JPH10218716A - Insect pest controlling material and its production - Google Patents

Abstract

PROBLEM TO BE SOLVED: To simply obtain the subject readily handleable material using an inexpensive material, capable of maintaining activity of a filamentous fungus for a long period of time, by including a filamentous fungus, an organic substrate and a porous substance. SOLUTION: (A) A filamentous fungus (e.g. Metarhizium anisopliae) is cultured in a culture base containing (B) an organic substrate (e.g. rice bran) which is preferably a plant substrate and (C) a porous substance (e.g. perlite) to give an insect pest controlling material having >=10<8> CFUs/g (CFU: colony forming unit) filamentous fungus spores in which the filamentous fungus spores are stuck to the pore of the component B. The ratio of the component C to the component B is 2:8 to 8:2, preferably 2:8 to 3:7, more preferably 3:7 by weight. The amount of water to be added to the culture base is preferably 20-60% the maximum water retention amount. Preferably culture temperature is 20-30 deg.C and the culture period is 2-5 weeks.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a pest control material and a method for producing the same, and more particularly, to a pest control material having a high pest control effect using natural enemy microorganisms (filamentous fungi), and capable of maintaining its activity and being stored for a long period of time. And its manufacturing method.

[0002]

2. Description of the Related Art Conventionally, microbial materials obtained by culturing filamentous fungi include those obtained by culturing filamentous fungi in a liquid medium, those obtained by culturing filamentous fungi in a solid powdered medium such as bran, and those using bran etc. as materials. (JP-B-7-37365), which have been used.

[0003]

However, among the above-mentioned filamentous fungal materials, those using a liquid medium have a problem in storage stability, while those using a powdered solid medium such as bran form a clump of material due to the formation of a hyphae bundle. Due to the formation, there is a problem in handling during production, application, and the like.

In the method using pellets such as bran, it is possible to increase the gas phase rate by using pellets and to suppress the formation of hyphal bundles. The grains become large. Since the filamentous fungal material needs to spread evenly on the soil surface or in the soil, it is preferable that the grain of the material be as small as possible.However, it is difficult to use the pellets to spread the material throughout the application on lawn or the like. ,
In order to pulverize and refine the pellets, a drying step is necessary, and the number of spores is reduced and spores are scattered.

[0005] Accordingly, the present invention provides a method for producing a spore having a filamentous fungus count of 10
Maintains ⁸ CFU / g or more (CFU: colony forming unit) and maintains the activity of filamentous fungi in the material for a long period of time. It is an object of the present invention to provide a simple material that does not go through a manufacturing process such as drying and pulverization using a simple material.

[0006]

SUMMARY OF THE INVENTION The present invention provides a pest control material containing a filamentous fungus, an organic substrate and a porous substance, and a method of culturing a filamentous fungus on a culture substrate containing an organic substrate and a porous substance. A method for producing a pest control material characterized by the following.

The filamentous fungus used as a natural enemy microorganism in the present invention is not particularly limited, but is preferably a fungus used for microbial pesticides, for example, Metarhiziu.
m) genus, genus Beauveria (eg Beauveria tenella, Beauveria bassiana) or Aschersonia (Aschersonia)
onia) genus (e.g., Ashersonia aleirodes)
hersonia aleyrodes)), more preferably a microorganism belonging to the genus Metarhizium, particularly Metarhizium anisoplia.
e) is used. Metarhidium Anisoprie (Metarhi
zium anisopliae) is not particularly limited, for example, Metarhizium anisopliae IFO
5940.

[0008] Organic substrates used in the present invention include plant substrates such as rice bran, soybean meal, bran, rapeseed meal, cottonseed meal, castor meal, and peach; steamed bone meal, dry blood, fish cake, hair meal, and skin An animal substance substrate such as flour, feather meal, crab and the like can be mentioned, and a vegetable substrate, particularly rice bran is preferred.

