JPWO2004074269A1 - TK-57-164A substance and TK-57-164B substance, their production process, and agricultural and horticultural fungicides containing them as active ingredients - Google Patents
TK-57-164A substance and TK-57-164B substance, their production process, and agricultural and horticultural fungicides containing them as active ingredients Download PDFInfo
- Publication number
- JPWO2004074269A1 JPWO2004074269A1 JP2005502755A JP2005502755A JPWO2004074269A1 JP WO2004074269 A1 JPWO2004074269 A1 JP WO2004074269A1 JP 2005502755 A JP2005502755 A JP 2005502755A JP 2005502755 A JP2005502755 A JP 2005502755A JP WO2004074269 A1 JPWO2004074269 A1 JP WO2004074269A1
- Authority
- JP
- Japan
- Prior art keywords
- substance
- agricultural
- culture
- present
- producing
- 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.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/20—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom three- or four-membered rings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/145—Fungal isolates
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D303/00—Compounds containing three-membered rings having one oxygen atom as the only ring hetero atom
- C07D303/02—Compounds containing oxirane rings
- C07D303/48—Compounds containing oxirane rings with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms, e.g. ester or nitrile radicals
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P15/00—Preparation of compounds containing at least three condensed carbocyclic rings
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Virology (AREA)
- Agronomy & Crop Science (AREA)
- Environmental Sciences (AREA)
- Biochemistry (AREA)
- Dentistry (AREA)
- General Engineering & Computer Science (AREA)
- Plant Pathology (AREA)
- Pest Control & Pesticides (AREA)
- Mycology (AREA)
- Biomedical Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Epoxy Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本発明は、下記の式（１）で表される新規なＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質並びにそれらの製造法及びそれらの少なくとも１つを有効成分とする農園芸用殺菌剤を提供する。本発明の新規化合物群は、畑、水田、果樹園、茶園、牧草地、芝生地などにおいて問題となる植物病害菌に対して優れた病原防除効果を示し、農園芸用殺菌剤としての利用が期待される。The present invention relates to a novel TK-57-164A substance and TK-57-164B substance represented by the following formula (1), a production method thereof, and an agricultural and horticultural fungicide containing at least one of them as an active ingredient I will provide a. The novel compound group of the present invention exhibits an excellent pathogen control effect against plant pathogens that are problematic in fields, paddy fields, orchards, tea gardens, pastures, lawns, etc., and can be used as an agricultural and horticultural fungicide. Be expected.
Description
本発明はＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質、並びにそれらの製造法及びそれらの少なくとも１つを有効成分とする農園芸用殺菌剤に関する。 The present invention relates to a TK-57-164A substance and a TK-57-164B substance, a production method thereof, and an agricultural and horticultural fungicide containing at least one of them as an active ingredient.
