JP3641492B2 - Soil disease control materials - Google Patents
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Description
【０００１】
【産業上の利用分野】
本発明は、エルビニア・カロトボーラ（Erwinia carotovora subsp. carotovora) による蔬菜軟腐病の発病を軽減し、且つ苗立枯れ病や芝ラージパッチ症などの土壌伝染性病原糸状菌に対し顕著な抑止力を有する土壌病害抑制資材に関する。
【０００２】
【従来の技術】
連作障害の内８割は病害が原因であり、さらにその中の８割が土壌病害であると言われている。
しかしながら、特産地形成等の組織的理由や農業資材や農業機械及び栽培技術が作物ごとに特化されたこと等の理由から、単一作物を同一圃場に連作することは益々多くなり、連作障害も激増している。
薬剤を使用することで土壌病害を克服し、連作を可能にしている例は多いが、薬剤連用に伴って病害菌が薬剤抵抗性を獲得したり、薬剤処理によって土壌微生物が無作為に殺菌され、土壌中で物質循環の主役として活躍したり、植物成長調節物質を生産している所謂有用菌と言われる微生物群も同時に死滅する等の問題がある。
さらに、健全な作物の根圏では、微生物が多様性と菌群同志の適当なバランスを維持していると言われ、部分殺菌は特定菌群を爆発的に増殖させる危険がある。
以上のような理由から、土壌微生物環境に悪影響がなく、且つ人体に対しても安全な土壌病害の生物的防除方法が待ち望まれている。
【０００３】
このような期待に応える資材として、バチルス菌・シュウドモナス菌等の細菌やストレプトマイセス菌などの放射菌、及びトリコデルマ菌・グリオクラディウム菌などの糸状菌を利用した資材が開発されている。
しかしながら、これらの殆どが土壌伝染性病原糸状菌に対して効果を発揮するものである。
蔬菜の栽培においてはリゾクトニア菌・バーティシリウム菌をはじめとする多くの糸状菌性病害と細菌性病害の軟腐病が同時に発生するため、軟腐病対策としてストレプトマイシンを葉面散布するのが一般的である。
ストレプトマイシンは葉面から吸収され、軟腐病を死滅させるが、同時に根圏のグラム染色陰性細菌に影響を及ぼす。連用によって耐性菌が誘導される危険もあり、高濃度では植害が認められる等の問題がある。
【０００４】
【発明が解決しようとする課題】
土壌伝染性病原糸状菌に対して抑止力を有する拮抗菌は多く知られているが、病原細菌に抑止力を有する拮抗菌については研究例も比較的少ない。
また、作物は播種期から収穫期まで多くの病害に罹病する危険性が有るため、生育全期間を健全に維持するためには多くの病原菌に抗菌活性を示す拮抗菌を利用した資材の開発が必要である。
本発明は、この課題に対して有効に対処するためになされたもので、エルビニア・カロトボーラ（Erwinia carotovora subsp. carotovora）を原因菌とする蔬菜軟腐病の発病を軽減し、且つ土壌伝染性病原糸状菌に対し顕著な抑止力を有する土壌病害抑制資材を提供したものである。
【０００５】
【課題を解決するための手段】
本発明者は、農業経営上問題となる主要な土壌伝染性病原糸状菌の多くに対して抗菌活性を有する拮抗菌の選抜について鋭意研究を重ね、リゾクトニア菌、フザリウム菌、アファノマイセス菌、バーティシリウム菌、ロゼリニア菌、ヘリコバジディウム菌に対して強い抑止力を有する菌株として、グリオクラディウム・ビレンス（Gliocladium virens）NT-10 株を見出した。
グリオクラディウム・ビレンス NT-10株は抗生物質グリオトキシン(gliotoxin) を高濃度に生産する菌株であり、従来知られている菌株を生産し得ない濃度のグリオトキシンを効率良く生産することを見出した。
さらに、本発明者はグリオトキシンの抗菌スペクトルを主要な土壌伝染性病原糸状菌とエルビニア・カロトボーラ（Erwinia carotovora subsp. carotovora）に限定して詳しく調べたところ、エルビニア・カロトボーラがグリオトキシン存在下では全く生育できないという新しい知見を得た。
【０００６】
そして、土壌改良資材と若干量の基質からなる培地でグリオクラディウム・ビレンス NT-10株を培養した土壌改良資材タイプの本発明資材と、培養物に有機質肥料原料を配合した肥料タイプの本発明資材が土壌病害を軽減し得ることを見出し、本発明の完成に至った。
【０００７】
本発明における微生物は、青森県上北郡六戸町の長芋連作土壌より分離した菌株である。この菌の土壌からの分離は、希釈平板法によって行った。すなわち、土壌１ｇを99mlの滅菌希釈水（トリスバッファー）に分散させ、同様の希釈水を用いて10倍及び 100倍に希釈し、各々１mlを滅菌シャーレに分取し、これにクロラムフェニコール含有のローズベンガル寒天培地10mlを流し込んで良く混和した後固化し、28℃で３日間培養して出現したコロニーの中から分離した。
【０００８】
▲１▼ 各培地における生育状況
オートミール寒天培地
生育旺盛（28.5℃ ３日間でｄ＝67mm）
気生菌糸は初め少なく、コロニーは白色を呈するが、後に綿毛状気生菌糸が豊富となる。
