JP6232602B2 - Soil infectious disease control agent and soil infectious disease control method using the same - Google Patents
Soil infectious disease control agent and soil infectious disease control method using the same Download PDFInfo
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- JP6232602B2 JP6232602B2 JP2013173440A JP2013173440A JP6232602B2 JP 6232602 B2 JP6232602 B2 JP 6232602B2 JP 2013173440 A JP2013173440 A JP 2013173440A JP 2013173440 A JP2013173440 A JP 2013173440A JP 6232602 B2 JP6232602 B2 JP 6232602B2
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- soil
- infectious disease
- disease control
- control agent
- bacteria
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Description
本発明は、土壌伝染性病害の原因となる植物病原糸状菌に対して拮抗作用を有する微生物を有効成分とする土壌伝染性病害防除剤、及びそれを用いて土壌伝染性病害の発病を予防又は防除する方法に関する。 The present invention is a soil infectious disease control agent comprising a microorganism having an antagonistic action against a phytopathogenic fungus causing a soil infectious disease as an active ingredient, and prevention or development of a soil infectious disease using the same. It relates to a method for controlling.
土壌伝染性病害は、土壌を介して伝播する農業上重要な伝染性の植物病害である。例えば、紫紋羽病菌(Helicobasidium mompa)を原因菌とする紫紋羽病は、草木植物及び木本植物を問わず、イネ科を除くほとんどの植物に感染し、病原性を示す多犯性の土壌伝染性病害である。日本や韓国等の東アジアを中心に発生し、農作物の極端な品質低下、収量減少等の多大な損害を与えている。また、紫紋羽病菌は、土壌中での生存期間が5〜20年にも及び、一度この菌に汚染されると無発病状態に改善するまで少なくとも5年以上、イネ科の輪作を行うか休閑地にする必要がある。 Soil-borne diseases are agriculturally important infectious plant diseases that are transmitted through soil. For example, purple crest rot, caused by Helicobasidium mompa, infects most plants, except grasses and woody plants, regardless of the plant and woody plant. It is a soil-borne disease. Occurring mainly in East Asia such as Japan and Korea, it causes great damage such as extreme quality degradation and yield reduction of crops. In addition, it has a life span of 5 to 20 years in the soil, and if it is contaminated with this fungus, will it rotate for at least 5 years before it improves to the absence of disease? Need to be fallow.
土壌伝染性病害の防除対策として、従来法では簡便で万能的な効果を示す臭化メチルを中心に、クロルピクリンやダゾメット等によって土壌消毒する化学的防除法が行われてきた。しかし、臭化メチルは2005年に原則使用廃止となり、2013年には全廃となった。そこで、臭化メチルに代わる新たな防除方法が検討されたが、クロルピクリンやダゾメット等を用いた化学防除方法は、臭化メチルと比較すると効果や使用面において問題が多く、臭化メチルの代替剤としては不十分であった。また、近年、化学農薬による防除法は、環境への負荷が大きい上に、作業者や食品への安全性という点で問題視されている。さらに、化学農薬は、土壌への施用によって標的とする土壌伝染性病害の原因菌以外の有用な微生物相にも重大な影響を及ぼし得る。それ故、化学的防除法に替わる安全で持続可能な土壌伝染性病害の防除法の開発が求められている。 As a control measure for soil infectious diseases, chemical control methods for disinfecting soil with chloropicrin, dazomet, etc. have been carried out mainly on methyl bromide, which has a simple and universal effect. However, methyl bromide was abolished in principle in 2005 and was abolished in 2013. Therefore, a new control method to replace methyl bromide was studied, but chemical control methods using chloropicrin, dazomet, etc. have more problems in terms of effect and use than methyl bromide, and are an alternative to methyl bromide. As it was insufficient. In recent years, the control method using chemical pesticides has been regarded as a problem in terms of safety to workers and foods as well as a large burden on the environment. In addition, chemical pesticides can also have a significant impact on useful microflora other than the causative organisms of soil infectious diseases targeted by application to soil. Therefore, there is a need for the development of safe and sustainable control methods for soil-borne diseases that replace chemical control methods.
上記のような状況の中で、植物の病原菌に対して拮抗作用を有する微生物(拮抗微生物)を病害の抑制に利用する微生物資材の研究が進められている。拮抗微生物とは、特定の微生物の増殖や活動を抑制する微生物の事である。無農薬農業等の生態系活用型(又は、環境保全型)農業への移行が求められている昨今において、拮抗微生物を微生物農薬として利用して、病原菌の増殖や活動を抑制し、結果的に植物に対する病害を軽減する技術は夢の技術といっても過言ではなく、その開発が切望されている(特許文献1)。 Under the circumstances as described above, research on microbial materials utilizing microorganisms having antagonistic action against plant pathogenic bacteria (antagonistic microorganisms) for disease control is underway. Antagonistic microorganisms are microorganisms that suppress the growth and activity of specific microorganisms. In recent years when there is a need to shift to ecosystem-based agriculture such as pesticide-free agriculture (or environmental conservation) agriculture, the use of antagonistic microorganisms as microbial pesticides suppresses the growth and activity of pathogenic bacteria. It is not an exaggeration to say that the technology for reducing diseases to plants is a dream technology, and its development is eagerly desired (Patent Document 1).
例えば、枯草菌バチルス・サブチリス(Bacillus subtilis)は、植物に病害をもたらす、ある種の病原菌と拮抗することが知られており、ナスやトマトの灰色かび病の防除剤として、日本では既に農薬登録されている。 For example, Bacillus subtilis (Bacillus subtilis) is known to antagonize certain pathogenic bacteria causing plant diseases, and has already been registered as a pesticide in Japan as a control agent for gray mold of eggplant and tomato. Has been.
また、特許文献2には、バチルス・サブチリス等のバチルス属に属する細菌の培養物から、胞子を乾燥重量で50重量%以上含むように調製した胞子画分を含有する農園芸植物の病害防除技術が開示されている。
Further,
さらに、特許文献3には、放線菌アミコラトプシス属(Amycolatopsis) A1菌株からなり、植物に対して病原性が無く、農業分野への適応が容易で、植物体に高い定着性を示し、糸状菌病害防除に優れた防除効果を発揮する糸状菌病害防除剤が開示されている。 Furthermore, Patent Document 3 is composed of the actinomycete Amycolatopsis A1 strain, has no pathogenicity to plants, is easily adaptable to the agricultural field, exhibits high colonization in plants, and is filamentous. A filamentous fungal disease control agent that exhibits an excellent control effect on fungal disease control is disclosed.
植物病原糸状菌を原因とする土壌伝染性病害に対して高い防除効果を有し、環境に対する負荷が少なく、安全で薬害のない微生物農薬を開発し提供すること、及びその微生物農薬を用いて土壌伝染性病害を効果的に防除する方法を開発することによって、作業労力やコストを低減し、安全な農産物を安定的に提供することである。 Develop and provide microbial pesticides that have a high control effect on soil-borne diseases caused by phytopathogenic fungi, have a low environmental impact, and are free from phytotoxicity. By developing a method to effectively control infectious diseases, the labor and cost can be reduced, and safe agricultural products can be provided stably.
