JP3898343B2 - Novel microorganism and method for controlling scarab beetles using the same - Google Patents
Novel microorganism and method for controlling scarab beetles using the same Download PDFInfo
- Publication number
- JP3898343B2 JP3898343B2 JP15036298A JP15036298A JP3898343B2 JP 3898343 B2 JP3898343 B2 JP 3898343B2 JP 15036298 A JP15036298 A JP 15036298A JP 15036298 A JP15036298 A JP 15036298A JP 3898343 B2 JP3898343 B2 JP 3898343B2
- Authority
- JP
- Japan
- Prior art keywords
- larvae
- strain
- microorganism
- bacillus
- present
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Description
【０００１】
【発明の属する技術分野】
本発明は、バチルス・ポピリエに属する新規微生物、その胞子を用いたコガネムシ科昆虫の防除方法および微生物製剤に関する。前記微生物は、植物害虫であるコガネムシ科幼虫に対して乳化病を誘発することができるので、コガネムシ科昆虫に対する微生物農薬として利用することができる。
【０００２】
【従来の技術】
従来、芝地、牧草地、農地、果樹園、庭園などにおいて植物害虫となっているコガネムシ科昆虫に対して、その防除には化学合成された農薬を用いることが一般的であった。しかし、環境問題が重要視される時代背景下、自然環境や人体への悪影響が懸念される化学農薬に代わって、環境保全に貢献することのできる安全性の高い生物的防除法が切望されている。
【０００３】
上記のような観点から、昆虫に対して殺虫性を有する微生物を用いた微生物農薬が開発されており、例えば、鱗翅目や蚊の幼虫に対して病原性を有するバチルス・チューリンゲンシス（Bacillus thuringiensis）の生菌又はその殺虫成分を農薬とするいわゆるＢＴ剤は代表的なものとして知られている。
【０００４】
コガネムシ科昆虫についても、乳化病に冒されたマメコガネ（Popillia japonica）幼虫から単離されたバチルス・ポピリエに属する微生物が、マメコガネ幼虫に対して乳化病を誘発することが知られており、既に、米国においては該微生物を用いた微生物製剤が市販されている。しかしながら、該微生物は日本で重要害虫となっているドウガネブイブイ（Anomala cuprea）に対しては効果がない（農業有用微生物−その利用と展望− 梅谷献二、加藤肇 ２３６頁 １９９０年）ため、我が国においてコガネムシ科幼虫防除剤として使用するには満足できるものではなかった。
【０００５】
ドウガネブイブイに対して殺虫性を有するバチルス属細菌としては、バチルス・チューリンゲンシスに属する微生物（バチルス・チューリンゲンシス・バー・ジャポネンシスＮ１４１株、特開平８−２２８７８３号公報）が知られているが、バチルス・ポピリエに属する微生物では知られていない。
【０００６】
【発明が解決しようとする課題】
本発明は、上記観点からなされたものであり、ドウガネブイブイをはじめとするコガネムシ科昆虫の防除に有効な微生物、及びその胞子を用いたコガネムシ科昆虫の防除方法及び微生物製剤を提供することを課題とする。
【０００７】
【課題を解決するための手段】
本発明者らは上記課題を解決すべく鋭意研究を重ねた結果、ドウガネブイブイやセマダラコガネ（Blitopertha orientalis）の幼虫に対する強い殺虫活性を有するバチルス・ポピリエに属する新規な微生物を見出し、この知見に基づいて本発明を完成するに至った。
【０００８】