５日以上の培養で緑色の無性胞子を形成するが少ない。
麦芽エキス寒天培地
生育旺盛（28.5℃ ３日間でｄ＝79mm）
気生菌糸は比較的多く、コロニーは初め白色、後に飴色を呈する。無性胞子を形成しない。
バレイショ・ブドウ糖寒天培地
生育旺盛（28.5℃ ３日間でｄ＝79mm）
気生菌糸綿毛状で豊富、後にコロニー中央から溶菌が始まり、コロニー裏面は飴色〜褐色となる。
５日以上の培養で緑色の無性胞子を形成するが極めて少なく、コロニー最端の気生菌糸上にのみ形成される。
サブロー寒天培地
生育やや旺盛（28.5℃ ３日間でｄ＝55mm）
気生菌糸は少なく、コロニーは白色〜僅かに飴色を呈する。無性胞子を形成しない。
【０００９】
▲２▼ 生理生態的性質
生育適温：25〜32℃
生育限界：10℃以下、40℃以上で生育しない
最適ｐＨ：５〜７
ｐＨ範囲：少なくとも３〜10の範囲で生育可能
【００１０】
▲３▼ 本発明菌株の種別
Trichoderma属及び Gliocladium属の種分類が記載されたK.H.DOMSH, W.GAMS AND T.H.ANDERSON:COMPENDIUM OF SOIL FUNGI, ACADEMIC PRESS, LONDON, U.K.VOLUME 1, 859pp. (1980)、およびM.A.RIFI:A REVISION OF GENUS TRICHODERMA, mycological Paper, No.116, COMMONWEALTH MYCOLOGICAL INSTITUTE, KEW, SURREY, U.K.56pp. (1969)におけるGliocladium virensに関する特徴を本発明菌株と比較した。
オートミール寒天培地での生育は極めて速く（20℃、５日間コロニー直径 5.8cm）、フィアライドは互いに狭い角度で輪生体を形成し、緑色の大きなボール状無性胞子塊を作る。
無性胞子は短楕円形 4.5-6×3.5-4 μm で、表面は滑面である。
これらの形態学的特徴が文献記載事項と一致することから、本発明における菌株はGliocladium virensと種別されるが、富栄養培地では高濃度のグリオトキシンを生産して自己溶菌する性質や、無性胞子の形成量が極めて少ない性質を有する点、天然物を基質とする固体培地では菌糸量が極めて多く、無性胞子形成が遅いなどの性質は同種の菌では認められない。このため、本発明における菌株は工業技術院生命工学工業技術研究所に受託番号 FERM P-14123 として受託されている。
本発明における変異株には、前記菌株が自然変異、または放射線あるいは化学薬品による突然変異され、高濃度のグリオトキシンを生産する菌株がある。
【００１１】
本発明の土壌病害抑制資材には、前記したように土壌改良資材タイプと肥料タイプとがある。このうち、本発明の土壌改良資材タイプ調製に用いられる土壌改良資材には、泥炭、ピートモス、バーミキュライト、ゼオライト、木炭等があり、基質には、麩、米糠、菜種油粕等がある。通常は、土壌改良資材７乃至９重量部と基質３乃至１重量部を混合して固体培地を調製し、グリオクラディウム・ビレンス NT-10株を接種し、20乃至30℃で７乃至10日間培養する。培養物は、そのまま或はこれに原料の土壌改良資材を混合して、顆粒状、タブレット状に成形したり、培養物を低温で乾燥粉末にしたりして土壌病害抑制資材とする。
また、肥料タイプ調製に用いる培養物は、土壌改良資材タイプと同様に培養し、得られた培養物に植物油粕類、米糠、魚粕、骨粉、皮粉等の有機質肥料原料を配合して調製する。培養物と有機質肥料原料との配合比は、特に限定されないが、本発明菌株が土壌中で土壌病害抑止力を発揮するためには、資材中の菌密度が高い方が望ましく、有機質原料は培養物重量の 100倍を上限とするのが理想的である。配合品に造粒助剤を添加し、顆粒状、タブレット状としたり、低温で乾燥して粉末状土壌病害抑制資材とする。
【００１２】
本発明の土壌病害抑制資材は、白菜の軟腐病、キュウリの苗立枯れ病、芝のラージパッチ症の発生の予防あるいは抑止に用いられる。使用量としては、土壌病害抑制資材を圃物あるいは芝10ａ当り10乃至 100kg（資材１ｇ当りグリオクラディウム・ビレンス生菌体或は胞子を104 乃至106 コロニーフォーミングユニット含有）を粉剤、顆粒剤あるいは液剤として散布することが望ましい。散布の時期は苗定植前であっても、あるいは上記症状が表れているとき、あるいは正常な発育がみられているときのいずれでもよい。
【００１３】
以下に実施例を示して本発明の効果を具体的に説明する。
【実施例１】
本発明資材に含有されるグリオクラディウム・ビレンス NT-10株(FERM P-14123)の代謝物質の抗菌活性について、蔬菜軟腐病菌と農業上問題とされる主要な土壌伝染性病原糸状菌を対象として調査した。
本発明資材は、ピートモス及び泥炭の混合物からなる担体９重量部に対して麸１重量部を配合した固体培地でグリオクラディウム・ビレンス NT-10株(FERM P-14123)を５日間培養して製造した。代謝物質は培養物 100ｇに酢酸エチル100ml を加えて抽出した後、濃縮してシリカゲルの薄層（メルク社製）で展開した。
