JP4287899B1 - Eggplant half body wilt inhibitor and method for suppressing eggplant half wilt disease - Google Patents
Eggplant half body wilt inhibitor and method for suppressing eggplant half wilt disease Download PDFInfo
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Abstract
【課題】 環境汚染・生態系破壊・人体への悪影響の懸念がある化学農薬を使用せずに、細菌を用いた生物資材でもって、ナスビのナス半身萎凋病を効果的に抑制するナス半身萎凋病抑制剤及びその抑制方法を提供する。
【解決手段】 バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）菌の培養物と、好熱性繊維素分解菌クロストリジュウム・サーモセルム（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ）ＳＫ５２２（受託番号 微工研条寄第３４５９号）と高熱性タンパク質分解菌サーマス・アクアティクス（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ）ＳＫ５４２（受託番号 微工研条寄第３３８２号）との共生培養物を混合した液状混合培養物に、ナス苗の根を浸漬してこれら菌を接種させた後にナス苗を畑土壌に植生する。
【選択図】 図１PROBLEM TO BE SOLVED: To effectively suppress eggplant half body wilt disease of eggplant by using biological materials using bacteria without using chemical pesticides that may cause environmental pollution, ecosystem destruction, or adverse effects on the human body A disease inhibitor and a method for suppressing the disease are provided.
SOLUTION: A culture of Bacillus thuringiensis N-029 strain (Accession number FERM AP-21647) and thermophilic fibrinolytic clostridium thermocellum (Accession number Clostridium thermocellum biovar (SK22) A liquid mixed culture obtained by mixing a symbiotic culture of Mikken Kenjo No. 3459) with a thermophilic proteolytic bacterium Thermus aquaticus biovar SK542 (Accession No. Mikuken Hojo No. 3382) After soaking the roots of the eggplant seedlings and inoculating these fungi, the eggplant seedlings are vegetated on the field soil.
[Selection] Figure 1
Description
本発明はナスビの収穫に大きな影響を与えるナス半身萎凋病の発生を、バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）を有効菌として用いて、又はこの菌と好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）と、高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）
とを６０℃以上の高熱性環境下で培養した共生培養物を混合して利用して、または、これら複数の培養物を炭酸カルシウム等の適当な担体に含有させた複合菌含有物を利用してナス半身萎凋病を抑制する技術に関する。本明細書・特許請求の範囲及び要約書中の細菌名の英文字の下線は、細菌名表記がイタリックであることを示す。
The present invention relates to the development of eggplant half body wilt that greatly affects the harvest of eggplant, using Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) as an effective bacterium, or Thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar SK522, Microtechnical Research Institute No. 3459) and thermophilic proteolytic bacteria Thermus aquaticus SK542 ( Thermus aquaticus biotechnological structure) No. 3382)
Are mixed with a symbiotic culture cultured in a high heat environment of 60 ° C. or higher, or a complex bacterium containing a plurality of these cultures in a suitable carrier such as calcium carbonate is used. The present invention relates to a technique for suppressing eggplant half body wilt. The underlined letter of the bacterial name in the present specification, claims and abstract indicates that the bacterial name is italicized.
現在、環境問題が大きな関心事となっている。色々な汚染物質による地球環境汚染が深刻化の度合いを深めているからである。農業分野は特にその危険性が指摘されている。それは農産物の増収を目的に化学肥料、化学農薬を多量に使用してきた過去の歴史の結末に他ならない。特に化学農薬が地球環境汚染、生態系の破壊、人体への悪影響等の元凶となっている事例も少なくなく、そのため、２００３年には農薬法の一部改正が施行されて、いくつかの化学農薬が登録を抹消された。土壌消毒剤として長年使用されてきた臭化メチルがオゾン層破壊物質として使用が禁止されたのはその一例である。このため化学農薬の使用量は年々減少傾向にある
最近の農業は地球環境保全に立脚した農業形態を目指している。有機農業への傾斜が強まっているのはこのためである。堆厩肥の活用や穀物や野菜などの害虫・病原菌に対する生物的防除の取組みなど、有機資材の活用を基本に、化学肥料や化学農薬などの化成品の使用を低減し、バランスの良い資材の施用で、健全で永続的に安定した営農は今や農業分野での基本理念となっている。
Currently, environmental issues are of great concern. This is because global environmental pollution caused by various pollutants is becoming more serious. The danger is particularly pointed out in the agricultural sector. That is nothing but the end of the past history of using large amounts of chemical fertilizers and chemical pesticides to increase the yield of agricultural products. In particular, there are many cases where chemical pesticides are the cause of global environmental pollution, ecosystem destruction, adverse effects on the human body, etc. Therefore, in 2003, a partial amendment of the Agricultural Chemicals Law was enacted, and some chemicals Agricultural chemicals have been deregistered. One example is the use of methyl bromide, which has been used as a soil disinfectant for many years, as an ozone depleting substance. For this reason, the amount of chemical pesticides used in recent agriculture, which has been decreasing year by year, aims at an agricultural form based on global environmental conservation. This is why the inclination to organic farming is growing. Application of well-balanced materials by reducing the use of chemical fertilizers and chemicals such as chemical pesticides based on the use of organic materials such as manure and biological control of pests and pathogens such as grains and vegetables Therefore, healthy, permanent and stable farming is now a basic philosophy in the field of agriculture.
このような中で、生物的防除剤への関心が高まっている。特にバチルス・チューリンジェンシス菌を活用した生物農薬の研究開発は世界的な規模で進められおり、実用化されたものも多い。しかし、これは殺虫剤としての活用が主で、細菌やカビなどの植物病原菌に対してはあまり開発が進んでいないのが現状である。 Under such circumstances, interest in biological control agents is increasing. In particular, research and development of biological pesticides using Bacillus thuringiensis has been carried out on a global scale, and many have been put into practical use. However, this is mainly used as an insecticide, and the development of phytopathogenic bacteria such as bacteria and fungi has not progressed much.
このような現状に鑑み、発明者らはバチルス・チューリンジェンシス菌を他の菌と複合させることにより、作物病害に対して防除的な効果を増強させ、且つ、土壌の化学的・物理的・生物的環境を改善し、地力増強、品質改善などの付随効果も期待される新しい土壌改良剤の発明に至った。 In view of such a current situation, the inventors have combined Bacillus thuringiensis with other bacteria to enhance the control effect against crop diseases, and the chemical / physical / The present invention has led to the invention of a new soil conditioner that improves the biological environment and is expected to have accompanying effects such as enhancement of geopower and quality improvement.
ナス半身萎凋病菌は前作で感染した状態で収穫し、完全な殺菌駆除をせず放置すると、残渣とともに土壌中で生存し続ける感染力の強い菌で、多くの作物に感染する。新しくナスの栽培を始めた時、前作の生存菌がナスの根から侵入し、導管を上昇して感染発病する。気温２０〜２４℃の比較的低温条件で多発することが多く、２８℃以上ではほとんど発病しない。病状は下葉に黒褐色の細長い筋条の病斑を生じ、次第に小葉の葉脈が葉周辺の縁から黄褐色に変わりやがて萎れ始める。症状が進行すると、下葉全体が灰緑色〜淡褐色となり青枯れ状に萎凋し枯れ死する。また、葉柄、果梗、クラウンの導管が褐変し、根は黒褐色に腐敗する。勿論収穫に多大な害をもたらす。 Eggplant half-wilt fungus is a highly infectious fungus that survives in the soil along with the residue if it is harvested in the state of infection in the previous crop and left without being completely disinfected. It infects many crops. When cultivation of eggplants is newly started, surviving bacteria from the previous crop invade from the roots of eggplants and raise the conduit to become infected. It often occurs frequently under relatively low temperature conditions of 20 to 24 ° C., and hardly occurs at 28 ° C. or higher. The pathological condition produces dark brown strips of lesions in the lower lobe, and the veins of the leaflets gradually turn yellowish brown from the edges around the leaves and begin to wither. As the symptom progresses, the entire lower lobe becomes grayish green to light brown, withering and withering and dying. In addition, the petiole, fruit stem, and crown ducts turn brown, and the roots rot to dark brown. Of course, it causes great harm to the harvest.
従来、植物及び果実における真菌感染または細菌感染による病害を抑制する方法として、抗菌活性を有するバチルス・チューリンジェンシス株ＡＱ５２（ＮＲＲＬ寄託番号Ｂ２１６１９）の菌の培養物を用いる方法が特表２００１−５２４８０６号公報で知られている。 Conventionally, as a method of suppressing diseases caused by fungal infection or bacterial infection in plants and fruits, a method using a culture of a bacterium of Bacillus thuringiensis strain AQ52 (NRRL deposit number B21619) having antibacterial activity is disclosed in JP-T-2001-524806. Known in the Gazette.
このバチルス・チューリンジェンシス株は、他の細菌株との組み合わせによってトマト植物の焼き枯れ病の抑制、ビートアワヨトウ幼虫の生育阻害を示す。しかしながらナス半身萎凋病に有効とまでは知られていない。また、このナス半身萎凋病を効果的に抑制する細菌は知られていなかった。
本発明が解決しようとする課題は従来の問題を解消し、ナス半身萎凋病を有効に抑制しうる細菌を用いたナス半身萎凋病抑制剤及び抑制方法を提供することにある。 The problem to be solved by the present invention is to solve the conventional problems and provide an eggplant half body wilt suppressor and method for suppressing eggplant half body wilt disease.