従来、多くの農園芸用殺菌剤が知られているが、薬剤抵抗性や安全性などの問題から、より有効で安全に使える薬剤が必要とされている。果樹、野菜、穀類などに発生する、うどんこ病、べと病、たんそ病、さび病、いもち病などの植物病原菌に対してこれまで各種の薬剤が使用されているが、薬剤抵抗性などの問題からそれらの薬剤の使用が禁止されたり、あるいは制限されてきている。したがってこれらの分野では従来の薬剤とは異なる新規な薬剤の出現が課題となってきており、本発明はこれらを解決することを目的とするものである。本発明は微生物が生産する新規なＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質並びにそれらの製造法及びそれらの少なくとも１つを有効成分とする農園芸用殺菌剤を提供する。 Conventionally, many agricultural and horticultural fungicides are known. However, due to problems such as drug resistance and safety, more effective and safe drugs are required. Various drugs have been used to date against plant pathogens such as powdery mildew, mildew, rot, rust, and blast that occur in fruit trees, vegetables, cereals, etc., but drug resistance, etc. Because of these problems, the use of these drugs has been prohibited or restricted. Accordingly, in these fields, the emergence of new drugs different from conventional drugs has become a problem, and the present invention aims to solve these problems. The present invention provides novel TK-57-164A and TK-57-164B substances produced by microorganisms, methods for producing them, and agricultural and horticultural fungicides containing at least one of them as an active ingredient.
農園芸用殺菌剤として利用できる新たな活性物質を、微生物の生産する培養物から探索したところ、本発明者らが新たに分離したＴＫ−５７株と命名したイサリア（Ｉｓａｒｉａ）属の子嚢菌の培養物中に植物病原菌に対し抗真菌活性を示す物質が生産されることを見出した。さらにこの抗真菌活性物質が下記の式（１）で表される化学構造を有することを見出し、新規な物質であることを確認して、本物質をＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質と命名した。
すなわち、本発明は下記の農園芸用殺菌活性物質ＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質を提供されるものである。
式（１）
また、本発明は、イサリア（Ｉｓａｒｉａ）属に属するＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質生産菌を培養し、その培養物からＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質を採取することを特徴とする上記に記載されているＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質の製造法を提供するものである。
さらに、上記に記載されているＴＫ−５７−１６４Ａ物質およびＴＫ−５７−１６４Ｂ物質の少なくとも１つを有効成分とする農園芸用殺菌剤およびイサリア（Ｉｓａｒｉａ）属に属する子嚢菌であって、上記のＴＫ−５７−１６４Ａ物質ならびにＴＫ−５７−１６４Ｂ物質を生産する特性を有ＴＫ−５７株またはその変異株に関する。
＜発明の実施の態様＞
本発明の第１の要旨とするところは、前記の式（１）で表され、下記の理化学的性状を有するＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質にある。
１．ＴＫ−５７−１６４Ａ物質の理化学的性状
（１）色および形状 無色固体
（２）マススペクトル Ｍ／Ｚ ４１０．２５４６（Ｍ＋Ｈ）＋
（３）分子式 Ｃ２２Ｈ３５ＮＯ６
（４）比旋光度［α］Ｄ −２１．３°（ｃ０．１０，ＣＨＣｌ３，２５℃）
（５）紫外部吸収スペクトル λｍａｘ（ＭｅＯＨ） ２１３ｎｍ
（６）赤外部吸収スペクトル νｍａｘ（ＫＢｒ） ｃｍ−１
３４００，３３００，２９２４，２８５３，２３５９，２３４１，１７４２，１６８４，１６４９，１６３２，１５３３，１４６０，１３６９，１３４４，１２７３，１１９８，１１５５，１１０１，１０８４，１００１，９７４，９３４，９１６，８４５，８３５，８１２
（７）ＨＰＬＣ カラム；Ｗａｔｅｒｓ Ｌ−ｃｏｌｕｍｎ ＯＤＳ １０φ×２５０ｍｍ、溶出溶媒；アセトニトリル−０．０５％ＴＦＡ（５２：４８）、流速；１．８ｍｌ／ｍｉｎ、検出；ＵＶ２５４ｎｍで保持時間４６．８４８分
（８）１Ｈ−ＮＭＲスペクトル；δ（ｐｐｍ）（ＣＤＣＬ３、４００ＭＨｚ）
０．８６（３Ｈ，ｔ，Ｊ＝６．８Ｈｚ），１．２０−１．３１（１２Ｈ，ｍ），１．４４（２Ｈ，ｍ），２．０３（１Ｈ，ｔ，Ｊ＝１２．７Ｈｚ），２．１８（２Ｈ，ｍ），２．２１（１Ｈ，ｂｒｓ），２．４９（１Ｈ，ｄｄｄ，Ｊ１＝１２．７Ｈｚ，Ｊ２＝４．９Ｈｚ，Ｊ３＝１．７Ｈｚ），３．１５（１Ｈ，ｄｄｄ，Ｊ１＝７．６Ｈｚ，Ｊ２＝３．４Ｈｚ，Ｊ３＝１．５Ｈｚ），３．５０（３Ｈ，ｓ），３．５２（１Ｈ，ｄｄ，Ｊ１＝３．６Ｈｚ，Ｊ２＝１．５Ｈｚ），３．６０（１Ｈ，ｄｄ，Ｊ１＝３．６Ｈｚ，Ｊ２＝１．２Ｈｚ），３．７４（１Ｈ，ｂｒｓ），３．９３（１Ｈ，ｄｄ，Ｊ１＝７．６Ｈｚ，Ｊ２＝１．２Ｈｚ），４．５０（１Ｈ，ｂｒｓ），４．８０（１Ｈ，ｍ），５．７７（１Ｈ，ｄｔ，Ｊ１＝１５．１Ｈｚ，Ｊ２＝１．５Ｈｚ），５．８３（１Ｈ，ｄ，Ｊ＝８．３Ｈｚ），６．８９（１Ｈ，ｄｔ，Ｊ１＝１５．１Ｈｚ，Ｊ２＝７．１Ｈｚ）
（９）１３Ｃ−ＮＭＲスペクトル；δ（ｐｐｍ）（ＣＤＣＬ３、１００ＭＨｚ）
１４．１（ｑ），２２．７（ｔ），２８．２（ｔ），２９．２−２９．７（４Ｃ，ｔ），３１．９（ｔ），３２．１（ｔ），３７．８（ｔ），４６．０（ｄ），５５．０（ｄ），５５．１（ｄ），５８．７（ｑ），６３．７（ｄ），７２．３（ｄ），７５．８（ｓ），７７．３（ｄ），１２２．９（ｄ），１４６．３（ｄ），１６５．４（ｓ），２０７．６（ｓ）
２．ＴＫ−５７−１６４Ｂ物質の理化学的性状
（１）色および形状 無色固体
（２）マススペクトル Ｍ／Ｚ ３８２．２２２７（Ｍ＋Ｈ）＋
（３）分子式 Ｃ２０Ｈ３１ＮＯ６
（４）比旋光度［α］Ｄ −１５．５°（ｃ０．１９，ＣＨＣｌ３，２５℃）
（５）紫外部吸収スペクトル λｍａｘ（ＭｅＯＨ） ２１３ｎｍ
（６）赤外部吸収スペクトル νｍａｘ（ＫＢｒ） ｃｍ−１
３４０２，３２９６，２９２６，２８５５，２３５９，２３４１，１７４２，１６８４，１６５１，１６３１，１５３１，１４５６，１３７１，１３４２，１２７３，１１９６，１１５５，１１０１，１０８４，１００１，９７４，９３４，９１６，８４５，８３３，８１２
（７）ＨＰＬＣ カラム；Ｓｅｎｓｈｕｐａｋ ＰＥＧＡＳＩＬ ＯＤＳ １０φ×２５０ｍｍ、溶出溶媒；アセトニトリル−０．０５％ＴＦＡ（５２：４８）、流速；１．８ｍｌ／ｍｉｎ、検出；ＵＶ２１０ｎｍで保持時間１７．５３１分
（８）１Ｈ−ＮＭＲスペクトル；δ（ｐｐｍ）（ＣＤＣＬ３、４００ＭＨｚ）
０．８８（３Ｈ，ｔ，Ｊ＝６．４Ｈｚ），１．２８（８Ｈ，ｍ），１．４３（２Ｈ，ｍ），２．０３（１Ｈ，ｔ，Ｊ＝１２．７Ｈｚ），２．１８（２Ｈ，ｍ），２．２１（１Ｈ，ｂｒｓ），２．４９（１Ｈ，ｄｄｄ，Ｊ１＝１２．７Ｈｚ，Ｊ２＝３．４Ｈｚ，Ｊ３＝１．９Ｈｚ），３．１５（１Ｈ，ｄｄｄ，Ｊ１＝７．８Ｈｚ，Ｊ２＝３．４Ｈｚ，Ｊ３＝１．９Ｈｚ），３．４９（３Ｈ，ｓ），３．５２（１Ｈ，ｄｄ，Ｊ１＝３．９Ｈｚ，Ｊ２＝１．５Ｈｚ），３．６０（１Ｈ，ｄｄ，Ｊ１＝３．９Ｈｚ，Ｊ２＝１．５Ｈｚ），３．７４（１Ｈ，ｂｒｓ），３．９３（１Ｈ，ｄｄ，Ｊ１＝７．８Ｈｚ，Ｊ２＝１．５Ｈｚ），４．５０（１Ｈ，ｂｒｓ），４．８０（１Ｈ，ｍ），５．７７（１Ｈ，ｄｔ，Ｊ１＝１５．１Ｈｚ，Ｊ２＝１．５Ｈｚ），５．８７（１Ｈ，ｄ，Ｊ＝８．３Ｈｚ），６．８９（１Ｈ，ｄｔ，Ｊ１＝１５．１Ｈｚ，Ｊ２＝７．１Ｈｚ）
（９）１３Ｃ−ＮＭＲスペクトル；δ（ｐｐｍ）（ＣＤＣＬ３、１００ＭＨｚ）