軟腐病菌に対しては、展開後のＴＬＣプレートを寒天培地に埋没させるバイオオートグラフィーの手法を用い、糸状菌に対しては病原菌の無性胞子又は栄養菌糸の懸濁液をクロマトスプレーで展開後のＴＬＣプレートに塗布するバイオオートグラフィーの手法を用い、菌の生育阻止帯形成を指標として各画分の抗菌活性を調査した。
結果は表１に示した通りで、Ｒｆ値0.35の還元型グリオトキシン画分とＲｆ値0.54のグリオトキシン画分が供試した全ての菌に強い抗菌活性を示した。
【００１４】
【表１】
【００１５】
【実施例２】
日本肥糧株式会社研究所（所在地：群馬県藤岡市）の白菜連作圃場において、本発明資材の軟腐病発病抑制効果を実証した。
圃場はクロールピクリンによる土壌消毒圃場と無消毒圃場の２箇所で行った。本発明資材は、ピートモスを主体とする培地にグリオクラディウム・ビレンスNT-10 株（FERM P-14123) を生育せしめた培養物９重量部と米糠１重量部を配合し、顆粒状に成形したものを供試した。
粒状固形肥料を全量元肥で施肥し、資材は苗定植前に10ａ当り 100kgを施用し耕耘した。供試圃場は白菜連作圃場であり、軟腐病菌は人為的に接種することなく自然発生に委ねた。栽培管理は慣行に従ったが、軟腐病防除のための農薬散布は行わなかった。
発病抑制効果の判定は、収穫期に白菜地際茎部の断面を調査し、褐変や腐敗の程度を４段階で評価し、発病度を下式により算出した。
発病度（％）＝(3・n3＋２・n2＋１・n1＋０・n0) ÷(3N)×100
n3：重症株数 n2：中症株数 n1：軽症株数 n0：健全株数
結果は表２に示したとおりで、本発明資材施用区では土壌消毒の有無に係らず、軟腐病の発病度は顕著に軽減された。
【００１６】
【表２】
【００１７】
【参考例】
1991年春に高麗芝のフェアーウェイでリゾクトニア菌によるラージパッチ症が発生した埼玉県大里郡岡部町に所在するゴルフ場において、本発明資材施用による回復試験を実施した。
すなわち1991年秋にラージパッチ症状のひどい部分に本発明資材を1ｍ2当り200ｇ表面撒布したところ、1992年春には太く活性の強いランナーが延び、ラージパッチ症は完全に回復した。
一方、本発明資材を施用しなかった部分で新たなラージパッチの発生が認められた。
更に、1992年から1994年にわたって、同ゴルフ場で発生したラージパッチに対して効果の確認試験を実施したところ、１m2当り100ｇの本発明資材を春又は秋に表面散布することにより、半年後には全てのラージパッチ症が回復することが実証された。
尚、試験に供試した本発明資材はピートモス及び泥炭の混合物からなる担体9重量部に対して麩１重量部を配合した培地でグリオクラディウム・ビレンスNT-10株（FERM P-14123）を10日間培養し、粒径2mm以下の顆粒状に成形し乾燥した物で、資材1g当りグリオクラディウム・ビレンスの胞子を106コロニーフォーミングユニット含有していた。
【００１８】
【実施例３】
本発明資材施用による苗立枯れ病発病抑制効果を実証した。
Rhizoctonia solani（AG-4）の人工汚染土壌に、本発明資材 (1)（土壌改良資材タイプ）又は本発明資材 (2)（有機質肥料タイプ）を土壌の重量に対して３％施用し、キュウリを栽培して苗立枯れ病の発病を調査した。
土壌、病原菌、資材は下記のとおりである。
土 壌：黒ボク土壌を主原料とする市販の園芸培土を用い、製品現物を非殺菌土壌として、蒸気殺菌処理を施して殺菌土壌として供試した。
病原菌：Rhizoctonia solani（AG-4）をピートモスと麸の混合培地で７日間培養し、土壌重量に対して0.05％接種して人工汚染土壌とした。
本発明資材 (1)：ピートモスと泥炭からなる担体８重量部に麸１重量部及び水１重量部を添加し高圧蒸気殺菌した培地に、グリオクラディウム・ビレンス NT-10株（RERM P-14123) を10日間培養して製造した。
本発明資材 (2)：本発明資材 (1)９重量部に麸１重量部を添加して製造した。
資材の施用量は土壌重量に対して３％とした。
キュウリ苗立枯れ病の罹病株率は表３に示した通りであり、殺菌土壌でも非殺菌土壌でも本発明資材の施用によって、苗立枯れ病は顕著に抑制された。
【００１９】
【表３】
[0001]
[Industrial application fields]
The present invention reduces the occurrence of sugar beet soft rot caused by Erwinia carotovora subsp. Carotovora, and has a remarkable deterrent against soil-borne pathogenic fungi such as seedling blight and turf large patch disease. It relates to soil disease control materials.
[0002]
[Prior art]
It is said that 80% of continuous cropping disorders are caused by diseases, and 80% of them are caused by soil diseases.