すなわち本発明は、（１）セマダラコガネ及びドウガネブイブイ に殺虫性を示すことを特徴とするバチルス・ポピリエ（Bacillus popilliae）に属する微生物、（２）バチルス・ポピリエ・セマダラ（Bacillus popilliae semadara）株（ＦＥＲＭ ＢＰ−８０６８）である（１）に記載の微生物、（３）上記（１）に記載の微生物をコガネムシ科昆虫に作用させることを特徴とするコガネムシ科昆虫の防除方法、（４）上記（１）に記載の微生物の胞子をコガネムシ科昆虫に作用させることを特徴とする（３）に記載の方法。（５）コガネムシ科昆虫がドウガネブイブイ 昆虫であることを特徴とする（３）に記載の方法。（６）コガネムシ科昆虫を防除するための微生物製剤であって、（１）に記載の微生物の胞子を含む微生物製剤、に関するものである。
【０００９】
本発明において、「コガネムシ科昆虫の防除」とは、コガネムシ科昆虫、特にコガネムシ科幼虫の駆除、及びコガネムシ科幼虫による植物虫害の予防及び改善をいう。
【００１０】
【発明の実施の形態】
以下、本発明を詳細に説明する。
【００１１】
＜１＞本発明の微生物本発明の微生物は、セマダラコガネ及びドウガネブイブイ に殺虫性を示すことを特徴とするバチルス・ポピリエに属する微生物である。該微生物として具体的には、バチルス・ポピリエ・セマダラ株が挙げられる。バチルス・ポピリエ・セマダラ株は、千葉県千葉市において乳化病に冒されたセマダラコガネ幼虫から、後記実施例に示すようにして単離された株である。バチルス・ポピリエ・セマダラ株は、平成１０年５月２１日より通商産業省工業技術院生命工学工業技術研究所（郵便番号３０５ 日本国茨城県つくば市東一丁目１番３号）に受託番号ＦＥＲＭ Ｐ−１６８１８の受託番号で寄託されている。その後、ブダペスト条約に基づく国際寄託に移管され、ＦＥＲＭ ＢＰ−８０６８の受託番号が付与された。
【００１２】
本発明のバチルス・ポピリエ・セマダラ株の細菌学的性質を表１に示す。この細菌学的性質、特に４０℃までの温度条件下で生育できる点、及びセマダラコガネに対し乳化病を誘発するという病原性から、本菌株はバチルス・ポピリエに属することを認めた。さらに、本菌株の病原スペクトルを調べたところ、ドウガネブイブイ幼虫に対しても殺虫効果を有することが認められ、従来公知のいずれのバチルス・ポピリエとも異なる新規な微生物であることが判明した。そこで、本菌株をバチルス・ポピリエ・セマダラ株と命名した。
【００１３】
＜２＞本発明のコガネムシ科昆虫の防除方法及び微生物製剤
本発明の微生物をコガネムシ科昆虫に作用させることにより、コガネムシ科昆虫を防除することができる。本発明の微生物をコガネムシ科昆虫に作用させることは、本発明の微生物の菌体又は胞子、好ましくは胞子を、コガネムシ科昆虫、好ましくは幼虫の体内に取り込ませることにより行われる。
【００１４】
本発明の方法は、コガネムシ科に属する昆虫に広く適用し得るが、特にセマダラコガネ及びドウガネブイブイに対して好適に適用することができる。
コガネムシ科昆虫体内に取り込ませるのは、本発明の微生物の菌体であってもよく、本発明の微生物が形成する胞子であってもよいが、胞子が好ましい。胞子は、例えば、次に示すようにして調製することができる。コガネムシ科幼虫、好ましくはセマダラコガネ幼虫あるいはドウガネブイブイ幼虫に本発明の微生物を取り込ませる。具体的には、本発明の微生物を前記幼虫の存在する飼育培土などに散布し、経口的に摂取させるか、又は、体液中に注射することにより注入する。この幼虫を好ましくは３週間〜４週間飼育し、該幼虫体内で本発明の微生物を増殖させる。幼虫体内で増やした胞子は、例えば、幼虫を切開あるいは穴をあけるなどして体液を採取し、得られた体液を例えば遠心分離又は濾過することによって、得ることができる。
【００１５】
本発明の方法において、胞子は、そのまま用いてもよく、あるいは必要に応じて水あるいは中性の緩衝液、例えばＰＢＳ緩衝液（ＮａＣｌ８ｇ、ＫＣｌ０．２ｇ、Ｎａ2ＨＰＯ4 １．４４ｇ、ＫＨ2ＰＯ4 ０．２４ｇ／１Ｌ、ｐＨ７．４）、リン酸緩衝液、又はＴｒｉｓ−ＨＣｌ緩衝液、好ましくはＰＢＳ緩衝液で洗浄してもよい。更に、胞子は、乾燥して粉末にしてもよいし、水あるいは上記緩衝液の懸濁液としてもよい。また、胞子は、微生物農薬等の微生物製剤に製造に通常用いられる担体、栄養剤等の成分と混合し、得られる組成物を微生物製剤として使用することもできる。さらに、この組成物にバチルス・チューリンゲンシスの菌体又はその殺虫成分を混合することによって、本発明の微生物と併用してもよく、コガネムシ幼虫に対してこれらの相乗作用が期待される。
【００１６】
本発明の微生物、胞子又は微生物製剤は、コガネムシ科幼虫を防除しようとする芝地、果樹園、農地などに土壌１ｍ2当たり胞子数が１×１０6個から１×１０14個、好ましくは１×１０9個から１×１０13個となるように散布して行う。その後、必要に応じて１ｍ2当たり１Ｌから２Ｌの散水し、及び／あるいは土壌を鍬こみ混和してもよい。懸濁液を用いる場合は、水あるいは前記中性緩衝液で１×１０6個／Ｌから１×１０14個／Ｌ、好ましくは１×１０9個／Ｌから１×１０13個／Ｌとなるように懸濁したものを用いることが好ましい。