かかる課題を解決した本発明の構成は、
１） バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）を有効菌とするナス半身萎凋病抑制剤
２） バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）を培養した培養物と、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）を６０℃以上の高熱性環境下で共生培養した共生培養物とを混合した培養混合物を有効成分とする、ナス半身萎凋病抑制剤
３） バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）の培養物と、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）の共生培養物とを、粉粒状担体に含有させた複合菌含有物を有効成分とする、ナス半身萎凋病抑制剤
４） 前記３）の担体が、粒度調整した炭酸カルシウム、消石灰、硫酸カルシウム、石灰岩粉、ドロマイト岩粉、貝化石粉末、カニ・エビ・シャコ・貝殻等の甲殻・貝殻粉末、ゼオライト、パーライト、バーミキュライト、珪藻土、木炭、くん炭、粉砕ピートモスのいずれか、あるいはこれらの混合物である、前記３）記載のナス半身萎凋病抑制剤
５） バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）の液状培養物に、ナス苗の根を浸漬後同ナス苗を土壌に植生する、ナス半身萎凋病抑制方法
６） バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）の培養物と、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）の共生培養物とを混合した液状培養物にナス苗の根を浸漬した後同ナス苗を畑土壌に植生する、ナス半身萎凋病抑制方法
にある。
The configuration of the present invention that solves this problem is as follows.
1) Eggplant half body wilt inhibitor 2 using Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) as an effective bacterium Bacillus thuringiensis N-029 strain ( (Certificate No. FERM AP-21647), thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar SK522, Mikokenjo No. 3459) and thermophilic proteolytic bacteria Thermus Aquatics SK542 (Thermus aquaticus biovar SK542, FERM BP-3382) symbiosis which was co-cultured with high heat resistance under the environment of more than 60 ℃ the And a mixed culture mixture active ingredient Yobutsu, and culture eggplant half body wilt inhibitor 3) Bacillus thuringiensis Jen cis (Bacillus thuringiensis) N-029 strain (accession number FERM AP-21647), thermophilic Fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar SK522, Microfabricated Kenjo No. 3459) and hyperthermogenic proteolytic fungus Thermus aquaticus SK542 ( Thermus aquaticus biovar No. 33) 4) The eggplant half body wilt disease inhibitor 4), wherein the carrier of 3) is a particle size-adjusted calcium carbonate, slaked lime, and potassium sulfate. Lucium, limestone powder, dolomite rock powder, shell fossil powder, shell / shellfish powder such as crab, shrimp, clam, shell, zeolite, perlite, vermiculite, diatomaceous earth, charcoal, charcoal, ground peat moss, or a mixture thereof The eggplant half body wilt inhibitor 5) as described in 3) above, after soaking roots of eggplant seedlings in a liquid culture of Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) Method for suppressing eggplant half body wilt disease by planting eggplant seedlings in soil 6) Culture of Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647), and thermophilic fibrinolytic fungi Lisium Thermoselm SK522 ( Clostridium thermocellum biovar SK522, Mikken Kenjo No. 3459) and a hyperthermolytic proteolytic thermus aquatics SK542 ( Thermus aquaticus biovar SK542, Mikken Kenjyo No. 3382) mixed liquid culture An eggplant seedling seedling root is dipped in a field soil, and then the eggplant seedling seedling is vegetated in a field soil.
本発明によれば、バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）を有効菌として利用することで、及びこの菌と好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）、及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）を複合菌として利用することでナス半身萎凋病を効果的に抑制することができた。また、その使用方法として別個に培養した上記３菌株の培養混合物、あるいは上記３菌株を別個に培養した培養物を粒度調整した炭酸カルシウム等の適当な担体に含有させて土壌又は栽培土に散布、又は漉き込むことによってナス半身萎凋病が発生し難い畑、土壌にできる。また、バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株の単独菌の培養物に、又は上記３菌株を別個に培養して混合した培養混合物にナス苗の根を浸した後、再度、畑や土壌に戻すことによってもナス半身萎凋病を抑制することができる。 According to the present invention, the Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) is used as an effective bacterium, and the bacterium and thermophilic fibrinolytic bacterium Clostridium thermocellum are used. SK522 ( Clostridium thermocellum biovar SK522, Mikokenjou No.3459) and hyperthermolytic proteolytic thermus aquatics SK542 ( Thermus aquaticus biovar SK542, Mikokenjoujou No.3382) It was possible to effectively suppress eggplant half body wilt. In addition, as a method of use, the culture mixture of the above three strains cultured separately, or the culture obtained by separately culturing the above three strains is contained in a suitable carrier such as calcium carbonate adjusted in particle size and sprayed on soil or cultivated soil, Or it can be made into a field and soil where eggplant half body wilt is less likely to occur by plowing. In addition, after soaking the roots of eggplant seedlings in a culture of a single strain of Bacillus thuringiensis N-029 or in a culture mixture obtained by separately culturing and mixing the above three strains, And eggplant half body wilt can be suppressed by returning to soil.
本発明のバチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）の培養物と、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）、及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）
の共生培養物を混合して、複合菌混合培養物として畑や土壌に散布するか、または前記３菌を別個に培養した培養物を炭酸カルシウム等の適当な担体に含有せしめて複合菌含有物として畑や土壌に散布することが最も望ましい。散布の時期としてナスを播種する１〜２週間前に直接畑や土壌に散布することが最も好ましい。
Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) of the present invention and thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar SK5 No. 3459), and hyperthermolytic proteolytic thermus aquatics SK542 ( Thermus aquaticus biovar SK542, Jikokenjo No. 3382)
Mixed with symbiotic cultures and sprayed on the field or soil as a mixed bacterial mixed culture, or a culture in which the three bacteria are cultured separately is contained in a suitable carrier such as calcium carbonate to contain the combined bacterial It is most desirable to spray it on fields and soils. It is most preferable to spray directly on the field or soil one to two weeks before sowing eggplant as the time of spraying.
以下、本発明の実施例１について詳しく説明する。
図１は、実施例１に用いたバチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）の細菌の寒天培地培養３日後の形状を示す説明図である。
図２は、実施例１で使用した抗菌活性試験用角型シャーレ寒天培地を示す説明図である。
図３は、実施例１のナス苗の菌液への接触要領を示す説明図である。
Hereinafter, Example 1 of the present invention will be described in detail.
FIG. 1 is an explanatory diagram showing the shape of bacteria of the Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) used in Example 1 after 3 days of agar culture.
FIG. 2 is an explanatory view showing the square petri dish agar medium for antibacterial activity test used in Example 1.
FIG. 3 is an explanatory view showing a contact point of the eggplant seedling of Example 1 to the bacterial solution.
図中、Ｐは４号ポット、Ｙは直径３０ｃｍ×深さ８ｃｍの円形状プラスチック容器、Ｎは根の先端を切ったナス苗、Ｓは４号ポットＰ内の培養土、Ｂはプラスチック容器Ｙ内の菌液、Ｋは抗菌活性試験用の長さ１４４ｍｍ×幅１０４ｍｍ×高さ１６ｍｍの角型シャーレ内の寒天培地、Ａは同培地に設けた内径８ｍｍで深さ５ｍｍの孔である。 In the figure, P is a No. 4 pot, Y is a circular plastic container having a diameter of 30 cm and a depth of 8 cm, N is an eggplant seedling with a root cut, S is a culture soil in the No. 4 pot P, and B is a plastic container Y The bacterial solution in the inside, K is an agar medium in a square petri dish of length 144 mm × width 104 mm × height 16 mm for antibacterial activity test, and A is a hole having an inner diameter of 8 mm and a depth of 5 mm provided in the medium.
（バチルス・チューリンジェンシスＮ−０２９株）
本発明に使用するバチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）の性状について説明する。
(Bacillus thuringiensis N-029 strain)
The properties of Bacillus thuringiensis N-029 strain (accession number FERM AP-21647) used in the present invention will be described.
生態学的性状
＊グラム染色 陽性
＊通性好気性
＊運動性 陽性
＊桿菌（単連鎖あり）
＊胞子形成（楕円形、芽胞位置 偏在）
＊胞子の横に不定形（いびつな球形、ダルマ型、ハート型のＰａｒａｓｐｏｒａｌ ｃｒｙｓｔａｌを作る（図１参照）
Ecological characteristics * Gram staining positive * facultative aerobic * motility positive * Neisseria gonorrhoeae (with single chain)
* Spore formation (oval, spore location unevenly distributed)
* Create an irregular shape (irregular sphere, dharma, heart-shaped parasporal crystal) next to the spore (see Fig. 1)
生化学的性状テストの結果は下記の表２のとおりである。 The results of the biochemical property test are shown in Table 2 below.
本発明に使用するバチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）の特性は下記の表３のとおりである。 The characteristics of Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) used in the present invention are as shown in Table 3 below.
本発明に使用するバチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）の寄託の情報
寄託所：独立行政法人産業技術総合研究所 特許生物寄託センター
受託番号：ＦＥＲＭ ＡＰ−２１６４７
分類学上の位置：バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）
識別のための表示：Ｎ−０２９
Information on deposit of Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) used in the present invention Depositary: National Institute of Advanced Industrial Science and Technology Patent Biological Depositary No .: FERM AP -21647
Taxonomic position: Bacillus thuringiensis
Indication for identification: N-029
（クロストリジュウム・サーモセルムＳＫ５２２菌株）
本発明に使用する好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）の性状について説明する。
(Clostridial thermocellum SK522 strain)
The properties of the thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar SK522, Meiko Kenjo No. 3459) used in the present invention will be described.
菌学的性質
旺盛なる繊維素分解力と弱又は微弱なるリグニン可溶化能を有する（随伴菌なしで単独では生育ができない）。
Mycological properties Has a vigorous fibrinolytic ability and weak or weak lignin solubilization ability (cannot grow alone without accompanying bacteria).
生態学的性状
＊グラム染色 陰性
＊嫌気性
＊直桿状（わずかに湾曲するものあり）。単独、ときどき２連。
＊運動性 見られない
＊胞子 末端に楕円、または円形の胞子を形成
Ecological properties * Gram staining Negative * Anaerobic * Straight (slightly curved) Single, sometimes twice.
* Mobility not seen * Spores Oval or circular spores formed at the ends
生理生化学的性質は次のとおりである。
酵素作用等
本菌は繊維素分解酵素を持ち、その酵素で繊維素の巨大分子の末端から分解していく。その性質を下記の表４に繊維素の基質を主体として示す。
Physiological and biochemical properties are as follows.
