JPH09194314A - Agent for controlling soil blight of plant of family solanaceae and controlling method - Google Patents
Agent for controlling soil blight of plant of family solanaceae and controlling methodInfo
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- JPH09194314A JPH09194314A JP8006693A JP669396A JPH09194314A JP H09194314 A JPH09194314 A JP H09194314A JP 8006693 A JP8006693 A JP 8006693A JP 669396 A JP669396 A JP 669396A JP H09194314 A JPH09194314 A JP H09194314A
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- soil
- disease
- tobacco
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Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、学名オクロバクト
ラム・アンスロピ（旧名：アクロモバクタ− Group V
d）に属する微生物を有効成分として含有するナス科植
物の土壌病害防除剤（特に、シュ−ドモナス・ソラナセ
アラムが原因となるタバコ立枯病及びナス科青枯病）並
びに該微生物を用いたナス科植物の土壌病害防除方法に
関する。TECHNICAL FIELD The present invention relates to the scientific name Ochrobactrum anthropi (former name: Achromobactor Group V).
Soil disease control agent for Solanaceae plants containing the microorganism belonging to d) as an active ingredient (particularly, tobacco wilt disease and Solanaceae wilt disease caused by Pseudomonas solanacearum) and Solanaceae using the microorganism The present invention relates to a method for controlling soil diseases of plants.
【０００２】[0002]
【従来の技術】植物病原細菌の１種であるシュ−ドモナ
ス・ソラナセアラム（Pseudomonas solanacearum）の寄
生によって起こるタバコ立枯病あるいはナス科植物青枯
病（以下本病と略す）は、タバコ、トマト、ナス、ピ−
マン等多くの作物で被害が多い。本病原細菌（以下立枯
病菌と略す）は、土壌中で長期間生存しやすく、いった
ん植物体に感染すると増殖が速いので、防除が極めて困
難であることから、作物病害の中でも難防除病害の一つ
とされている。2. Description of the Related Art Tobacco wilt caused by parasitism of Pseudomonas solanacearum, which is one of plant pathogenic bacteria, or bacterial wilt of Solanaceae (hereinafter abbreviated as "this disease") is produced in tobacco, tomato, Eggplant, pee
Many crops such as mankind are damaged. This pathogenic bacterium (hereinafter abbreviated to bacterial wilt) is easy to survive in the soil for a long period of time and grows rapidly once it infects the plant body. Therefore, it is extremely difficult to control. It is supposed to be one.
【０００３】現在用いられている本病の防除対策は、耕
種的防除方法として抵抗性品種の利用、有機物の施用、
土壌の耕うんなどがあげられているが、安定した防除効
果を示さないことが多い。また、土壌くん蒸剤、例えば
クロルピクリンや臭化メチルなどの薬剤が化学的防除方
法として用いられている。しかしながら、近年、これら
の薬剤の使用は、刺激臭による公害の発生、環境汚染お
よびオゾン層の破壊の原因となることや、土壌中の病原
菌だけでなく有用な微生物までも死滅させることから、
より安全で効果の高い防除方法が求められてきた。[0003] Currently used control measures against this disease include the use of resistant varieties, application of organic substances,
Tillage of soil is mentioned, but it often does not show a stable control effect. In addition, soil fumigants such as chloropicrin and methyl bromide are used as chemical control methods. However, in recent years, the use of these agents causes pollution due to irritating odor, causes environmental pollution and destroys the ozone layer, and kills not only pathogenic bacteria in the soil but also useful microorganisms,
A safer and more effective control method has been demanded.
【０００４】一方、自然の土壌中には、多くの微生物が
存在し、お互いに影響を及ぼし合いながら生態系を形成
している。これらの微生物の中には、病原菌に対して拮
抗作用を示す微生物が多数存在することが明らかになっ
ており、安全で効果的な防除方法を提供するために、こ
れらの拮抗性の土壌微生物を用いて本病を防除する試み
が広く行われている。その例を示すと、立枯病に対して
は、シュ−ドモナス・プチ−ダ（Pseudomonas putida）
を用いる方法（日本植物病理学会報 (1990) 56巻：40
4）、シュ−ドモナス・フルオレセンス（Pseudomonas f
luorescens）を用いる方法（Revista de Microbiologia
(1989) Vol.20:18-26）、弱病原性のシュ−ドモナス・
ソラナセアラム バクテリオシン産生菌株（Pseudomona
s solanacearum）を用いる方法（特開平1-16579号公
報）等があげられる。On the other hand, many microorganisms exist in natural soil and form an ecosystem by influencing each other. It has been revealed that many of these microorganisms have antagonistic effects against pathogenic bacteria, and in order to provide a safe and effective control method, these antagonistic soil microorganisms are used. Attempts to control this disease by using it have been widely made. As an example, Pseudomonas putida can be used for the wilt disease.
Method (Japanese Journal of Plant Pathology (1990) 56:40
4), Pseudomonas f
luorescens) method (Revista de Microbiologia
(1989) Vol.20: 18-26), weakly pathogenic Pseudomonas
Solanacearum bacteriocin producing strain (Pseudomona
s solanacearum) (Japanese Patent Application Laid-Open No. 1-16579).