In the present invention, a porous substance is used as a culture substrate in addition to the organic substrate. As a result, the use of inexpensive materials to suppress the formation of filamentous fungal hyphae (lumps of hyphae) formed in the material by culturing a single material such as bran and obtaining a gas phase rate equal to or higher than that of pellets Can be. In addition, the particles of the material are small without being crushed as in the case of using pellets, and the activity of the filamentous fungi in the material can be maintained for a long period of time. Examples of the porous substance used in the present invention include perlite, isolite, zeolite, vermiculite, charcoal, coral sand, and preferably perlite.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION The pest control material of the present invention can be produced by culturing filamentous fungi on a culture substrate containing an organic substrate and a porous substance. By culturing in the presence of the porous substance as described above, the filamentous fungal hypha grows so as to cover the porous substance, the filamentous fungal spores adhere to the pores of the porous substance (FIG. 1), and the pest control. The number of filamentous fungal spores in the material can be at least 10 ⁸ CFU / g. The number of filamentous fungal spores in the pest control material of the present invention is usually 10 ⁸ to 10 ¹⁰
CFU / g. By simply mixing the porous material after the culture, the filamentous fungi are not incorporated into the porous material (FIG. 2), and the number of filamentous fungal spores in the pest control material is 10 ⁸ CF
U / g or more cannot be achieved, and the pest control effect becomes insufficient.

The weight ratio of the porous substance to the organic substrate is preferably from 2: 8 to 8: 2, more preferably from 2: 8 to 3: 7, and most preferably 3: 7. When the use ratio of the porous substance is 1: 9 or less by weight ratio of the porous substance and the organic substrate, a mycelial bundle is formed, a lump is formed, and application unevenness at the time of application and unevenness of the effect occur. Grinding is required. As a combination of the porous substance and the organic substrate, perlite and a vegetable substrate, particularly perlite and rice bran are preferable.

In order to maintain a sufficient spore formation rate and to omit the drying step after culturing, the amount of water added to the culture substrate is preferably 20 to the maximum water retention amount.
60%, more preferably about 30%. The culture temperature is preferably 20 to 30 ° C, more preferably around 25 ° C, and the culture period is preferably 2 to 5 weeks, more preferably 4 weeks.

[0013]

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 Relationship between Organic Substrate and Spore Count An organic substrate necessary for the growth of bacteria in a material was examined. Twelve types of organic substrates were examined: steamed bone meal, dry blood, fish meal, hair meal, skin powder, feather meal, crab, and crab, and botanicals such as rice bran, soybean meal, bran, rapeseed meal, and castor meal. It is. For each, perlite was mixed as a porous substance so that the ratio of perlite: organic substrate (weight ratio) was 5: 5, water was added so as to be 60% of the maximum water retention, and the mixture was filled in a culture bag and sterilized in an autoclave. Then, 10 ⁴ spores of Metalolidium anisoprie IFO 5940 were inoculated per 1 g of the material, and cultured at 25 ° C. After 3 weeks of culturing, the number of spores in the material was counted by direct microscopy using a microscope. As a result, good spore formation was observed in all conditions for the plant material substrate, and for steamed bone meal and dry blood in the animal material substrate. (Table 1). Among them, rice bran was considered to be the most suitable as a substrate for the culture substrate, since the formation of spores was most favorable when rice bran was used as the substrate and the cheapest material for culturing.

[0015]

[Table 1]

Example 2 Relationship between Mixing Ratio of Porous Material and Organic Substrate and Number of Spores The mixing ratio of the porous material (here, using perlite) and the organic substrate (here, using rice bran) was determined. By changing the mixing conditions, the mixing conditions under which the formation of spores was highest while suppressing the formation of hyphal bundles were determined. A mixture of perlite and rice bran at a weight ratio of 0:10 to 10: 0, 60% of the maximum water retention was added, and autoclaved, metalolidium anisoprie IFO 5940 spores were used as material 1
Culture was performed at 25 ° C. by inoculating 10 ⁴ cells per g. After culturing for 3 weeks, the formation of hyphal bundles was confirmed and the number of filamentous fungal spores in the material was counted. As a result, the formation of hyphal bundles was suppressed as the ratio of perlite increased, but the number of spores tended to decrease (FIG. 3). When the use ratio of perlite is 1: 9 or less by weight of perlite and rice bran, a mycelial bundle is formed, a lump is formed, and application unevenness at the time of application and non-uniformity of the effect occur. there were. The number of spores in the material
If it is 10 ⁸ CFU / g or more, the number of spores showing insecticidal activity can be maintained when the material is actually applied to the soil. Therefore, the mixing ratio of perlite and rice bran in the culture substrate is 2% by weight. : 8 to 8: 2 is desirable.