Enzyme action, etc. This fungus has a fibrin-degrading enzyme, which decomposes from the end of the fibrin macromolecule. The properties are shown in Table 4 below mainly using a fibrous substrate.
生産物試験
また、本菌は繊維素や他の物質を旺盛に分解して様々な産生物質を作る。下記の表５にそれらを示す。
Product test In addition, the bacterium actively decomposes fibrin and other substances to produce various product substances. They are shown in Table 5 below.
本菌の生育条件を下記の表６に示す。 The growth conditions of this bacterium are shown in Table 6 below.
本菌の特性は下記の表７のとおりである。 The characteristics of this bacterium are as shown in Table 7 below.
本発明に使用する好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）に関する寄託の情報
寄託所：微工研条寄（現：特許生物寄託センター）
受託番号：第３４５９号（ＦＥＲＭ ＢＰ−３４５９）
分類学上の位置：好熱性繊維素分解菌クロストリジュウム・サーモセルム（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ）
識別のための表示：ＳＫ−５２２
Deposit information regarding thermophilic fibrinolytic bacterium Clostridium thermocellum biovar SK522 (Mikken Kenjo No. 3459) used in the present invention Depositary: Miken Kenjo (currently: Patent Biological Depositary Center) )
Accession Number: No. 3459 (FERM BP-3459)
Taxonomic position: thermophilic fibrinolytic bacterium Clostridium thermocellum
Display for identification: SK-522
（サーマス・アクアティクスＳＫ５４２菌株）
本発明に使用する高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）の性状について説明する。
(Thermus Aquatics SK542 strain)
The properties of the thermophilic protein-degrading bacterium Thermus Aquatics SK542 ( Thermus aquaticus biovar SK542, Meiko Kenjo No. 3382) used in the present invention will be described.
菌学的性質
強いタンパク質分解力を持ち、黄色色素（カロチノイド系色素）を産生する。また、好熱性繊維素分解菌との共生培養によって、好熱性繊維素分解菌のリグニン可溶化能を顕著に増強する。
Mycological properties It has a strong proteolytic ability and produces yellow pigments (carotenoid pigments). Moreover, the lignin solubilization ability of a thermophilic fibrinolytic bacterium is remarkably enhanced by symbiotic culture with a thermophilic fibrinolytic bacterium.
生態学的性状
＊グラム染色 陰性
＊絶対好気性
＊長桿状、０．４〜０．６×３．０〜５．０μ（培養が古くなると糸状、長さ２０〜１３０μ）
＊鞭毛なし
＊運動性 見られない
＊内生胞子 なし
＊増殖 活発。ジェネレーションタイム２０〜５０分。
Ecological properties * Gram staining negative * Absolute aerobic * Long rod shape, 0.4-0.6 × 3.0-5.0μ (when the culture becomes old, filamentous, length 20-130μ)
* No flagella * Mobility not seen * Endospores None * Proliferation Active. Generation time 20-50 minutes.
生理生化学的性質は次のとおりである。 Physiological and biochemical properties are as follows.
本菌の様々な物質（酵素）に対する作用状況を下記の表８に示す。
酵素作用等
Table 8 below shows the action of this bacterium on various substances (enzymes).
Enzyme action
生産物試験
本菌の生産物試験の結果を下記の表９に示す。
Product test The results of the product test of this fungus are shown in Table 9 below.
生育条件
本菌の生育条件を下記の表１０に示す。
Growth conditions The growth conditions of this bacterium are shown in Table 10 below.
本菌の特性は下記の表１１のとおりである。 The characteristics of this bacterium are as shown in Table 11 below.
本発明に使用する高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）に関する寄託の情報
寄託所：微工研条寄（現：特許生物寄託センター）
受託番号：第３３８２号（ＦＥＲＭ ＢＰ−３３８２）
分類学上の位置：高熱性タンパク質分解菌サーマスアクアティックス（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ）
識別のための表示：ＳＫ−５４２
Deposit information regarding the thermophilic proteolytic bacterium Thermus Aquatics SK542 used in the present invention ( Thermus aquaticus biovar SK542, Mikenkenjo No. 3382) Depositary: Mikenkenjo (currently: Patent Biological Depositary Center)
Accession Number: No. 3382 (FERM BP-3382)
Taxonomic position: Thermus aquaticus ( Thermus aquaticus )
Display for identification: SK-542
菌の分離と培養
バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）のスクリーニング方法
試験に供する多くのバチルス・チューリンジェンシス菌を収集するため、日本各地で２１５個の土壌サンプルを採取した。その土壌サンプルから定法に従って６３株のバチルス・チューリンジェンシス菌を分離した。それらのバチルス・チューリンジェンシス菌を使って、ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）との抗菌活性試験を実施したところ、ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）に抗菌活性を示すバチルス・チューリンジェンシス菌が７菌株見出された。その中で東京都荒川区荒川自然公園で採取した土壌から分離したバチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）が最も強い抗菌活性を示したことからこの菌だけをスクリーニングしたものである。
以下、バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）のスクリーニング方法について説明する。
Bacterial isolation and culture Screening method for Bacillus thuringiensis N-029 strain (Accession number FERM AP-21647) In order to collect a large number of Bacillus thuringiensis bacteria to be tested Soil samples were collected. From the soil sample, 63 strains of Bacillus thuringiensis were isolated according to a conventional method. Using these Bacillus thuringiensis Jen cis bacteria, was carried antimicrobial activity test of the eggplant body half wilt pathogens (Verticillium dahliae NBRC9765), Bacillus Turin exhibiting antimicrobial activity eggplant half body wilt pathogens (Verticillium dahliae NBRC9765) Seven strains of Gensis were found. Among them, Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) isolated from soil collected at Arakawa Nature Park in Arakawa-ku, Tokyo showed the strongest antibacterial activity. Only screened.
The following describes the screening method of Bacillus thuringiensis Jen cis (Bacillusthuringiensis) N-029 strain (accession number FERM AP-21647).
バチルス・チューリンジェンシス菌の分離
サンプル土壌１ｇを容量５０ｍｌの遠心チューブに入れ、９ｍｌの滅菌水を加えてボルテックミキサーで１分間攪拌後1分間静置して、その上澄水５ｍｌを容量１５ｍｌの遠心チューブに分取し、６５℃のインキュベーターに３０分間静置して殺菌した。この液から１ｍｌを容量１５ｍｌの遠心チューブに分取し、滅菌水でこの液の１０３倍希釈溶液、１０４倍希釈溶液、１０５倍希釈溶液を作製し、その希釈溶液をそれぞれ２００μlずつ分取し、バチルス・チューリンジェンシス寒天培地（細菌用魚肉エキス１０ｇ、ポリペプトン１０ｇ、塩化ナトリウム２ｇ、寒天２０ｇを蒸留水に溶解し、全体を１リットルとしてオートクレーブ滅菌して作製）に塗抹し、インキュベーターで２８℃で３日間培養した。培養後、出現したコロニーを滅菌した竹串で一部取り、位相差顕微鏡下で１０００倍に拡大してクリスタルの有無を観察した。およそ視界内に８割以上のクリスタルが形成されている菌がバチルス・チューリンジェンシス菌であることから、６３菌株についてこれを確認した。
Isolation of Bacillus thuringiensis 1 g of sample soil was placed in a 50 ml centrifuge tube, 9 ml of sterilized water was added, stirred for 1 minute with a vortex mixer and allowed to stand for 1 minute, and 5 ml of the supernatant water was centrifuged at a capacity of 15 ml. The sample was collected in a tube and sterilized by standing in a 65 ° C. incubator for 30 minutes. 1 ml of this solution is dispensed into a centrifuge tube having a capacity of 15 ml, and 10 3 times diluted solution, 10 4 times diluted solution, and 10 5 times diluted solution of this solution are prepared with sterilized water, and 200 μl of each diluted solution is dispensed. Take a Bacillus thuringiensis agar medium (bacteria fish extract 10g, polypeptone 10g, sodium chloride 2g, agar 20g dissolved in distilled water, autoclave sterilized as a whole) and smear in an incubator. The cells were cultured at 28 ° C. for 3 days. After culturing, a part of the emerged colony was taken with a sterilized bamboo skewer and magnified 1000 times under a phase contrast microscope to observe the presence or absence of crystals. Since the bacteria in which 80% or more of the crystals are formed in the visual field are Bacillus thuringiensis, this was confirmed for 63 strains.
バチルス・チューリンジェンシス菌ストック菌液の作製
これら６３菌株のコロニーから、それぞれ滅菌した竹串でバチルス・チューリンジェンシス菌を寒天培地に画線培養し、単コロニーを分離した。その単コロニーから1白金耳を１ｍｌ滅菌水に懸濁し、うち２００μlをバチルス・チューリンジェンシス寒天培地に植菌し７日間培養した。
このようにして培地表面に生成した菌体をスパーテルで掻き取り、その菌体重量を測定、その重量の３倍の滅菌水に懸濁した菌液を作製、これをバチルス・チューリンジェンシス菌ストック菌液とし、合計６３菌株のストック菌液を作製した。
Preparation of Bacillus thuringiensis stock bacterial solution From these 63 colonies, Bacillus thuringiensis was streaked on an agar medium with sterilized bamboo skewers to isolate single colonies. One platinum loop from the single colony was suspended in 1 ml of sterilized water, 200 μl of which was inoculated on a Bacillus thuringiensis agar medium and cultured for 7 days.
The bacterial cells generated on the surface of the medium in this way are scraped with a spatula, the weight of the bacterial cells is measured, and a bacterial solution suspended in sterile water three times the weight is prepared. This is the Bacillus thuringiensis stock. A total of 63 stock bacterial solutions were prepared as the bacterial solution.