【０００５】[0005]
【発明が解決しようとする課題】上記のシュ−ドモナス
・プチ−ダあるいはシュ−ドモナス・フルオレセンスを
用いた例では、これらの拮抗細菌が立枯病菌に対して培
地上で抗菌活性を示し、温室内の短期実験で発病抑制効
果が認められている。しかし、汚染畑では防除効果が認
められなかったり、栽培後期に防除効果が著しく低下す
る例がほとんどで、栽培期間の長いナス科植物に使用し
て後期まで満足する防除効果を示すものは、今のところ
認められていない。In the examples using Pseudomonas putida or Pseudomonas fluorescens described above, these antagonistic bacteria show antibacterial activity on the medium against wilt disease bacteria. , A short-term experiment in a greenhouse has been shown to have a disease suppressing effect. However, in most cases, the control effect is not observed in contaminated fields, or the control effect remarkably decreases in the latter period of cultivation, and the one that shows satisfactory control effect until the latter period when used for solanaceous plants with a long cultivation period is now However, it is not recognized.
【０００６】また、弱病原性の立枯病菌でバクテリオシ
ン産生菌であるＯＭ２菌株を用いた例では、処理した菌
株の根部への定着に18℃以上の温度条件が必要なことが
明らかになった（日本植物病理学会報 (1989)55:51
1）。一般のタバコ畑の移植時は３月にあたり、この温
度条件を満たせないことから、処理した菌が減少しやす
く、そのために本病の防除効果が低いあるいは不安定に
なることが認められた。以上のように、現時点では、タ
バコ根に栽培後期まで安定して定着し、本病に対して安
定して高い防除効果を示す実用性のある微生物は見い出
されていない。本発明は、従来の上記欠点を克服し、環
境や生態系に支障を及ぼすことが無く、ナス科植物の土
壌病害（特に、タバコ立枯病及びナス科植物青枯病）に
対して、その栽培後期まで安定して高い防除効果を示す
ナス科植物の土壌病害防除剤及びその防除方法を提供す
るためになされたものである。In addition, in the case of using the OM2 strain, which is a weakly pathogenic wilt bacterium and a bacteriocin-producing bacterium, it became clear that a temperature condition of 18 ° C. or higher is required for the roots of the treated strain to settle. (Japanese Journal of Plant Pathology (1989) 55:51
1). It was confirmed that the temperature was not satisfied at the time of transplantation of a general tobacco field in March, and thus the treated bacteria were likely to be reduced, and thus the control effect of the disease was low or unstable. As described above, at the present time, no practical microorganisms have been found that are stably established in tobacco roots until the late stage of cultivation and stably show a high control effect against this disease. The present invention overcomes the above-mentioned drawbacks of the related art, does not impair the environment and the ecosystem, and prevents soil diseases of Solanaceae plants (particularly tobacco wilt disease and Solanaceae wilt disease). The present invention has been made to provide a soil disease controlling agent for Solanaceae plants which shows a stable and high controlling effect until the latter stage of cultivation, and a controlling method thereof.
【０００７】[0007]
【課題を解決するための手段】本発明者らは、さらに研
究を進め、従来までの研究に使用されている細菌とは異
なり、本病防除に有効な微生物を分離した。すなわち、
タバコ根部から分離したオクロバクトラム・アンスロピ
に属する微生物が、植物体に悪影響を及ぼさず、タバコ
栽培後期までタバコ根部に安定して定着し、温室、野外
いずれの実験においても本病防除効果が高いことを発見
し、この知見に基づいて、本発明を成すに至った。即
ち、本発明の構成を下記に示す。[Means for Solving the Problems] The present inventors have further advanced the research and isolated a microorganism effective for controlling the present disease, unlike the bacteria used in the previous studies. That is,
Microorganisms belonging to Ochrobactrum anthropi isolated from the tobacco root did not adversely affect the plant body, stably settled on the tobacco root until the late stage of tobacco cultivation, and it is highly effective in controlling this disease in both greenhouse and field experiments. The present invention was discovered and based on this finding, the present invention was accomplished. That is, the constitution of the present invention is shown below.
【０００８】（１） ナス科植物の土壌病害を防除する
性質を有するオクロバクトラム・アンスロピ（Ochrobac
trum anthropi）に属する微生物及び／又はその培養物
を有効成分として含有することを特徴とするナス科植物
の土壌病害防除剤。 （２） オクロバクトラム・アンスロピ（Ochrobactrum
anthropi）に属する微生物が、オクロバクトラム・ア
ンスロピＴＲＢ１９菌株である前記（１）に記載のナス
科植物の土壌病害防除剤。 （３） 土壌病害が、シュードモナス・ソラナセアラム
（Pseudomonas solanacearum）が原因で起こるタバコ立
枯病及びナス科植物青枯病である前記（１）又は（２）
に記載のナス科植物の土壌病害防除剤。(1) Ochrobactorum anthropi (Ochrobac) having the property of controlling soil diseases of Solanaceae plants
A soil disease controlling agent for Solanaceae plants, which comprises a microorganism belonging to trum anthropi) and / or a culture thereof as an active ingredient. (2) Ochrobactrum
The agent for controlling soil diseases of solanaceous plants according to (1), wherein the microorganism belonging to anthropi) is Ochrobactrum anthropi TRB19 strain. (3) The soil disease is tobacco wilt caused by Pseudomonas solanacearum or wilt disease of solanaceae (1) or (2)