【００１７】
【実施例】
以下、本発明の実施例によりさらに具体的に説明する。
【００１８】
【実施例１】
バチルス・ポピリエ・セマダラ株の単離・同定
千葉県千葉市に所在する千葉県農業試験場芝草試験園場おいて１９９７年の夏、セマダラコガネ幼虫が大発生した。高密度となった該幼虫個体群の中には、体が乳白色に変化しているものが多数発見され、そのほとんどが２週間以内に死亡するという現象が観察された。この乳化病に冒されたセマダラコガネ幼虫から以下に示す方法で病原菌株を単離した。即ち、乳白色になり死亡した幼虫に注射針で穴をあけ、血体腔から乳白色の体液を回収した。
【００１９】
上記体液から遠心分離により沈殿画分を採取した。顕微鏡観察により、この画分には、細菌の胞子が含まれていると推定された。この画分を６０℃で１５分間熱処理した後、ＭＹＰＧＰプレート（ミュラー−ヒントンブロス(Mueller-Hinton broth)１０ｇ、酵母エキス１０ｇ、Ｋ2ＨＰＯ4 ３ｇ、Ｃ3Ｈ3Ｏ3Ｎａ１ｇ、グルコース０．５ｇ、寒天２０ｇ／１Ｌ）に繰り広げ、３０℃で、嫌気状態で７日間培養した。培地上に形成したコロニーを分離し、再び前記と同様にして培養する操作を繰り返すことにより純粋菌株を分離した。
上記のようにして分離された菌株の細菌学的性質を調べた結果を表１に示す。
【００２０】
【表１】
表１に示す細菌学的性質から、本菌株はバチルス・ポピリエに属することを認めた。バージェイズ・マニュアル・オブ・デターミネイティブ・バクテリオロジー（Bargeys Manual of Determinative Bacteriology）に記載されているバチルス・ポピリエの菌学的性質によれば、形態的性質は長さ：１．３〜５．２μｍ、幅：０．５〜０．８μｍの桿菌であり、生育温度が２０〜３５℃である点以外は、表１と一致する。尚、バージェイズ・マニュアル・オブ・デターミネイティブ・バクテリオロジーには、３％ＮａＣｌ存在下での生育については記載されていない。
一方、実施例３及び実施例４に示すように、上記菌株はセマダラコガネやドウガネブイブイなどのコガネムシ科幼虫に対し乳化病を誘発するという病原性を有していた。ドウガネブイブイ幼虫に対しても殺虫効果を有する点で、本菌株は従来公知のいずれのバチルス・ポピリエとは異なる新規な微生物であり、バチルス・ポピリエ・セマダラ株と命名した。
【００２２】
【実施例２】
バチルス・ポピリエ・セマダラ株の胞子画分の製造
腐葉土と砂を３：１の割合で混合した飼育培土を直径６ｃｍのプラスチックカップに２０ｇを入れ、乳化病に感染していた幼虫から採取した胞子を１×１０3個程度散布した。セマダラコガネ３令幼虫をカップに入れ、２５℃で飼育した。幼虫の体が乳白色を帯びてきた時点で体表面を１％次亜塩素酸ナトリウム液で殺菌し、注射針で穴をあけ体液を採取した。該体液から１００００rpm、５分間の遠心分離により胞子を回収した。胞子をＰＢＳ緩衝液で２回洗浄し、１×１０9個／ｍｌとなるように調整し、胞子画分とした。
【００２３】
寄託菌株から胞子を調製する際は、幼虫の体液中に注射により栄養細胞を１頭当たり約１００個程度注入し、３０℃で２週間〜４週間飼育し、乳白色になった幼虫の体表面に穴をあけ体液を採取すればよい。
【００２４】
【実施例３】
本発明の微生物のセマダラコガネ幼虫に対する殺虫活性
腐葉土と砂を３：１の割合で混合した飼育培土を直径６ｃｍのプラスチックカップ２０個に各２０ｇずつ入れ、胞子数が１×１０7個となるように実施例２で得た胞子画分を散布した。同様にして胞子数が１×１０7個となるようなカップも２０個用意した。ぞれぞれのカップにセマダラコガネ３令幼虫を１頭ずつ、更にサツマイモ小片を１つずつ入れ、２５℃で４０日間飼育しながら死亡個体数を調べた。また、死亡幼虫体内に胞子が形成されているかどうか体液を顕微鏡で調べた。
【００２５】
結果を表２に示す。胞子を１×１０7個／カップ散布した区では、３０日目以降４０％の幼虫の死亡が確認された。１×１０8個／カップ散布した区においては散布後１４日目から乳白色になった個体が観察され、散布後３０日目で１００％の幼虫が死亡し、６０％が体内に胞子を形成していた。
【００２６】
【表２】
【実施例４】
本発明の微生物のドウガネブイブイ幼虫に対する殺虫活性