１４．１（ｑ），２２．６（ｔ），２８．２（ｔ），２９．１（２Ｃ，ｔ），３１．７（ｔ），３２．１（ｔ），３７．８（ｔ），４６．０（ｄ），５５．０（ｄ），５５．１（ｄ），５８．６（ｑ），６３．７（ｄ），７２．３（ｄ），７５．８（ｓ），７７．３（ｄ），１２２．９（ｄ），１４６．３（ｄ），１６５．４（ｓ），２０７．６（ｓ）
本発明の第２の要旨とするところは、イサリア属に属するＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質生産菌を培養し、その培養物からＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質を採取することを特徴とする、ＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質の製造法である。
本発明に使用されるＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質生産菌の一例としては、本発明者らにより新たに分離され、以下のように独立行政法人産業技術総合研究所特許生物寄託センターに、ＦＥＲＭ ＢＰ−０８６００として国際寄託されているイサリア属ＴＫ−５７株が挙げられる。
▲１▼寄託機関： 国際寄託機関、独立行政法人産業技術総合研究所特許生物寄託センター
住所： 日本国茨城県つくば市東１丁目１番地１ 中央第６（郵便番号３０５−８５６６）
▲２▼寄託日： 原寄託日：２００２年８月６日
移管請求日：２００４年１月２７日（２００２年８月６日）に寄託されたＦＥＲＭ Ｐ−１８９６３号より移管）
▲３▼寄託番号： ＦＥＲＭ ＢＰ−０８６００
このＴＫ−５７株の菌学的性状は以下の通りである。
１．ＴＫ−５７株の菌学的性状
（１）コロニーの性状
ツァペック酵母エキス寒天培地上で生育は良好で、２５℃、１０日間の培養でコロニーの直径は７５〜７８ｍｍに達する。 白色〜淡黄白色、綿毛状、平坦、やや密な菌糸層からなり、分生子を疎らに形成する。裏面は黄白色となる。酵母エキス寒天培地上で生育は良好で、２５℃、１０日間の培養でコロニーの直径は３５〜４２ｍｍに達する。白色、綿毛状、平坦、やや密な菌糸層からなり、分生子を疎らに形成する。裏面は黄土色となる。ポテトデキストロース寒天培地上で生育は良好で、２５℃、１０日間の培養でコロニーの直径は４０〜４２ｍｍに達する。黄白色、綿毛状、平坦、密な菌糸層からなり、分生子を豊富に形成する。コロニー表面に透明の抽出液を生じる。裏面は黄白色となる。３７℃の培養では、どの培地上でも生育しない。
（２）形態的性状
菌糸は一部束状になる。フィアライドはアンプル形もしくは細い円筒形、菌糸の側面から単生し、先端が彎曲する場合がある。分生子は、長円形〜円筒形、滑面、大きさ８〜１２×４〜６μｍである。
以上の菌学的性状よりＴＫ−５７株を、イサリア属に属すると同定した。なお、ＴＫ−５７株は、他の糸状菌と同様に、その性状が変化することがある。したがって、例えばＴＫ−５７株に由来する突然変異株、形質接合体または遺伝子組換え体であっても、ＴＫ−５７−１６４Ａ物質およびＴＫ−５７−１６４Ｂ物質を生産するものは全て本発明に使用できる。なお、本発明で用いられるＴＫ−５７−１６４Ａ物質およびＴＫ−５７−１６４Ｂ物質生産菌は、これらの微生物に限定されるものではない。ＴＫ−５７−１６４Ａ物質およびＴＫ−５７−１６４Ｂ物質を生産する能力を有している菌であればＴＫ−５７−１６４Ａ物質およびＴＫ−５７−１６４Ｂ物質生産菌としていずれを用いてもよい。
１．ＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質生産菌の培養法
イサリア属であるＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質生産菌は、通常の微生物が利用しうる栄養物を含有する培地で培養する。栄養源としては、炭素源としては例えば、グルコース、水飴、デキストリン、澱粉、糖蜜、動・植物油などを使用でき、窒素源としては例えば、大豆粉、小麦胚芽、コーン・スティーブ・リカー、綿実粕、肉エキス、ペプトン、酵母エキス、硫酸アンモニウム、硝酸アンモニウム、尿素などを使用でき、その他必要に応じて、ナトリウム、カリウム、カルシウム、マグネシウム、コバルト、塩素、燐酸、硫酸およびその他のイオンを生成し得る無機塩類を添加することができる。また菌の生育を助け、ＴＫ−５７−１６４Ａ物質の生産を促進するような有機物・無機物を適当に添加することができる。
培養方法としては、好気的条件で、２０〜３０℃が最適である。１５〜２３日間程度の培養で、培養を停止し、培養物から目的物質の単離精製を行う。
２．ＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質の精製法
ＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質は、その理化学的性状を利用して、通常の分離手段、例えば、溶媒抽出法、イオン交換樹脂法、吸着または分配カラムクロマト法、ゲルろ過法、透析法、沈殿法などを、単独であるいは適宜組み合わせて精製することができる。例えば、ＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質は、培養菌体からアセトン−水、アセトン、メタノール、酢酸エチルなどで抽出される。ＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質を精製するには、シリカゲルやＯＤＳ系修飾シリカゲルを用いた吸着・分配クロマトを用いることができる。
本発明のＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質は各種の植物病害菌に対して優れた防除効果を示し、農園芸用殺菌剤として利用することができる。
その代表例は、ウリ類うどんこ病、べと病、たんそ病、灰色かび病、コムギ赤さび病、麦類うどんこ病、イネいもち病などである。
本発明化合物であるＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質を、農園芸用殺菌剤の有効成分として用いる場合は、そのまま用いても良いが、通常は適当な固体担体、液体担体、ガス状担体、界面活性剤、分散剤その他の製剤用補助剤と混合して乳剤、液剤、懸濁剤、水和剤、粉剤、粒剤、錠剤、油剤、エアゾール、フロワブル剤等の任意の剤型にして使用する。
固体担体としては、例えばタルク、ベンナイト、クレー、カオリン、ケイソウ土、バーミキュライト、ホワイトカーボン、炭酸カルシウム等があげられる。
液体担体としては、例えばメタノール、ｎ−ヘキサノール、エチレングリコール等のアルコール類、アセトン、メチルエチルケトン、シクロヘキサノン等のケトン類、ｎ−ヘキサン、ケロシン、灯油等の脂肪族炭化水素類、トルエン、キシレン、メチルナフタレン等の芳香族炭化水素類、ジエチルエーテル、ジオキサン、テトラヒドロフラン等のエーテル類、酢酸エチル等のエステル類、アセトニトリル、イソブチロニトリル等のニトリル類、ジメチルホルムアミド、ジメチルアセトアミド等の酸アミド類、ダイズ油、綿実油等の植物油類、ジメチルスルホキシド、水等があげられる。また、ガス担体としてはＬＰＧ、空気、窒素、炭酸ガス、ジメチルエーテル等があげられる。
乳化、分散、展着等のための界面活性剤、分散剤としては、例えばアルキル硫酸エステル類、アルキルスルホン酸塩類、アリールスルホン酸塩類、ポリオキシアルキレンアルキルエーテル類、ポリオキシアルキレンアリールエーテル類、多価アルコールエステル類、リグニンスルホン酸塩等が用いられる。
更に、製剤の性状を改善するための補助剤としては、例えばカルボキシメチルセルロース、アラビアガム、ポリエチレングリコール、ステアリン酸カルシウム等が用いられる。
上記の担体、界面活性剤、分散剤、及び補助剤は、必要に応じて各々単独で、あるいは組み合わせて用いられる。
これらの製剤中の有効成分の含有量は、乳剤で通常１−７５重量％、粉剤では通常０．３−２５重量％、水和剤では通常１−９０重量％、粒剤では通常０．５−１０重量％が適当である。