2. A soil disease control agent for Solanaceae plants according to.
【０００９】（４） 前記（１）〜（３）のいずれか１
つに記載のナス科植物の土壌病害防除剤を、ナス科植物
の根部、栽培地及び／又はその土壌に導入する工程を含
むことを特徴とするナス科植物の土壌病害防除方法。 （５） 土壌病害が、シュードモナス・ソラナセアラム
（Pseudomonas solanacearum）が原因で起こるタバコ立
枯病及びナス科植物青枯病である前記（４）に記載のナ
ス科植物の土壌病害防除方法。(4) Any one of the above (1) to (3)
5. A method for controlling soil diseases of Solanaceae plants, which comprises the step of introducing the soil disease controlling agent for Solanaceae plants described in 1) into the roots, cultivated areas and / or soil of the Solanaceae plants. (5) The method for controlling soil diseases of Solanaceae plants according to (4), wherein the soil diseases are bacterial wilt of tobacco and Solanaceae wilt caused by Pseudomonas solanacearum.
【００１０】アクロモバクタ−属細菌に関する文献とし
ては、アクロモバクタ−属細菌を供試して、フサリウム
(Fusarium)、リゾクトニア(Rhizoctonia) 、コレトリカ
ム(Colletotricum) のおこすワタの病害を防除する方法
が検討されている（Plant Disease Reporter(1976) 60-
5:371-373）。しかし、これまで発明者が知る限り、オ
クロバクトラム・アンスロピに属する細菌が植物の病
害、特に細菌病の防除に用いられた例はみられない。従
って、本発明は、オクロバクトラム・アンスロピを使用
して、タバコ立枯病、ナス科青枯病を防除しようとする
ものである。[0010] As a literature on the bacterium belonging to the genus Achromobacter, a bacterium belonging to the genus Achromobacter was tested, and Fusarium
(Fusarium), Rhizoctonia, and Colletotricum (Colletotricum) have been studied for controlling the diseases of cotton (Plant Disease Reporter (1976) 60-
5: 371-373). However, as far as the inventor knows, there have been no cases where bacteria belonging to Ochrobactrum anthropi have been used for controlling plant diseases, particularly bacterial diseases. Therefore, the present invention is intended to control tobacco wilt disease and Solanaceae wilt disease by using Ochrobactrum anthropi.
【００１１】[0011]
【発明の実施の態様】本発明において、「オクロバクト
ラム・アンスロピに属する微生物」をナス科植物の根
部、栽培地及び／又はその土壌に存在させることで、ナ
ス科植物の土壌病害（特に、タバコ立枯病及びナス科植
物青枯病）を防除する性質を有する。更に、オクロバク
トラム・アンスロピに属するこのような性質を有する微
生物は、試験すべき菌株を植物の根部、栽培地及び／又
はその土壌に存在させ、ナス科植物の土壌病害を防除す
る性質を有すると認められる菌株を自然界または公知の
オクロバクトラム・アンスロピに属する微生物の中から
選抜することによって、再現性良く入手することができ
る。このような選抜法としては、後述する実施例１の方
法及びこれらと同等と認められる方法が挙げられる。以
下の記述によって本発明が限定的に解釈されないことを
前提として、上記微生物によってナス科植物の土壌病害
が防除できる理由は、上記微生物が植物の根に定着し、
土壌の病原菌の植物体への侵入を防ぐとともに、植物体
に該土壌の病原菌に対する抵抗性を誘導するのではない
かと考えられる。BEST MODE FOR CARRYING OUT THE INVENTION In the present invention, the presence of a "microorganism belonging to Ochrobactrum anthropi" in the roots of solanaceous plants, cultivated areas and / or soils thereof causes soil diseases of the solanaceous plants (particularly tobacco plants). Blight and wilt of Solanaceae plant). Furthermore, the microorganism having such properties belonging to Ochrobactrum anthropi is recognized as having the property of controlling soil diseases of solanaceous plants by allowing the strain to be tested to exist in the root of the plant, the cultivation area and / or the soil thereof. The strain can be obtained with good reproducibility by selecting it from the microorganisms belonging to the natural world or known Ochrobactrum anthropi. Examples of such selection methods include the method of Example 1 described below and the methods recognized as equivalent thereto. Assuming that the present invention is not limitedly interpreted by the following description, the reason why the soil disease of Solanaceae plants can be controlled by the microorganisms is that the microorganisms are rooted in the roots of plants,
It is considered that it may prevent the invasion of soil pathogens into the plant and induce the plant resistance to the pathogens in the soil.
【００１２】本発明において、上記性質を有するオクロ
バクトラム・アンスロピに属する微生物としては、具体
的には、後述する実施例において記載されているオクロ
バクトラム・アンスロピＴＲＢ１９菌株（以下、「ＴＲ
Ｂ１９菌株」という）が挙げられる。In the present invention, as the microorganism belonging to Ochrobactrum anthropi having the above-mentioned properties, specifically, the Ochrobactrum anthropi TRB19 strain (hereinafter referred to as “TR
"B19 strain").
【００１３】本発明において、「防除」とは、病害の予
防のみならず、病害の除去をも含む意味で用いるものと
する。また、「根部」とは、植物を栽培した場合に土壌
中または水耕液中にあって水分や栄養分の吸収を行う部
分をいうものとする。本発明の土壌病害防除剤及び防除
方法が適用可能なナス科植物としては、例えばタバコ、
ナス、トマト、ピーマン、ジャガイモ、トウガラシ、ペ
ピーノ等が挙げられ、立枯病菌の宿主植物には適用可能
である。In the present invention, "control" means not only the prevention of disease but also the removal of disease. In addition, the “root portion” means a portion in the soil or in the hydroponic solution that absorbs water and nutrients when the plant is cultivated. Examples of the Solanaceae plants to which the soil disease control agent and the control method of the present invention are applicable include, for example, tobacco,
Eggplants, tomatoes, bell peppers, potatoes, capsicum, pepino and the like can be mentioned, and they are applicable to host plants of wilt disease fungus.