(Example 3) Relationship between the amount of water added and the number of spores The rate of addition of water added to the culture substrate was examined. When the moisture content of the culture substrate exceeds 60% of the maximum water retention,
Although the growth of filamentous fungi in the material is sufficient, a drying step of the material is required to improve the handling after production. Therefore, the water content in the culture medium was adjusted to 5, 10, 20, 30, 40, 50 or 60% of the maximum water retention in the culture medium, and the water content on spore formation was examined.

As the filamentous fungus, Metalolidium anisoprie IFO 5940 was used, and its spores were added to 10 ⁴ per gram of the material.
The cells were inoculated and cultured at 25 ° C for 3 weeks. As a result, even if the added water content is reduced to 30% of the maximum water retention in the culture substrate, 60%
%, And it was confirmed that the drying step after culturing can be omitted by reducing the added water content to 30% of the maximum water retention in the culture substrate (FIG. 4). . Therefore, the amount of water to be added to the culture substrate is preferably 20 to 60% of the maximum water retention, and more preferably 30%.

Example 4 Preferred Production Example YPMG (3 g of yeast extract, 5 g of peptone, 1.5 g of meat extract,
Glucose 10g, agar 18g, water 1L, pH7.2) 5mm of metalolidium anisoprie IFO 5940 cultured on a plate medium
× 5 mm sections were inoculated into 50 ml of YPMG liquid medium, and cultured in a rotary incubator at 25 ° C. for 4 days to prepare a bacterial solution for inoculation containing 10 ⁸ or more spores per ml.

The culture substrate for inoculation of the bacterial solution is prepared by adding rice bran, bran, or a mixture of both to the perlite, so that the ratio of perlite to the plant substrate is 3: 7 by weight. To 500 g of the mixture, add water at 30% of the maximum water holding capacity of the base material, fill in a bag capable of aseptic air circulation capable of withstanding autoclave sterilization at 121 ° C, and mix for 60 minutes. Autoclave sterilization was performed. A bacterial solution cultured in a liquid medium was inoculated to a sterilized culture substrate, and cultured in a 30 ° C. incubator for 4 weeks.

A material obtained using rice bran as a plant substrate (the number of filamentous fungal spores in the material is 9.18 log CFU / g, and the number of filamentous fungal spores per gram of porous substance (pearlite) is 9.1.
6 log CFU) were used in the following test examples.

(Test Example 1) Insecticidal activity test of the material against scarab beetle larvae The material produced in Example 4 (using rice bran as a plant substrate) was used to test the insecticidal activity against the beetle larvae of the species Scarabaeidae. For the test, second-instar larvae of Douganebuui, grown on humus without metalolidium, were used. 10 ⁵ , 1 metallidium spores per gram
The materials adjusted to be 0 ⁶ , 10 ⁷ , and 10 ⁸ were mixed at a ratio of 1 g to 100 g of the humus, and the number of spores per gram of the humus was 10 ³ , 10 ⁴ , 10 ⁵ , and 10 ⁶ . Twenty larvae of the Douganebuui were released. As a control, the larvae of Douganebuui were also released to the humus soil to which the above-mentioned materials were not added, and raised at 25 ° C. for 3 weeks. Table 2 shows the results. Under the conditions of 10 ⁴ , 10 ⁵ , and 10 ⁶ spores per gram of mulch, the death of the larvae of Douganebuui by the infection with metallidium was confirmed.
^{For 6} , 90% of the larvae of the Douganebuui died from metalolidium infection.