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、光合成細菌であるロドシュードモナス属(Rhodopseudomonas)細菌と土壌細菌であるバチルス属(Bacillus)細菌を組み合わせて施用した時に、その複合作用により土壌伝染性病害の原因菌である植物病原糸状菌に対して極めて高い拮抗性と肥料効果を示すことを見出し、本発明を完成させるに至った。本発明は、上記知見に基づくもので、以下を提供する。 As a result of intensive studies to solve the above-mentioned problems, the present inventors have obtained a combined action when a photosynthetic bacterium, Rhodopseudomonas bacterium and a soil bacterium, Bacillus bacterium are applied in combination. Has found that it exhibits extremely high antagonistic properties and fertilizer effects against phytopathogenic fungi, which are the causative agents of soil-borne diseases, and has completed the present invention. The present invention is based on the above findings and provides the following.
(1)植物病原糸状菌に対する土壌伝染性病害防除剤の有効成分として使用するためのロドシュードモナス属細菌。
(2)ロドシュードモナス属細菌が受託番号FERM BP-19431又はFERM BP-19432の細菌である、(1)に記載のロドシュードモナス属細菌。
(3)植物病原糸状菌に対する土壌伝染性病害防除剤の有効成分として使用するためのバチルス属細菌。
(4)バチルス属細菌が受託番号NITE BP-01678又はNITE BP-01679の細菌である、(3)に記載のバチルス属細菌。
(5)光合成細菌及びバチルス属細菌を有効成分として含む植物病原糸状菌に対する土壌伝染性病害防除剤。
(6)前記光合成細菌がロドシュードモナス属細菌である、(5)に記載の土壌伝染性病害防除剤。
(7)ロドシュードモナス属細菌及びバチルス属細菌の混合比率が1:1〜1000:1である、(6)に記載の土壌伝染性病害防除剤。
(8)前記ロドシュードモナス属細菌が受託番号FERM BP-19431又はFERM BP-19432の細菌である、(6)又は(7)に記載の土壌伝染性病害防除剤。
(9)前記バチルス属細菌が受託番号NITE BP-01678又はNITE BP-01679の細菌である、(5)〜(8)のいずれかに記載の土壌伝染性病害防除剤。
(10)植物病原糸状菌がヘリコバシディウム属(Helicobasidium)菌、ロゼリニア属(Rosellinia)菌、フザリウム属(Fusarium)菌、ピシウム属(Pythium)菌、ピレノカエタ属(Pyrenochaeta)菌、モノスポラスカス属(Monosporascus)菌、バーティシリウム属(Verticillium)菌、リゾクトニア属(Rhizoctonia)菌、プラスモディオフォラ属(Plasmodiophora)菌、フィトソフラ属(Phitophthora)菌、及びスクレロチウム属(Sclerotium)菌からなる群から選択される、(5)〜(9)のいずれかに記載の土壌伝染性病害防除剤。
(11)土壌伝染性病害が、紋羽病、根こぶ病、苗立枯病、萎凋病、根腐病、白絹病、及びフィトソフラ疫病からなる群から選択される、(5)〜(10)のいずれかに記載の土壌伝染性病害防除剤。
(12)(5)〜(11)のいずれかに記載の土壌伝染性病害防除剤を土壌又は培地に施用する、土壌伝染性病害防除方法。
(13)前記土壌が対象植物の栽植用土壌である、(12)に記載の土壌伝染性病害防除方法。
(14)対象植物の植え付け前に施用する、(13)に記載の土壌伝染性病害防除方法。
(1) A Rhodopseudomonas bacterium for use as an active ingredient of a soil infectious disease control agent against phytopathogenic fungi.
(2) The Rhodopseudomonas bacterium according to (1), wherein the Rhodopseudomonas bacterium is a bacterium having an accession number of FERM B P-19431 or FERM B P-19432.
(3) A Bacillus bacterium for use as an active ingredient of a soil infectious disease control agent against phytopathogenic fungi.
(4) The Bacillus bacterium according to (3), wherein the Bacillus bacterium is a bacterium having an accession number of NITE B P-01678 or NITE B P-01679.
(5) A soil infectious disease control agent for phytopathogenic fungi containing photosynthetic bacteria and Bacillus bacteria as active ingredients.
(6) The soil infectious disease control agent according to (5), wherein the photosynthetic bacterium is a genus Rhodopseudomonas.
(7) The soil infectious disease control agent according to (6), wherein the mixing ratio of Rhodopseudomonas bacteria and Bacillus bacteria is 1: 1 to 1000: 1.
(8) The soil infectious disease control agent according to (6) or (7), wherein the Rhodopseudomonas bacterium is a bacterium having an accession number of FERM B P-19431 or FERM B P-19432.
(9) The soil infectious disease control agent according to any one of (5) to (8), wherein the Bacillus bacterium is a bacterium having an accession number of NITE B P-01678 or NITE B P-01679.
(10) The phytopathogenic fungi are Helicobasidium, Rosellinia, Fusarium, Pythium, Pyrenochaeta, Monosporacus (Monosporascus), Verticillium, Rhizoctonia, Plasmodiophora, Phitophthora, and Sclerotium The soil infectious disease control agent according to any one of (5) to (9).
(11) The soil infectious disease is selected from the group consisting of coat feather disease, root-knot disease, seedling blight disease, wilt disease, root rot disease, silkworm disease, and phytosofra disease. ) The soil infectious disease control agent according to any one of the above.
(12) A soil infectious disease control method, wherein the soil infectious disease control agent according to any one of (5) to (11) is applied to soil or a culture medium.
(13) The soil infectious disease control method according to (12), wherein the soil is soil for planting a target plant.
(14) The soil infectious disease control method according to (13), which is applied before planting the target plant.
本発明のロドシュードモナス属細菌又はバチルス属細菌によれば、植物病原糸状菌の増殖や活動を効率的に抑制することができる。 According to the Rhodopseudomonas or Bacillus bacteria of the present invention, the growth and activity of phytopathogenic fungi can be efficiently suppressed.
本発明の土壌伝染性病害防除剤によれば、植物に対する病害性がなく、植物病原糸状菌を原因とする土壌伝染性病害に対して高い防除効果を示し、また環境に対する負荷が少なく、安全で薬害のない微生物農薬を提供することができる。 According to the soil infectious disease control agent of the present invention, there is no disease to plants, it exhibits a high control effect on soil infectious diseases caused by phytopathogenic fungi, and has a low environmental impact and is safe. A microbial pesticide free from phytotoxicity can be provided.
本発明の土壌伝染性病害防除方法によれば、本発明の土壌伝染性病害防除剤を用いて土壌伝染性病害を効果的に防除することができる。またそれによって、労力やコストを低減し、安全な農産物を安定的に提供することができる。 According to the soil infectious disease control method of the present invention, the soil infectious disease can be effectively controlled using the soil infectious disease control agent of the present invention. Moreover, it can reduce labor and cost, and can provide safe agricultural products stably.
1.土壌伝染性病害防除剤
1−1.概要と定義
本発明の第1の態様は、植物病原糸状菌に対する土壌伝染性病害防除剤である。本態様の土壌伝染性病害防除剤は、特定の細菌を有効成分として、その細菌の占有作用及び拮抗作用によって標的とする植物病原糸状菌の増殖や活動を抑制することができる。それ故、所望の植物に施用することで、その植物の植物病原糸状菌による病害を軽減することができる。
1. 1. Soil infectious disease control agent 1-1. Outline and Definition The first aspect of the present invention is a soil infectious disease control agent for phytopathogenic fungi. The soil infectious disease control agent of this aspect can suppress the growth and activity of phytopathogenic fungi targeted by specific bacteria as an active ingredient through the occupying action and antagonism of the bacteria. Therefore, the disease caused by the phytopathogenic fungus of the plant can be reduced by applying it to a desired plant.
