores sprayed, a spore sac obtained by preparing a plate having the same composition as the plate P shown in Example 3 and culturing it under the same conditions was used.

The remaining five were tested as controls without sporangia. In each cup, one third-stage larva of the moss, Semaphora kogane, was placed one by one, bred in an incubator at 25 ° C. for 40 days, the number of dead individuals was examined, and the presence of emulsified disease spores in the dead bodies was examined with a microscope.

Table 4 shows the results of the insecticidal activity of the extracorporeal sporangia of the insect against Scutellaria cricket. By day 30, all larvae had died, and two of the five larvae that had died formed spores of emulsified disease in the body.

[0064]

[Table 4]

Example 5 Three cups prepared in the same manner as in Example 4 were prepared. Then put 2nd instar larvae of Douganebuui in the incubator at 25 ° C.
After breeding for 0 days, the number of dead animals was examined, and the presence of emulsified disease spores in the dead bodies was examined under a microscope. Table 5 shows the results of the insecticidal activity test of the sporangia produced by the production method of the present invention with respect to Douganebuui. By day 30, 67% of the larvae had died, and one out of two dead larvae
The head had formed a sporocyst of emulsified disease in the body.

[0066]

[Table 5]

[0067]

Industrial Applicability The present invention provides a Bacillus popirie spore that can obtain a large amount of Bacillus popirie spores containing spores and parasporal bodies using an artificial medium without using larvae of Scarabaeidae. It is possible to provide an industrial method for producing a sac, an excellent control agent for a scarabid insect that causes a great deal of damage to the roots of turf, agricultural and horticultural crops, trees, and the like, and a method for controlling the sac as an active ingredient. .

[Brief description of the drawings]

FIG. 1 is a schematic diagram of a Bacillus popirie sporangia containing spores and a parasporal body.

FIG. 2 shows that in Example 1, the concentration of activated carbon added was 0.1% by weight.
5 is a photomicrograph of a sporangium of a semaphora strain obtained in the case of FIG.

FIG. 3 is a graph showing the relationship between the activated carbon concentration in a medium and the sporangialization rate. The vertical axis represents the sporangialization rate (%), and the horizontal axis represents the activated carbon concentration (% by weight) in the medium.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) // (C12N 1/20 (C12N 1/20 C12R 1:07) C12R 1:07) (72) Inventor Hideharu Nishihashi 2-23-9 Osakidai, Sakura City, Chiba Prefecture (72) Inventor Azusa Fujiya 256-3, Kamiichichi, Mutsawa-machi, Nagao-gun, Chiba Prefecture (72) Inventor Makoto Hasegawa 486-E, Shiinazakicho, Midori-ku, Chiba-shi, Chiba Prefecture −503 (72) Masao Tanaka, Inventor 6-11-12 Takahama, Mihama-ku, Chiba-shi, Chiba Pref. (72) Tomoko Yokoyama 5-16-1-501, Isobe, Mihama-ku, Chiba-shi, Chiba F-term (reference) 4B065 AA15X BB02 BB15 BB23 BB26 BB29 BC13 BC41 CA48 CA60 4H011 AC01 BB21 DC11

Claims (10)

[Claims] 1. A spore of Bacillus popirie containing a spore and a parasporal body, wherein the bacterium belonging to Bacillus popirie is cultured in a medium containing 0.01% to 0.5% by weight of activated carbon. How to make the capsule. 2. The method according to claim 1, wherein the ratio of the number of sporangia to the total number of bacteria, which is represented by the sporangialization rate of the formula 1, is 0.1% or more. (Formula 1) Sporulation rate (%) = (spore number) / (total number of bacteria) × 100 3. The method according to claim 1, wherein the concentration of glucose contained in the medium is 0.5.
3. The composition according to claim 1, wherein the content is not more than 01% by weight.
The production method described in 1. 4. The method according to claim 1, wherein the sporangia containing the spores and the parasporal body has insecticidal properties against insects of the order Scarabaeidae. 5. The production method according to claim 4, wherein the insect of the family Scarabaeidae is Scutellaria cruzi. 6. The production method according to claim 1, wherein the culture is a solid culture. 7. The method according to claim 6, wherein the medium contains agar, polypeptone, yeast extract and phosphate. 8. The bacterium belonging to Bacillus popiliae is Bacillus popiriae semadara (FERM P-1681).
8) The method according to any one of claims 1 to 7, wherein 9. A control of an insect of the order Scarabaeidae, comprising a spore of Bacillus popirie containing a spore and a parasporal body produced by the method of any one of claims 1 to 8 as an active ingredient. Agent. 10. A Scarabaeid insect, wherein a Bacillus popirie sporangia containing a spore and a parasporal body produced by the method according to any one of claims 1 to 8 is allowed to act on a Scarabaeid insect. Control method.