【００１４】本発明における上記オクロバクトラム・ア
ンスロピの培養は、特別な培養基を用いる必要がなく、
培地の種類および培養条件を含めて任意のものでありう
る。培地としては、キングＢ培地（J. Lab. Clin. Med.
(1954) vol.44, 301-307）、Ｍ５２３培地（Phytopatho
logy(1954) vol.44, 969-976）あるいは肉エキス培地な
ど一般的な培地が挙げられる。また、液体培地以外に寒
天入りの斜面培地及び平板培地等を用いてもよい。それ
ら培養によって増殖させ、所望の菌体量を得ることがで
きる。The culture of Ochrobactrum anthropi in the present invention does not require the use of a special culture medium,
It may be arbitrary, including the type of medium and culture conditions. As the medium, King B medium (J. Lab. Clin. Med.
(1954) vol.44, 301-307), M523 medium (Phytopatho
logy (1954) vol.44, 969-976) or a general medium such as meat extract medium. In addition to liquid medium, agar-containing slant medium, plate medium, etc. may be used. The cells can be grown by these cultures to obtain a desired amount of cells.
【００１５】培地の炭素源としては、本発明による微生
物が同化しうるあらゆるものが利用できる。具体的に
は、グルコース、ガラクトース、ラクトース、アラビノ
ース、マンノース、麦芽エキス澱粉加水分解物などの糖
の他に、該微生物が利用し得る各種の合成または天然炭
素源がある。窒素源にしても同様に、ペプトン、肉エキ
ス、酵母エキス等の有機窒素含有物をはじめ、該微生物
が利用しうる各種の合成または天然物が利用可能であ
る。微生物培養の定法に従って、食塩、リン酸塩などの
無機塩類、カルシウム、マグネシウム、鉄などの金属の
塩類、ビタミン、アミノ酸などの微量栄養源も必要に応
じて添加することができる。As the carbon source of the medium, any carbon source that the microorganism of the present invention can assimilate can be used. Specifically, in addition to sugars such as glucose, galactose, lactose, arabinose, mannose, and malt extract starch hydrolyzate, there are various synthetic or natural carbon sources that can be utilized by the microorganism. Similarly, as a nitrogen source, various synthetic or natural products usable by the microorganism can be used, including organic nitrogen-containing products such as peptone, meat extract, and yeast extract. Inorganic salts such as sodium chloride and phosphate, salts of metals such as calcium, magnesium and iron, and trace nutrients such as vitamins and amino acids can be added, if necessary, according to conventional methods for culturing microorganisms.
【００１６】培養は、振とう培養、静置培養、通気培養
などの好気的条件下で行なうことができる。培養温度
は、２５〜３０℃、好ましくは２５〜２８℃、培地のｐ
Ｈは、ｐＨ５〜８、好ましくはｐＨ６〜７、培養期間
は、１〜４日間、好ましくは２〜３日間が適当である。
本発明による土壌病害防除剤（以下単に「防除剤」とも
いう）において、微生物が生菌として適用される場合、
上記微生物を水１ｍｌあたり１０6 〜１０10個、好まし
くは１０7 〜１０9 個の濃度で、苗を本畑に移植する７
日前から移植後の１ヵ月までの間に植物体の根部に１回
〜複数回、浸漬処理、灌注処理または灌流処理等をする
ことによって適用されることが好ましい。浸漬処理によ
る場合には、浸漬時間は３０分〜３時間、好ましくは１
時間〜１時間３０分である。また灌注処理または灌流処
理による場合には、移植前苗１株あたり５〜２０ｍｌ、
移植後は苗１株あたり５０〜２００ｍｌを適用すること
が好ましい。また、微生物の培養物として適用される場
合には、菌体を依然として含んだものとして適用される
のが好ましく、その適用時期及び適用量は上記生菌の場
合に準じて適宜決定される。The culture can be carried out under aerobic conditions such as shaking culture, static culture, aeration culture and the like. The culture temperature is 25 to 30 ° C., preferably 25 to 28 ° C.
H is pH 5 to 8, preferably pH 6 to 7, and the culture period is 1 to 4 days, preferably 2 to 3 days.
In the soil disease controlling agent according to the present invention (hereinafter also simply referred to as “controlling agent”), when the microorganism is applied as a live bacterium,
The above-mentioned microorganisms are transplanted to the main field at a concentration of 10 6 to 10 10 , preferably 10 7 to 10 9 per 1 ml of water 7.
It is preferable that the root portion of the plant is applied once to a plurality of times from the day before to one month after the transplantation by performing a dipping treatment, an irrigation treatment, a perfusion treatment, or the like. In the case of the dipping treatment, the dipping time is 30 minutes to 3 hours, preferably 1
Time is 1 hour and 30 minutes. In the case of irrigation or perfusion treatment, 5 to 20 ml per seedling before transplantation,
After transplantation, it is preferable to apply 50 to 200 ml per seedling. Further, when it is applied as a culture of a microorganism, it is preferably applied as still containing bacterial cells, and its application time and application amount are appropriately determined according to the case of the above-mentioned viable bacteria.