本明細書において「土壌伝染性病害防除剤」とは、土壌伝染性病害の発病の予防及び防除を目的とする微生物農薬をいう。 As used herein, “soil-borne disease control agent” refers to a microbial pesticide for the purpose of preventing and controlling the occurrence of soil-borne disease.
「微生物農薬」とは、生存状態の微生物を有効成分として、その微生物の性質や活性等を利用し、主として植物に病害をもたらす標的生物の増殖抑制、活動抑制、生育阻害、又は致死を目的とした農薬をいう。本明細書において「微生物」とは、通常、肉眼での認識が困難な微小生物、例えば、細菌(バクテリア;真正細菌)や菌をいう。菌(Fungi)は、菌界に属する真核生物群であり、真菌又は糸状菌とも称される。菌は、酵母のような単細胞真核微生物、又は肉眼での認識が比較的困難な糸状菌(カビを含む)若しくはきのこのような多細胞真核微生物を含む。 “Microbial pesticides” are intended for the purpose of inhibiting the growth of target organisms, suppressing the activity, inhibiting growth, or killing the target organisms that cause disease to plants by using the properties and activities of the microorganisms as active ingredients. Refers to pesticides. In the present specification, the “microorganism” generally refers to a micro-organism that is difficult to recognize with the naked eye, such as bacteria (bacteria; eubacteria) or fungi. Fungi is a eukaryotic group belonging to the fungal kingdom, and is also called a fungus or a filamentous fungus. The fungus includes a unicellular eukaryotic microorganism such as yeast, or a multicellular eukaryotic microorganism such as a filamentous fungus (including mold) or a mushroom that is relatively difficult to recognize with the naked eye.
本明細書において「土壌伝染性病害」とは、土壌又は栽植培地(液体培地を含む)を介して伝播される植物病原糸状菌の感染によって植物に発症する病害をいう。例えば、紋羽病(紫紋羽病及び白紋羽病を含む)、根こぶ病、苗立枯病(イネ苗立枯病を含む)、萎凋病(トマト萎凋病及びナス半身萎凋病を含む)、根腐病、白絹病及びフィトソフラ疫病が挙げられる。紋羽病、特に紫紋羽病は、本態様の土壌伝染性病害防除剤による予防、防除の対象として好ましい。 As used herein, “soil infectious disease” refers to a disease that develops in a plant by infection with a phytopathogenic fungus transmitted through soil or a planting medium (including a liquid medium). For example, including coat rot (including purple coat rot and white coat rot), root-knot, seedling blight (including rice seed blight), wilt (including tomato wilt and eggplant half wilt) ), Root rot, white silkworm, and phytosofla plague. Hermaphroditis, particularly purple herb disease, is preferred as an object of prevention and control with the soil infectious disease control agent of this embodiment.
本明細書において「植物病原糸状菌」とは、本態様の土壌伝染性病害防除剤の標的となる微生物であって、生きた植物に感染し、前記土壌伝染性病害をもたらす糸状菌をいう。本態様の植物病原糸状菌としては、例えば、ヘリコバシディウム属(Helicobasidium)菌、ロゼリニア属(Rosellinia)菌、フザリウム属(Fusarium)菌、ピシウム属(Pythium)菌、ピレノカエタ属(Pyrenochaeta)菌、モノスポラスカス属(Monosporascus)菌、バーティシリウム属(Verticillium)菌、リゾクトニア属(Rhizoctonia)菌、プラスモディオフォラ属(Plasmodiophora)菌、フィトソフラ属(Phitophthora)菌、又はスクレロチウム属(Sclerotium)菌が挙げられる。中でもヘリコバシディウム属菌は、本態様の土壌伝染性病害防除剤の好適な標的となり得る。ヘリコバシディウム属菌は、背着性でフェルト状の子実体を宿主植物の株元などに形成し、その表面で、湾曲した円筒状の担子器と無色、卵形〜楕円形で基部が突出した形状を示す担子胞子を形成することが知られている。現在、分類学上の有効種としてH.モンパ(H. mompa)、H.ブレビッソニイ(H. brebissonii(若しくはH.パープレウム;H. purpureum))、及びH.ロンギスポルム(H. longisporum)が同定されており、いずれも本態様の土壌伝染性病害防除剤の標的となり得る。中でも、紫紋羽病の原因菌であるH.モンパは、特に好適である。 In the present specification, the “phytopathogenic fungus” refers to a microorganism that is a target of the soil infectious disease control agent of this embodiment and that infects a living plant and causes the soil infectious disease. Examples of the plant pathogenic filamentous fungi of this embodiment include, for example, Helicobasidium bacteria, Roselinia bacteria, Fusarium bacteria, Pythium bacteria, Pyrenochaeta bacteria, Monosporascus bacteria, Verticillium bacteria, Rhizoctonia bacteria, Plasmodiophora bacteria, Phitophthora bacteria, or Sclerotium bacteria Can be mentioned. Among them, Helicobacterium can be a suitable target for the soil infectious disease control agent of this embodiment. Helicobacterium genus forms a backed, felt-like fruiting body on the host plant stock, etc., and on its surface, a curved cylindrical basidiode and a colorless, oval to oval base It is known to form basidiospores having a protruding shape. Currently, H. mompa, H. brebissonii (or H. purpureum), and H. longisporum have been identified as taxonomic effective species. Both can be targets of the soil infectious disease control agent of this embodiment. Among these, H. mompa, which is a causative bacterium of purple coat feather disease, is particularly suitable.
1−2.構成
1−2−1.組成
本態様の土壌伝染性病害防除剤は、必須成分である有効成分と、選択成分である担体を含む。
A.有効成分
本態様の土壌伝染性病害防除剤は、少なくとも2種の異なる細菌、すなわち、光合成細菌とバチルス属細菌を有効成分として含む。以下、それぞれの細菌について具体的に説明をする。
1-2. Configuration 1-2-1. Composition The soil infectious disease control agent of this embodiment includes an active ingredient that is an essential ingredient and a carrier that is a selective ingredient.
A. Active ingredient The soil infectious disease control agent of this embodiment contains at least two different bacteria, that is, photosynthetic bacteria and Bacillus bacteria as active ingredients. Hereinafter, each bacterium will be specifically described.