ドウガネブイブイの２令幼虫を用い、実施例３と同様の方法で行った。ただし、飼育培土として腐葉土と火山灰土を１：１の割合で混合したものを使用した。
【００２８】
結果を表３に示す。胞子を１×１０7個／カップ散布した区では、３０日目以降１５％の幼虫の死亡が確認された。１×１０8個／カップ散布した区においては散布後１８日目から乳白色になった個体が視察され、散布後４０日目で９５％の幼虫が死亡し、５５％が体内に胞子を形成していた。
【００２９】
【表３】
【実施例５】
ドウガネブイブイ幼虫防除による芝被害の軽減
芝を生やした面積約５００ｃｍ2のポットを用意し、その土中にドウガネブイブイ２令幼虫を１ポット当たり１０頭ずつ埋めた。該ポットに、実施例２で得た胞子を１．２５×１０10個／ｍ2あるいは１．２５×１０11個／ｍ2となるように散布した。また、胞子を散布しない区も対照として設けた。それぞれの区における芝の被害度を３０日間にわたって観察した。試験は各区４連で行い、被害度を平均値で表４に示した。
【００３１】
【表４】
【発明の効果】
本発明の微生物は、コガネムシ科幼虫に対し致死性の乳化病を誘発するため、菌体、特に胞子を幼虫に作用させることによってコガネムシ科幼虫を防除し、芝、牧草、果樹、農園芸植物などを該害虫の被害から保護することができる。特に本微生物は、バチルス・ポピリエに属する微生物としては、日本で重要害虫となっているドウガネブイブイの幼虫に対しても殺虫作用を有することが示された初めての菌株であり、よって我が国におけるその使用効果は非常に大きい。その上、自然環境への悪影響や人体への毒性はほとんどなく、本発明の防除方法は地球環境保全にも貢献する優れた防除方法である。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a novel microorganism belonging to Bacillus popilie, a method for controlling a scarab beetle insect using the spore, and a microorganism preparation. Since the microorganism can induce an emulsifying disease on a plant beetle larvae, it can be used as a microbial pesticide against a beetle insect.
[0002]
[Prior art]
Conventionally, it has been common to use chemically synthesized pesticides for the control of Scarabaeidae insects that are plant pests in turf, pasture, farmland, orchards, gardens and the like. However, under the era when environmental issues are regarded as important, instead of chemical pesticides that are concerned about adverse effects on the natural environment and the human body, highly safe biological control methods that can contribute to environmental conservation are eagerly desired. Yes.
[0003]
From the above viewpoint, microbial pesticides using microorganisms having insecticidal properties against insects have been developed. For example, Bacillus thuringiensis having pathogenicity against lepidoptera and mosquito larvae A so-called BT agent using a living bacterium or an insecticidal component thereof as an agrochemical is known as a representative one.
