JP5561637B2 - Antibacterial / sterilization technology using mushroom volatile antibacterial substances - Google Patents
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Description
本発明は、きのこの揮発性抗菌物質を用いた防菌・除菌技術に関する。 The present invention relates to antibacterial and antibacterial techniques using mushroom volatile antibacterial substances.
現在使用されている殺菌剤の多くは、安全性の面から選択性の高い殺菌剤が主流で抗菌スペクトラムが狭く、殺菌剤に対する耐性菌の出現などの問題も生じている。微生物が生産する抗菌物質は、すでに抗生物質として農薬や医薬等に利用されているが、そのほとんどは非揮発性の物質である。しかし、非揮発性の抗菌物質は、作物や環境に残留し、環境汚染を引き起こす可能性があり、安全性の面から問題が多い。一方、揮発性の抗菌物質は、通気の少ない施設栽培など使用条件は限定されるものの効果が隅々まで行き渡り、通気によって揮発性抗菌物質が容易に拡散、消失することから、安全面、健康面および環境面からも優れた特徴を持つと考えられる。特に、きのこ類、中でも食用きのこの揮発性物質は安全性が高いといえる。 Many of the currently used bactericides are mainly selective bactericides from the viewpoint of safety, have a narrow antibacterial spectrum, and problems such as the emergence of bacteria resistant to the bactericides have arisen. Antimicrobial substances produced by microorganisms are already used as antibiotics in agricultural chemicals and medicines, but most of them are non-volatile substances. However, non-volatile antibacterial substances remain in crops and the environment and may cause environmental pollution, which is problematic in terms of safety. On the other hand, although volatile antibacterial substances are limited in terms of use conditions such as facility cultivation with low ventilation, the effects spread to every corner, and volatile antibacterial substances diffuse and disappear easily by aeration. It is also considered to have excellent characteristics from the environmental aspect. In particular, volatile substances of mushrooms, especially edible mushrooms, can be said to be highly safe.
きのこ類が生産する揮発性物質については、これまで医学・薬学の分野で研究が行われており、ヒトの健康増進などに関与する幾つかの生理活性物質が報告されている。しかし、抗菌作用に着目した研究としては、菌類が生産する揮発性抗菌物質による病害防除の例として、コムギ葉上から得た拮抗菌類Irpex lacteusによるうどんこ病防除などが報告されているのみである（非特許文献１）。現在に至るまで、食用きのこを含め、広範なきのこ類により生産される揮発性抗菌物質生産を利用した病害防除の報告は皆無である。 The volatile substances produced by mushrooms have been studied in the fields of medicine and pharmacology, and some physiologically active substances involved in promoting human health have been reported. However, studies focusing on antibacterial activity have only reported powdery mildew control by the antagonistic fungus Irpex lacteus obtained from wheat leaves as an example of disease control by volatile antibacterial substances produced by fungi (Non-Patent Document 1). To date, there are no reports of disease control using volatile antibacterial substances produced by a wide range of mushrooms, including edible mushrooms.
きのこ類、特に食用きのこから生産される揮発性抗菌物質を植物の病害防除に利用することが本発明の課題であった。 It was an object of the present invention to use volatile antibacterial substances produced from mushrooms, particularly edible mushrooms, for plant disease control.
本発明者らは、上記課題を解決せんと鋭意研究を行った結果、ある種のきのこから発せられる揮発性物質が植物病原菌の防除に有用であることを見出し、本発明を完成させるに至った。 As a result of diligent research to solve the above problems, the present inventors have found that volatile substances emitted from certain types of mushrooms are useful for controlling plant pathogens, and have completed the present invention. .
すなわち、本発明は、以下のものを提供する：
（１）Piptoporus属のきのこ、Mycoleptodonoides属のきのこ、Climacodon属のきのこ、Gloiothele属のきのこ、Lopharia属のきのこ、Microporus属のきのこ、Scytinostroma属のきのこ、Neolentinus属のきのこ、Hebeloma属のきのこ、Ischnoderma属のきのこ、Laetiporus属のきのこ、Daedaleopsis属のきのこ、Steccherinum属のきのこ、Trametes属のきのこ、Boidinia属のきのこ、Cymatoderma属のきのこ、Scopuloides属のきのこ、Ceriporia属のきのこおよびPleurotus属のきのこからなる群より選択される１種またはそれ以上のきのこの菌糸体、子実体、またはその廃菌床から発せられる揮発性物質を植物に適用することを特徴とする、植物病原菌の防除方法；
（２）きのこが食用きのこである（１）記載の方法；
（３）シロカイメンタケ（Piptoporus soloniensis）、ブナハリタケ（Mycoleptodonoides aitchisonii）、エゾハリタケ（Climacodon septentrionalis）、キヌシブカワタケ（Gloiothele citrina）、オリーブウロコタケ（Lopharia spadicea）、ウチワタケ（Microporus affinis）、チチイロコウヤクタケ（Scytinostroma galactinum）、ヤケイロコウヤクタケ（Scytinostroma portentosum）、マツオウジ（Neolentinus lepideus）、ナガエノスギタケ（Hebeloma radicosum）、ヤニタケ（Ischnoderma resinosum）、ヒラフスベ（Laetiporus versisporus）、エゴノキタケ（Daedaleopsis styracina）、ニセニクハリタケ（Steccherinum murashinskyi）、チチイロアミタケ（Trametes lactinea）、ヒダウロコタケ（Cymatoderma lamellatum）、ウスヒラタケ（Pleurotus pulmonarius）、ヒラタケ（Pleurotus ostreatus）、エリンギ（Pleurotus eryngii）、Trametes scabrosa、Boidinia lacticolor、Scopuloides hydnoidesおよびCeriporia lacerataからなる群より選択される１種またはそれ以上のきのこの菌糸体、子実体またはその廃菌床から発せられる揮発性物質を用いる、（１）記載の方法。；
（４）きのこがブナハリタケまたはウスヒラタケである（３）記載の方法；
（５）ウチワタケ、チチイロコウヤクタケ、チチイロアミタケおよびヒダウロコタケからなる群より選択される１種または２種のきのこの菌糸体または子実体から発せられる揮発性物質を用いて、植物病原菌類の菌糸生育を阻害することにより植物病原菌の防除を行う、（３）記載の方法；
（６）オリーブウロコタケ、ヤニタケ、ヒラフスベおよびCeriporia lacerataからなる群より選択される１種またはそれ以上のきのこの菌糸体または子実体から発せられる揮発性物質を用いて、植物病原細菌の増殖を阻害することにより植物病原菌の防除を行う、（３）記載の方法；
（７）ブナハリタケ、オリーブウロコタケ、チチイロコウヤクタケ、マツオウジ、ヤケイロコウヤクタケおよびヤニタケからなる群より選択される１種またはそれ以上のきのこの菌糸体または子実体から発せられる揮発性物質を用いて、植物病原菌類の胞子の発芽を阻害することにより植物病原菌の防除を行う、（３）記載の方法；
（８）きのこがブナハリタケである（７）記載の方法；