これらの製剤は、そのまま用いても良いが、場合によっては希釈して用いる。また、これらの製剤は他の殺菌剤、殺虫剤、殺ダニ剤、除草剤、植物成長調節剤、肥料等と混合して用いることもできる。When a new active substance that can be used as an agricultural and horticultural fungicide was searched for from a culture produced by microorganisms, the Ascaria spp. Of the genus Isaria, which was named TK-57 strain newly isolated by the present inventors, was found. It has been found that substances exhibiting antifungal activity against phytopathogenic fungi are produced in the culture. Further, it was found that this antifungal active substance has a chemical structure represented by the following formula (1), and was confirmed to be a novel substance. This substance was designated as TK-57-164A substance and TK-57- It was named 164B substance.
That is, the present invention provides the following agricultural and horticultural bactericidal active substances TK-57-164A and TK-57-164B.
Formula (1)
Further, the present invention cultivates a TK-57-164A substance and a TK-57-164B substance-producing bacterium belonging to the genus Isaria, and the TK-57-164A substance and the TK-57-164B substance are obtained from the culture. A method for producing the TK-57-164A substance and the TK-57-164B substance described above, which is characterized by being collected.
Further, an agricultural and horticultural fungicide containing at least one of the TK-57-164A substance and the TK-57-164B substance described above and an ascomycete belonging to the genus Isaria, Of TK-57-164A substance and TK-57-164B substance or a mutant thereof.
<Aspect of the Invention>
The first gist of the present invention resides in the TK-57-164A substance and the TK-57-164B substance represented by the above formula (1) and having the following physicochemical properties.
1. Physicochemical properties of TK-57-164A substance (1) Color and shape Colorless solid (2) Mass spectrum M / Z 410.2546 (M + H) +
(3) Molecular formula C 22 H 35 NO 6
(4) Specific rotation [α] D −21.3 ° (c0.10, CHCl 3 , 25 ° C.)
(5) UV absorption spectrum λmax (MeOH) 213 nm
(6) Infrared absorption spectrum νmax (KBr) cm −1
3400, 3300, 2924, 2853, 2359, 2341, 1742, 1684, 1649, 1632, 1533, 1460, 1369, 1344, 1273, 1198, 1155, 1101, 1084, 1001, 974, 934, 916, 845, 835 812
(7) HPLC column; Waters L-column ODS 10φ × 250 mm, elution solvent: acetonitrile-0.05% TFA (52:48), flow rate: 1.8 ml / min, detection; UV 254 nm, retention time 46.848 minutes ( 8) 1H-NMR spectrum; δ (ppm) (CDCL 3 , 400 MHz)
0.86 (3H, t, J = 6.8 Hz), 1.20-1.31 (12H, m), 1.44 (2H, m), 2.03 (1H, t, J = 12.7 Hz) ), 2.18 (2H, m), 2.21 (1H, brs), 2.49 (1H, ddd, J 1 = 12.7 Hz, J 2 = 4.9 Hz, J 3 = 1.7 Hz), 3.15 (1H, ddd, J 1 = 7.6 Hz, J 2 = 3.4 Hz, J 3 = 1.5 Hz), 3.50 (3H, s), 3.52 (1H, dd, J 1 = 3.6 Hz, J 2 = 1.5 Hz), 3.60 (1H, dd, J 1 = 3.6 Hz, J 2 = 1.2 Hz), 3.74 (1H, brs), 3.93 (1H, dd, J 1 = 7.6Hz, J 2 = 1.2Hz), 4.50 (1H, brs), 4.80 (1H, m), 5.77 (1H, dt, J 1 = 5.1Hz, J 2 = 1.5Hz), 5.83 (1H, d, J = 8.3Hz), 6.89 (1H, dt, J 1 = 15.1Hz, J 2 = 7.1Hz)
(9) 13 C-NMR spectrum; δ (ppm) (CDCL 3 , 100 MHz)
14.1 (q), 22.7 (t), 28.2 (t), 29.2-29.7 (4C, t), 31.9 (t), 32.1 (t), 37. 8 (t), 46.0 (d), 55.0 (d), 55.1 (d), 58.7 (q), 63.7 (d), 72.3 (d), 75.8 (S), 77.3 (d), 122.9 (d), 146.3 (d), 165.4 (s), 207.6 (s)
2. Physicochemical properties of TK-57-164B substance (1) Color and shape Colorless solid (2) Mass spectrum M / Z 382.2227 (M + H) +
(3) Molecular formula C 20 H 31 NO 6
(4) Specific rotation [α] D −15.5 ° (c0.19, CHCl 3 , 25 ° C.)