[0004]
As for the beetle insects, it is known that a microorganism belonging to Bacillus popilie isolated from the larvae of Populus japonica affected by the emulsification disease induces the emulsification disease against the beetle larvae, In the United States, a microbial preparation using the microorganism is commercially available. However, the microorganisms are not effective against Anomala cuprea, which is an important pest in Japan (Agricultural useful microorganisms-their utilization and prospects-Seiji Umeya, Satoshi Kato, page 236, 1990). It was unsatisfactory for use as a control of larvae.
[0005]
As bacteria belonging to the genus Bacillus having insecticidal activity against Douganebububu, microorganisms belonging to Bacillus thuringiensis (Bacillus thuringiensis bar Japonensis N141 strain, JP-A-8-287883) are known. It is not known for microorganisms belonging to Popilie.
[0006]
[Problems to be solved by the invention]
The present invention has been made from the above viewpoints, and it is an object of the present invention to provide a microorganism effective for controlling Scarabaeidae insects such as Douganebububu, a method for controlling Scarabaeidae insects using the spores, and a microorganism preparation. To do.
[0007]
[Means for Solving the Problems]
As a result of intensive research aimed at solving the above-mentioned problems, the present inventors have found a novel microorganism belonging to Bacillus popilie having a strong insecticidal activity against larvae of the mosquitoes and Blitopertha orientalis. The invention has been completed.
[0008]
That is, the present invention relates to (1) a microorganism belonging to Bacillus popilliae characterized in that it exhibits insecticidal activity against cedar and pupae, and (2) a Bacillus popilliae semadara strain ( FERM ). BP-8068 ), the microorganism according to (1), (3) a method for controlling a scarab beetle, which comprises causing the microorganism according to (1) to act on a scarab beetle, (4) the above (1) The method according to (3), wherein the spore of the microorganism according to (4) is allowed to act on a scarab beetle insect. (5) The method according to (3), wherein the scarab beetle insect is a Douganebububu insect. (6) A microbial preparation for controlling Scarabaeidae insects, comprising a microbial spore according to (1).
[0009]
In the present invention, the term “control of scarab beetles” refers to the control of scarab beetles, in particular the scarab beetles, and the prevention and improvement of plant insect damage by scarabs.
[0010]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, the present invention will be described in detail.
[0011]
<1> Microorganism of the Present Invention The microorganism of the present invention is a microorganism belonging to Bacillus popilie, characterized by its insecticidal activity against cedar and pupae. Specific examples of the microorganism include a Bacillus popilie cemadara strain. The Bacillus popilie cemadara strain is a strain isolated from the larvae of the scallop larvae affected by the emulsification disease in Chiba City, Chiba Prefecture, as shown in the Examples below. From May 21, 1998, Bacillus popilie cemadara strain has been entrusted to the Institute of Biotechnology, Institute of Industrial Science and Technology, Ministry of International Trade and Industry (Postal Code 305 1-3, Higashi 1-chome, Tsukuba City, Ibaraki, Japan). It is deposited with a deposit number of -16818. Later, the deposit was transferred to an international deposit based on the Budapest Treaty. The accession number of BP-8068 was given.
[0012]
Table 1 shows the bacteriological properties of the Bacillus popilie sedara strain of the present invention. From this bacteriological property, in particular, the ability to grow under temperature conditions up to 40 ° C., and the pathogenicity of inducing emulsifying disease against the scallop, it was confirmed that this strain belongs to Bacillus popilie. Further, when the pathogenic spectrum of this strain was examined, it was found that it has an insecticidal effect against moth larvae, and it was found to be a novel microorganism different from any conventionally known Bacillus popilie. Therefore, this strain was named the Bacillus popilie cemadara strain.
[0013]
<2> Method of controlling Scarabaeidae insects and microbial preparations of the present invention By causing the microorganism of the present invention to act on Scarabaeidae insects, Scarabaeidae insects can be controlled. The action of the microorganism of the present invention on a scarab beetle insect is carried out by incorporating the cells or spores, preferably spores, of the microorganism of the present invention into the body of a scarab beetle insect, preferably a larva.
[0014]
Although the method of the present invention can be widely applied to insects belonging to the family Scarabaeidae, it can be suitably applied particularly to the stag beetle and the stag beetle.