【００１７】更に、本発明の好ましい態様によれば、上
記微生物またはその培養物を土壌に混和または撹拌散布
したのち、ナス科植物を移植してもよい。この場合の混
和または撹拌散布の時期およびその量は、移植の１ヶ月
前から直前の間に土壌１ｇ当たり菌体１０5 個以上とな
るように混和するのが好ましい。Furthermore, according to a preferred embodiment of the present invention, the Solanaceae plant may be transplanted after the above-mentioned microorganism or its culture is mixed in the soil or stirred and sprayed. In this case, it is preferable that the time and amount of mixing or stirring / spraying are mixed so as to be 10 5 cells or more per 1 g of soil between one month before and immediately before transplantation.
【００１８】本発明において、オクロバクトラム・アン
スロピの培養物とは、その微生物の培養物の培養懸濁
液、生菌、培養ろ液、またはその土壌病害防除に有効な
成分の抽出液をいうものとする。In the present invention, the culture of Ochrobactrum anthropi means a culture suspension of a culture of the microorganism, viable bacteria, a culture filtrate, or an extract of an ingredient effective for controlling soil diseases. To do.
【００１９】本発明による土壌病害防除剤は、いわゆる
担体と組み合わされて、農薬組成物とされてもよい。好
ましい担体の例としては、所望によりｐＨ緩衝液を加え
た水溶性溶媒、スキムミルクなどの保護剤とともに凍結
乾燥後タルクなどの助剤を加えた粉末剤、顆粒剤、並び
にバーミキュライトなどの多孔質体等が挙げられる。本
発明の土壌病害防除剤は、液体、粉末、錠剤、シート等
のいずれの形態をも採ることができる。本発明の土壌病
害防除剤は、他の有効成分と組み合わされてもよい。他
の有効成分は特に限定されるものではないが、例えば殺
虫剤、除草剤、殺菌剤等が挙げられる。また、本発明の
土壌病害防除剤は、土壌改良材、堆肥、肥料等と組み合
わせた組成物とされてもよい。土壌病害防除剤として処
方された際の微生物およびその培養物の量、さらには適
用時期及び適用量は上記生菌の場合に準じて適宜決定さ
れる。The soil disease controlling agent according to the present invention may be combined with a so-called carrier to form an agrochemical composition. Examples of preferable carriers include a water-soluble solvent optionally added with a pH buffer, a protective agent such as skim milk, and a powder agent including an auxiliary agent such as talc after freeze-drying, a granule agent, and a porous body such as vermiculite. Is mentioned. The soil disease controlling agent of the present invention can take any form of liquid, powder, tablets, sheets and the like. The soil disease controlling agent of the present invention may be combined with other active ingredients. Other active ingredients are not particularly limited, and examples thereof include insecticides, herbicides, fungicides and the like. Further, the soil disease controlling agent of the present invention may be a composition in combination with a soil conditioner, compost, fertilizer and the like. The amount of the microorganism and the culture thereof when formulated as a soil disease controlling agent, as well as the application time and the application amount, are appropriately determined according to the case of the above-mentioned viable bacteria.
【００２０】[0020]
【実施例】以下に実施例をあげて本発明の内容を説明す
るが、本発明はこの実施例により限定されるものではな
い。 実施例１ 本発明のオクロバクトラム・アンスロピの分離・選抜方
法について述べる。本発明者らは、全国のタバコ畑から
タバコ根部を採取した。そして、タバコ根面あるいは根
圏から数多くの細菌株を分離した。具体的には、健全な
タバコ根部を掘り取り、ハサミで根部を切り離し、付着
している土壌を振り落とした。約１ｃｍの長さに切断し
た根を10mlの滅菌蒸留水に入れ、ミキサ−で攪拌した
後、得られた懸濁液を１白金耳取り、キングＢ培地（プ
ロテオ−スペプトンＮｏ３ 20.0g, リン酸二カリウム
1.5g, 硫酸マグネシウム 1.5g, グリセリン 10.0m
l,蒸留水 1,000ml：J. Lab. Clin. Med(1954)Vol.44:3
01〜307）上に画線した。28℃、３日間培養後、得られ
た単一のコロニ−を-80℃のフリ−ザ−で保存した。こ
のようにして、根面に良く定着すると考えられる細菌を
得た。EXAMPLES The contents of the present invention will be described below with reference to examples, but the present invention is not limited to these examples. Example 1 A method for separating and selecting Ochrobactrum anthropi of the present invention will be described. The present inventors collected tobacco roots from tobacco fields nationwide. Then, many bacterial strains were isolated from tobacco roots or rhizosphere. Specifically, a healthy tobacco root was dug, the root was cut with scissors, and the attached soil was shaken off. Roots cut to a length of about 1 cm were placed in 10 ml of sterilized distilled water, stirred with a mixer, 1 platinum loop of the resulting suspension was taken, and King B medium (Proteo-peptone No3 20.0 g, phosphoric acid was used). Dipotassium
1.5g, magnesium sulfate 1.5g, glycerin 10.0m
l, 1,000 ml of distilled water: J. Lab. Clin. Med (1954) Vol.44: 3
01-307) The above was drawn. After culturing at 28 ° C for 3 days, the obtained single colony was stored in a freezer at -80 ° C. In this way, a bacterium believed to be well-established on the root surface was obtained.