（９）ブナハリタケ、オリーブウロコタケ、ヤケイロコウヤクタケ、ヤニタケ、ニセニクハリタケ、チチイロアミタケおよびScopuloides hydnoidesからなる群より選択される１種またはそれ以上のきのこの菌糸体または子実体から発せられる揮発性物質を用いて、植物病原菌類の感染を阻害することにより植物病原菌の防除を行う、（３）記載の方法；
（１０）きのこがブナハリタケである（９）記載の方法；
（１１）ウスヒラタケ、ヒラタケおよびエリンギからなる群より選択される１種またはそれ以上のきのこの菌糸体または子実体から発せられる揮発性物質を用いて、植物病原菌類の胞子発芽あるいは感染を阻害することにより植物病原菌の防除を行う、（３）記載の方法；
（１２）きのこがウスヒラタケである（１１）記載の方法；
（１３）ウスヒラタケ、ヒラタケおよびエリンギからなる群より選択される１種またはそれ以上のきのこの廃菌床から発せられる揮発性物質を用いて、植物病原菌類の胞子発芽あるいは感染を阻害することにより植物病原菌の防除を行う、（３）記載の方法；
（１４）きのこがウスヒラタケである（１３）記載の方法；
（１５）きのこの菌糸体または子実体または廃菌床と植物とが閉鎖系に置かれる（１）〜（１４）のいずれかに記載の方法。
That is, the present invention provides the following:
(1) Mushrooms of the genus Piptoporus, Mushrooms of the genus Mycoleptodonoides, Mushrooms of the genus Climacodon, Mushrooms of the genus Gloiothele, Mushrooms of the genus Lopharia, Mushrooms of the genus Scytinostroma, Mushrooms of the genus Neolentinus, derma of the genus Isch Mushrooms, mushrooms of the genus Laetiporus, mushrooms of the genus Daedaleopsis, mushrooms of the genus Steccherinum, mushrooms of the genus Trametes, mushrooms of the genus Boidinia, mushrooms of the genus Cymatoderma, mushrooms of the genus Scopuloides, mushrooms of the genus Ceriporia A method for controlling a phytopathogenic fungus, which comprises applying to a plant a volatile substance emanating from one or more selected mycelium, fruiting bodies, or waste beds thereof;
(2) The method according to (1), wherein the mushroom is an edible mushroom;
(3) white sponge bamboo (Piptoporus soloniensis), Bunaharitake (Mycoleptodonoides aitchisonii), Ezoharitake (Climacodon septentrionalis), Kinushibukawatake (Gloiothele citrina), olive scales data Ke (Lopharia spadicea), Uchiwatake (Microporus affinis), Chi Chi Iroko Uyaku bamboo (Scytinostroma galactinum), Scytinostroma portentosum, pine eel (Neolentinus lepideus), Nagano radicosum, shinitake (Ischnoderma resinosum), hirafusbe (Laetiporus versisporus), Egoch Trametes lactinea, Cymatoderma lamellatum, Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus eryngii, Trametes scabrosa, Boidinia lactico The method according to (1), wherein one or more mushroom mycelium selected from the group consisting of lor, Scopuloides hydnoides and Ceriporia lacerata, a volatile substance emitted from a fruit body or a waste fungus bed thereof is used. ;
(4) The method according to (3), wherein the mushroom is Beech Haritake or Ushiratake;
(5) The mycelial growth of phytopathogenic fungi using volatile substances emitted from mycelia or fruit bodies of one or two mushrooms selected from the group consisting of prickly shoots, prickly berries The method according to (3), wherein phytopathogenic fungi are controlled by inhibiting;
(6) Inhibiting the growth of phytopathogenic bacteria using a volatile substance emitted from one or more mycelium or fruit bodies of one or more mushrooms selected from the group consisting of olive mushrooms, mushrooms, hirafusube and ceriporia lacerata The method according to (3), wherein the plant pathogen is controlled by
(7) Using a volatile substance emitted from one or more mycelia or fruit bodies of one or more mushrooms selected from the group consisting of Beech Haritake, Olive scale, Mushroom, Matsuouji, Yakeiro, and Mushroom The method according to (3), wherein phytopathogenic fungi are controlled by inhibiting germination of phytopathogenic fungi spores;
(8) The method according to (7), wherein the mushroom is Beech Haritake;
(9) Use of a volatile substance emitted from one or more mycelium or fruiting bodies of mushrooms selected from the group consisting of beech haritake, olive mushroom, yake mushroom mushroom, mushroom mushroom, moss mushroom, chichiiroamitake and Scopuloides hydnoides And controlling the phytopathogenic fungi by inhibiting infection with phytopathogenic fungi;
(10) The method according to (9), wherein the mushroom is Beech Haritake;
(11) Inhibiting spore germination or infection of phytopathogenic fungi using volatile substances emitted from one or more mushroom mycelium or fruiting bodies selected from the group consisting of Ushiratake, Oyster mushrooms and eringi The method according to (3), wherein plant pathogens are controlled by:
(12) The method according to (11), wherein the mushroom is Ushiratake;
(13) A plant by inhibiting spore germination or infection of phytopathogenic fungi using a volatile substance emitted from one or more mushroom waste beds selected from the group consisting of Ushiratake, Oyster mushrooms and Elingi The method according to (3), wherein the pathogen is controlled;
(14) The method according to (13), wherein the mushroom is Ushiratake;
(15) The method according to any one of (1) to (14), wherein the mycelium or fruit body or the waste fungus bed of the mushroom and the plant are placed in a closed system.