The microorganisms of the present invention may be incorporated into the insects of the scarab beetle, and the spores formed by the microorganisms of the present invention may be used, but spores are preferred. For example, the spore can be prepared as follows. The microorganism of the present invention is incorporated into a larvae of the family Scarabaeidae, preferably a larvae of the stag beetle or pupae. Specifically, the microorganisms of the present invention are sprayed on a rearing medium or the like where the larvae are present and ingested orally or injected into a body fluid. The larvae are preferably raised for 3 to 4 weeks, and the microorganisms of the present invention are propagated in the larvae. The spore increased in the larvae can be obtained, for example, by collecting the body fluid by incising or making a hole in the larvae and centrifuging or filtering the obtained body fluid.
[0015]
In the method of the present invention, the spore may be used as it is, or if necessary, water or a neutral buffer such as PBS buffer (NaCl 8 g, KCl 0.2 g, Na 2 HPO 4 1.44 g, KH 2 PO 4 0.24 g / 1 L, pH 7.4), phosphate buffer, or Tris-HCl buffer, preferably PBS buffer. Furthermore, the spore may be dried to a powder, or may be a suspension of water or the above buffer solution. In addition, the spore can be used as a microbial preparation by mixing the spore with components such as a carrier and a nutrient that are usually used in a microbial preparation such as a microbial pesticide. Furthermore, by mixing a Bacillus thuringiensis microbial cell or an insecticidal component thereof with this composition, it may be used in combination with the microorganism of the present invention, and these synergistic effects are expected against the scarab beetle larvae.
[0016]
The microorganism, spore or microbial preparation of the present invention has a spore count of 1 × 10 6 to 1 × 10 14 per 1 m 2 of soil, such as turf, orchard, farmland, etc. to control Scarabaeidae larvae, preferably 1 Sprinkling is performed so that × 10 9 to 1 × 10 13 . Thereafter, if necessary, 1 to 2 L of water per 1 m 2 may be sprinkled and / or the soil may be mixed and mixed. When a suspension is used, it is 1 × 10 6 cells / L to 1 × 10 14 cells / L, preferably 1 × 10 9 cells / L to 1 × 10 13 cells / L with water or the neutral buffer. It is preferable to use a suspended material.
[0017]
【Example】
Hereinafter, the present invention will be described in more detail with reference to examples.
[0018]
[Example 1]
Isolation and identification of Bacillus popilie sedara strains In the summer of 1997, a large larvae of the scallop larvae emerged at the Chiba Prefectural Agricultural Experiment Station in Chiba City, Chiba Prefecture. Among the larvae populations that became dense, many were found to have milky white color, and most of them died within 2 weeks. A pathogenic strain was isolated from the larvae of the scallop larvae affected by this emulsification disease by the following method. That is, a hole was made in the larvae that became milky white and died with an injection needle, and milky white body fluid was collected from the blood body cavity.
[0019]
The precipitate fraction was collected from the body fluid by centrifugation. By microscopic observation, it was estimated that this fraction contained bacterial spores. After heat-treating this fraction at 60 ° C. for 15 minutes, MYPGP plate (Mueller-Hinton broth 10 g, yeast extract 10 g, K 2 HPO 4 3 g, C 3 H 3 O 3 Na 1 g, glucose 0.5 g And agar 20 g / 1 L) and cultured at 30 ° C. in an anaerobic state for 7 days. A pure strain was isolated by repeating the operation of isolating colonies formed on the medium and culturing again in the same manner as described above.
The results of examining the bacteriological properties of the strains isolated as described above are shown in Table 1.
[0020]
[Table 1]
From the bacteriological properties shown in Table 1, it was confirmed that this strain belongs to Bacillus popilie. According to the bacteriological properties of Bacillus popilie described in the Bargeys Manual of Determinative Bacteriology, the morphological properties are length: 1.3-5. It is the same as Table 1 except that it is a koji mold having a width of 2 μm and a width of 0.5 to 0.8 μm and the growth temperature is 20 to 35 ° C. Incidentally, the Barjays Manual of Detergent Bacteriology does not describe the growth in the presence of 3% NaCl.
On the other hand, as shown in Example 3 and Example 4, the strain had a pathogenicity of inducing emulsifying diseases against Scarabaeidae larvae such as the stag beetle and Douganebububu. This strain is a novel microorganism different from any conventionally known Bacillus popilie in that it also has an insecticidal effect against the moth larvae, and was named Bacillus popilie cemadara strain.