【００２１】そして１）細菌自体が作物に対して病原性
あるいは悪影響を与えない、２）立枯病菌による病気の
発生を圃場で栽培後期まで効果的に抑制する、などの性
質を有している有用な菌株を温室あるいは圃場試験で選
抜した。具体的には、植物体への影響及び効果の評価法
として、タバコ（品種：ＢＹ４）の８〜９枚苗を供試
し、細菌株をキングＢ液体培地で培養後、培養液をタバ
コ苗の根部に灌注接種し、土壌を入れた４寸鉢に移植し
た。さらに、立枯病菌液を移植苗の株元に土壌灌注接種
後、30℃に設定した温室内で定期的に立枯病の発生につ
いて調査し、防除効果の高い有用な細菌株を選抜した。And, 1) bacteria themselves do not cause pathogenicity or adverse effects on crops, and 2) effectively suppress the occurrence of diseases caused by the wilt fungus in the field until the latter stage of cultivation. Useful strains were selected in greenhouse or field trials. Specifically, as an evaluation method of effects and effects on plants, 8 to 9 tobacco (cultivar: BY4) seedlings were tested, the bacterial strain was cultivated in King B liquid medium, and then the culture solution was added to the tobacco seedlings. The roots were irrigated and inoculated, and then transplanted in a 4 inch pot containing soil. Furthermore, after inoculating the seedlings of the transplant seedlings with soil irrigation, the occurrence of wilting disease was regularly investigated in a greenhouse set at 30 ° C, and useful bacterial strains with high control effect were selected.
【００２２】次に、温室内で高い防除効果を有する菌株
を、キングＢ液体培地で培養後、遠心集菌し（×10,000
g,15分）、滅菌水中に109CFU/mlの濃度で懸濁した後、
タバコ（品種：つくば２号）の９枚苗根部に灌注接種
し、立枯病菌の汚染畑（汚染菌濃度１０3 ＣＦＵ／１ｇ
乾土）に移植した。その後、タバコ栽培後期まで発病調
査を行ない、後期まで効果が持続する細菌株のみを最終
的に選抜した。その結果、栃木県小山市のタバコ（品
種：ＢＹ４）の根内から有用細菌TRB19菌株を得た。TRB
19菌株の細菌学的性質を第１表に示す。Then, the strain having a high controlling effect in a greenhouse is cultured in King B liquid medium and then collected by centrifugation (× 10,000).
g, 15 minutes), after suspending in sterile water at a concentration of 10 9 CFU / ml,
9 seedlings of tobacco (variety: Tsukuba No. 2) were irrigated and inoculated, and the field contaminated with wilting fungus (concentration of contaminating bacteria 10 3 CFU / 1 g
Transplanted to dry soil). After that, the disease was investigated until the latter stage of tobacco cultivation, and only the bacterial strains whose effects persisted until the latter stage were finally selected. As a result, a useful bacterium TRB19 strain was obtained from the root of tobacco (variety: BY4) in Oyama City, Tochigi Prefecture. TRB
The bacteriological properties of the 19 strains are shown in Table 1.
【００２３】[0023]
【表１】 [Table 1]
【００２４】本細菌TRB19菌株を、Bergey,s manual of
determinative bacteriology ９版(1994)より、オクロ
バクトラム・アンスロピ（Ochrobactrum anthropi）と
同定した。This bacterium, TRB19 strain, was subjected to Bergey, s manual of
From determinative bacteriology 9th edition (1994), it was identified as Ochrobactrum anthropi.
【００２５】実施例２ TRB19をキングＢ液体培地で28℃、２日間振とう培養し
た後、109cfu/mlに調整した。タバコ苗（ブライトイエ
ロー４）は、36本植えの塩化ビニール製のポットで栽培
した９葉苗を20本ずつ供試した。菌培養液の接種は、株
当たり10mlを株元に灌注接種した。接種したタバコ苗
は、12時間、28℃の温室内で栽培した後、直径12cmの素
焼鉢に肥土を使用して移植し、30℃の温室内で栽培し
た。移植２週間後に、立枯病菌液（106CFU/ml）を株当
たり20mlずつ株元に灌注接種した。対照として、キング
Ｂ液体培地を同様にタバコ株元に灌注接種したタバコを
供試した。接種２週間後に立枯病の発病状況を調査し
た。発病程度は、第２表が示すように０から５までの６
段階とし、以下の式により平均罹病指数を求め、防除率
を算出した。また、発病率は、発病株数を供試本数で割
った値に１００をかけることにより算出した。Example 2 TRB19 was cultured in King B liquid medium at 28 ° C. for 2 days with shaking, and then adjusted to 10 9 cfu / ml. As tobacco seedlings (Bright Yellow 4), 20 9-leaf seedlings cultivated in a 36-planted vinyl chloride pot were used. For inoculation of the bacterial culture, 10 ml per strain was irrigated and inoculated to the original strain. The inoculated tobacco seedlings were cultivated in a greenhouse at 28 ° C for 12 hours, then transplanted using a fertilizer in a clay pot with a diameter of 12 cm, and cultivated in a greenhouse at 30 ° C. Two weeks after the transplantation, 20 ml of the bacterial wilt disease solution (10 6 CFU / ml) was irrigated and inoculated into each strain source. As a control, tobacco was similarly inoculated with King B liquid medium by irrigating the tobacco strain. Two weeks after the inoculation, the occurrence status of the wilt disease was investigated. The degree of illness is 6 from 0 to 5 as shown in Table 2.