本発明により、広範囲の植物病原菌に有効で、安全性が高く、環境汚染の少ない植物病原菌防除方法が提供される。 The present invention provides a method for controlling phytopathogenic fungi that is effective against a wide range of phytopathogenic fungi, is highly safe, and has little environmental pollution.
本発明は、１の態様において、Piptoporus属のきのこ、Mycoleptodonoides属のきのこ、Climacodon属のきのこ、Gloiothele属のきのこ、Lopharia属のきのこ、Microporus属のきのこ、Scytinostroma属のきのこ、Neolentinus属のきのこ、Hebeloma属のきのこ、Ischnoderma属のきのこ、Laetiporus属のきのこ、Daedaleopsis属のきのこ、Steccherinum属のきのこ、Trametes属のきのこ、Boidinia属のきのこ、Cymatoderma属のきのこ、Scopuloides属のきのこ、Ceriporia属のきのこおよびPleurotus属からなる群より選択される１種またはそれ以上のきのこの菌糸体、子実体、またはその廃菌床から発せられる揮発性物質を植物に適用することを特徴とする、植物病原菌の防除方法を提供する。 In one aspect, the present invention provides a mushroom of the genus Piptoporus, a mushroom of the genus Mycoleptodonoides, a mushroom of the genus Climacodon, a mushroom of the genus Gloiothele, a mushroom of the genus Lopharia, a mushroom of the genus Microporus, a mushroom of the genus Scytinostroma, a mushroom of the genus Neolentinus, Genus mushrooms, mushrooms of the genus Ischnoderma, mushrooms of the genus Laetiporus, mushrooms of the genus Daedaleopsis, mushrooms of the genus Steccherinum, mushrooms of the genus Trametes, mushrooms of the genus Cyididerma, mushrooms of the genus Cymatoderma, mushrooms of the genus Scopuloides A method for controlling a phytopathogenic fungus characterized by applying to a plant one or more mushroom mycelium selected from the group consisting of a genus, a fruiting body, or a volatile substance emanating from its waste fungus bed provide.
本発明の植物病原菌の防除方法に使用しうるきのこは、植物病原菌の防除を達成しうる揮発性物質を生産するものであれば、いずれのきのこであってもよい。なお、本明細書において「きのこ」あるいは「きのこ類」は、真菌類に属し、子実体を形成して子実体に胞子を生じさせるものをいう。きのこの大部分は担子菌門または子のう菌門に属している。 The mushroom that can be used in the method for controlling phytopathogenic fungi of the present invention may be any mushroom as long as it produces a volatile substance that can achieve control of phytopathogenic fungi. In the present specification, “mushroom” or “mushrooms” refers to those belonging to fungi and forming a fruiting body to produce spores in the fruiting body. Most mushrooms belong to the Basidiomycota or Ascomycota.
上で述べたように、生産される揮発性物質の安全性の面から、食用きのこの菌糸体、子実体、または廃菌床を用いることが好ましい。好ましい食用きのこの例としては、アイタケ、アミガサタケ、アミタケ、アラゲキクラゲ、アラゲコベニチャワンタケ、アワタケ、アワビタケ、アンズタケ、イモタケ、ウラベニホテイシメジ、エノキタケ、エリンギ、オウギタケ、オニフスベ、カラカサタケ、カンゾウタケ、キクラゲ、キシメジ、キツネタケ、キヌガサタケ、クギタケ、コノミタケ、サケツバタケ、シイタケ、シバフタケ、ショウゲンジ、ショウロ、シロオオハラタケ、シロカノシタ、シロキクラゲ、スギヒラタケ、スッポンタケ、セイヨウショウロ、セイヨウタマゴタケ、タマキクラゲ、タマゴタケ、タモギタケ、チチタケ、チャホウキタケモドキ、冬虫夏草、ツクリタケ、ナメコ、ナラタケ、ヌメリイグチ、ノボリリュウ、ハエトリシメジ、バカマツタケ、ハタケシメジ、ハナイグチ、ハラタケ、ハルシメジ、ヒラタケ、フクロタケ、ブナシメジ、ブナハリタケ、ベニナギナタタケ、ホンシメジ、マイタケ、マツタケ、ムキタケ、ムラサキシメジ、ヤマドリタケ、ヤマドリタケモドキ、ヤマブシタケ、ユキワリなどが挙げられるが、これらに限定されない。 As described above, it is preferable to use an edible mushroom mycelium, fruit body, or waste fungus bed from the viewpoint of safety of volatile substances to be produced. Examples of preferred edible mushrooms include agaric mushroom, agaricus mushroom, agaric mushroom, agaric jellyfish, agarico mushroom, agaric mushroom, agaric mushroom, chanterelle mushrooms, agaricus mushroom, enokitake mushroom, eringi, ougitake mushroom, agaric mushroom, Fox, mushroom, octopus mushroom, cynotake, salmon mushroom, shitake mushroom, shibatatake mushrooms, mushrooms, mushrooms, mushroom mushrooms, mushroom , Tsukuritake, Nameko, Naratake, Numeriiguchi, Nobori Ryu, Fly swordfish, Bakamatsutake, Hatakeshime , Suillus grevillei, Agaricus, Harushimeji, Pleurotus, straw mushroom, Bunashimeji, Bunaharitake, Beni Naginata bamboo, hon-shimeji, maitake, matsutake, Panellus Serotinus, purple shimeji, penny bun, Boletus reticulatus, Hericium erinaceus, but such calocybe gambosa include, but are not limited to.