(5) UV absorption spectrum λmax (MeOH) 213 nm
(6) Infrared absorption spectrum νmax (KBr) cm −1
3402, 3296, 2926, 2855, 2359, 2341, 1742, 1684, 1651, 1631, 1531, 1456, 1371, 1342, 1273, 1196, 1155, 1101, 1084, 1001, 974, 934, 916, 845, 833 812
(7) HPLC column; Senshupak PEGASIL ODS 10φ × 250 mm, elution solvent: acetonitrile-0.05% TFA (52:48), flow rate: 1.8 ml / min, detection; UV 210 nm, retention time 17.531 minutes (8) 1H-NMR spectrum; δ (ppm) (CDCL 3 , 400 MHz)
0.88 (3H, t, J = 6.4Hz), 1.28 (8H, m), 1.43 (2H, m), 2.03 (1H, t, J = 12.7Hz), 2. 18 (2H, m), 2.21 (1H, brs), 2.49 (1H, ddd, J 1 = 12.7 Hz, J 2 = 3.4 Hz, J 3 = 1.9 Hz), 3.15 ( 1H, ddd, J 1 = 7.8 Hz, J 2 = 3.4 Hz, J 3 = 1.9 Hz), 3.49 (3H, s), 3.52 (1H, dd, J 1 = 3.9 Hz) J 2 = 1.5 Hz), 3.60 (1H, dd, J 1 = 3.9 Hz, J 2 = 1.5 Hz), 3.74 (1H, brs), 3.93 (1H, dd, J 1 = 7.8Hz, J 2 = 1.5Hz) , 4.50 (1H, brs), 4.80 (1H, m), 5.77 (1H, dt, J 1 = 15.1Hz J 2 = 1.5Hz), 5.87 ( 1H, d, J = 8.3Hz), 6.89 (1H, dt, J 1 = 15.1Hz, J 2 = 7.1Hz)
(9) 13 C-NMR spectrum; δ (ppm) (CDCL 3 , 100 MHz)
14.1 (q), 22.6 (t), 28.2 (t), 29.1 (2C, t), 31.7 (t), 32.1 (t), 37.8 (t) 46.0 (d), 55.0 (d), 55.1 (d), 58.6 (q), 63.7 (d), 72.3 (d), 75.8 (s), 77.3 (d), 122.9 (d), 146.3 (d), 165.4 (s), 207.6 (s)
The second gist of the present invention is that a TK-57-164A substance and a TK-57-164B substance-producing bacterium belonging to the genus Isaria are cultured, and the TK-57-164A substance and TK-57- are cultured from the culture. A method for producing a TK-57-164A substance and a TK-57-164B substance, characterized in that 164B substance is collected.
As an example of the TK-57-164A substance and the TK-57-164B substance-producing bacterium used in the present invention, it has been newly isolated by the present inventors, and is as follows. The depository center includes the Isalia TK-57 strain that has been deposited internationally as FERM BP-08600.
(1) Depositary Institution: International Depositary, National Institute of Advanced Industrial Science and Technology, Patent Biological Depositary Center Address: 1-1, Higashi 1-chome, Tsukuba City, Ibaraki, Japan 6th (Zip 305-8586)
(2) Deposit date: Original deposit date: August 6, 2002
Request date for transfer: Transfer from FERM P-18963 deposited on January 27, 2004 (August 6, 2002)
(3) Deposit number: FERM BP-08600
The mycological properties of the TK-57 strain are as follows.
1. Bacteriological properties of TK-57 strain (1) Colony properties Growth is good on zapek yeast extract agar medium, and the colony diameter reaches 75 to 78 mm when cultured at 25 ° C. for 10 days. It consists of white to pale yellowish white, fluffy, flat, slightly dense mycelium layer, and conidia are formed sparsely. The back side is yellowish white. The growth is good on the yeast extract agar medium, and the colony diameter reaches 35 to 42 mm after culturing at 25 ° C. for 10 days. It consists of white, fluffy, flat, and somewhat dense mycelium layers that form conidia loosely. The back is ocher. The growth is good on potato dextrose agar medium, and the colony diameter reaches 40 to 42 mm after culturing at 25 ° C. for 10 days. It consists of yellowish white, fluffy, flat, dense mycelium layer and forms abundant conidia. A clear extract is produced on the surface of the colony. The back side is yellowish white. In the culture at 37 ° C., it does not grow on any medium.
(2) Morphological properties Mycelium is partly bundled. The phialide is ampoule-shaped or thin cylindrical, grows from the side of the mycelium, and the tip may be bent. The conidia are oval to cylindrical, smooth, and have a size of 8 to 12 × 4 to 6 μm.
From the above mycological characteristics, the TK-57 strain was identified as belonging to the genus Isaria. In addition, the property of TK-57 strain may change in the same manner as other filamentous fungi. Therefore, for example, all mutants, zygotes or gene recombinants derived from the TK-57 strain that produce the TK-57-164A substance and the TK-57-164B substance are used in the present invention. it can. The TK-57-164A substance and TK-57-164B substance-producing bacteria used in the present invention are not limited to these microorganisms. Any bacteria may be used as the TK-57-164A substance and the TK-57-164B substance-producing bacteria as long as they have the ability to produce the TK-57-164A substance and the TK-57-164B substance.
1. Method for culturing TK-57-164A substance and TK-57-164B substance-producing bacteria The TK-57-164A substance and TK-57-164B substance-producing bacteria belonging to the genus Isaria contain nutrients that can be used by ordinary microorganisms. Incubate in the culture medium. As a nutrient source, for example, glucose, starch syrup, dextrin, starch, molasses, animal / vegetable oil, etc. can be used as a carbon source, and as a nitrogen source, for example, soybean flour, wheat germ, corn steve liquor, cottonseed meal , Meat extract, peptone, yeast extract, ammonium sulfate, ammonium nitrate, urea, etc. Other inorganic salts that can produce sodium, potassium, calcium, magnesium, cobalt, chlorine, phosphoric acid, sulfuric acid and other ions as required Can be added. In addition, organic substances and inorganic substances that assist the growth of the bacteria and promote the production of the TK-57-164A substance can be appropriately added.