The average morbidity index was calculated by the following formula, and the control rate was calculated. The disease incidence was calculated by dividing the number of diseased strains by the number of test specimens and multiplying by 100.
【００２６】[0026]
【表２】 [Table 2]
【００２７】[0027]
【数１】 [Equation 1]
【００２８】[0028]
【表３】 [Table 3]
【００２９】第３表の結果から明らかなように対照区よ
りもTRB19処理区の方が発病率が有意に低かった（ｔ検
定，１％水準）。平均罹病指数も処理区の方が有意に低
く（ｔ検定，１％水準）、立枯菌接種２週間後の防除率
は、62.3％であった。As is clear from the results shown in Table 3, the incidence of disease was significantly lower in the TRB19 treated group than in the control group (t-test, 1% level). The average morbidity index was also significantly lower in the treated section (t-test, 1% level), and the control rate 2 weeks after the inoculation of dead bacteria was 62.3%.
【００３０】実施例３ TRB19をキングＢ液体培地で28℃、２日間振とう培養し
た後、遠心集菌（10,000×g,10分間）し、滅菌水中に懸
濁した（109CFU/ml）。実施例２と同様の方法でタバコ
を栽培し、TRB19菌懸濁液を同様に接種した。実施例２
と同様の方法で立枯病菌を接種し、２週間後に発病を調
査した。対照として、滅菌蒸留水を接種したタバコを供
試した。Example 3 TRB19 was cultured in King B liquid medium at 28 ° C. for 2 days with shaking, and then the cells were collected by centrifugation (10,000 × g, 10 minutes) and suspended in sterile water (10 9 CFU / ml). . Tobacco was cultivated in the same manner as in Example 2, and the TRB19 bacterial suspension was similarly inoculated. Example 2
In the same manner as above, the bacterial wilt disease was inoculated, and two weeks later, the onset of disease was investigated. Tobacco inoculated with sterile distilled water was used as a control.
【００３１】[0031]
【表４】 [Table 4]
【００３２】第４表の結果から明らかなように対照区よ
りもTRB19処理区の方が発病率が有意に低かった（ｔ検
定，５％水準）。平均罹病指数も処理区の方が有意に低
く（ｔ検定，１％水準）、立枯病菌接種２週間後の防除
率は、64％であった。As is clear from the results of Table 4, the TRB19-treated group had a significantly lower disease incidence than the control group (t-test, 5% level). The average morbidity index was also significantly lower in the treated section (t-test, 1% level), and the control rate 2 weeks after inoculation of the wilt disease bacteria was 64%.
【００３３】実施例４ TRB19をキングＢ液体培地で28℃、２日間振とう培養し
た後、遠心集菌（10,000×g,10分間）し、滅菌水中に懸
濁した（109CFU/ml）。菌濃度は、キングＢ平板培地に
よって確認した。タバコ苗（つくば２号）は、36本植え
の塩化ビニール製のポットで栽培し、本畑に移植する大
きさの９葉苗を供試した。菌懸濁液の接種は、以下の処
理によって実施した。 （１）TRB19移植前処理：移植１日前に菌懸濁液（109CF
U/ml)を株当たり10ml株元灌注接種した後に本畑に移植
した。 （２）TRB19移植前処理＋土寄時処理：（１）の様に移
植したタバコ株に、（１）と同様の方法で培養、遠心集
菌した菌体を蒸留水に懸濁し（108CFU/ml）、土寄時に
株当たり200mlずつ土壌灌注接種した。 （３）対照区：滅菌蒸留水を移植前に同様の方法で処理
したタバコ苗を移植した。Example 4 TRB19 was cultured in King B liquid medium at 28 ° C. for 2 days with shaking, and then the cells were collected by centrifugation (10,000 × g, 10 minutes) and suspended in sterile water (10 9 CFU / ml). . The bacterial concentration was confirmed by King B plate medium. Tobacco seedlings (Tsukuba No. 2) were cultivated in a 36-plant vinyl chloride pot, and 9-leaf seedlings of a size to be transplanted to the main field were tested. The bacterial suspension was inoculated by the following treatment. (1) TRB19 pre-transplantation treatment: One day before transplantation, the bacterial suspension (10 9 CF
U / ml) was pre-irrigated with 10 ml per strain and then transplanted to the main field. (2) TRB19 pre-transplantation treatment + soiling treatment: Tobacco strains transplanted as in (1) were cultivated and centrifuged in the same manner as in (1) to suspend the cells in distilled water (10 8 CFU / ml), and 200 ml of each soil was irrigated at the time of soil donation. (3) Control group: Tobacco seedlings treated with the same method as above before transplanting sterile distilled water were transplanted.
【００３４】移植前処理したタバコ苗は、12時間、28℃
の温室内で栽培した後、立枯病菌汚染畑（103CFU/1g乾
土)に、４月１４日移植した。土寄時の処理は、５月２
０日に実施した。移植後、通常の耕作方法によって栽培
し、定期的に立枯病の発病を調査した。Tobacco seedlings that had been pre-transplanted were treated at 28 ° C for 12 hours.