本発明の１の好ましい態様において、Piptoporus属のきのこ、Mycoleptodonoides属のきのこ、Climacodon属のきのこ、Gloiothele属のきのこ、Lopharia属のきのこ、Microporus属のきのこ、Scytinostroma属のきのこ、Neolentinus属のきのこ、Hebeloma属のきのこ、Ischnoderma属のきのこ、Laetiporus属のきのこ、Daedaleopsis属のきのこ、Steccherinum属のきのこ、Trametes属のきのこ、Boidinia属のきのこ、Cymatoderma属のきのこ、Scopuloides属のきのこ、Ceriporia属のきのこおよびPleurotus属からなる群より選択される１種またはそれ以上のきのこの菌糸体、子実体、または廃菌床を用いる。より好ましくは、シロカイメンタケ（Piptoporus soloniensis）、ブナハリタケ（Mycoleptodonoides aitchisonii）、エゾハリタケ（Climacodon septentrionalis）、キヌシブカワタケ（Gloiothele citrina）、オリーブウロコタケ（Lopharia spadicea）、ウチワタケ（Microporus affinis）、チチイロコウヤクタケ（Scytinostroma galactinum）、ヤケイロコウヤクタケ（Scytinostroma portentosum）、マツオウジ（Neolentinus lepideus）、ナガエノスギタケ（Hebeloma radicosum）、ヤニタケ（Ischnoderma resinosum）、ヒラフスベ（Laetiporus versisporus）、エゴノキタケ（Daedaleopsis styracina）、ニセニクハリタケ（Steccherinum murashinskyi）、チチイロアミタケ（Trametes lactinea）、ヒダウロコタケ（Cymatoderma lamellatum）、ウスヒラタケ（Pleurotus pulmonarius）、ヒラタケ（Pleurotus ostreatus）、エリンギ（Pleurotus eryngii）、Trametes scabrosa、Boidinia lacticolor、Scopuloides hydnoidesおよびCeriporia lacerataからなる群より選択される１種またはそれ以上のきのこの菌糸体、子実体、または廃菌床から発せられる揮発性物質を用いる。さらに好ましくは、ブナハリタケまたはウスヒラタケの菌糸体、子実体または廃菌床から発せられる揮発性物質を用いる。上記の好ましいきのこ類は例示であり、これらに限定されるものではない。
In one preferred embodiment of the present invention, a mushroom of the genus Piptoporus, a mushroom of the genus Mycoleptodonoides, a mushroom of the genus Climacodon, a mushroom of the genus Gloiothele, a mushroom of the genus Lopharia, a mushroom of the genus Microporus, a mushroom of the genus Scytinostroma, a mushroom of the genus Neolentinus, Genus mushrooms, mushrooms of the genus Ischnoderma, mushrooms of the genus Laetiporus, mushrooms of the genus Daedaleopsis, mushrooms of the genus Steccherinum, mushrooms of the genus Trametes, mushrooms of the genus Cyididerma, mushrooms of the genus Cymatoderma, mushrooms of the genus Scopuloides One or more mushroom mycelium, fruiting bodies, or waste fungus beds selected from the group consisting of a genus are used. More preferably, white sponge bamboo (Piptoporus soloniensis), Bunaharitake (Mycoleptodonoides aitchisonii), Ezoharitake (Climacodon septentrionalis), Kinushibukawatake (Gloiothele citrina), olive scale data Ke (Lopharia spadicea), Uchiwatake (Microporus affinis), Chi Chi Iroko Uyaku bamboo ( Scytinostroma galactinum), Scytinostroma portentosum, pine mushroom (Neolentinus lepideus), Nagano radicosum, schizarder dinosai (Nelentin) (Trametes lactinea), Cymatoderma lamellatum, Pleurotus pulmonarius, Pleurotus ostreatus, Pleurotus eryngii, Trametes scabrosa, B Volatile substances emitted from one or more mushroom mycelium, fruiting bodies, or waste fungi selected from the group consisting of oidinia lacticolor, Scopuloides hydnoides and Ceriporia lacerata are used. More preferably, a volatile substance emitted from the mycelium, fruit body or waste fungus bed of Bunaharitake or Ushiratake is used. The above preferred mushrooms are examples, and are not limited thereto.
本発明の方法により防除されうる植物病原菌はいずれのものであってもよく、菌類、細菌を問わず、特に限定されないが、植物病原菌類としては、ナシ黒斑病菌（Alternaria alternata Japanese pear pathotype）、キャベツ黒斑病菌（Alternaria brassicae）、キャベツ黒すす病菌（Alternaria brassicicola）キュウリ炭疽病菌（Colletotrichum lagenarium）、トマト褐色輪紋病菌（Corynespora cassiicola）、キュウリ灰色疫病菌（Phytophthora capsici）、カンキツ褐色腐敗病菌（Phytophthora palmivora）、植物病原細菌としてはイネ白葉枯病菌（Xanthomonas oryzae pv. oryzae）、イネ内穎褐変病菌（Pantoea ananatis）、イネもみ枯細菌病菌（Burkholderia glumae）などが挙げられるが、これらに限定されない。 The plant pathogenic bacteria that can be controlled by the method of the present invention may be any, regardless of the fungus or bacteria, and are not particularly limited, but the plant pathogenic fungi include pear black spot fungus (Alternaria alternata Japanese pear pathotype), Cabbage black spot fungus (Alternaria brassicae), cabbage black soot fungus (Alternaria brassicicola) cucumber anthracnose fungus (Colletotrichum lagenarium), tomato brown rot fungus (Corynespora cassiicola), cucumber gray plague fungus (Phytophthora capsici), citrus brown rot fungus palmivora) and phytopathogenic bacteria include, but are not limited to, rice white leaf blight fungus (Xanthomonas oryzae pv. oryzae), rice brown rot fungus (Pantoea ananatis), rice blight blight fungus (Burkholderia glumae), and the like.