(1)光合成細菌
光合成細菌とは、光合成を行う細菌で、紅色硫黄細菌、紅色非硫黄細菌、緑色硫黄細菌及び緑色非硫黄細菌が含まれる。光合成細菌は、本態様の土壌伝染性病害防除剤に必須の有効成分であり、その種類は、特に限定はしないが、好ましくは紅色非光合成細菌であり、特に好ましくはロドシュードモナス属(Rhodopseudomonas)細菌である。例えば、R.ファエカリス(R. faecalis)、R.パルステリス(R. palustris)、R.レノバセンシス(R. rhenobacensis)等が含まれる。好適なロドシュードモナス属細菌として、R. ファエカリスに近縁の Rhodopseudomonas A株(受託番号FERM BP-19432)及びRhodopseudomonas B株(受託番号FERM BP-19431)が挙げられる。中でも、Rhodopseudomonas A株(受託番号FERM BP-19432)は特に好ましい。Rhodopseudomonas A株の菌学的性質は、耐アルカリ性で、バクテリアクロロフィルa、ノイロスポレン、リオコピンを有する。Rhodopseudomonas B株の菌学的性質は、A株と同様に耐アルカリ性で、バクテリアクロロフィルa、ノイロスポレンを有する。本明細書において「耐アルカリ性」とは、主にpH 8〜10で最大増殖速度を持つ性質をいう。Rhodopseudomonas A株及びB株は、特許公報3699987号に詳述されており、また現在は、独立行政法人製品評価技術基盤機構(292-0818日本国千葉県木更津市かずさ鎌足2-5-8 120号室)に寄託されている。有効成分として2種以上の光合成細菌、例えば、Rhodopseudomonas A株及びRhodopseudomonas B株を含んでいてもよい。
(1) Photosynthesis bacteria Photosynthesis bacteria are bacteria which perform photosynthesis, and include red sulfur bacteria, red non-sulfur bacteria, green sulfur bacteria, and green non-sulfur bacteria. The photosynthetic bacterium is an essential active ingredient for the soil infectious disease control agent of the present embodiment, and the type thereof is not particularly limited, but is preferably a red non-photosynthetic bacterium, and particularly preferably a Rhodopseudomonas bacterium. It is. For example, R. faecalis, R. palustris, R. rhenobacensis and the like are included. Suitable Rhodopseudomonas bacteria include Rhodopseudomonas A strain (accession number FERM B P-19432) and Rhodopseudomonas B strain (accession number FERM B P-19431) closely related to R. faecalis. Among them, Rhodopseudomonas A strain (Accession No. FERM B P-19432) is particularly preferable. The mycological properties of Rhodopseudomonas A strain are alkali-resistant and have bacterial chlorophyll a, neurosporene and lycopene. The bacteriological properties of Rhodopseudomonas B strain are alkali-tolerant like strain A and have bacterial chlorophyll a and neurosporene. As used herein, “alkali resistance” refers to a property having a maximum growth rate mainly at pH 8-10. The Rhodopseudomonas A and B strains are described in detail in Japanese Patent Publication No. 3699987, and are currently incorporated by the National Institute of Technology and Evaluation (2-0-8 Kazusa Kamashi, Kisarazu City, Chiba Prefecture 292-0818) No. Room). As an active ingredient, two or more kinds of photosynthetic bacteria, for example, Rhodopseudomonas A strain and Rhodopseudomonas B strain may be contained.
(2)バチルス属細菌
バチルス属(Bacillus)細菌とは、芽胞を形成する偏性好気性又は通性好気性のグラム陽性桿菌をいう。例えば、B.エアロフィルス(B. aerophilus)、B.シアメンシス(B. siamensis)、B.サブチリス(B.subtilis)、B.モジャバンシス(B. mojavensis)、及びB.アミロリクエファシエンス(B. amyloliquefaciens)等が含まれる。
(2) Bacillus bacteria Bacillus bacteria refer to obligate aerobic or facultative aerobic Gram-positive rods that form spores. For example, B. aerophilus, B. siamensis, B. subtilis, B. mojavensis, and B. amyloliquefaciens ) Etc. are included.
バチルス属細菌は、光合成細菌と共に本態様の土壌伝染性病害防除剤に必須の有効成分である。バチルス属細菌の種類は、特に限定はしないが、B. エアロフィルス又は16S rRNA遺伝子分類においてB. エアロフィルスに近縁の種が好ましい。例えば、図1に示すBacillus AUT-8株(受託番号NITE BP-01678)又はBacillus AUT-9株(受託番号NITE BP-01679)は好適である。これらのバチルス属細菌株は、2013年8月2日付で、独立行政法人製品評価技術基盤機構(292-0818日本国千葉県木更津市かずさ鎌足2-5-8 120号室)に受領されている。有効成分として2種以上のバチルス属細菌、例えば、B. AUT-8及びB. AUT-9を含んでいてもよい。 A Bacillus bacterium is an active ingredient essential for the soil infectious disease control agent of this embodiment together with photosynthetic bacteria. The type of bacteria belonging to the genus Bacillus is not particularly limited, but a species closely related to B. aerophilus in the B. aerophilus or 16S rRNA gene classification is preferable. For example, the Bacillus AUT-8 strain (Accession Number NITE B P-01678) or Bacillus AUT-9 strain (Accession Number NITE B P-01679) shown in FIG. 1 is suitable. These bacteria belonging to the genus Bacillus were received by the National Institute of Technology and Evaluation (2-0-8, Kazusa Kamashi, Kisarazu-shi, Chiba 292-0818) on August 2, 2013. . As an active ingredient, two or more kinds of bacteria belonging to the genus Bacillus, for example, B. AUT-8 and B. AUT-9 may be contained.
(3)混合比率等
本態様の土壌伝染性病害防除剤は、上述したように、少なくとも光合成細菌及びバチルス属細菌を有効成分として、それらを生存状態で含有する。本態様の土壌伝染性病害防除剤における光合成細菌及びバチルス属細菌の混合比率は、両細菌が包含されていれば特に限定はしないが、好ましくは光合成細菌:バチルス属細菌が1:1〜10000:1である。
(3) Mixing ratio, etc. As described above, the soil infectious disease control agent of this aspect contains at least photosynthetic bacteria and Bacillus bacteria as active ingredients and contains them in a living state. The mixing ratio of the photosynthetic bacteria and Bacillus bacteria in the soil infectious disease control agent of this embodiment is not particularly limited as long as both bacteria are included, but preferably the photosynthetic bacteria: Bacillus bacteria are 1 : 1 to 10000: 1.
光合成細菌及びバチルス属細菌は、土壌伝染性病害防除剤の施用時に混合状態にあればよく、施用前、例えば、保存過程において両細菌を必ずしも混合状態にしていなくてもよい。また、本明細書において「生存状態」とは、細菌が代謝及び/又は分裂、増殖が可能な状態をいう。それ故、バチルス属細菌における芽胞も発芽力を有する限り生存状態に含まれる。ただし、土壌伝染性病害防除剤は、死亡した光合成細菌及び/又はバチルス属細菌を含んでいても構わない。 The photosynthetic bacterium and the Bacillus genus bacterium need only be in a mixed state at the time of application of the soil infectious disease control agent, and the two bacteria may not necessarily be in a mixed state before application, for example, during the preservation process. In this specification, the “viable state” refers to a state in which bacteria can metabolize and / or divide and proliferate. Therefore, spores in Bacillus bacteria are also included in the living state as long as they have germination power. However, the soil infectious disease control agent may contain dead photosynthetic bacteria and / or Bacillus bacteria.
B.担体
本態様の土壌伝染性病害防除剤は、有効成分である光合成細菌及びバチルス属細菌の生存及び生理活性を阻害又は抑制しない範囲において農薬製剤上許容可能な担体を含むことができる。
B. Carrier The soil infectious disease control agent of the present embodiment can contain a carrier that is acceptable in an agrochemical formulation as long as it does not inhibit or suppress the survival and physiological activity of photosynthetic bacteria and Bacillus bacteria which are active ingredients.
「農薬製剤上許容可能な担体」とは、土壌伝染性病害防除剤の施用を容易にし、光合成細菌及びバチルス属細菌の生存や植物病原糸状菌に対する拮抗作用又は抑制作用を維持する物質及び/又は土壌伝染性病害防除剤の作用速度を制御する物質であって、土壌若しくは水質等の環境、及び/又は動物、特にヒト、に対する有害性がないか若しくは低い物質をいう。本態様の土壌伝染性病害防除剤における担体としては、例えば、賦形剤が挙げられる。賦形剤は、粉砕天然鉱物、粉砕合成鉱物、乳化剤、分散剤及び界面活性剤等を含む。 “Agrochemically acceptable carrier” means a substance that facilitates the application of a soil-borne disease control agent, maintains the survival of photosynthetic bacteria and Bacillus bacteria, and antagonism or suppression of phytopathogenic fungi, and / or A substance that controls the action rate of a soil infectious disease control agent and that is not harmful or low in the environment such as soil or water quality and / or animals, particularly humans. Examples of the carrier in the soil infectious disease control agent of this embodiment include excipients. Excipients include ground natural minerals, ground synthetic minerals, emulsifiers, dispersants, surfactants, and the like.