[0023]

[Table 2]

(Test Example 2) Control Test for Sweet Potato At the laboratory level (Test Example 1), the insecticidal activity of the pest control material of the present invention against scarab beetle larvae was confirmed.
Next, an actual field-level control test was conducted. The target crop was sweet potato, and the material produced in Example 4 (using rice bran as a plant substrate) was applied for the purpose of controlling the damage of sweet potato by the scarab beetle, Douganebuui. Test group and area, District 1 4.54m × 5 m 24.75 m ² in triplicate, ridges number strains 45cm in furrow 90cm in 5 ridges, the cultivation was carried out in black and white double-Mulch. Regarding the material application date, application rate and method, on May 16, the material was applied to the soil surface in an amount of 100 g / m ² , and after application, the material was mixed into the soil by a tractor. Regarding the method of applying the materials, two types of application were performed: the entire application in the test plot and the application in the ridge. As a control plot, a non-treatment plot and a field where a conventional pesticide was applied by a visit were provided, and sweet potatoes were planted and a test was started. The number of metalloid bacteria in soil was measured before and immediately after application of the material, and the soil was sampled during the harvest survey in August and October, and the number of metalloid bacteria was counted using a selective medium. ² CFU /
Although the amount was less than g soil, the bacterial count of 10 ⁵ to 10 ⁶ CFU / g soil was maintained from immediately after application to the time of harvest survey. With regard to the judgment of the control of sweet potato damage by the larvae of the larvae of the larvae, sweet potatoes were dug at the time of harvesting, and the controllable effect of the metallidium material was determined by obtaining the harmful area of the larvae of the sweet potatoes and the surface of the sweet potatoes that were harmed by the larvae of the larvae. Table 3 shows the results. Regarding the degree of damage to sweet potato, the application material applied area of the Example material applied area showed a very high control effect in both the whole treatment and the in-ridge treatment compared to the non-treated area and the applied pesticide-built area. It was shown that sweet potato was effective in controlling the larvae of the larvae of the sweet potato.

[0025]

[Table 3]

(Test Example 3) Control Test for Golf Course Turf A test was performed using the fairway of a golf course as a test section.
The turf variety was Noshiba, and the beetle to be controlled was Semaphora kogane. The test plot was provided with a plot in which the material produced in Example 4 (using rice bran as a vegetable substrate) was applied, an untreated plot as a control plot, and a plot in which a conventional pesticide, MEP emulsion (trade name Sumithion) was applied. The application of the materials was done in August,
The application rate of the material is 10 metarhidium bacteria per gram of soil.
^The material was sprayed on a turf surface at a rate of 100 g per 1 m ² so as to have a concentration of ⁵ to 10 ⁶ CFU, rubbed lightly, and then sufficiently watered. The soil was sampled before and immediately after the application of the material and every two weeks, and the number of metalloid bacteria in the soil was counted. For judging the effectiveness of the mosquito larvae against turf damage, the soil with a diameter of 15 cm and a depth of 20 cm was sampled using a hole cutter in each test plot before and every two weeks after application, and the number of larvae in the soil was counted. Was done by doing. Regarding the number of metarhidium bacteria in the soil, it was less than 10 ² / g soil before application, but it became an order of 10 ⁵ to 10 ⁶ CFU / g soil after application of the example materials, and the bacterial count was maintained thereafter. Continued (FIG. 5). In FIG. 5, the larva density correction index represents the number of larvae in the treated group as a ratio to the larva number in the untreated group, taking the number of larvae in the untreated group as 100. The lower the value, the higher the effect. The number of larvae in the soil before the treatment was very high, around 80 before treatment, and in the untreated plot, about 80 larvae of the larvae in the untreated area were also confirmed (Table 1). 4). In the material application plots and the MEP emulsion treatment plots, which are conventional pesticides, the number of larvae of Semaphoridae in the soil showed a decreasing tendency after the treatment. Eventually, the number of larvae of the mosquitoes in the soil was about half that of the untreated group in the group applied with the example material and the group treated with the MEP emulsion. The materials of the examples also showed a high control effect against turf damage caused by larvae of the mosquitoes.