本明細書において「揮発性物質」あるいは「揮発性抗菌物質」は、常温常圧で揮発性を有し、大気中で気体状となる物質をいう。本発明は、揮発性抗菌物質というこれまでの殺菌剤とは異なった新しい視点からの病害防除技術の可能性を検討したことにおいて意義が大きいものである。上述のごとく、揮発性の抗菌物質は、通気の少ない施設栽培、例えば、室内や温室など使用条件は限定されるものの効果が隅々まで行き渡り、通気によって揮発性抗菌物質が容易に拡散、消失し、残留しないことから、安全面、健康面および環境面からも優れた特徴を持つと考えられる。特に、食用きのこの揮発性物質は安全性が高いといえる。 In the present specification, the “volatile substance” or “volatile antibacterial substance” refers to a substance that is volatile at normal temperature and pressure and is gaseous in the atmosphere. The present invention has a great significance in examining the possibility of a disease control technique from a new viewpoint different from conventional fungicides called volatile antibacterial substances. As described above, volatile antibacterial substances can be used in greenhouse cultivation where there is little ventilation, for example, indoors and greenhouses, although the conditions of use are limited. Since it does not remain, it is considered to have excellent characteristics from the viewpoints of safety, health and environment. In particular, edible mushroom volatile substances are safe.
本発明の方法においては、きのこの菌糸体または子実体、あるいはきのこを菌床栽培して収穫した後に得られる廃菌床から発せられる揮発性物質を植物に適用して植物病原菌の防除を行う。とりわけ、食用きのこの廃菌床は大量に廃棄されており、これを再利用することは環境にも優しい技術といえる。揮発性物質は植物病原菌の防除効果を有するものであればよい。植物病原菌の「防除」は、例えば、植物病原菌の増殖、菌糸生育、胞子の形成または発芽、感染などを阻害、抑制または停止させて、植物病原菌による植物への害を防止、抑制、排除、あるいは植物病原菌に感染した植物を治癒することを含む。 In the method of the present invention, phytopathogenic fungi are controlled by applying to a plant a volatile substance emitted from a mycelium or fruit body of a mushroom or a waste fungus bed obtained after harvesting mushrooms by cultivation. In particular, waste mushroom beds for edible mushrooms are discarded in large quantities, and reusing them is an environmentally friendly technology. Any volatile substance may be used as long as it has an effect of controlling plant pathogens. “Control” of plant pathogens, for example, inhibits, suppresses or stops plant pathogenic fungi, mycelial growth, spore formation or germination, infection, etc. Includes healing of plants infected with phytopathogenic fungi.
本発明の１の好ましい態様において、ウチワタケ、チチイロコウヤクタケ、チチイロアミタケおよびヒダウロコタケからなる群より選択される１種または２種のきのこの菌糸体または子実体から発せられる揮発性物質を用いて、植物病原菌類の菌糸生育を阻害することにより植物病原菌の防除を行うことができる。 In one preferred embodiment of the present invention, a volatile substance emanating from a mycelium or fruiting body of one or two mushrooms selected from the group consisting of prickly shoots, mushrooms, mushrooms, and mushrooms is used, Plant pathogens can be controlled by inhibiting mycelial growth of pathogenic fungi.
本発明の１の好ましい態様において、オリーブウロコタケ、ヤニタケ、ヒラフスベおよびCeriporia lacerataからなる群より選択される１種またはそれ以上のきのこの菌糸体または子実体から発せられる揮発性物質を用いて、植物病原細菌の増殖を阻害することにより植物病原菌の防除を行うことができる。 In one preferred embodiment of the present invention, a volatile substance emanating from one or more mycelium or fruiting bodies of one or more mushrooms selected from the group consisting of olive mushrooms, mushrooms, hirafusube and ceriporia lacerata is used to Plant pathogens can be controlled by inhibiting the growth of pathogenic bacteria.
本発明の１の好ましい態様において、ブナハリタケ、オリーブウロコタケ、チチイロコウヤクタケ、マツオウジ、ヤケイロコウヤクタケおよびヤニタケからなる群より選択される１種またはそれ以上のきのこの菌糸体または子実体から発せられる揮発性物質を用いて、植物病原菌類の胞子の発芽を阻害することにより植物病原菌の防除を行うことができる。この好ましい態様において、きのこがブナハリタケであることが、より好ましい。 In one preferred embodiment of the present invention, it originates from one or more mycelium or fruiting bodies of one or more mushrooms selected from the group consisting of beech haritake, olive mushroom, chichiiro mushroom, pine mushroom, mushroom mushroom, and mushroom. It is possible to control phytopathogenic fungi by inhibiting spore germination of phytopathogenic fungi using the volatile substances obtained. In this preferred embodiment, it is more preferred that the mushroom beech beech.
本発明の１の好ましい態様において、ブナハリタケ、オリーブウロコタケ、ヤケイロコウヤクタケ、ヤニタケ、ニセニクハリタケ、チチイロアミタケおよびScopuloides hydnoidesからなる群より選択される１種またはそれ以上のきのこの菌糸体または子実体から発せられる揮発性物質を用いて、植物病原菌類の感染を阻害することにより植物病原菌の防除を行うことができる。この好ましい態様において、きのこがブナハリタケであることが、より好ましい。 In one preferred embodiment of the present invention, the sprout or fruiting body of one or more mushroom mycelium or fruiting bodies selected from the group consisting of beech haritake, olive mushrooms, mushroom mushrooms, mushrooms, moss mushrooms, mushrooms and Scopuloides hydnoides. It is possible to control phytopathogenic fungi by inhibiting infection with phytopathogenic fungi using the volatile substances obtained. In this preferred embodiment, it is more preferred that the mushroom beech beech.
本発明の１の好ましい態様において、ウスヒラタケ、ヒラタケおよびエリンギからなる群より選択される１種またはそれ以上のきのこの菌糸体または子実体、から発せられる揮発性物質を用いて、植物病原菌類の感染を阻害することにより植物病原菌の防除を行うことができる。この好ましい態様において、ウスヒラタケの廃菌床を用いることが、より好ましい。 In one preferred embodiment of the present invention, infection with phytopathogenic fungi using volatile substances emanating from one or more mycelium or fruiting bodies of one or more mushrooms selected from the group consisting of oyster mushrooms, oyster mushrooms and eryngii It is possible to control plant pathogens by inhibiting. In this preferred embodiment, it is more preferable to use the waste fungus bed of Ushiratake.