粉砕天然鉱物には、例えば、カオリン、クレイ、タルク及びチョークが挙げられる。
粉砕合成鉱物には、例えば、高分散シリカ及びシリケートが挙げられる。
Examples of ground natural minerals include kaolin, clay, talc and chalk.
Examples of pulverized synthetic minerals include highly dispersed silica and silicate.
乳化剤には、非イオン性乳化剤やアニオン性乳化剤(例えば、ポリオキシエチレン脂肪アルコールエーテル、アルキルスルホネート及びアリールスルホネート)が挙げられる。
分散剤としては、例えば、リグノ亜硫酸廃液及びメチルセルロースが挙げられる。
Emulsifiers include nonionic emulsifiers and anionic emulsifiers (eg, polyoxyethylene fatty alcohol ethers, alkyl sulfonates and aryl sulfonates).
Examples of the dispersant include lignosulfite waste liquor and methylcellulose.
界面活性剤としては、例えば、リグノスルホン酸、ナフタレンスルホン酸、フェノールスルホン酸、ジブチルナフタレンスルホン酸のアルカリ金属塩、アルカリ土類金属塩及びアンモニウム塩、アルキルアリールスルホネート、アルキルスルフェート、アルキルスルホネート、脂肪アルコールスルフェート、脂肪酸及び硫酸化脂肪アルコールグリコールエーテル、さらに、スルホン化ナフタレン及びナフタレン誘導体とホルムアルデヒドの縮合物、ナフタレン又はナフタレンスルホン酸とフェノール及びホルムアルデヒドの縮合物、ポリオキシエチレンオクチルフェニルエーテル、エトキシル化イソオクチルフェノール、オクチルフェノール、ノニルフェノール、アルキルフェニルポリグリコールエーテル、トリブチルフェニルポリグリコールエーテル、トリステアリルフェニルポリグリコールエーテル、アルキルアリールポリエーテルアルコール、アルコール及び脂肪アルコール/エチレンオキシドの縮合物、エトキシル化ヒマシ油、ポリオキシエチレンアルキルエーテル、エトキシル化ポリオキシプロピレン、ラウリルアルコールポリグリコールエーテルアセタール、ソルビトールエステル、リグノ亜硫酸廃液、及びメチルセルロースが挙げられる。 Examples of the surfactant include lignosulfonic acid, naphthalenesulfonic acid, phenolsulfonic acid, dibutylnaphthalenesulfonic acid alkali metal salt, alkaline earth metal salt and ammonium salt, alkylaryl sulfonate, alkyl sulfate, alkyl sulfonate, fat Alcohol sulfates, fatty acids and sulfated fatty alcohol glycol ethers, sulfonated naphthalene and naphthalene derivatives and formaldehyde condensates, naphthalene or naphthalenesulfonic acid and phenol and formaldehyde condensates, polyoxyethylene octylphenyl ether, ethoxylated isoforms Octylphenol, octylphenol, nonylphenol, alkylphenyl polyglycol ether, tributylphenyl polyglycol Ether, tristearyl phenyl polyglycol ether, alkylaryl polyether alcohol, alcohol and fatty alcohol / ethylene oxide condensate, ethoxylated castor oil, polyoxyethylene alkyl ether, ethoxylated polyoxypropylene, lauryl alcohol polyglycol ether acetal, sorbitol Examples include esters, lignosulfite waste liquors, and methylcellulose.
本発明の土壌伝染性病害防除剤は、農薬製剤上許容可能な担体を1以上包含することが可能である。また、この他に、有効成分の細菌に影響しない範囲において、他の薬理作用を有する有効成分、すなわち、除草剤、殺菌剤、殺虫剤、肥料(例えば、尿素、硝酸アンモニウム、過リン酸塩)を包含することもできる。 The soil infectious disease control agent of the present invention can include one or more carriers that are acceptable in agricultural chemical formulations. In addition to this, active ingredients having other pharmacological actions, that is, herbicides, fungicides, insecticides, fertilizers (for example, urea, ammonium nitrate, and superphosphates), as long as the active ingredients do not affect bacteria. It can also be included.
1−2−2.剤形
本発明の土壌伝染性病害防除剤の剤形は、有効成分である光合成細菌及びバチルス属細菌を生存状態で保持し得ることができれば、特に限定はしない。例えば、各細菌を適当な溶液に懸濁した液体状態、固体状態又はその組み合わせとすることができる。液体状態の場合、光合成細菌及び/又はバチルス属細菌を適切な溶液に懸濁したものであればよい。適切な溶液としては、例えば、水(滅菌水、脱イオン水、超純水を含む)、生理食塩水、バッファー(リン酸緩衝液、炭酸緩衝液を含む)、その細菌の培地が挙げられる。固体状態の場合、例えば、顆粒状態、粉末状態、ゲルのような半固体状態が挙げられる。これらの具体例として、液剤、粉剤、粒剤、育苗培土剤等の剤形が含まれる。
1-2-2. Dosage Form The form of the soil infectious disease control agent of the present invention is not particularly limited as long as it can retain photosynthetic bacteria and Bacillus bacteria as active ingredients in a living state. For example, it can be in a liquid state, a solid state, or a combination thereof in which each bacterium is suspended in an appropriate solution. In the case of a liquid state, what is necessary is just to suspend a photosynthetic bacterium and / or a Bacillus bacterium in an appropriate solution. Suitable solutions include, for example, water (including sterilized water, deionized water, and ultrapure water), saline, buffers (including phosphate buffer and carbonate buffer), and bacterial cultures thereof. In the case of a solid state, for example, a granular state, a powder state, and a semi-solid state such as a gel can be mentioned. Specific examples of these include dosage forms such as liquids, powders, granules, and seedling culture media.
1−2−3.含有量
本発明の土壌伝染性病害防除剤の所定量あたりにおける光合成細菌及びバチルス属細菌の含有量は、各細菌の種類やその組み合わせ、施用対象植物の種類、剤形、及び施用方法等の諸条件によって異なる。通常は、本発明の土壌伝染性病害防除剤を施用する際に光合成細菌及びバチルス属細菌が植物病原糸状菌に対する拮抗作用又は抑制作用を発揮する上で十分な量を含んでいることが好ましい。この含有量は、当該分野の技術常識の範囲において光合成細菌及びバチルス属細菌が施用後に所定の体積あたりの土壌中に所望の存在量となるように各条件を勘案し、決定すればよい。一例として、本態様の土壌伝染性病害防除剤における光合成細菌及びバチルス属細菌の含有量は、両細菌の濃度として106〜1011cfu/gの範囲にあればよい。この場合、必要に応じて施用時に、水、生理食塩水、バッファー等で10〜1000倍に希釈することもできる。
1-2-3. Content The content of photosynthetic bacteria and Bacillus bacteria per predetermined amount of the soil infectious disease control agent of the present invention includes various types of bacteria and combinations thereof, types of plants to be applied, dosage forms, application methods, and the like. Varies depending on conditions. Usually, when applying the soil infectious disease control agent of the present invention, it is preferable that the photosynthetic bacteria and Bacillus bacteria contain a sufficient amount to exert an antagonistic action or inhibitory action against phytopathogenic fungi. This content may be determined in consideration of each condition so that photosynthetic bacteria and Bacillus bacteria can be in a desired amount in the soil per predetermined volume after application within the scope of common general technical knowledge in the field. As an example, the content of photosynthetic bacteria and Bacillus bacteria in the soil infectious disease control agent of this embodiment may be in the range of 10 6 to 10 11 cfu / g as the concentration of both bacteria. In this case, it can be diluted 10 to 1000 times with water, physiological saline, buffer, or the like, as needed, at the time of application.