[0027]

[Table 4]

(Test Example 4) Material Storage Test A long-term storage test was performed using the material produced in Example 4 (using rice bran as a plant substrate). Regarding the storage conditions, a study was made on the case where the material was stored aseptically and the case where the culture bag was opened and stored aseptically under two kinds of temperature conditions of room temperature and 5 ° C., respectively. Save 1
The number of bacteria of metalolidium in the material was counted every month for one year. Table 5 shows the results. Under all storage conditions, it was confirmed that the number of metalloid bacteria in the material maintained the number of bacteria immediately after production even after one year.

[0029]

[Table 5]

[0030]

Industrial Applicability According to the present invention, it is possible to provide at low cost a material in which filamentous fungi are effective for controlling pest larvae, and the material for controlling pests of the present invention has a long term activity. It is possible to maintain and control pest larvae.

[Brief description of the drawings]

FIG. 1 is a photograph showing the morphology of an organism when the pest control material of the present invention is observed under magnification with a microscope.

FIG. 2 is a photograph showing the morphology of an organism when a pest control material obtained by mixing a porous substance after culturing is observed with a microscope under magnification.

FIG. 3 is a graph showing the relationship between the mixing ratio of rice bran and perlite and the number of spores.

FIG. 4 is a diagram showing the relationship between the amount of water added and the number of spores.

FIG. 5 is a diagram showing the relationship between the number of metalloid bacteria and the number of larvae in soil.

────────────────────────────────────────────────── ───

[Procedure amendment]

[Submission date] February 12, 1997

[Procedure amendment 1]

[Document name to be amended] Drawing

[Correction target item name] Fig. 1

[Correction method] Change

[Correction contents]

FIG.

[Procedure amendment 2]

[Document name to be amended] Drawing

[Correction target item name] Figure 2

[Correction method] Change

[Correction contents]

FIG. 2

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Katsunori Noguchi 5-5511 Namiki, Tsuchiura-shi, Ibaraki Katakura Chikarin Co., Ltd. Tsukuba Research Institute (72) Inventor Tadahiko Kameyama 1-2-3 Otemachi, Chiyoda-ku, Tokyo No. Katakura Tickerin Co., Ltd.

Claims (14)

[Claims] 1. A pest control material containing a filamentous fungus, an organic substrate, and a porous substance. 2. The pest control material according to claim 1, wherein the filamentous fungus is incorporated in the porous substance. The number of filamentous fungal spores in the pest control material is 10 ⁸ CF.
The insect pest controlling material according to claim 1, which is not less than U / g. 4. The number of filamentous fungal spores in the pest control material is 10 ⁸ or more.
The pest control material according to claim 1, wherein the material is 10 ¹⁰ CFU / g. 5. The pest control material according to claim 1, wherein the filamentous fungus is Metarhizium anisopliae. 6. The pest control material according to claim 1, wherein the weight ratio of the porous substance to the organic substrate is from 2: 8 to 8: 2. 7. The pest control material according to claim 1, wherein the porous substance is perlite, and the organic substrate is a plant substrate. 8. The pest control material according to claim 7, wherein the plant substrate is rice bran. 9. A method for producing a pest control material, comprising culturing filamentous fungi on a culture substrate containing an organic substrate and a porous substance. 10. The method according to claim 9, wherein the filamentous fungus is Metarhizium anisopliae. 11. The method according to claim 9, wherein the weight ratio of the porous substance to the organic substrate is from 2: 8 to 8: 2. 12. The method according to claim 9, wherein the porous substance is perlite, and the organic substrate is a plant substrate. 13. The vegetable substrate is rice bran.
The manufacturing method as described. 14. The production method according to claim 9, wherein water of 20 to 60% of the maximum water retention is added to the culture substrate.