本発明の１の好ましい態様において、ウスヒラタケ、ヒラタケおよびエリンギからなる群より選択される１種またはそれ以上のきのこの廃菌床から発せられる揮発性物質を用いて、植物病原菌類の感染を阻害することにより植物病原菌の防除を行うことができる。この好ましい態様において、ウスヒラタケの廃菌床を用いることが、より好ましい。 In one preferred embodiment of the present invention, a volatile substance emanating from a waste bed of one or more mushrooms selected from the group consisting of oyster mushrooms, oyster mushrooms and eryngii is used to inhibit infection of phytopathogenic fungi Therefore, it is possible to control plant pathogens. In this preferred embodiment, it is more preferable to use the waste fungus bed of Ushiratake.
上述の本発明の好ましい態様は例示であり、これらの態様に限定されない。また、本発明の好ましい態様を２つ以上組み合わせてもよい。 The preferred embodiments of the present invention described above are illustrative and are not limited to these embodiments. Two or more preferred embodiments of the present invention may be combined.
本発明における、揮発性物質の植物への適用は、きのこの菌糸体または子実体、あるいはその廃菌床から発せられる揮発性物質が、植物病原菌と接触するような態様であればよい。きのこの菌糸体または子実体、あるいはその廃菌床を植物の近傍に配置することが好ましい。例えば、きのこの菌糸体または子実体、あるいはその廃菌床を、植物が生えている土壌に置く、あるいは植物の周囲に置く、吊すなどにより、揮発性物質を植物病原菌に適用することができる。きのこの菌糸体、子実体、または廃菌床を細かくほぐす、あるいは切断して、植物近傍に適用してもよい。きのこの菌糸体または子実体、あるいはその廃菌床を適当な容器に入れて保持してもよい。これらの適用の態様は例示であり、限定的なものではない。 In the present invention, the volatile substance may be applied to the plant as long as the volatile substance emitted from the mycelium or fruit body of the mushroom or the waste fungus bed comes into contact with the phytopathogenic fungus. It is preferable to arrange the mycelium or fruit body of the mushroom or the waste fungus bed in the vicinity of the plant. For example, a volatile substance can be applied to a phytopathogenic fungus by placing a mycelium or fruit body of a mushroom or a waste microbial bed thereof on soil where the plant grows, or surrounding or hanging the plant. Mushroom mycelium, fruiting bodies or waste fungus beds may be loosened or cut and applied to the vicinity of the plant. The mycelium or fruit body of mushrooms or the waste fungus bed thereof may be held in an appropriate container. These aspects of application are exemplary and not limiting.
揮発性抗菌物質は、効果が隅々まで行き渡り、通気によって揮発性抗菌物質が容易に拡散、消失することから、安全面、健康面および環境面からも優れた特徴を持つ。揮発性抗菌物質のこのような特性を考えると、きのこの菌糸体または子実体、あるいはその廃菌床と植物を閉鎖系に置いて、植物病原菌の防除を行うことが好ましい。閉鎖系の使用により、植物周囲の揮発性抗菌物質の濃度を高く保つことができ、閉鎖系を一部あるいは全部解除できるようにしておけば、植物周囲の揮発性抗菌物質の濃度をコントロールすることも容易である。閉鎖系の例としては、フード、テント、ハウス、温室、植物工場、農作物収穫後の貯蔵倉庫（貯蔵病害の防除）、農作物出荷時のコンテナー（市場病害の防除）などが挙げられるが、これらに限定されない。 Volatile antibacterial substances have excellent characteristics from the viewpoints of safety, health and the environment because the volatile antibacterial substances are effective in every corner and the volatile antibacterial substances diffuse and disappear easily by ventilation. Considering such characteristics of the volatile antibacterial substance, it is preferable to control the phytopathogenic fungi by placing the mycelium or fruit body of the mushroom or the waste fungus bed and the plant in a closed system. By using a closed system, the concentration of volatile antibacterial substances around the plant can be kept high, and if the closed system can be partially or completely released, the concentration of volatile antibacterial substances around the plant should be controlled. Is also easy. Examples of closed systems include food, tents, houses, greenhouses, plant factories, storage warehouses after crop harvest (control of stored diseases), and containers at the time of crop shipment (control of market diseases). It is not limited.
また、本発明は、一般家屋、病院、公共施設などの閉鎖室内の防菌・除菌にも使用することができる。 The present invention can also be used for sterilization and sterilization in closed rooms such as general houses, hospitals, and public facilities.
本明細書において上で定義した以外の用語は、当該分野において通常に認識されている意味に解され、例えば、農業、農薬、菌学、生化学などの分野の教科書、辞書、論文等に使用されている意味に解される。 Terms other than those defined above in this specification are understood to have the meanings commonly recognized in the field, and are used, for example, in textbooks, dictionaries, articles, etc. in fields such as agriculture, agricultural chemicals, mycology, and biochemistry. It is understood in the meaning that is.
後で説明する実施例における主な実験材料および実験方法について説明する。本発明の実施における材料および方法は、ここで説明するものに限定されるものではない。 Main experimental materials and experimental methods in Examples described later will be described. The materials and methods for practicing the invention are not limited to those described herein.