As a culture method, 20-30 degreeC is optimal on aerobic conditions. The culture is stopped for about 15 to 23 days, and the target substance is isolated and purified from the culture.
2. Purification method of TK-57-164A substance and TK-57-164B substance TK-57-164A substance and TK-57-164B substance can be obtained by using conventional physicochemical properties, for example, by conventional separation means such as solvent extraction method. The ion exchange resin method, the adsorption or distribution column chromatography method, the gel filtration method, the dialysis method, the precipitation method and the like can be purified alone or in appropriate combination. For example, TK-57-164A substance and TK-57-164B substance are extracted from cultured cells with acetone-water, acetone, methanol, ethyl acetate and the like. In order to purify the TK-57-164A substance and the TK-57-164B substance, adsorption / distribution chromatography using silica gel or ODS modified silica gel can be used.
The TK-57-164A substance and the TK-57-164B substance of the present invention exhibit an excellent control effect against various plant diseases and can be used as agricultural and horticultural fungicides.
Typical examples are powdery mildew, downy mildew, anthracnose, gray mold, wheat rust, wheat powdery mildew, and rice blast.
When the TK-57-164A substance and the TK-57-164B substance, which are the compounds of the present invention, are used as active ingredients of agricultural and horticultural fungicides, they may be used as they are, but usually a suitable solid carrier, liquid carrier, Arbitrary agents such as emulsions, liquids, suspensions, wettable powders, powders, granules, tablets, oils, aerosols, flowables, etc. mixed with gaseous carriers, surfactants, dispersants and other formulation adjuvants Use as a mold.
Examples of the solid carrier include talc, bennite, clay, kaolin, diatomaceous earth, vermiculite, white carbon, calcium carbonate and the like.
Examples of the liquid carrier include alcohols such as methanol, n-hexanol and ethylene glycol, ketones such as acetone, methyl ethyl ketone and cyclohexanone, aliphatic hydrocarbons such as n-hexane, kerosene and kerosene, toluene, xylene and methylnaphthalene. Aromatic hydrocarbons such as diethyl ether, dioxane and tetrahydrofuran, esters such as ethyl acetate, nitriles such as acetonitrile and isobutyronitrile, acid amides such as dimethylformamide and dimethylacetamide, soybean oil , Vegetable oils such as cottonseed oil, dimethyl sulfoxide, water and the like. Examples of the gas carrier include LPG, air, nitrogen, carbon dioxide gas, dimethyl ether and the like.
Examples of surfactants and dispersants for emulsification, dispersion, spreading, etc. include alkyl sulfates, alkyl sulfonates, aryl sulfonates, polyoxyalkylene alkyl ethers, polyoxyalkylene aryl ethers, many A monohydric alcohol ester, a lignin sulfonate, etc. are used.
Furthermore, as an auxiliary agent for improving the properties of the preparation, for example, carboxymethyl cellulose, gum arabic, polyethylene glycol, calcium stearate and the like are used.
The above carriers, surfactants, dispersants, and adjuvants may be used alone or in combination as necessary.
The content of the active ingredient in these preparations is usually 1-75% by weight for emulsions, usually 0.3-25% by weight for powders, 1-90% by weight for wettable powders, and usually 0.5% for granules. -10% by weight is suitable.
These preparations may be used as they are, but may be diluted in some cases. These preparations can also be used by mixing with other fungicides, insecticides, acaricides, herbicides, plant growth regulators, fertilizers and the like.
以下に本発明の製造例、試験例を示すが、本発明はこれらの例示のみに限定されるものではない。 Although the manufacture example and test example of this invention are shown below, this invention is not limited only to these illustrations.
１００ｍｌ容量の大試験管に１０８液体培地（グルコース１．０％，スターチ２．０％，ペプトン０．５％，イーストエキス０．５％，炭酸カルシウム０．４％，ｐＨ７．０）１０ｍｌを分注し、オートクレーブにて滅菌処理（１２０℃ ３０分）を行った後、ＳＤＹ培地（グルコース２．０％，ペプトン１．０％，イーストエキス１．０％，寒天２．０％，ｐＨ６．０）のシャーレ上で生育させておいたＴＫ−５７の菌体を、カギ型白金耳にて１ｃｍ四方に切り取り培地に植菌した。２７℃の好気的環境で、３日間振盪培養（２００回転／分）して、第１種培養とした。
次に５００ｍｌ容量の三角フラスコに１０８液体培地１００ｍｌを分注し、滅菌処理（１２０℃３０分）の後、第１種培養液５ｍｌを植菌した。２７℃好気的環境で、３日間振盪培養（２００回転／分）して、第２種培養液とした。
５００ｍｌ容量の三角フラスコ７５本に生産培地（玄米３０％，イーストエキス０．０７％，酒石酸ナトリウム０．０３％，Ｋ２ＨＰＯ４ ０．０３％、ｐＨ無調整）を５０ｍｌ分注し、滅菌処理（１２０℃３０分）後、第２種培養液５ｍｌを植菌した。火炎滅菌したガラス棒でよくまぜ、２４℃２１日間静置培養を行った。生育状態は良好で、培地表面に純白の菌糸を形成した。