病原菌の培養
ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）をポテトデキストロースブロス培養液（DIFCO社製PDB培地２４ｇを蒸留水に溶解して全体を１リットルにしてオートクレーブ滅菌して作製）に１白金耳植菌し、１５０ｒｐｍ、２４℃で１４日間振盪培養した。このようにして得られた菌液を、滅菌した２重ガーゼで濾過して短菌糸菌液を作製、それをもとに、短菌糸菌液１０１倍、１０２倍、１０３倍希釈溶液を作製した。この菌液各２００μｌを滅菌コンラージでポテトデキストロースブロス寒天培地（DIFCO社製PDA培地３９ｇを蒸留水に溶解して全体を１リットルにしてオートクレーブ滅菌して作製）に塗抹し、２４℃にて３日間静置培養した後、菌数を測定した。その結果、ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）の短菌糸菌液中の菌数は約４×１０３／ｍｌであった。菌数が所定量だったため、この短菌糸菌液の原液をそのまま試験に供した。
Cultivation of pathogenic fungus Eggplant half-wilt pathogenic fungus ( Verticillium dahlia NBRC9765) in potato dextrose broth culture solution (prepared by autoclav sterilization by dissolving 24g of DIFCO PDB medium in distilled water) And cultured with shaking at 150 rpm and 24 ° C. for 14 days. The thus obtained bacterial solution, sterile double gauze produce filtered to short hyphae bacterial liquid, on the basis of it, the short hyphae bacterial suspension 10 x 1, 10 2 times, 10 3 fold dilutions Was made. 200 μl of each bacterial solution is smeared onto a potato dextrose broth agar medium (dissolved in 39 g of DIFCO PDA medium in distilled water and sterilized in an autoclave to make 1 liter) and sterilized at 3 ° C. for 3 days. After static culture, the number of bacteria was measured. As a result, the number of bacteria in the short mycelia solution of eggplant half body wilt pathogenic bacteria ( Verticillium dahlia NBRC9765) was about 4 × 10 3 / ml. Since the number of bacteria was a predetermined amount, this stock solution of short mycelia was used for the test as it was.
抗菌活性試験用混合培地
抗菌活性試験で使用した混合培地の組成、及び作成法を下記の表１２に示す。この培地はバチルス・チューリンジェンシス菌とナス半身萎凋病病原菌が丁度拮抗する培地組成である。角型シャーレ（長さ１４４ｍｍ×幅１０４ｍｍ×深さ１６ｍｍ）に下記の表１２に示す混合培地を４０ｍｌ注入して固化せしめた寒天培地に金属製のペニシリンカップ（外径８ｍｍ×長さ１０ｍｍ）で径８ｍｍ×深さ５ｍｍの孔Ａを２８個穿った抗菌活性試験用寒天培地Ｋを作製し、抗菌活性試験に供した（図２参照）。
Mixed Medium for Antibacterial Activity Test The composition of the mixed medium used in the antibacterial activity test and the preparation method are shown in Table 12 below. This medium has a medium composition in which Bacillus thuringiensis and eggplant half body wilt pathogens antagonize. In a square petri dish (length 144 mm x width 104 mm x depth 16 mm), 40 ml of the mixed medium shown in Table 12 below was injected and solidified on an agar medium solidified with a metal penicillin cup (outer diameter 8 mm x length 10 mm). An agar medium K for antibacterial activity test having 28 holes A having a diameter of 8 mm and a depth of 5 mm was prepared and used for the antibacterial activity test (see FIG. 2).
抗菌活性の判定方法
抗菌活性試験方法は、図２に示す孔Ａにバチルス・チューリンジェンシス菌液とナス半身萎凋病病原菌菌液を所定量混合して充填し、所定温度、所定時間培養して、どちらの菌がより強く増殖しているかで当該バチルス・チューリンジェンシス菌の抗菌活性の強弱を判定するものである。バチルス・チューリンジェンシス菌が強い抗菌活性があると、バチルス・チューリンジェンシス菌特有の茶褐色のコロニーが大きく現れ、逆にナス半身萎凋病病原菌の増殖が強い（逆に言えば、バチルス・チューリンジェンシス菌の抗菌活性がない）場合にはナス半身萎凋病病原菌特有の白色の菌糸が大きく増殖して現れる。それによってバチルス・チューリンジェンシス菌のナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）に対する抗菌活性の強弱が判定でき、最終的に最強のバチルス・チューリンジェンシス菌をスクリーニングすることができる。これらは全て目視による観察によって判定した。
Method for determining antibacterial activity The test method for antibacterial activity is to fill a hole A shown in FIG. 2 with a predetermined amount of Bacillus thuringiensis bacterial solution and eggplant half body wilt pathogen bacterial solution, and to incubate at a predetermined temperature for a predetermined time. The strength of the antibacterial activity of the Bacillus thuringiensis is determined based on which of the bacteria is growing stronger. When Bacillus thuringiensis has strong antibacterial activity, a brownish brown colony peculiar to Bacillus thuringiensis appears large, and conversely, the growth of pathogens of eggplant half body wilt is strong (conversely, Bacillus thuringiensis When there is no antibacterial activity of cis bacteria, white hyphae peculiar to eggplant half-wilt disease pathogens appear to grow greatly. Thereby, the strength of the antibacterial activity of the Bacillus thuringiensis bacterium against vermicillium dahlia NBRC9765 can be determined, and finally the strongest Bacillus thuringiensis bacterium can be screened. These were all determined by visual observation.
抗菌活性試験
２１５個のサンプル土壌から分離した６３菌株のバチルス・チューリンジェンシス菌のストック菌液を使って、それぞれの１００倍希釈溶液を作製し、その２５μｌと、短菌糸濃度を１００個／２５μlに調整したナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）菌液２５μlを混合し、図２に示す抗菌活性試験用寒天培地Ｋの孔Ａへ充填し、２４℃にて３日間培養した。培養後の孔中で、どの菌がより強く増殖しているかを目視観察した。全部で６３菌株のバチルス・チューリンジェンシス菌株を試験した結果、ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）に対して７菌株のバチルス・チューリンジェンシス菌が抗菌活性を有していることが分かった。その中でバチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）が半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）を強く抑制して増殖し、その褐色のコロニーを極めて大きく拡大していたことから最大の抗菌活性を有すると判定し、バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）をナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）に対して最も大きな抑制効果がある有効菌としてスクリーニングした。
Antibacterial activity test Using a stock bacterial solution of 63 strains of Bacillus thuringiensis isolated from 215 sample soils, a 100-fold diluted solution of each was prepared, 25 μl thereof, and a short hypha concentration of 100/25 μl. 25 μl of a solution of eggplant half body wilt disease pathogenic bacteria ( Verticillium dahliae NBRC9765) was mixed, filled into the hole A of the agar medium K for antibacterial activity test shown in FIG. 2, and cultured at 24 ° C. for 3 days. It was visually observed which bacteria were growing more strongly in the holes after the culture. As a result of testing a total of 63 Bacillus thuringiensis strains, it was found that 7 strains of Bacillus thuringiensis had antibacterial activity against the eggplant half body wilt pathogenic bacterium ( Verticillium dahliae NBRC9765) . Among them, Bacillus thuringiensis N-029 strain (accession number FERM AP-21647) proliferates by strongly suppressing the half body wilt pathogenic bacterium ( Verticillium dahlia NBRC9765), and the brown colony is greatly expanded. It was determined that it has the maximum antibacterial activity, and Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) was isolated against eggplant half body wilt pathogenic bacterium ( Verticillium dahllia NBRC9765) Screened as effective bacteria with the greatest inhibitory effect.
ナス半身萎凋病ポット試験
抗菌活性試験によりナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）に対して最も抗菌活性が大きく表れたバチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）について、単独で使用した場合と他の菌と複合して使用した場合とでナス半身萎凋病の抑制効果にどのような差異があるかを、実際の土（培養土）でナスを栽培してのポット試験で比較検討した。
Eggplant half body wilt pot test Bacillus thuringiensis N-029 strain (Accession number FERM AP-21647) which showed the most antibacterial activity against the eggplant half body wilt pathogen ( Verticillium dahlia NBRC9765) by the antibacterial activity test ) About the difference in the inhibitory effect of eggplant half body wilt disease when used alone and in combination with other bacteria, cultivated eggplant in the actual soil (cultured soil) All pot tests were compared.
ポット試験用菌液の作製
ポット試験に先立って、試験に必要なそれぞれの菌液を作製した。
Preparation of bacterial solution for pot test Prior to the pot test, each bacterial solution required for the test was prepared.
ＢＴ菌菌液
バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）をバチルス・チューリンジェンシス菌培養液( 魚肉エキス１０ｇ、ポリペプトン１０ｇを蒸留水に溶解して１０００ｃｃとし、オートクレーブ滅菌したもの)８ｍｌに植菌し、２８℃で、１２０rpm、１８時間、振盪培養で前培養（少量培養）した。その後、この培養液を８００ｍｌにスケールアップして２４時間、１２０rpm、２８℃で振盪培養してＢＴ菌菌液とした。
BT fungus solution Bacillus thuringiensis N-029 strain (Accession number FERM AP-21647) was dissolved in distilled water with 10 g of Bacillus thuringiensis culture solution (fish meat extract 10 g, polypeptone 10 g) , Autoclaved), inoculated into 8 ml, and precultured (small amount culture) by shaking culture at 28 ° C. and 120 rpm for 18 hours. Thereafter, this culture solution was scaled up to 800 ml and cultured with shaking at 120 rpm and 28 ° C. for 24 hours to obtain a BT fungus solution.