1−3.効果
本態様の土壌伝染性病害防除剤によれば、光合成細菌(特にロドシュードモナス属細菌)及びバチルス属細菌を有効成分として、植物病原糸状菌の増殖や活動を効率的に抑制することができる。
1-3. Effect According to the soil infectious disease control agent of this embodiment, it is possible to efficiently suppress the growth and activity of phytopathogenic fungi using photosynthetic bacteria (particularly Rhodopseudomonas bacteria) and Bacillus bacteria as active ingredients.
本発明の土壌伝染性病害防除剤は、植物に対する毒性や病原性がなく、植物病原糸状菌を原因とする土壌伝染性病害に対して高い防除効果を有する。また、自然界に存在する細菌を有効成分とする微生物農薬であることから安全性が高く、化学農薬と比較して環境に対する影響が小さい。土壌施用しても人畜への危険性や、環境汚染や農作物への残留等の問題もない。それ故、消費者に安全で薬害のない農作物を提供することができる。また、化学農薬と比較して標的である植物病原糸状菌の耐性菌出現率が非常に低い。 The soil infectious disease control agent of the present invention is not toxic or pathogenic to plants, and has a high control effect against soil infectious diseases caused by phytopathogenic fungi. In addition, since it is a microbial pesticide containing bacteria that exist in nature as an active ingredient, it is highly safe and has less impact on the environment than chemical pesticides. Even if it is applied to the soil, there is no danger to human livestock, no problems such as environmental pollution and residue on agricultural products. Therefore, it is possible to provide consumers with safe and harmless crops. Moreover, the resistant bacteria appearance rate of the phytopathogenic filamentous fungus which is a target is very low compared with a chemical pesticide.
また、本発明の土壌伝染性病害防除剤は、植物活性効果も併せ持つ。
本発明の土壌伝染性病害防除剤によれば、土壌中の微生物活性(FDA加水分解活性)が向上し、微生物密度が高まることから、元来土壌中に棲息する有益な土壌微生物が活性化され、その微生物コミュニティー(細菌叢)が植物病原糸状菌に対して抑制的に働き、植物における土壌伝染性病害の発病を抑える環境ができると考えられる。
Moreover, the soil infectious disease control agent of this invention also has a plant activity effect.
According to the soil infectious disease control agent of the present invention, microbial activity (FDA hydrolysis activity) in soil is improved and microbial density is increased, so that beneficial soil microorganisms originally inhabiting in soil are activated. It is considered that the microbial community (bacteria flora) works against phytopathogenic fungi and suppresses the occurrence of soil-borne diseases in plants.
2.土壌伝染性病害防除方法
2−1.概要と定義
本発明の第2の態様は、土壌伝染性病害防除方法である。本態様の防除方法は、第1態様に記載の土壌伝染性病害防除剤を土壌又は培地に施用して、対象植物における土壌伝染性病害の発生を予防又は防除することを特徴とする。
2. 2. Soil-borne disease control method 2-1. Outline and Definition The second aspect of the present invention is a soil infectious disease control method. The control method according to this aspect is characterized in that the soil infectious disease control agent according to the first aspect is applied to soil or a medium to prevent or control the occurrence of soil infectious diseases in the target plant.
本明細書において「対象植物」とは、土壌伝染性病害防除剤の施用対象となる植物である。対象植物は、植物病原糸状菌の感染によって土壌伝染性病害を発病し得る植物であれば、その種類は問わない。被子植物又は裸子植物のいずれであってもよい。被子植物の場合、双子葉植物又は単子葉植物を含む。単子葉類植物では、例えば、イネ科(Poaceae)植物が挙げられる。また、双子葉類植物であれば、ヒルガオ科(Convolvulaceae)、バラ科(Rosaceae)植物、セリ科(Apiaceae)、ナス科(Solanaceae)植物、ユリ科(Liliaceae)、マメ科(Fabaceae)植物、ウリ科(Cucurbitaceae)植物、アブラナ科(Brassicaceae)植物等が挙げられる。 In the present specification, the “target plant” is a plant to be applied with a soil infectious disease control agent. The target plant is not limited as long as it is a plant that can cause soil-borne diseases by infection with phytopathogenic fungi. Either angiosperms or gymnosperms may be used. In the case of angiosperms, dicotyledonous plants or monocotyledonous plants are included. Monocotyledonous plants include, for example, Poaceae plants. For dicotyledonous plants, Convolvulaceae, Rosaceae, Apiaceae, Solanaceae, Liliaceae, Fabaceae, cucumber, and cucumber Examples include Cucurbitaceae plants and Brassicaceae plants.
具体的な対象植物は、土壌伝染性病害を引き起こす植物病原糸状菌の種類によって変わり得るが、例えば、植物病原糸状菌が紋羽病、特に紫紋羽病であれば、サツマイモ(Ipomoea batatas)、ニンジン(Daucus carota subsp. sativus)、ジャガイモ(Solanum tuberosum)、アスパラガス(Asparagus spp)、リンゴ(Malus pumila)、ナシ(Pyrus pyrifolia var. culta、P. bretschneideri、P. communis)等が挙げられる。 The specific target plant may vary depending on the type of phytopathogenic fungus causing the soil infectious disease. For example, if the phytopathogenic fungus is a coat rot, particularly purple potato, sweet potato (Ipomoea batatas), Examples include carrot (Daucus carota subsp. Sativus), potato (Solanum tuberosum), asparagus (Asparagus spp), apple (Malus pumila), pear (Pyrus pyrifolia var. Culta, P. bretschneideri, P. communis).
2−2.方法
2−2−1.施用工程
本態様の土壌伝染性病害防除方法は、施用工程を含む。
本明細書において「施用工程」とは、第1態様の土壌伝染性病害防除剤を土壌又は培地に施用する工程をいう。ここでいう「土壌」は、対象植物の生育が可能な土壌であれば特に制限はしない。通常は、適当な養分(窒素、リン、カリウム等)を含み、適切なpH値を有する対象植物の栽植用土壌が利用される。土壌の場所は、問わない。播種又は苗の植え付け前の本圃土壌又は育苗土壌のいずれであってもよい。また、「培地」は、人工的に調製した対象植物の栽植用培地をいう。寒天培地のような固体培地であってもよいし、液体培地であってもよい。培地の例として、例えば、隔離ベッド、根域制限ポット又は苗床が挙げられる。培地の組成は、当該分野で公知の培地組成でよい。適用作物の種類等によって適宜選択することができる。
2-2. Method 2-2-1. Application process The soil infectious disease control method of this aspect includes an application process.
In this specification, "application process" means the process of applying the soil infectious disease control agent of a 1st aspect to soil or a culture medium. The “soil” here is not particularly limited as long as the target plant can grow. Usually, soil for planting target plants containing appropriate nutrients (nitrogen, phosphorus, potassium, etc.) and having an appropriate pH value is used. The place of soil is not ask | required. Either the main soil or seedling soil before sowing or seedling planting may be used. In addition, “medium” refers to an artificially prepared medium for planting a target plant. It may be a solid medium such as an agar medium or a liquid medium. Examples of the culture medium include, for example, an isolation bed, a root zone restriction pot, or a nursery. The medium composition may be a medium composition known in the art. It can be selected as appropriate depending on the type of crop to be applied.