培養終了後、フラスコ１本につき水３０ｍｌを加えて全培養物を取り出し、これにアセトン１０Ｌを加えて３時間撹拌した後、抽出した。抽出液をセライトを用いて吸引ろ過し、濾液のアセトンを減圧下留去し、黄色を帯びた茶色の濃縮液を得た。
この濃縮液（ｐＨ５．７）を、等量の酢酸エチルで５回抽出し、抽出液を無水硫酸ナトリウムで乾燥後、減圧下溶媒を留去して暗褐色の油状の培養抽出物８．２ｇを得た。
シリカゲル６０（７０−２３０ｍｅｓｈ、ＡＳＴＭ）１６０ｇをカラム管（６．６φ×７３ｃｍ）に充填し、培養抽出物を付し、クロロホルム−メタノール（６０：１〜４０：１）で活性物質の溶出を行った。さらにシリカゲルクロマト（シリカゲル６０、クロロホルム−メタノール（８０：１〜６０：１）に付し、粗精製物２６８ｍｇを得た。この粗精製物をＯＡＳＩＳ ＨＬＢ固相抽出カートリッジ（６ｇ／３５ｍｌＬＰ，粒径６０μｍ、Ｗａｔｅｒｓ）に付し、メタノール−水（８０：２０）で溶出した画分を高速液体クロマトグラフィー（ＨＰＬＣ）（カラム；ｓｅｎｓｈｕｐａｋ ＰＥＧＡＳＩＬ ＯＤＳ １０φ×２５０ｍｍ、溶出溶媒；アセトニトリル−０．０５％ＴＦＡ（５５：４５〜１００：０）、流速；１．８ｍｌ／ｍｉｎ、検出；ＵＶ２５４ｎｍ）、および（カラム；Ｗａｔｅｒｓ Ｌ−ｃｏｌｕｍｎ ＯＤＳ １０φ×２５０ｍｍ、溶出溶媒；アセトニトリル−０．０５％ＴＦＡ（５２：４８）、流速；１．８ｍｌ／ｍｉｎ、検出；ＵＶ２５４ｎｍ）で溶出しＴＫ−５７−１６４Ａ物質を３．６ｍｇ得た。
また、メタノール−水（７０：３０）で溶出した画分を高速液体クロマトグラフィー（ＨＰＬＣ）（カラム；ｓｅｎｓｈｕｐａｋ ＰＥＧＡＳＩＬ ＯＤＳ １０φ×２５０ｍｍ、溶出溶媒；アセトニトリル−０．０５％ＴＦＡ（５２：４８）、流速；１．８ｍｌ／ｍｉｎ、検出；ＵＶ２１０ｎｍ）で溶出しＴＫ−５７−１６４Ｂ物質を３．１ｍｇ得た。
試験例
キュウリべと病（Ｐｓｅｕｄｏｐｅｒｏｎｏｓｐｏｒａ ｃｕｂｅｎｓｉｓ）
培養土を入れたプラスチック製ポットに播種した後、１０日程度育苗し子葉が完全に展開したキュウリ苗（品種：四葉）を供試植物とした。ＴＫ−５７−１６４Ａ物質の６６７ｐｐｍに調整した薬液をスプレーガンを用いて３ポットあたり５ｍｌ散布処理し、風乾させた後、あらかじめべと病を発病させていたキュウリ葉より採集、調整した分生胞子懸濁液（１〜５×１０６個／ｍｌ）を均一に噴霧接種し、２１℃の湿室内に２４時間静置した。その後夜間１８℃、日中２２℃の人工気象室に移して発病させ、接種７日後に子葉での発病面積を調査し、下記に示す基準で指数を与え、下記の式によって発病度、防除価を算出した。
指数 ０：病斑なし
１：数個の病斑
２：発病面積が葉面積の１／４未満
３：発病面積が葉面積の１／４以上１／２未満
４：発病面積が葉面積の１／２以上３／４未満
５：発病面積が葉面積の３／４以上
発病度＝Σ（指数×該当株数）／（調査株数×５）×１００
防除価＝（１−処理区の発病度／無処理区の発病度）×１００
その結果、ＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質はともに１００％の防除価を示した。10 ml of 108 liquid medium (glucose 1.0%, starch 2.0%, peptone 0.5%, yeast extract 0.5%, calcium carbonate 0.4%, pH 7.0) in a large test tube with a capacity of 100 ml After sterilization (120 ° C. for 30 minutes) in an autoclave, SDY medium (glucose 2.0%, peptone 1.0%, yeast extract 1.0%, agar 2.0%, pH 6.0) The TK-57 cells grown on the petri dish were cut into 1 cm squares using a key-shaped platinum ear and inoculated into the culture medium. The first seed culture was performed by shaking culture (200 rpm) for 3 days in an aerobic environment at 27 ° C.
Next, 100 ml of 108 liquid medium was dispensed into a 500 ml Erlenmeyer flask, and after sterilization (120 ° C. for 30 minutes), 5 ml of the first seed culture was inoculated. The culture medium was shaken for 3 days (200 rpm) in an aerobic environment at 27 ° C. to obtain a second type culture solution.
Dispense 50 ml of production medium (brown rice 30%, yeast extract 0.07%, sodium tartrate 0.03%, K 2 HPO 4 0.03%, pH unadjusted) into 75 500 ml Erlenmeyer flasks and sterilize After (120 ° C. for 30 minutes), 5 ml of the second kind culture solution was inoculated. The mixture was thoroughly mixed with a flame-sterilized glass rod and cultured at 24 ° C. for 21 days. The growth state was good, and pure white mycelium was formed on the surface of the medium.
After completion of the culture, 30 ml of water was added to each flask to remove the whole culture, 10 L of acetone was added thereto, and the mixture was stirred for 3 hours and then extracted. The extract was subjected to suction filtration using celite, and acetone in the filtrate was distilled off under reduced pressure to obtain a yellowish brown concentrate.
This concentrated solution (pH 5.7) was extracted five times with an equal amount of ethyl acetate, and the extract was dried over anhydrous sodium sulfate, and then the solvent was distilled off under reduced pressure to obtain 8.2 g of a dark brown oily culture extract. Got.
160 g of silica gel 60 (70-230 mesh, ASTM) is packed into a column tube (6.6φ × 73 cm), a culture extract is attached, and the active substance is eluted with chloroform-methanol (60: 1 to 40: 1). It was. Furthermore, it was subjected to silica gel chromatography (silica gel 60, chloroform-methanol (80: 1 to 60: 1) to obtain 268 mg of a crude product. This crude product was obtained from an OASIS HLB solid phase extraction cartridge (6 g / 35 ml LP, particle size 60 μm). , Waters), and the fraction eluted with methanol-water (80:20) was subjected to high performance liquid chromatography (HPLC) (column; senshupak PEGASIL ODS 10φ × 250 mm, elution solvent: acetonitrile-0.05% TFA (55 : 45 to 100: 0), flow rate; 1.8 ml / min, detection; UV254 nm), and (column; Waters L-column ODS 10φ × 250 mm, elution solvent; acetonitrile-0.05% TFA (52:48), Flow rate; 1.8 ml / min, detection; UV Eluted TK-57-164A material was obtained 3.6mg at 54 nm).