ＳＫ菌菌液
好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）、及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）を共生Ｅ培地（リン酸水素二カリウム５ｇ、尿素２ｇ、硫安２ｇ、ペプトン５ｇ、酵母エキス５ｇ、セルロース１５ｇ、炭酸カルシウム２ｇを蒸留水に溶解して１リットルとしてオートクレーブ滅菌して作製）で７日間培養した培養液をＳＫ菌菌液とした。
SK fungus solution Thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar SK522, Meiko Kenjo No. 3459), and thermophilic proteolytic fungus Thermus aquaticus SK542 ( Thermus aquaticus SK54) Kenjo No. 3382) was symbiotic with E medium (5g dipotassium hydrogen phosphate, 2g urea, 2g ammonium sulfate, 5g peptone, 5g yeast extract, 15g cellulose, 2g calcium carbonate in distilled water and autoclaved as 1 liter. The culture solution cultivated for 7 days was used as the SK fungus solution.
病原菌菌液
ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）を前記のポテトデキストロースブロス液体培地（ＰＤＢ培地）６００ｍｌに植菌し、１５０ｒｐｍ、１４日振盪培養した。この液を二重ガーゼで２回濾過し、濾液を遠心機で３０００rpm、５分、４℃で遠心し、集菌後、２回滅菌水で遠心洗浄した。その後、ペレットの湿重を測定し、５．００ｇであったため、１０倍量の滅菌水５０ｍｌに懸濁し、ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）原液とした。菌原液の菌数を前記のポテトデキストロースブロス寒天培地（ＰＤＡ培地）で測定した結果、菌数２×１０７ｓｐｏ/ｍｌであったので、滅菌水で希釈して１×１０６ｓｐｏ／ｍｌに調整した液を作製、この病原菌菌液をポット試験用ナスへの病原菌の接種源とした。ｓｐｏは胞子数の単位である。
これらの菌液はポット試験前に全てを作製、準備して試験に供した。
Pathogenic fungus solution Eggplant half-wilt pathogenic fungus ( Verticillium dahlia NBRC9765) was inoculated into 600 ml of the potato dextrose broth liquid medium (PDB medium) and cultured with shaking at 150 rpm for 14 days. This solution was filtered twice with double gauze, and the filtrate was centrifuged with a centrifuge at 3000 rpm, 5 minutes, 4 ° C., collected and centrifuged twice with sterilized water. Thereafter, the wet weight of the pellet was measured, and it was 5.00 g. Therefore, the pellet was suspended in 50 ml of 10-fold sterilized water to prepare a stock solution of eggplant half body wilt pathogen ( Verticillium dahlia NBRC9765). As a result of measuring the number of bacteria in the stock solution with the above-mentioned potato dextrose broth agar medium (PDA medium), the number of bacteria was 2 × 10 7 spo / ml, so it was diluted with sterilized water to 1 × 10 6 spo / ml. An adjusted solution was prepared, and this pathogen fungus solution was used as an inoculation source of the pathogen to the eggplant for pot test. spo is a unit of the number of spores.
All of these bacterial solutions were prepared and prepared before the pot test.
ポット試験
前記で作製した菌液を使用してポット試験を実施した。ポット試験の手順を日程に従って説明する。
Pot test A pot test was carried out using the bacterial solution prepared above. The pot test procedure will be explained according to the schedule.
ポット試験用ナス苗の育苗(平成２０年４月８日〜６月１８日)
タキイ種まき培土（
野菜・草花全般）をオートクレーブで滅菌し、３×３×５ｃｍ１２８マスの苗箱に敷き詰め、４月８日に播種、５月１９日に清新産業株式会社製のＪＡくみあい園芸培養土をオートクレーブで滅菌した滅菌培養土を充填した４号ポットＰに移植し、本葉５〜６葉期になるまで育苗したものを使用した。
Raising eggplant seedlings for pot test (April 8th-June 18th, 2008)
Takii seed sowing medium (
Vegetables and flowers) are sterilized in an autoclave and spread in a 3 x 3 x 5 cm 128 square seedling box. Seeded on April 8 and sterilized on May 19th by JA Kumiai Horticultural Cultivated Soil by Seishin Sangyo Co., Ltd. The plant was transplanted to No. 4 pot P filled with the sterilized cultured soil and grown up to the 5th to 6th leaf stage.
バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）、及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）、及びナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）の接種(平成２０年６月１８、１９日) Bacillus thuringiensis N-029 strain (accession number FERM AP-21647), thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar SK522, Microtechnical Institute No. 34 Inoculation with hyperthermolytic protease Thermus aquatics SK542 ( Thermus aquaticus biovar SK542, Meiko Kenjo No. 3382) and eggplant half body wilt pathogen ( Verticillium dahlia NBRC9765) (June 18, 19th, 2008)
ナス苗根部の先端切断
ナス苗を４号ポットＰより取り出し、水道水で根を洗浄し、根の先端を約５分の１、アルコールで滅菌したハサミで切断した。
Tip cuttings of eggplant seedling roots Eggplant seedlings were taken out from No. 4 pot P, the roots were washed with tap water, and the tips of the roots were cut with scissors sterilized with alcohol about 1/5.
ナス苗根部のＢＴ菌菌液への接触
ＢＴ菌菌液２００ｃｃをプラスチック製の円形のプラスチック容器Ｙ（直径３０ｃｍ×深さ８ｃｍ）に入れ、根の先端を切ったナス苗Ｎの根全体を円形のプラスチック容器ＹのＢＴ菌菌液に１０分間浸した後、再び４号ポットＰに戻して、滅菌した新しい培養土をポット上端まで充填し、十分散水した（図３参照）。
翌日、再びナス苗Ｎを４号ポットＰより取り出し、前日の手順に従ってナス苗根部を別のプラスチック製の円形のプラスチック容器Ｙ（直径３０ｃｍ×深さ８ｃｍ）に入れた、ナス半身萎凋病病原菌菌液２００ｃｃに１０分間浸し、再度４号ポットＰに戻し、新しい培養土Ｓをポット上端まで充填し、十分散水した。試験の概要を図３に示す。
Contact of the eggplant seedling root with the BT fungus solution 200 cc of the BT fungus solution is placed in a plastic circular plastic container Y (diameter 30 cm × depth 8 cm), and the whole root of the eggplant seedling N cut from the tip of the root is circular. After immersing in the BT bacterium solution of the plastic container Y for 10 minutes, it was returned to the No. 4 pot P, filled with sterilized fresh culture soil up to the upper end of the pot, and fully dispersed (see FIG. 3).
The next day, the eggplant seedling N is again taken out from No. 4 pot P, and the eggplant seedling root is placed in another plastic circular plastic container Y (diameter 30 cm × depth 8 cm) according to the procedure of the previous day. It was immersed in 200 cc of the solution for 10 minutes, returned to the No. 4 pot P again, filled with fresh culture soil S to the upper end of the pot, and fully dispersed water. The outline of the test is shown in FIG.
ナス苗根部のＳＫ菌菌液への接触
ＳＫ菌菌液２００ｃｃを図３の円形のプラスチック容器Ｙに入れ、根の先端を切ったナス苗Ｎの根全体を円形のプラスチック容器Ｙの菌液に１０分間浸した後、再び４号ポットＰに戻して、滅菌した新しい培養土Ｓをポット上端まで充填し、十分散水した。
翌日、再びナス苗Ｎを４号ポットＰより取り出し、前日の手順に従ってナス苗根部を図３の円形のプラスチック容器Ｙに入れた、ナス半身萎凋病病原菌菌液２００ｃｃに１０分間浸し、再度４号ポットＰに戻し、滅菌した新しい培養土Ｓをポット上端まで充填し、十分散水した。
Contacting the eggplant seedling root with the SK fungus solution 200 cc of the SK fungus solution is placed in the circular plastic container Y of FIG. After soaking for 10 minutes, it was returned to the No. 4 pot P again, and sterilized fresh culture soil S was filled up to the upper end of the pot and fully dispersed.
The next day, the eggplant seedling N is again taken out from the No. 4 pot P, and the eggplant seedling root is immersed in 200 cc of eggplant half body wilt pathogenic bacteria solution in the circular plastic container Y of FIG. The pot was returned to the pot P, and sterilized fresh culture soil S was filled up to the upper end of the pot and fully dispersed.
ＢＴＳＫ混合菌液の作製、及びナス苗根部のＢＴＳＫ混合菌液への接触
図３の円形のプラスチック容器ＹにＢＴ菌菌液１００ｃｃと、ＳＫ菌菌液１００ｃｃを入れてよく混合し、ＢＴＳＫ菌液を作製した。この混合菌液２００ｃｃに先端を切ったナス苗Ｎの根全体を１０分間浸した後、再び４号ポットＰに戻して、滅菌した新しい培養土Ｓをポット上端まで充填し、十分散水した。
翌日、再びナス苗Ｎを４号ポットＰより取り出し、前日の手順に従ってナス苗根部を図３の円形のプラスチック容器Ｙに入れた、ナス半身萎凋病病原菌菌液２００ｃｃに１０分間浸し、再度４号ポットＰに戻し、滅菌した新しい培養土Ｓをポット上端まで充填し、十分散水した。
Preparation of BTSK mixed bacterial solution and contact of eggplant seedling root with BTSK mixed bacterial solution 100 cc of BT bacterial bacterial solution and 100 cc of SK bacterial bacterial solution are placed in the circular plastic container Y of FIG. Was made. The whole root of eggplant seedling N cut off at the tip of this mixed bacterial solution 200 cc was soaked for 10 minutes, and then returned to No. 4 pot P again, and sterilized fresh culture soil S was filled up to the upper end of the pot and fully dispersed with water.
The next day, the eggplant seedling N is again taken out from the No. 4 pot P, and the eggplant seedling root is immersed in 200 cc of eggplant half body wilt pathogenic bacteria solution in the circular plastic container Y of FIG. The pot was returned to the pot P, and sterilized fresh culture soil S was filled up to the upper end of the pot and fully dispersed.