[0022]
[Example 2]
Manufacture of the spore fraction of Bacillus popilie cemadara strain 20g of breeding soil mixed with humus and sand in a ratio of 3: 1 was put into a 6cm diameter plastic cup, and spores collected from larvae infected with emulsifying disease were collected. About 1 × 10 3 were sprayed. The third instar larvae were placed in a cup and raised at 25 ° C. When the larva body became milky white, the body surface was sterilized with 1% sodium hypochlorite solution, a hole was made with an injection needle, and the body fluid was collected. Spores were collected from the body fluid by centrifugation at 10,000 rpm for 5 minutes. Spores were washed twice with PBS buffer and adjusted to 1 × 10 9 cells / ml to obtain a spore fraction.
[0023]
When preparing spores from the deposited strain, about 100 vegetative cells are injected by injection into the larva body fluid, and raised at 30 ° C. for 2 to 4 weeks. A bodily fluid may be collected by making a hole.
[0024]
[Example 3]
20 g each of 20-cm plastic cups having a diameter of 3: 1 mixed with an insecticidal active humus and sand mixed with microbial larvae of the microorganism of the present invention in a ratio of 3: 1 so that the number of spores becomes 1 × 10 7. The spore fraction obtained in Example 2 was sprayed. Similarly, 20 cups having 1 × 10 7 spores were prepared. Each cup was filled with one third instar larvae and one small piece of sweet potato, and the number of dead individuals was examined while rearing at 25 ° C. for 40 days. In addition, the body fluid was examined under a microscope to determine whether spores were formed in the dead larvae.
[0025]
The results are shown in Table 2. In the group where 1 × 10 7 spores / cup were sprayed, 40% of larval deaths were confirmed after the 30th day. In the area where 1 × 10 8 pieces / cup was sprayed, individuals that became milky white were observed from the 14th day after spraying, 100% of the larvae died 30 days after spraying, and 60% formed spores in the body. It was.
[0026]
[Table 2]
[Example 4]
Insecticidal activity against the moth larvae of the microorganism of the present invention The second larvae of the moth moth were used in the same manner as in Example 3. However, a mixture of humus and volcanic ash soil at a ratio of 1: 1 was used as rearing culture soil.
[0028]
The results are shown in Table 3. In the group sprayed with 1 × 10 7 spores / cup, 15% of the larvae were confirmed after the 30th day. In the area sprayed with 1 × 10 8 / cup, milky white individuals were observed from the 18th day after spraying, 95% of the larvae died 40 days after spraying, and 55% formed spores in the body. It was.
[0029]
[Table 3]
[Example 5]
Mitigation of turf damage by controlling larvae larvae: Pots with an area of approximately 500 cm 2 were prepared, and 10 larvae of 2 larvae were buried in the soil. The spores obtained in Example 2 were sprayed on the pot so as to be 1.25 × 10 10 cells / m 2 or 1.25 × 10 11 cells / m 2 . In addition, a section where no spores were sprayed was also provided as a control. The degree of turf damage in each ward was observed over 30 days. The test was carried out in quadruplicate in each ward, and the degree of damage is shown in Table 4 as an average value.
[0031]
[Table 4]
【The invention's effect】
The microorganism of the present invention induces a fatal emulsification disease against the larvae of the scarab beetle, so that the larvae, especially spores are controlled by causing the larvae to act on the larvae, grass, grass, fruit trees, agricultural and horticultural plants, etc. Can be protected from damage by the pests. In particular, this microorganism is the first strain that has been shown to have an insecticidal action against the larvae of Doganebuibu, which is an important pest in Japan, as a microorganism belonging to Bacillus popilie. Is very big. In addition, there is almost no adverse effect on the natural environment and no toxicity to the human body, and the control method of the present invention is an excellent control method that contributes to the preservation of the global environment.