土壌伝染性病害防除剤の施用方法は、土壌伝染性病害の種類や適用作物の種類等によって適宜選択することができる。例えば、本発明の土壌伝染性病害防除剤を、対象植物の栽植用土壌、好ましくは対象植物の根群域の相当する部分(例えば畝部)、又は土壌伝染性病害の発病程度の高い部分に混和、散布、又は潅注などにより施用すればよい。 The application method of the soil infectious disease control agent can be appropriately selected according to the type of soil infectious disease, the type of applied crop, and the like. For example, the soil infectious disease control agent of the present invention is applied to soil for planting the target plant, preferably a portion corresponding to the root group of the target plant (for example, a heel), or a portion having a high degree of occurrence of a soil infectious disease. What is necessary is just to apply by mixing, spraying, or irrigation.
なお、必要に応じて、堆厩肥等を土壌伝染性病害防除剤と併用してもよい。また補足すべき肥料成分があれば各種成分の化成肥料や土壌改良剤および有機質肥料等との併用も可能である。 In addition, you may use compost manure etc. together with a soil infectious disease control agent as needed. In addition, if there are fertilizer components to be supplemented, they can be used in combination with various components of chemical fertilizers, soil conditioners, organic fertilizers, and the like.
2−2−2.施用時期
本態様の土壌伝染性病害防除方法において、第1態様の土壌伝染性病害防除剤の施用時期は、限定はしないが、対象作物の植え付け前、例えば、播種前又は定植前が好ましい。土壌伝染性病害防除剤を植え付け前に予め施用しておくことで、有効成分である光合成細菌及びバチルス属細菌が土壌で増殖し、占有作用によって植物病原糸状菌の侵入を防ぎ、また既に存在する植物病原糸状菌を拮抗作用によって排除することで、対象作物の植物病原糸状菌による感染率を低減し、土壌伝染性病害の発病を効率的に予防又は防除できるからである。施用した土壌に対象植物を植え付ける時期は、土壌伝染性病害防除剤の施用後2日後〜3週間後、好ましくは10日後〜2週間後である。
2-2-2. Application time In the soil infectious disease control method of this embodiment, the application time of the soil infectious disease control agent of the first embodiment is not limited, but is preferably before planting the target crop, for example, before sowing or before planting. By applying a soil infectious disease control agent in advance before planting, photosynthetic bacteria and Bacillus bacteria, which are active ingredients, grow in the soil, prevent the invasion of phytopathogenic fungi by occupancy, and already exist This is because by eliminating phytopathogenic fungi by antagonism, the infection rate of the target crops by phytopathogenic fungi can be reduced, and the occurrence of soil-borne diseases can be efficiently prevented or controlled. The timing for planting the target plant in the applied soil is 2 days to 3 weeks after application of the soil infectious disease control agent, preferably 10 days to 2 weeks later.
2−2−3.施用量
本態様の土壌伝染性病害防除方法において、第1態様の土壌伝染性病害防除剤の施用量は、剤形、施用方法や施用時期、対象植物の種類等によって異なることから、条件によって適宜調整すればよい。一例として、土壌伝染性病害防除剤が液剤の場合には、土壌10アールあたり、0.5〜20トン、好ましくは1〜10トンの施用量で使用すればよい。
2-2-3. Application rate In the soil infectious disease control method of this aspect, the application rate of the soil infectious disease control agent of the first aspect varies depending on the dosage form, application method, application time, type of target plant, etc. Adjust it. As an example, when the soil infectious disease control agent is a liquid agent, it may be used at an application rate of 0.5 to 20 tons, preferably 1 to 10 tons per 10 ares of soil.
2−2−4.施用回数
本態様の土壌伝染性病害防除方法において、第1態様の土壌伝染性病害防除剤の施用回数は、制限はしない。対象植物の植え付け前に施用した場合には、原則として1回で足りる。しかし、対象植物の植え付け前後に関わらず、複数回施用することで土壌伝染性病害防除効果は増加する。したがって、土壌病害抑止土壌を形成させるには、土壌伝染性病害防除剤の施用回数を増やすことが好ましい。また、光合成細菌は、有機肥料としての効果が知られている(北村 博、1984、光合成細菌、347-349)。したがって、通常の堆厩肥と組み合わせることで、土壌の微生物相を一層向上させることが可能となる。その結果、植物病原糸状菌による土壌伝染性病害の防除効果がさらに向上する。その点からも、土壌伝染性病害防除剤の施用回数は多い方が好ましい。
2-2-4. Number of applications In the soil infectious disease control method of this embodiment, the number of applications of the soil infectious disease control agent of the first embodiment is not limited. If applied before planting the target plant, in principle it is sufficient. However, the soil infectious disease control effect increases by applying multiple times regardless of before and after planting of the target plant. Therefore, in order to form soil disease control soil, it is preferable to increase the application frequency of a soil infectious disease control agent. Photosynthetic bacteria are known to be effective as organic fertilizers (Hiro Kitamura, 1984, photosynthetic bacteria, 347-349). Therefore, it becomes possible to improve the microflora of soil further by combining with normal manure. As a result, the effect of controlling soil-borne diseases caused by phytopathogenic fungi is further improved. From this point, it is preferable that the number of application times of the soil infectious disease control agent is larger.
2−3.効果
本態様の土壌伝染性病害防除方法によれば、所望の植物を植え付ける土壌等に土壌伝染性病害防除剤を施用することによって土壌伝染性病害を効果的に防除することができる。またそれによって、労力やコストを低減し、消費者には安全な農産物を安定的に提供することができる。
2-3. Effect According to the soil infectious disease control method of this embodiment, the soil infectious disease can be effectively controlled by applying the soil infectious disease control agent to the soil or the like in which a desired plant is planted. Moreover, it can reduce labor and cost, and can stably provide safe agricultural products to consumers.
以下、実施例によって本発明を具体的に説明するが、これらの実施例は本発明の範囲を何ら限定するものではない。
<土壌伝染性病害防除剤による植物病原糸状菌の拮抗性試験>
(材料及び方法)
1.使用微生物
植物病原糸状菌として紫紋羽病菌を用いた。紫紋羽病菌は、鹿児島県鹿屋市有明町の紫紋羽病に感染したサツマイモ畑の土壌から採取した。具体的には、前記サツマイモ畑の土1gを水1mLに希釈して、その懸濁液をオートミール培地(オートミール7.5%(w/v))に塗布する希釈平板法を用いて、25℃にて分離した。
EXAMPLES Hereinafter, the present invention will be specifically described with reference to examples, but these examples do not limit the scope of the present invention.
<Competitiveness test of phytopathogenic fungi with soil-borne disease control agent>
(Materials and methods)
1. Microorganisms used As a plant pathogenic filamentous fungus, purple crest fungus was used. The purple wing fungus was collected from the soil of a sweet potato field infected with purple wing in Ariake-cho, Kanoya City, Kagoshima Prefecture. Specifically, 1 g of the soil of the sweet potato field was diluted to 1 mL of water, and the suspension was applied to an oatmeal medium (oatmeal 7.5% (w / v)) at 25 ° C. separated.