ナス苗根部の化学農薬希釈液への接触
病害の発生に対して効果の比較のためにタケダ園芸製の化学農薬ベンレート水和剤（粉末）の１０００倍液を使用した。調整したナス半身萎凋病病原菌菌液を図３の円形のプラスチック容器Ｙに２００ｃｃ入れ、この菌液に根の先端を切ったナス苗Ｎの根全体を１０分間浸した後、再び４号ポットＰに戻して、滅菌した新しい培養土Ｓをポット上端まで充填し、十分散水した。
翌日、ポットより再びナス苗Ｎを掘り起こして取り出し、図３の円形のプラスチック容器Ｙにベンレート水和剤の１０００倍液を２００ｃｃ入れ、このベンレート希釈液に根の先端を切ったナス苗Ｎの根全体を１０分間浸した後、再び４号ポットＰに戻して、滅菌した新しい培養土Ｓをポット上端まで充填し、十分散水した。
Contact of eggplant seedling root with chemical pesticide dilution solution For comparison of the effect on the occurrence of disease, a 1000 times solution of chemical pesticide benrate wettable powder (powder) manufactured by Takeda Horie was used. 200 cc of the adjusted eggplant half-wilt disease pathogen fungus solution is placed in the circular plastic container Y of FIG. 3, and the whole root of the eggplant seedling N with the root tip cut off is immersed in this fungus solution for 10 minutes. The sterilized fresh culture soil S was filled up to the upper end of the pot and fully dispersed with water.
The next day, the eggplant seedling N is dug up and taken out from the pot again, and 200 cc of a 1000-fold solution of benrate wettable powder is placed in the circular plastic container Y of FIG. After immersing the whole for 10 minutes, it was returned to the No. 4 pot P again, filled with sterilized fresh culture soil S up to the upper end of the pot, and fully dispersed water.
これらの試験の概要を下記の表１３に示す。 A summary of these tests is shown in Table 13 below.
表中○はそれぞれの菌の使用、×は不使用を表す。各試験区の内容はブランク区、バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）菌区（ＢＴ菌区）、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）、及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）共生菌区（ＳＫ菌区）、バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）と、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）、及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）の混合菌区（ＢＴＳＫ菌区）、化学農薬ベンレート水和剤（粉末）の１０００倍液施用区（化学農薬区）、ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）区（病原菌区）の６試験区で、各試験区について1区画５ポット（１ポットにナス苗１株栽培）ずつを試験した。各試験区のナス苗はブランク区を除いて防除のための菌液に接触させた後、前述で調整したナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）菌液に浸したが、化学農薬区のみは通常の化学農薬施用の慣例に従って、ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）菌液に浸した翌日に化学農薬ベンレートの１０００倍液に接触させて試験した。 In the table, ○ indicates the use of each bacterium, and × indicates non-use. The contents of each test plot are blank, Bacillus thuringiensis N-029 strain (accession number FERM AP-21647) strain (BT strain), thermophilic fibrinolytic clostridium thermocellum SK522 ( Clostridium thermocellum biovar SK522, Mikenkenjou No. 3459), and thermophilic proteolytic thermus aquaticus SK542 ( Thermus aquaticus biovar SK542, Mikkokenjou No.3382) Bacillus thuringiensis Jen cis (Bacillus thuringiensis) N-029 strain (the accession number FERM AP-21647), thermophilic cellulose decomposing bacteria clostripain Jewelery · Samoserumu SK522 (Clostridium thermocellum biovar SK522, FERM BP-3459), and thermophilic protein-degrading bacterium Thermus aquaticus SK542 (Thermus aquaticus biovar SK542, FERM BP-3382) mixed bacteria District ( BTSK fungus group), chemical pesticide benrate wettable powder (powder) 1000 times liquid application group (chemical pesticide group), eggplant half body wilt pathogenic fungus ( Verticillium dahlia NBRC9765) group (pathogen group) Each section was tested in 5 pots (1 eggplant seedling cultivation per pot). The eggplant seedlings in each test group were contacted with the bacterial solution for control except for the blank group, and then immersed in the eggplant half body wilt pathogenic fungus ( Verticillium dahliae NBRC9765) prepared as described above. According to the usual practice of chemical pesticide application, the test was carried out by contacting with 1000 times the chemical pesticide benrate the next day after soaking in the solution of eggplant half body wilt pathogen ( Verticillium dahlia NBRC9765).
１．判定(平成２０年７月１０日)
接種より、２１日後の7月１０日にそれぞれのナス苗の葉を１枚ずつ病徴調査を実施、それぞれの葉の黄変・褐変、枯死程度により、下記の表１４に示すような判定基準に従って０〜４の発病指数で病徴を判定し、各株ずつそれを集計して下記の計算式に従って発病度、及び防除価を算出した。
1. Judgment (July 10, 2008)
On July 10th, 21 days after the inoculation, a symptom survey was conducted on each leaf of each eggplant seedling, and the criteria shown in Table 14 below were determined according to the degree of yellowing, browning, and death of each leaf. Symptoms were determined with a disease index of 0 to 4 according to the above, and each strain was counted and the disease severity and control value were calculated according to the following formula.
発病度＝Σ(発病指数別株数×指数)／(４×調査株数)×１００
防除価＝１００−（当該菌区発病度／病原菌区発病度）×１００
下記の表１５に試験結果を示す。
Disease severity = Σ (number of strains by disease index × index) / (4 × number of strains) × 100
Control value = 100− (degree of disease occurrence / degree of disease) × 100
Table 15 below shows the test results.
試験結果
ブランク区（試験番号０）はこの試験区で栽培した５ポットのナス苗の根部が菌との接触の全くない試験区で、発病は全く見られなかった。試験土壌が健全な土壌であったことが分かる。
一方、ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）菌液のみにナス苗を接触させた病原菌区（試験番号５）は６５．２の発病度を示した。当該病原菌がナス苗に対して強い感染力を持っていたことがわかる。
これに対してナス半身萎凋病を抑制する効果を目的にＢＴ菌液、ＳＫ菌液、ＢＴＳＫ菌液にナス苗を接触させて、しかる後にナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）に接触させた各試験区は使用した材料によって発病の抑制効果にバラツキがあった。この試験方法はナス苗がナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）に感染する前に有効菌を施用して、ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）の感染発病を防止する効果を見るために実施した試験方法である。
その結果、ＢＴ菌区（試験番号１）は、抗菌活性試験でナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）に対して強い抗菌活性を持つことが明らかになったバチルス・チューリンジェンシス菌だけの試験区で、ポット試験でも４７．１の防除価が得られた。
また、ＳＫ菌区（試験番号２）は好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）、及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）を共生培養して得られた共生複合菌の培養液を使用した試験区で、３０．７の防除価が得られた。ＳＫ共生菌は植物の生育を促し、強い体質の植物を成長させるのに大きな効果を持つ共生菌で、植物を強壮にすることにより耐病性を付与する働きを持つ。その結果、バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）を使用したＢＴ菌区には及ばないものの、３０．７の防除価を示した。
Test results The blank group (test number 0) was a test group in which the roots of 5 pot eggplant seedlings grown in this test group had no contact with the fungus, and no disease was observed. It can be seen that the test soil was healthy.
On the other hand, the pathogenic fungus group (test number 5) in which the eggplant seedlings were contacted only with the bacterial solution of eggplant half body wilt disease ( Verticillium dahlia NBRC9765) showed a disease severity of 65.2. It can be seen that the pathogen had a strong infectivity against eggplant seedlings.
On the other hand, eggplant seedlings were brought into contact with the BT bacterial solution, SK bacterial solution, and BTSK bacterial solution for the purpose of suppressing eggplant half-wilt disease, and then contacted with eggplant half-wilt disease germ ( Verticillium dahliae NBRC9765). Each test group had variations in the disease control effect depending on the materials used. This test method by applying an effective bacteria before the eggplant seedlings infected eggplant half body wilt pathogens (Verticillium dahliae NBRC9765), in order to see the effect of preventing infection pathogenesis eggplant half body wilt pathogens (Verticillium dahliae NBRC9765) This is the test method performed.
As a result, the BT fungus group (Test No. 1) was tested only for Bacillus thuringiensis, which showed that the antibacterial activity test showed strong antibacterial activity against the eggplant half body wilt pathogen ( Verticillium dahliae NBRC9765). The control value of 47.1 was also obtained in the pot in the section.
In addition, the SK fungus group (Test No. 2) is a thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar SK522; A control value of 30.7 was obtained in a test section using a culture solution of a symbiotic complex obtained by co-culturing Thermus aquaticus biovar SK542, Mikken Kenjo No. 3382). The SK symbiotic fungus is a symbiotic fungus that promotes the growth of a plant and has a great effect on growing a strong plant, and has a function of imparting disease resistance by strengthening the plant. As a result, the control value of 30.7 was shown although it did not reach the BT fungal area using Bacillus thuringiensis N-029 strain (Accession number FERM AP-21647).
しかし、これらの有効菌を個別に使用した試験結果（試験番号１、２）は、確かに病原菌区に比較して発病率は抑えられたものの、ベンレート水和剤の１０００倍液を施用した化学農薬区（試験番号４）の防除価８０．７に比べると防除価は半分以下でそれほど大きな効果には至っていない。これに比較してバチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）菌と好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）、及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）共生菌を混合して使用した試験区（試験番号３、ＢＴＳＫ菌区）は防除価が６８．７で、化学農薬区（ベンレート１０００倍液施用）には及ばないものの、ＢＴ菌区（防除価４７．１）、ＳＫ菌区（防除価３０．７）よりも高い防除価を示した。バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）と、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）、及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）共生菌との複合的な相乗効果によりかなり発病度が低下することが明らかとなった。
However, the test results (
ここでバチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）が、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）、及び高熱性タンパク質分解菌サーマス・アクアティクスＳＫ５４２（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２、微工研条寄第３３８２号）共生菌との混合により、ナス半身萎凋病病原菌（Ｖｅｒｔｉｃｉｌｌｉｕｍ ｄａｈｌｉａｅ ＮＢＲＣ９７６５）に対して強い抗菌活性を持つ結果となった、これらの複合菌の相乗効果について述べる。 Here, Bacillus thuringiensis N-029 strain (Accession No. FERM AP-21647) is a thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 (Crostridium thermocellum biovar SK522, Koken SK ), And the hyperthermolytic proteolytic thermus aquatics SK542 ( Thermus aquaticus biovar SK542, Meiko Kenjo No. 3382) symbiotic bacteria, it is antibacterial against Verticillium dahlia NBRC9765 The synergistic effect of these complex bacteria that resulted in activity will be described.