After being cultivated in the greenhouse, the plant was transplanted to a field contaminated with bacterial wilt disease (10 3 CFU / 1 g dry soil) on April 14th. The processing at the time of the donation May 2
It was carried out on the 0th day. After the transplantation, the plants were cultivated by the usual cultivation method, and the onset of wilt disease was regularly investigated.
【００３５】[0035]
【表５】 [Table 5]
【００３６】第５表の結果から明らかなように、対照区
よりもTRB19菌懸濁液処理区の方が発病率が有意に低く
（５％水準）、防除率も栽培後期まで安定していた。ま
た、処理区では移植前処理区よりも移植前＋土寄時処理
区の効果の方が防除効果が高かった。調査本数のばらつ
きは、ウイルス病による被害のために各々の試験区で欠
株が生じたためである。As is clear from the results shown in Table 5, the TRB19 bacterial suspension treatment group had a significantly lower disease incidence (5% level) than the control group, and the control rate was stable until the latter stage of cultivation. . Moreover, in the treated plots, the control effect of the pre-transplantation + soil-during treatment plots was higher than that of the pre-transplantation treatment plots. The variation in the number of surveys was due to the lack of strains in each test plot due to the damage caused by the viral disease.
【００３７】[0037]
【発明の効果】本発明の防除剤および防除方法は、環境
や生態系に支障を及ぼすことが無く、ナス科植物の土壌
病害、特にタバコ立枯病及びナス科植物青枯病に対し
て、その栽培後期まで安定して高い防除効果を示すこと
ができる。EFFECTS OF THE INVENTION The control agent and control method of the present invention have no adverse effects on the environment or the ecosystem, and against soil diseases of Solanaceae plants, particularly tobacco wilt disease and Solanaceae wilt disease, It is possible to stably exhibit a high control effect until the latter stage of cultivation.
Claims (5)
有するオクロバクトラム・アンスロピ（Ochrobactrum a
nthropi）に属する微生物及び／又はその培養物を有効
成分として含有することを特徴とするナス科植物の土壌
病害防除剤。1. Ochrobactrum a that has the property of controlling soil diseases of Solanaceae plants.
A soil disease control agent for Solanaceae plants, which comprises a microorganism belonging to nthropi) and / or a culture thereof as an active ingredient.
actrum anthropi）に属する微生物が、オクロバクトラ
ム・アンスロピＴＲＢ１９菌株である請求項１に記載の
ナス科植物の土壌病害防除剤。2. Ochrob anthropi
The soil disease controlling agent for Solanaceae according to claim 1, wherein the microorganism belonging to actrum anthropi) is Ochrobactrum anthropi TRB19 strain.
アラム（Pseudomonas solanacearum）が原因で起こるタ
バコ立枯病及びナス科植物青枯病である請求項１又は２
に記載のナス科植物の土壌病害防除剤。3. The soil disease is tobacco bacterial wilt caused by Pseudomonas solanacearum, or wilt disease of Solanaceae plant, which is caused by Pseudomonas solanacearum.
2. A soil disease control agent for Solanaceae plants according to.
ス科植物の土壌病害防除剤を、ナス科植物の根部、栽培
地及び／又はその土壌に導入する工程を含むことを特徴
とするナス科植物の土壌病害防除方法。4. A method comprising the step of introducing the soil disease control agent for Solanaceae plants according to any one of claims 1 to 3 into roots, cultivated areas and / or soils of Solanaceae plants. For controlling soil diseases of solanaceous plants.
アラム（Pseudomonas solanacearum）が原因で起こるタ
バコ立枯病及びナス科植物青枯病である請求項４に記載
のナス科植物の土壌病害防除方法。5. The method for controlling soil diseases of solanaceous plants according to claim 4, wherein the soil diseases are tobacco wilt disease and solanaceous plant wilt disease caused by Pseudomonas solanacearum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP8006693A JPH09194314A (en) | 1996-01-18 | 1996-01-18 | Agent for controlling soil blight of plant of family solanaceae and controlling method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP8006693A JPH09194314A (en) | 1996-01-18 | 1996-01-18 | Agent for controlling soil blight of plant of family solanaceae and controlling method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| JPH09194314A true JPH09194314A (en) | 1997-07-29 |
Family
ID=11645433
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP8006693A Pending JPH09194314A (en) | 1996-01-18 | 1996-01-18 | Agent for controlling soil blight of plant of family solanaceae and controlling method |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH09194314A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998022572A1 (en) * | 1996-11-22 | 1998-05-28 | Pioneer Hi-Bred International, Inc. | Moniliformin detoxification compositions and methods |
| WO2009145074A1 (en) | 2008-05-29 | 2009-12-03 | 日本たばこ産業株式会社 | Bacterium capable of reducing heavy metal content in plant |
-
1996
- 1996-01-18 JP JP8006693A patent/JPH09194314A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998022572A1 (en) * | 1996-11-22 | 1998-05-28 | Pioneer Hi-Bred International, Inc. | Moniliformin detoxification compositions and methods |
| WO2009145074A1 (en) | 2008-05-29 | 2009-12-03 | 日本たばこ産業株式会社 | Bacterium capable of reducing heavy metal content in plant |
| US8383390B2 (en) | 2008-05-29 | 2013-02-26 | Japan Tobacco Inc. | Bacteria that reduce content of heavy metals in plant |
| EP2578675A1 (en) | 2008-05-29 | 2013-04-10 | Japan Tobacco Inc. | Bacteria that reduce content of heavy metals in plant |
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