However, due to organizational reasons such as the formation of special production areas and the fact that agricultural materials, agricultural machinery, and cultivation techniques have been specialized for each crop, continuous cropping of a single crop on the same field has become more and more difficult. Has also increased dramatically.
In many cases, the use of chemicals overcomes soil diseases and enables continuous cropping, but with the continuous use of chemicals, pathogenic bacteria acquire drug resistance, and soil microorganisms are randomly sterilized by chemical treatment. However, there are problems such as the fact that so-called useful bacteria that play an active role in material circulation in soil and produce plant growth regulators are also killed at the same time.
Furthermore, in healthy rhizospheres of microorganisms, microorganisms are said to maintain an appropriate balance between diversity and fungi, and partial sterilization has the risk of exploding specific fungi.
For these reasons, a biological control method for soil diseases that has no adverse effect on the soil microbial environment and is safe for the human body is awaited.
[0003]
As materials that meet such expectations, materials using bacteria such as Bacillus bacteria and Pseudomonas bacteria, radioactive bacteria such as Streptomyces bacteria, and filamentous fungi such as Trichoderma bacteria and Gliocladium bacteria have been developed.
However, most of these are effective against soil-borne pathogenic fungi.
In the cultivation of sugar beet, many fungal diseases such as Rhizoctonia and Verticillium bacteria and soft rot of bacterial diseases occur at the same time, so it is common to spray streptomycin as a countermeasure against soft rot. is there.
Streptomycin is absorbed from the foliar surface and kills soft rot, but at the same time affects gram-staining negative bacteria in the rhizosphere. There is a danger that resistant bacteria are induced by continuous use, and there is a problem that plant damage is observed at a high concentration.
[0004]
[Problems to be solved by the invention]
Many antagonistic bacteria having a deterring power against soil-borne pathogenic fungi are known, but there are relatively few studies on antagonistic bacteria having a deterring power against pathogenic bacteria.