１．供試きのこ類
本研究では、鳥取大学農学部附属菌類きのこ遺伝資源研究センターに保存されている多数の木材腐朽性きのこ類の中から芳香臭のある菌株を選抜して使用した（表１）。使用に際しては、ジャガイモ煎汁寒天培地（ＰＤＡ培地）で保存された菌株の菌糸片を、ＰＤＡを入れたシャーレ（９ｃｍ）に植え付け、２５℃暗下で１週間静置培養した。また、食用きのことして、菌床栽培されたヒラタケ（Pleurotus ostreatus）、ウスヒラタケ（Pleurotus pulmonarius）、エリンギ（Pleurotus eryngii）およびヤマブシタケ（Hericium erinaceum）のきのこ（子実体）および廃菌床を使用した。
２．供試病原菌および植物
揮発性抗菌物質の検定には、表２に示す各種植物病原菌類および細菌を使用した。使用に際しては、ＰＤＡ培地で保存された病原菌類の菌糸片および細菌のコロニーを、ＰＤＡの入ったシャーレ（９ｃｍ）に植え付け、２５℃暗下で菌類は１週間、細菌は３日間静置培養した。また、病原菌類の場合（Phytophthora属菌は除く）は、胞子を得るために保存菌株の菌糸片をＶ８ジュース寒天培地（Ｖ８培地）（キャベツ黒斑病菌ではローズベンガル添加）の入ったシャーレ（９ｃｍ）に植え付け、２０℃で約１ヶ月間ＢＬＢランプ照射下に静置して培地上に胞子を形成させた。なお、キュウリ炭疽病菌はPDY培地（酵母エキス添加PDA）の入ったシャーレ（９ｃｍ）に植え付け、２５℃暗下に約２週間静置し培地上に胞子を形成させた。培地上に形成された胞子を蒸留水（ＤＷ）に懸濁し、遠心分離（８００ｘｇ、５分）により数回洗浄して、胞子濃度を５ｘ１０５個／ｍｌに調製後、実験に使用した。
2. Various phytopathogenic fungi and bacteria shown in Table 2 were used for the test of test pathogens and plant volatile antibacterial substances. In use, mycelia and bacterial colonies of pathogenic fungi preserved in PDA medium were planted in a petri dish (9 cm) containing PDA, and the fungi were allowed to stand for 1 week and bacteria for 3 days in the dark at 25 ° C. In the case of pathogenic fungi (except Phytophthora spp.), In order to obtain spores, the mycelium of the conserved strain is petri dish (9 cm with Rose Bengal added in V8 juice agar medium (V8 medium) (cabbage black spot fungus)) And allowed to stand under BLB lamp irradiation at 20 ° C. for about 1 month to form spores on the medium. The cucumber anthracnose was planted in a petri dish (9 cm) containing a PDY medium (yeast extract-added PDA) and allowed to stand in the dark at 25 ° C. for about 2 weeks to form spores on the medium. Spores formed on the medium were suspended in distilled water (DW), washed several times by centrifugation (800 × g, 5 minutes), adjusted to a spore concentration of 5 × 10 5 cells / ml, and then used for experiments.
一方、病原菌類のキャベツ黒すす病菌（Ｏ−２６４菌株）およびトマト褐色輪紋病菌（ＬＣ９３０２０菌株）の宿主植物として、それぞれ植物育苗室で栽培したキャベツ（品種：初秋）およびトマト（品種：桃太郎）を使用した。
３．きのこ培養菌糸体における揮発性抗菌物質生産の検定
きのこ培養菌糸体における揮発性抗菌物質生産の検定法の概要を図１に、スキームを図２に示す。ＰＤＡ培地上で生育させた各種きのこ類菌株の菌糸片をＰＤＡ培地の入った大型シャーレ（直径１５ｃｍ）の３箇所に植え付け、２５℃暗下で培養して培地全面に菌糸を生育させた後、大型シャーレの上下を逆転（菌糸体上面）した。植物病原菌類および細菌の生育に対する抗菌活性をみるために、病原菌類の菌糸片（６ｍｍ）をＰＤＡ培地上に置床したシャーレ（５ｃｍ）または病原細菌コロニーをＰＤＡ培地上に塗布したシャーレ（５ｃｍ）を大型シャーレ内に置いた。大型シャーレをテープで密封し、２５℃暗下に置き、病原菌類の場合は１週間後の菌糸生育を、病原細菌の場合は３日後のコロニー増殖を測定した。また、病原菌類の場合には、胞子発芽および宿主植物への感染に対する抗菌活性をみるため、胞子懸濁液を噴霧したスライドグラスおよび胞子懸濁液を噴霧した宿主切取り葉を、培地全面に各種きのこ類菌株の菌糸を生育させた大型シャーレの水で湿らせたペーパータオルの上に置き、テープで密封して２５℃暗下に静置して、２４時間後にスライドグラス上の胞子発芽率を、４８時間後に宿主切取り葉における病斑数を調べた。
3. Test of production of volatile antibacterial substances in mushroom culture mycelium FIG. 1 shows an outline of a test method for production of volatile antibacterial substances in mushroom culture mycelium, and FIG. After hyphae of various mushroom strains grown on PDA medium were planted in three places on a large petri dish (diameter 15 cm) containing PDA medium and cultured in the dark at 25 ° C., mycelia were grown on the entire surface of the medium. The petri dish was turned upside down (upper surface of the mycelium). In order to check the antibacterial activity against the growth of phytopathogenic fungi and bacteria, a petri dish (5 cm) on which a mycelium piece (6 mm) of pathogenic fungi is placed on a PDA medium or a petri dish (5 cm) coated with a pathogenic bacterial colony on a PDA medium. Placed in a large petri dish. A large petri dish was sealed with a tape and placed in the dark at 25 ° C. In the case of pathogenic fungi, mycelial growth after 1 week was measured, and in the case of pathogenic bacteria, colony growth after 3 days was measured. In the case of pathogenic fungi, in order to check the antibacterial activity against spore germination and infection to host plants, various types of slide glass sprayed with spore suspension and host cut leaves sprayed with spore suspension were applied to the entire surface of the medium. Place it on a paper towel moistened with water from a large petri dish on which the mycelium of the mushroom strain was grown, seal with tape and leave it in the dark at 25 ° C. After 24 hours, the spore germination rate on the slide glass was 48 The number of lesions on the host cut leaves was examined after time.