土壌伝染性病害防除剤の一方の有効成分であるロドシュードモナス属細菌には、特許公報3699987号に記載の受託番号FERM BP-19432で示されるロドシュードモナス A株を用いた。ロドシュードモナス A株は、特許公報3699987号に記載の培養条件、すなわち、基本培地1:0.03%(w/v) KH2PO4, 0.03%(w/v)K2HPO4, 0.1%(w/v)NH4Cl, 0.02%(w/v)MgCl2・6H2O, 0.02%(w/v) NaCl, 0.005%(w/v) CaCl2・2H2O, 0.005%(w/v) Yeast extract, 0.005%(w/v) Na2S2O3・5H2O, 0.01%(w/v) D.L.Malic acid, 0.01%(w/v) Sodium Acetate, 0.1% Growth factor Solution(0.0001%(w/v) Vitamine B2, 0.0001%(w/v) Pyridoxine Hydrochloride, 0.0003%(w/v) p-Aminobenzoic Acid, 0.0005%(w/v) D-Biotin), 0.1% Trace Element Solution(2%(w/v) EDTA-2Na, 2%(w/v) FeSO4・7H2O, 0.1%(w/v) H3BO3, 0.1%(w/v) ZnCl2, 0.1%(w/v) MnCl・4H2O) (pH7.8)で25℃にて培養した。 The Rhodopseudomonas bacterium is one of the active ingredients of the soil-borne disease control agent was used Rhodopseudomonas A strain represented by accession number FERM B P-19432 described in Japanese Patent Publication 3,699,987. Rhodopseudomonas strain A is cultured under the conditions described in Japanese Patent Publication No. 3699987, ie, basic medium 1: 0.03% (w / v) KH 2 PO 4 , 0.03% (w / v) K 2 HPO 4 , 0.1% (w / v) NH 4 Cl, 0.02% (w / v) MgCl 2 · 6H 2 O, 0.02% (w / v) NaCl, 0.005% (w / v) CaCl 2 · 2H 2 O, 0.005% (w / v) Yeast extract, 0.005% (w / v) Na 2 S 2 O 3・ 5H 2 O, 0.01% (w / v) DLMalic acid, 0.01% (w / v) Sodium Acetate, 0.1% Growth factor Solution ( 0.0001% (w / v) Vitamine B2, 0.0001% (w / v) Pyridoxine Hydrochloride, 0.0003% (w / v) p-Aminobenzoic Acid, 0.0005% (w / v) D-Biotin), 0.1% Trace Element Solution (2% (w / v) EDTA-2Na, 2% (w / v) FeSO 4・ 7H 2 O, 0.1% (w / v) H 3 BO 3 , 0.1% (w / v) ZnCl 2 , 0.1 % (W / v) MnCl · 4H 2 O) (pH 7.8) at 25 ° C.
土壌伝染性病害防除剤の他方の有効成分であるバチルス属細菌株には、鹿児島県鹿屋市有明町のサツマイモ畑の土壌より分離したバチルス属細菌株及び鹿児島大学が保有するバチルス属細菌株(コード株名AUT-5、-7、-8、-9、-10、及び-11)を用いた。これらのバチルス属細菌株は、16S rRNA遺伝子分類によれば、いずれもバチルス エアロフィルスと同種又は極めて近縁な種である。本実施例で用いたバチルス属細菌株は、いずれもNB液体培地(1%(w/v) Polypepton, 0.5 %(w/v) Meat Extract, 0.2 %(w/v) NaCl, pH7.0)で25℃にて培養した。 The other active ingredient of the soil infectious disease control agent, Bacillus spp., Is Bacillus spp. Isolated from the soil of sweet potato fields in Ariake-cho, Kanoya City, Kagoshima Prefecture, and the Bacillus spp. Strain names AUT-5, -7, -8, -9, -10, and -11) were used. According to the 16S rRNA gene classification, these Bacillus genus bacterial strains are all the same species or very closely related to Bacillus aerophilus. All the Bacillus strains used in this example were NB liquid medium (1% (w / v) Polypepton, 0.5% (w / v) Meat Extract, 0.2% (w / v) NaCl, pH 7.0) Incubated at 25 ° C.
紫紋羽病菌、ロドシュードモナス A株、及び各バチルス属細菌株を拮抗性試験の2週間前に上記条件でそれぞれ培養した。 Purple rot fungus, Rhodopseudomonas A strain, and each Bacillus genus bacterial strain were cultured under the above conditions two weeks before the antagonistic test.
続いて、ロドシュードモナス A株及び各バチルス属細菌株の培養液を10μL採取して混合した混合液をディッシュ内の前記オートミール培地上に配置した直径約6mmのろ紙プレートに共接種した。コントロールとして、それぞれ一方の菌株のみを前記オートミール培地上に配置した同サイズのろ紙プレートに接種した。各紙プレートの上に紫紋羽菌の菌糸をくり抜いて得た直径約5mmの寒天片を置き、25℃の暗所下で培養した。培養5日後、7日後、及び10日後に各ディッシュを撮影し、紫紋羽菌の菌糸の成長を検証した。 Subsequently, 10 μL of a culture solution of Rhodopseudomonas strain A and each Bacillus genus bacterial strain was collected and mixed, and co-inoculated on a filter paper plate having a diameter of about 6 mm placed on the oatmeal medium in the dish. As a control, only one strain was inoculated on the same size filter paper plate placed on the oatmeal medium. On each paper plate, an agar piece having a diameter of about 5 mm obtained by hollowing out the mycelium of purple crest fungus was placed and cultured in the dark at 25 ° C. Each dish was photographed after 5 days, 7 days, and 10 days of culture to verify the growth of mycelium of purple clover.
(結果)
表1及び図2に拮抗性試験の結果を示す。
(result)
Table 1 and FIG. 2 show the results of the antagonistic test.
図2には、処理番号1(図2A,B)、4(図2D)、5(図2F)、10(図2C)、及び11(図2E)のみを示している。図2において紫紋羽病菌が増殖したディッシュ(図2B、D、F)は、伸長した赤紫の菌糸により、暗い色を呈している。つまり、この結果は、ロドシュードモナス A株単独又はバチルス細菌株単独では、紫紋羽病菌の増殖や活性を十分に抑制できないことを示している。一方、ロドシュードモナス A株、及びバチルス属細菌株のうちAUT-8又はAUT-9を混合して組み合わせた場合(図2C、E)には、紫紋羽病菌がほとんど増殖できず、培養0日目(図2A)とほぼ同様の色を呈していることが示された。 FIG. 2 shows only process numbers 1 (FIGS. 2A and 2B), 4 (FIG. 2D), 5 (FIG. 2F), 10 (FIG. 2C), and 11 (FIG. 2E). In FIG. 2, the dish (FIGS. 2B, D, and F) in which purple coat fungus has grown has a dark color due to the elongated red-purple hyphae. In other words, this result indicates that Rhodopseudomonas strain A alone or Bacillus bacteria strain alone cannot sufficiently suppress the growth and activity of purple rot fungus. On the other hand, when AUT-8 or AUT-9 was mixed and combined among Rhodopseudomonas strain A and Bacillus genus strains (Fig. 2C, E), purple herring fungus was hardly able to grow and cultured for 0 days. It was shown that it had almost the same color as the eyes (FIG. 2A).
この結果から、ロドシュードモナス属細菌とバチルス属細菌の少なくとも2種類の細菌を有効成分として含むことにより、植物病原糸状菌の増殖や活性を抑制し、対象植物の土壌伝染性病害を予防又は防除できることが明らかとなった。 From this result, by containing at least two kinds of bacteria, Rhodopseudomonas bacteria and Bacillus bacteria, as active ingredients, it is possible to suppress the growth and activity of phytopathogenic fungi and prevent or control soil-borne diseases of the target plant Became clear.
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