ＳＫ共生菌の相乗効果
嫌気性の好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ
ＳＫ５２２、微工研条寄第３４５９号）と、絶対好気性の高熱性タンパク質分解菌サーマス・アクアティックス（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２ 受託番号ＦＥＲＭ ＢＰ−３３８２）との共生培養の特殊効果について説明する。
好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）と高熱性タンパク質分解菌サーマス・アクアティックス（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２ 受託番号ＦＥＲＭ ＢＰ−３３８２）の高温環境下での共生培養物は、嫌気性の好熱性繊維素分解菌ＳＫ５２２菌株単独ではほとんど不可能な天然の難分解性有機物の分解発酵を顕著に増強する。以下、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ
ＳＫ５２２、微工研条寄第３４５９号）をＳＫ５２２、絶対好気性の高熱性タンパク質分解菌サーマス・アクアティックス（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２ 受託番号ＦＥＲＭ ＢＰ−３３８２）をＳＫ５４２と略記する。
Synergistic effect of SK symbiotic bacteria Anaerobic thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar
The special effects of co-cultivation with SK522, Jikoken Hoyori No. 3459) and the absolute aerobic hyperthermolytic proteolytic thermus aquatics ( Thermus aquaticus biovar SK542 accession number FERM BP-3382) will be described.
Thermophilic cellulose degrading bacteria Kurosuto Rijuumu-Samoserumu SK522 of (Clostridium thermocellum biovar SK522, FERM BP-3459) and a high fever protein-degrading bacterium Thermus Aquatics (Thermus aquaticus biovar SK542 accession number FERM BP-3382) A symbiotic culture under a high temperature environment remarkably enhances the degradation fermentation of natural persistent organic substances, which is almost impossible with the anaerobic thermophilic fibrinolytic SK522 strain alone. Hereinafter, thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar)
SK522, Meiko Kenjo No. 3459) is abbreviated as SK522, an absolute aerobic hyperthermolytic proteolytic thermus aquatics ( Thermus aquaticus biovar SK542 accession number FERM BP-3382).
まず、ＳＫ５４２菌株の増殖はＳＫ５２２菌株の欠如するタンパク分解活性を補充する。しかも、イナワラ、ムギワラ、モミガラ、牧草等の堆・厩肥をはじめ廃木材、都市下水汚泥等の難分解性有機物の分解発酵は、すべて固体発酵の状態で、かなり通気のよい環境下に置かれている。そして、初期の段階におけるＳＫ５４２菌株の増殖は発酵過程にある有機物を高温嫌気的環境へと変移せしめ、これによって通常の自然環境ではなかなか単独での増殖が困難なＳＫ５２２菌株の自然界での生育が可能となる。 First, growth of the SK542 strain supplements the proteolytic activity that the SK522 strain lacks. Moreover, the decomposition fermentation of indegradable organic matter such as compost and manure such as inowara, wheat straw, rice straw, and pasture, waste wood, and municipal sewage sludge is all in the state of solid fermentation in a fairly well-ventilated environment. Yes. The growth of the SK542 strain at the initial stage changes the organic matter in the fermentation process to a high temperature anaerobic environment, which makes it possible to grow the SK522 strain in nature, which is difficult to grow alone in a normal natural environment. It becomes.
リグニンの可溶化
ここで、特に注目されることは、ＳＫ５４２菌株によってＳＫ５２２菌株のリグニン可溶化能が顕著に増強されることである。難分解性有機物は、実際には繊維素がリグニン、その他の有機成分と強く結合した状態で存在する。こうした天然のリグノセルロースは繊維素分解力の旺盛なるＳＫ５２２菌株であっても、その単独施用ではほとんど分解が不可能である。リグニンの化学構造は、未だ完全に明らかにされていないが、ベンゼン環に炭素数３つの側鎖を持つフェニルプロパンが基本単位になって、この単位体がパーオキシダーゼによって触媒されるラジカル反応によりランダムに三次元的に重合した高分子化合物である。微生物の分解に対する抵抗がきわめて強く、きわめて分解が困難であることはよく知られている。ところが、本願発明で使用するＳＫ５２２菌株は弱または微弱ではあるが、リグニンを可溶化する。そして、このリグニン可溶化能が、ＳＫ５４２菌株と共生培養することによって、たとえば、イナワラリグニンの５０％以上が１０日前後の短日時において可溶化されることが分かっている。その作用効果は５０〜８０℃以上という高温期段階で発揮され、分解発酵へその腐植化が急速に進行する。
Here, it is particularly noted that the lignin solubilization ability of the SK522 strain is significantly enhanced by the SK542 strain. The hardly decomposable organic substance actually exists in a state where the fiber is strongly bound to lignin and other organic components. Even if such natural lignocellulose is the SK522 strain with a strong fibrinolytic ability, it cannot be almost decomposed by its single application. The chemical structure of lignin has not yet been fully clarified, but phenylpropane having a side chain with 3 carbon atoms on the benzene ring is the basic unit, and this unit is randomly generated by a radical reaction catalyzed by peroxidase. It is a polymer compound polymerized three-dimensionally. It is well known that the resistance to microbial degradation is extremely strong and extremely difficult to degrade. However, the SK522 strain used in the present invention is weak or weak, but solubilizes lignin. And it is known that 50% or more of Inawara lignin is solubilized in a short time around 10 days, for example, by co-culturing this lignin solubilizing ability with SK542 strain. The effect is exhibited at a high temperature stage of 50 to 80 ° C. or higher, and the humification progresses rapidly to decomposition fermentation.
黄色色素の産生
ＳＫ５４２菌株をはじめ、サーマス属（ｇｅｎｕｓ Ｔｈｅｒｍｕｓ）の産生する黄色色素はカロチノイド系不溶性色素である。菌体中のこの黄色色素が他の微生物等による分解を受け、水溶性の低分子量体のものとなり、植物体に吸収され、必要部位に移行し、丁度都合のよい前駆物質となる。すなわち、このような黄色色素が、ほかの溶菌した細胞内容物や分泌物、それに発酵生産物等とともに根茎葉部位の増大繁茂を促すだけでなく、花芽の形成、着花、果実の肥大等の生殖生長の代謝系に深く関与しており、果実の味、色沢、貯蔵性等品質向上に大きな役割をもつ。
Production of yellow pigments Yellow pigments produced by the genus Thermus, including the SK542 strain, are carotenoid insoluble pigments. This yellow pigment in the fungus body is decomposed by other microorganisms, becomes a water-soluble low molecular weight substance, is absorbed by the plant body, migrates to a necessary site, and becomes just a convenient precursor. That is, such a yellow pigment not only promotes the growth of rhizomes with other lysed cell contents and secretions, and fermentation products, but also forms flower buds, flowering, fruit enlargement, etc. It is deeply involved in the metabolic system of reproductive growth and plays a major role in improving the quality of fruits such as taste, color and storage.
微生物生態系の混合複合化
また、この黄色色素を含むＳＫ５４２菌株の菌体を施用すると、それを基質として繁殖する一般の従属栄養微生物、土壌有効菌等の増殖を促す。さらに重要なことは、堆肥の製造現場、あるいは自然界の植物体の腐植化の過程では、ＳＫ５２２菌株のような好熱性微生物が、すぐに増殖活性化するわけではない。当初は、一般の常温性従属栄養微生物が増殖し、これらの作用による発酵熱が蓄積されて品温の上昇に役立つものであり、単一菌株の好熱性繊維素分解菌や少数の微生物のみが、天然の難分解性有機資材の腐植化に関与するわけではない。微生物フローラの混合複合化、多様化が必要で、様々なタイプの有効菌が多く、さらに「エサ（基質）」も適度にあるという複合的内容が望ましい。こうして、難分解性有機物の高温分解は、ＳＫ５２２菌株を中心とする細菌フローラがまず形成され、その腐植化過程が単純な構成成分の変化だけでなく、きわめて複雑な微生物フローラの相互作用やその変遷等が深い関わりをもって最も効率よく進行するのである。
Mixed complexation of microbial ecosystems When the SK542 strain containing this yellow pigment is applied, it promotes the growth of general heterotrophic microorganisms, soil effective bacteria, etc. that propagate using the SK542 strain as a substrate. More importantly, thermophilic microorganisms such as the SK522 strain are not immediately proliferated and activated in the compost production site or in the process of humification of natural plants. Initially, ordinary thermophilic heterotrophic microorganisms proliferate, and the heat of fermentation due to these actions accumulates to help increase the product temperature. Only a single strain of thermophilic fibrinolytic bacteria and a few microorganisms It is not involved in the humification of natural persistent organic materials. It is desirable to have a complex content in which mixed microbial flora needs to be mixed and diversified, there are many different types of effective bacteria, and there is also a moderate amount of “food (substrate)”. In this way, high-temperature decomposition of persistent organic substances, bacterial flora centered on SK522 strain is first formed, and the humus process is not only a simple component change, but also the interaction and transition of extremely complex microbial flora Etc. proceed most efficiently with deep involvement.