４．食用きのこの子実体および廃菌床における揮発性抗菌物質生産の検定
食用きのこの子実体および廃菌床における揮発性抗菌物質生産の検定法のスキームを図３に示す。食用きのこの子実体および廃菌床それぞれ３０ｇ（生重）をシャーレ（直径９ｃｍ）に入れ、シャーレの蓋をせずに密封プラスチック容器（横２２ｃｍ、幅１６ｃｍ、高さ６ｃｍ）内に置いた。さらに、植物病原菌類の胞子懸濁液を噴霧したスライドグラスおよび胞子懸濁液を噴霧した宿主切取り葉を、食用きのこの子実体および廃菌床それぞれ３０ｇ（新鮮重）を入れたプラスチック容器内の水で湿らせたペーパータオルの上に置き、プラスチック容器を密封し２５℃暗下に静置して、２４時間後にスライドグラス上の胞子発芽率を、４８時間後に宿主切取り葉における病斑形成を調べた。
4). Test of volatile antibacterial substance production in edible mushroom fruit body and waste fungus bed FIG. 3 shows a scheme of a test method of volatile antibacterial substance production in edible mushroom fruit body and waste fungus bed. 30 g (fresh weight) of each edible mushroom fruit body and waste fungus bed were placed in a petri dish (diameter 9 cm) and placed in a sealed plastic container (width 22 cm, width 16 cm, height 6 cm) without a petri dish lid. Furthermore, a slide glass sprayed with a spore suspension of a phytopathogenic fungus and a host cut leaf sprayed with a spore suspension were placed in a plastic container containing 30 g (fresh weight) of edible mushroom fruit bodies and waste fungus beds. Place it on a paper towel moistened with water, seal the plastic container and let it stand in the dark at 25 ° C. After 24 hours, the spore germination rate on the slide glass was examined, and after 48 hours, the lesion formation in the host cut leaves was examined. .
以下に実施例を示して本発明をさらに具体的かつ詳細に説明するが、実施例はあくまでも本発明の例示説明であり、本発明を限定するものではない。 EXAMPLES The present invention will be described more specifically and in detail below with reference to examples. However, the examples are merely illustrative of the present invention and do not limit the present invention.
１．各種きのこ類の菌糸体における揮発性抗菌物質生産
鳥取大学農学部附属菌類きのこ遺伝資源研究センターに保存されている芳香臭のあるきのこ菌株の培養菌糸体を用いて揮発性抗菌物質生産の有無を調べた。各種植物病原菌類の菌糸生育に対して供試菌株の多くが顕著ではないものの菌糸生育を阻害し、中でも３３８２４菌株（ウチワタケ）および３３８６３菌株（チチイロコウヤクタケ）、３４６９９菌株（チチイロアミタケ）、３４８５８菌株（ヒダウロコタケ）は、Phytophthora属菌を除く供試病原菌類すべてに抗菌活性を示した（表３、図４）。また、３４６３４菌株はPhytophthora属の２菌株に抗菌活性を示した（表３）。
一方、各種植物病原細菌の増殖に対しても、供試菌株の多くの菌糸体が顕著ではないものの増殖を阻害し、中でも１０１０８菌株（オリーブウロコタケ）、３４１４９菌株（ヤニタケ）、３４２２８菌株（ヒラフスベ）および３５０９９菌株はほとんどの供試病原細菌の増殖を阻害した（表４、図４）。なお、病原細菌の中でＣＴＢ１１３５菌株（イネ内穎褐変病菌）は、ほとんどのきのこの菌株において顕著な阻害を受けなかった。
次に、植物病原菌類の胞子発芽に対する阻害は、多くのきのこ類の菌糸体において認められ、中でも、１００９９菌株（ブナハリタケ）、１０１０８菌株（オリーブウロコタケ）、３３８６３（チチイロコウヤクタケ）、３３８８３（マツオウジ）、３３８８５（ヤケイロコウヤクタケ）および３４１４９（ヤニタケ）の菌糸体は、供試したほとんどの病原菌類胞子の発芽を阻害し、特に１００９９菌株（ブナハリタケ）ではいずれの胞子も全く発芽しなかった（表５、図４）。また、植物病原菌類の感染に対する阻害は、多くのきのこ類の菌糸体において認められ、中でも、１００９９菌株（ブナハリタケ）、１０１０８菌株（オリーブウロコタケ）、３３８８５菌株（ヤケイロコウヤクタケ）、３４１４９菌株（ヤニタケ）、３４２３８（ニセニクハリタケ）、３４６９９菌株（チチイロアミタケ）および３４８６５菌株の菌糸体は、検定したキャベツ黒すす病菌およびトマト褐色輪紋病菌の感染を阻害し、特に１００９９菌株（ブナハリタケ）は顕著な阻害活性を示した（表６、図４）。
以上の結果から、芳香臭のあるきのこ菌株の菌糸体の多くは、検定法の違いによって抗菌活性は異なるものの、揮発性抗菌物質を生産することが明らかとなった。その中で、食用きのこである１００９９菌株（ブナハリタケ）は、病原細菌の増殖にはほとんど効果を示さなかったが、病原菌類の胞子発芽や感染に対しては顕著な阻害活性を示した。 From the above results, it has been clarified that many mycelia of mushroom strains with an aromatic odor produce volatile antibacterial substances, although the antibacterial activity differs depending on the assay method. Among them, the edible mushroom 10099 strain (Bunaharitake) showed little effect on the growth of pathogenic bacteria, but showed remarkable inhibitory activity against spore germination and infection of pathogenic fungi.
２．各種食用きのこ類の子実体および廃菌床における揮発性抗菌物質生産
培養菌糸体による顕著な揮発性抗菌物質の生産がみられたブナハリタケは食用きのこであることから、菌床栽培の行われているヒラタケ、ウスヒラタケ、エリンギおよびヤマブシタケの４種の食用きのこの子実体および廃菌床を用いて揮発性抗菌物質生産の検定を行った）。その結果（表７）、子実体では、ウスヒラタケ、ヒラタケおよびエリンギが胞子発芽および宿主植物への感染を阻害し、中でもウスヒラタケの活性が顕著であった。なお、ヤマブシタケはほとんど活性を示さなかった。一方、廃菌床では、ウスヒラタケのみが胞子発芽および宿主植物への感染を顕著に阻害し、ヒラタケおよびエリンギの廃菌床は若干の活性を示した。
本発明は、農業分野、園芸分野、植物病理学の研究などにおいて利用可能である。 The present invention can be used in the fields of agriculture, horticulture, plant pathology, and the like.
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