Claims (5)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP15036298A JP3898343B2 (en) | 1998-05-29 | 1998-05-29 | Novel microorganism and method for controlling scarab beetles using the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP15036298A JP3898343B2 (en) | 1998-05-29 | 1998-05-29 | Novel microorganism and method for controlling scarab beetles using the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH11332556A JPH11332556A (en) | 1999-12-07 |
| JP3898343B2 true JP3898343B2 (en) | 2007-03-28 |
Family
ID=15495343
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP15036298A Expired - Fee Related JP3898343B2 (en) | 1998-05-29 | 1998-05-29 | Novel microorganism and method for controlling scarab beetles using the same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP3898343B2 (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002293708A (en) * | 2001-03-30 | 2002-10-09 | Dainippon Ink & Chem Inc | Bacillus popilliae, method for controlling insect of family scarabaeidae by using the same and controlling agent |
| US7033993B2 (en) | 2001-04-13 | 2006-04-25 | Dainippon Ink And Chemicals, Inc. | Polypeptide having larvae growth inhibiting or insecticidal effect on scarabaeidae insects |
| EP1277763B1 (en) * | 2001-07-04 | 2006-12-20 | Dainippon Ink And Chemicals, Inc. | Polypeptides having larvae growth inhibiting or insecticidal effect on scarabaeidae insects and polynucleotides encoding the same |
| US7319028B2 (en) * | 2003-12-09 | 2008-01-15 | Dainippon Ink And Chemicals, Inc. | Process for producing sporangia of Bacillus popilliae, control agent, and controlling method |
| JP2007223973A (en) * | 2006-02-24 | 2007-09-06 | Certis Japan Kk | Granular preparation for controlling scarabaeidae insect pest, method for producing the preparation, and method for controlling scarabaeidae insect pest by using the preparation |
| CN109957534B (en) * | 2019-04-26 | 2020-07-03 | 沧州市农林科学院 | Bacillus subtilis and application thereof |
-
1998
- 1998-05-29 JP JP15036298A patent/JP3898343B2/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| JPH11332556A (en) | 1999-12-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR101334742B1 (en) | A novel Bacillus subtilis strain and use thereof for protecting the rust of plant roots | |
| JP3612071B2 (en) | Use of Streptomyces WYEC108 to control plant pathogens | |
| TWI302942B (en) | Bacillus sp.d747 strain, agent for controlling plant diseases and agent for controlling pests using the same, and controlling method using the same | |
| JP3428658B2 (en) | Antimicrobial microbial preparation, its production method and treatment method | |
| KR20180118266A (en) | Microbial pesticides against thrips | |
| HU220582B1 (en) | Composition and method for controlling plant diseases | |
| KR101680638B1 (en) | New microorganism Beauveria bassiana FG274 and Microbial control agent for the prevention of Spodoptera exigua larva | |
| JP3898343B2 (en) | Novel microorganism and method for controlling scarab beetles using the same | |
| Nagtzaam et al. | Efficacy of Talaromyces flavus alone or in combination with other antagonists in controlling Verticillium dahliae in growth chamber experiments | |
| KR100294023B1 (en) | Bacteria for disease prevention of crops, microorganisms containing them and uses thereof | |
| JP3665295B2 (en) | Microbial preparation for biological control using novel Trichoderma microbial strain and method for producing the same | |
| KR20020031961A (en) | Streptomyces kasugaensis GBA-0927 | |
| CN106244480B (en) | One plant of false Grignon anthropi and its application for preventing and treating plant nematode | |
| MX2014001251A (en) | Bionematicide composition and method for controlling phytopathogenic nematodes using the same. | |
| KR101389975B1 (en) | culture medium for preventing disease and promoting growth of plant, manufacturing method of microbial agent using the same | |
| JP3132195B2 (en) | New microorganism and plant disease control agent | |
| Abada et al. | Bacterial bioagents and compost as two tools for management of eggplant Fusarium wilt | |
| TW200825182A (en) | Novel bacillus bacterium strain capable of degrading/volume-reducing plant residue | |
| KR100314323B1 (en) | Bacillus sp. GB-017 KFCC-11070 | |
| KR100319135B1 (en) | Microbiological preparation inhibiting plant diseases | |
| JP2021020864A (en) | Antibacterial agents, agrochemicals, and methods of controlling plant infectious diseases by microorganisms | |
| JP4337193B2 (en) | Doganebuibu control method and control agent | |
| JP2001149066A (en) | Method for producing sporangium of bacillus popilliae | |
| KR102670639B1 (en) | Metarhizium anisopliae 432 strain for controlling bulb mite and uses thereof | |
| KR100458765B1 (en) | Bacillus thuringiensis strain for controlling dipteran pests and method for producing biological pesticide using it |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20040311 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20060919 |
|
| A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20061113 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20061205 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20061221 |
|
| R150 | Certificate of patent or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20100105 Year of fee payment: 3 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110105 Year of fee payment: 4 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120105 Year of fee payment: 5 |
|
| FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130105 Year of fee payment: 6 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
| LAPS | Cancellation because of no payment of annual fees |