Further, the fraction eluted with methanol-water (70:30) was subjected to high performance liquid chromatography (HPLC) (column: senshupak PEGASIL ODS 10φ × 250 mm, elution solvent: acetonitrile-0.05% TFA (52:48), flow rate. Elution at 1.8 ml / min, detection; UV 210 nm) to obtain 3.1 mg of TK-57-164B substance.
Test example Cucumber downy mildew (Pseudoperonospora cubensis)
After sowing in a plastic pot containing culture soil, a cucumber seedling (cultivar: four-leaf) in which the seedlings were grown for about 10 days and the cotyledons were completely developed was used as a test plant. The TK-57-164A substance adjusted to 667 ppm is sprayed with 5 ml per 3 pots, air-dried, then collected and adjusted from cucumber leaves that had previously caused downy mildew. The suspension (1-5 × 10 6 cells / ml) was uniformly spray-inoculated and allowed to stand in a humid chamber at 21 ° C. for 24 hours. Then, move to an artificial meteorological room at 18 ° C during the night and 22 ° C during the day to investigate the disease, and after 7 days of inoculation, investigate the diseased area in the cotyledons and give an index according to the criteria shown below. Was calculated.
Index 0: No lesion 1: Several lesions 2: Disease area is less than 1/4 of leaf area 3: Disease area is 1/4 or more and less than 1/2 of leaf area 4: Disease area is 1 of leaf area / 2 or more but less than 3/4 5: Disease area is 3/4 or more of leaf area Disease severity = Σ (index × number of strains) / (number of strains surveyed × 5) × 100
Control value = (1—the severity of the treated area / the severity of the untreated area) × 100
As a result, both the TK-57-164A substance and the TK-57-164B substance showed a control value of 100%.
本発明のＴＫ−５７−１６４Ａ物質及びＴＫ−５７−１６４Ｂ物質は上記のように植物病害菌に対して優れた防除効果を示し、農園芸用殺菌剤としての利用が期待される。The TK-57-164A substance and the TK-57-164B substance of the present invention exhibit an excellent control effect against plant diseases as described above, and are expected to be used as agricultural and horticultural fungicides.
Claims (4)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2003042038 | 2003-02-20 | ||
| JP2003042038 | 2003-02-20 | ||
| PCT/JP2004/001862 WO2004074269A1 (en) | 2003-02-20 | 2004-02-19 | Substances tk-57-164a and tk-57-164b, process for producing the same and agricultural/horticultural bactericides containing the same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPWO2004074269A1 true JPWO2004074269A1 (en) | 2006-06-01 |
Family
ID=32905338
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2005502755A Pending JPWO2004074269A1 (en) | 2003-02-20 | 2004-02-19 | TK-57-164A substance and TK-57-164B substance, their production process, and agricultural and horticultural fungicides containing them as active ingredients |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JPWO2004074269A1 (en) |
| WO (1) | WO2004074269A1 (en) |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5626198A (en) * | 1979-08-08 | 1981-03-13 | Takara Shuzo Co Ltd | Preparation of antitumor substance |
| JPH01104087A (en) * | 1987-03-17 | 1989-04-21 | Yoshitomi Pharmaceut Ind Ltd | Production of immunosuppressive substance |
-
2004
- 2004-02-19 JP JP2005502755A patent/JPWO2004074269A1/en active Pending
- 2004-02-19 WO PCT/JP2004/001862 patent/WO2004074269A1/en active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| WO2004074269A1 (en) | 2004-09-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR940010036B1 (en) | Process for preparing macrocyclic compound | |
| KR100754836B1 (en) | Serratia plymuthica A21-4, biological control method of Phytophthora diseases using the A21-4 and antifungal compound produced by the A21-4 | |
| JP2006124280A (en) | Agent for improving germination rate of plant seed | |
| KR102131411B1 (en) | Composition and method for controlling plant diseases using vulculic acid isolated from Paraboremia adianticola SFC20150402-M24 strain | |
| JPWO2004074269A1 (en) | TK-57-164A substance and TK-57-164B substance, their production process, and agricultural and horticultural fungicides containing them as active ingredients | |
| JP5694708B2 (en) | A-87774 compounds or salts thereof, processes for producing them, and agricultural chemicals containing them as active ingredients | |
| CN110093383A (en) | A kind of application of Benzofurantone compound as disinfectant use in agriculture | |
| KR101044273B1 (en) | Formulation for controlling plant diseases containing bikaverin and method for controlling plant diseases using same | |
| JP3605432B2 (en) | Novel antibiotic AB5366, its production method and its use | |
| JPS6053597B2 (en) | New antibiotic N-461 substance, its manufacturing method, and agricultural and horticultural fungicides containing it as an active ingredient | |
| KR100991298B1 (en) | Biotic pesticide using Arthrobacter zoysiae with inhibition activity of germinaton and growth of seeds | |
| JPH0398593A (en) | Production of antibiotic 5'-o-sulfurmoyltubercidin and its use | |
| KR100790314B1 (en) | Salaceyin from streptomyces laceyi ms53 strain effective against plant pathogens and preparation method thereof | |
| JPH1112280A (en) | New antibiotic ab5529, its production and insecticide | |
| JPS6322583A (en) | Si-4228 based substance and germicide containing said substance as active ingredient | |
| JPH0559078A (en) | Antibiotic substance nba-2006 | |
| JPH0748348A (en) | New antibiotic substance ab4015-b and ab4015-l, their production and use | |
| JPH0522514B2 (en) | ||
| JPH06277084A (en) | New antibiotic ab4063-a and b, their production and use thereof | |
| JPH1045662A (en) | New antibiotic ab5362-a,-b and-c and their production and use | |
| JP2004073114A (en) | New microorganism, soil disease injury controlling agent and method for controlling soil disease injury | |
| JPH0859612A (en) | Antibiotics ab4015-a1, a2 and a3 and their production and agricultural/horticultural germicide | |
| JP2000239269A (en) | New compound f-15507 and its production | |
| JP2001302671A (en) | New compound mu-118 | |
| KR20000056783A (en) | Novel Compound Having Antifungal Activity Produced from Myxococcus stipitatus and Agricultural Antifungal Agent Containing the Same |