このように、好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）と、高熱性タンパク質分解菌サーマス・アクアティックス（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２ 受託番号ＦＥＲＭ ＢＰ−３３８２）の高温環境下での共生培養により増殖した共生菌は植物繊維素の軟らかいセルロース部位は勿論、難分解性のリグニンをも高効率で分解するために土壌に残留せる繊維素の分解効率を高め、腐植を増大せしめ、土壌地力を増強するのに大きな役割を果たす。
また、前述しているように高熱性タンパク質分解菌サーマス・アクアティックス（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２ 受託番号ＦＥＲＭ ＢＰ−３３８２）の産生するカロチノイド系の黄色色素は他の微生物等による分解を受け、ほかの溶菌した細胞内容物や分泌物、それに発酵生産物等とともに植物の自立強壮体質への向上に大きな役割をもつ。即ち、病気に罹患することが少ない植物体を育成する力を持っているのである。
Thus, the thermophilic fibrinolytic bacterium Clostridium thermocellum SK522 ( Clostridium thermocellum biovar SK522, Meiko Kenjo No. 3459) and the thermophilic proteolytic bacterium Thermus aquaticus No. In BP-3382), symbiotic bacteria grown by co-cultivation under high temperature environment are not only soft cellulose parts of plant fibrin but also fibrin remaining in the soil in order to decompose highly degradable lignin with high efficiency. It plays a major role in increasing efficiency, increasing humus, and enhancing soil strength.
In addition, as mentioned above, the carotenoid yellow pigment produced by thermus aquaticus biovar SK542 accession number FERM BP-3382 is subject to degradation by other microorganisms, etc. Along with lysed cell contents and secretions, fermented products, etc., it plays a major role in improving the self-sustaining tonic nature of plants. In other words, it has the power to grow plants that are less likely to suffer from diseases.
バチルス・チューリンジェンシス菌の他の菌との相乗効果
このように好熱性繊維素分解菌クロストリジュウム・サーモセルムＳＫ５２２（Ｃｌｏｓｔｒｉｄｉｕｍ ｔｈｅｒｍｏｃｅｌｌｕｍ ｂｉｏｖａｒ ＳＫ５２２、微工研条寄第３４５９号）と、高熱性タンパク質分解菌サーマス・アクアティックス（Ｔｈｅｒｍｕｓ ａｑｕａｔｉｃｕｓ ｂｉｏｖａｒ ＳＫ５４２ 受託番号ＦＥＲＭ ＢＰ−３３８２）の高温環境下での共生培養により増殖した共生菌による植物体そのものの基礎体力の増強と相まって、バチルス・チューリンジェンシス（Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）Ｎ−０２９株（受託番号 ＦＥＲＭ ＡＰ−２１６４７）菌によるナス半身萎凋病の抑制効果は一層増強される。
Synergistic effect of Bacillus thuringiensis with other bacteria Thus, thermophilic fibrinolytic bacterium Clostridium thermocellum biovar SK522, Mikken Kenjo No. 3459, and thermophilic proteolytic bacteria Combined with the enhancement of the basic physical strength of the plant body by the symbiotic bacteria grown by the symbiotic culture of Thermus aquatics ( Thermus aquaticus biovar SK542 accession number FERM BP-3382) in a high temperature environment, Bacillus thuringiensis ( Bacillus thuringiensis) ) The inhibitory effect on eggplant half body wilt caused by N-029 strain (Accession No. FERM AP-21647) is further enhanced.
ここで、生物的防除がいかに環境保全的な特性を持ちながら農業の健全化に寄与しているか、その特性が何に由来するものであるか、そのために生物的防除はどのような要件を満たさなければならないかを述べる。 Here, how biological control contributes to the health of agriculture while having environmentally friendly characteristics, and what the characteristics originate from, and therefore what requirements does biological control meet? Describe what must be done.
化学農薬は即効性があり、病害菌に対して劇的な効果があり、安価で使い易いという、農業従事者にとって極めて大きな利点を有する。しかし、その利点が逆に弊害をも生み出す欠点にもなっている。そして、それは地球環境破壊、生態系の破壊という人類の生存にとって致命的な欠点である。化学農薬は害虫や病原菌の壊滅を劇的にするために毒性の強い人工的な薬剤を含んでいる場合が多く、その特殊性の故にオゾン層を破壊して地球環境の危険性を増幅したり、また、一般の生物にもその毒性が強く作用する場合が多く、生態系の破壊という危険な因子を内包している。そのことは即、我々人類の生命の存亡にかかわる重大な因子となる場合がしばしばである。
また、化学農薬を使い続けることによって、有害生物、有害病害菌とともにミミズなどの有効生物、有効微生物までも絶滅しかねない弊害を持ち、圃場そのものの疲弊につながる危険性が大きい。最近のあいつぐ化学農薬の規制はこのような化学農薬の危険な実態が根底にある。我々はこのような実態に深く思いを廻らし、今こそ恒久的に地球環境保全、生態系の破壊防止のために安全な農業資材の使用という命題に全力を傾注しなければならない。
Chemical pesticides have immediate benefits, have a dramatic effect on disease-causing bacteria, and have tremendous advantages for farmers, being cheap and easy to use. However, the advantage is also a disadvantage that also causes adverse effects. And it is a fatal defect for the survival of mankind, the destruction of the global environment and the destruction of the ecosystem. Chemical pesticides often contain highly toxic artificial drugs to dramatically destroy pests and pathogens, and because of their peculiarities, they destroy the ozone layer and amplify the dangers of the global environment. In addition, the toxicity of many common organisms often acts strongly, and it contains a dangerous factor that destroys the ecosystem. That is often a critical factor in the immediate existence of our lives.
In addition, by continuing to use chemical pesticides, harmful organisms such as earthworms as well as harmful organisms and harmful pests can be extinct, and there is a high risk of exhausting the field itself. The recent regulation of chemical pesticides is based on the dangerous nature of such chemical pesticides. We have to think deeply about this situation, and now we must concentrate our efforts on the proposition of using safe agricultural materials in order to permanently preserve the global environment and prevent destruction of the ecosystem.
このような実態を背景として化学農薬の使用は年々減少しており、それに代替する資材として、生物的防除剤への関心が高まっている。その理由として生物的防除剤は有害生物のみに作用し、生態系を破壊しない、地球環境保全的な特性を持つからである。その作用も化学農薬のように即効性があるわけでなく、また効果も緩慢で、遅効性である。しかし、この柔らかな効果がかえって地球環境保全的な利点を持つ。しかも、それを長く使い続けることによって圃場には有効菌が増殖し、その有効菌の働きで圃場は次第に地力を増強し、有害生物、有害菌の少ない、病気の出ない健全な土壌となる。もし、生物的防除剤に化学農薬並みの激烈な効果が認められたら、それは他の生物や人間への悪影響の懸念を払拭しきれないため化学農薬並みに徹底的な影響調査が必要とされている。化学農薬はその効果が激烈なものがＡランク（ほとんどの病害菌が死滅するが動物や人間にも害がある場合がある）、効果が柔らかいものがＢランク（効果は若干弱いが、動物や人間に対する害が少ない）とされており、生物的防除剤はＣランクくらいが最も適切（効果は化学農薬よりは弱いが、動物や人間には全く無害である）とされている。 Against this background, the use of chemical pesticides is decreasing year by year, and interest in biological control agents is increasing as an alternative material. The reason for this is that biological control agents act only on pests and do not destroy ecosystems and have global environmental conservation properties. Its action is not as fast as chemical pesticides, and its action is slow and slow. However, this soft effect has the advantage of conserving the global environment. In addition, by continuing to use it for a long time, effective bacteria grow in the field, and by the action of the effective bacteria, the field gradually strengthens the ground strength, and it becomes a healthy soil free of diseases and pests. If a biological control agent has a dramatic effect similar to that of a chemical pesticide, it cannot eliminate the concern of adverse effects on other organisms and humans. Yes. Chemical pesticides are highly effective in A rank (most of the fungus will be killed but may also be harmful to animals and humans), and soft in effect are B rank (although the effect is slightly weak, animals and It is said that the C-rank is the most appropriate biological control agent (the effect is weaker than chemical pesticides, but completely harmless to animals and humans).
本発明が化学農薬の防除価より若干小さいのは生物的防除剤の宿命であり、むしろ、他の生物的防除剤の非力な効果に比べたら、化学農薬に迫るくらいの防除結果が得られており、極めて有効な生物的防除剤と言えるであろう。
生物的防除効果としてはこれくらいの効果が最適であることは言うまでもない。
It is the fate of biological control agents that the present invention is slightly smaller than the control value of chemical pesticides. Rather, compared to the ineffective effects of other biological control agents, the control results that are close to chemical pesticides are obtained. It is a very effective biological control agent.
Needless to say, this is the optimal biological control effect.
その意味で本発明は「化学農薬ほどの劇的な効果はないが、その効果はＣランクくらいで、長く使い続けることにより効果を持続させ、動物や人間に無害で生態系を破壊しない最も妥当な生物的防除剤的効能を持った、土壌改良剤としては新しいタイプの発明と言える。 In this sense, the present invention is “not as dramatic as chemical pesticides, but the effect is about C rank, and the effect is sustained by continuing to use for a long time. It is harmless to animals and humans and does not destroy the ecosystem. It can be said that this is a new type of soil amendment agent having a biological control agent effect.
本発明のナス半身萎凋病抑制剤は、今回はナス半身萎凋病だけにその効果を確認したが、まだ試験を実施していない他の多くの作物病害を抑制する可能性を秘めている。最終的には、他のＢＴ菌や抗菌性微生物を添加することにより、リスクの大きい化学農薬の代替剤として環境保全的特性を持った万能薬的な作物病害抑制剤としての展開が期待できる。 The eggplant half wilt disease inhibitor of the present invention has been confirmed to be effective only for eggplant half wilt disease this time, but has the potential to suppress many other crop diseases that have not yet been tested. Eventually, by adding other BT bacteria and antibacterial microorganisms, it can be expected to develop as an all-purpose crop disease inhibitor with environmental conservation properties as a substitute for high-risk chemical pesticides.
Ａ 孔
Ｂ 菌液
Ｋ 寒天培地
Ｎ ナス苗
Ｐ ４号ポット
Ｓ 培養土
Ｙ プラスチック容器
A hole B fungus solution K agar medium N eggplant seedling P No. 4 pot S culture soil Y plastic container
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