In addition, since crops are at risk of suffering from many diseases from the sowing season to the harvest season, the development of materials using antagonistic bacteria that have antibacterial activity against many pathogenic bacteria is necessary to maintain a healthy overall growth period. is necessary.
The present invention has been made in order to effectively deal with this problem, to reduce the onset of vegetable crops soft rot caused bacteria of Erwinia carotovora (Erwinia carotovora subsp. Carotovora), and soil-borne pathogenic filamentous The present invention provides a soil disease control material having a remarkable deterrent against bacteria.
[0005]
[Means for Solving the Problems]
The present inventor has conducted extensive research on selection of antagonistic bacteria having antibacterial activity against many of the major soil-borne pathogenic fungi that are problematic in agricultural management, and Rhizoctonia, Fusarium, Aphanomyces, Verticillium Gliocladium virens NT-10 was found as a strain having strong deterrence against Bacteria, Roselinia and Helicobacterium.
Is Gliocladium & Birensu NT-10 strain is a strain producing antibiotic gliotoxin the (gliotoxin) at a high concentration, to efficiently produce a gliotoxin concentrations not produce conventionally known strain I found it.
Furthermore, the present inventors have revealed that investigated by limiting the antibacterial spectrum of gliotoxin major soil-borne pathogenic fungi and Erwinia carotovora (Erwinia carotovora subsp. Carotovora), Erwinia carotovora are all in the presence gliotoxin New knowledge that it cannot grow was obtained.
[0006]
And this invention material of soil improvement material type that cultured Gliocladium bilens NT-10 strain in a medium consisting of soil improvement material and a small amount of substrate, and fertilizer type book that contains organic fertilizer raw material in the culture The inventors have found that the inventive material can reduce soil diseases and have completed the present invention.
[0007]
The microorganism in the present invention is a strain isolated from Nagatoro continuous cropping soil in Rokunohe, Kamikita-gun, Aomori Prefecture. Separation of this fungus from soil was performed by the dilution plate method. In other words, 1 g of soil was dispersed in 99 ml of sterilized diluted water (Tris buffer), diluted 10-fold and 100-fold with the same diluted water, and 1 ml each was taken into a sterilized petri dish, and chloramphenicol was added thereto. The resulting rose bengal agar medium (10 ml) was poured, mixed well, solidified, and separated from the colonies that emerged after culturing at 28 ° C. for 3 days.
[0008]
(1) Growth status in each medium
Oatmeal agar medium Vigorous growth (d = 67mm at 28.5 ℃ for 3 days)
At first there are few aerial hyphae and the colonies are white, but later become abundant with fluffy aerial hyphae.
A green asexual spore is formed by culturing for 5 days or more, but little.
Malt extract agar medium Vigorous growth (d = 79mm at 28.5 ℃ for 3 days)
The aerial hyphae are relatively large, and the colonies are initially white and later amber. Does not form asexual spores.
Potato / glucose agar medium Vigorous growth (d = 79mm at 28.5 ℃ for 3 days)
Aerobic mycelium fluffy and abundant, later lysis starts from the center of the colony, and the back side of the colony is amber to brown.
A green asexual spore is formed by culturing for 5 days or more, but it is very little and is formed only on the aerial hyphae at the end of the colony.
Sabouraud agar <br/> Slight growth (28.5 ℃, 3 days d = 55mm)
The aerial hyphae are few and the colonies are white to slightly amber. Does not form asexual spores.
[0009]
(2) Physiological and ecological properties Suitable growth temperature: 25-32 ° C
Growth limit: 10 ° C or less, optimum pH that does not grow above 40 ° C: 5-7
pH range: can grow in the range of at least 3-10
(3) Type of strain of the present invention
Trichoderma genus and Gliocladium spp classification KHDOMSH described, W.GAMS AND THANDERSON:. COMPENDIUM OF SOIL FUNGI, ACADEMIC PRESS, LONDON, UKVOLUME 1, 859pp (1980), and MARIFI: A REVISION OF GENUS TRICHODERMA, mycological Paper , No. 116, COMMONWEALTH MYCOLOGICAL INSTITUTE, KEW, SURREY, UK 56pp. (1969), characteristics of Gliocladium virens were compared with the strain of the present invention.
Growth on oatmeal agar is very fast (20 ° C, 5 days colony diameter 5.8cm), and phialides form a ring at a narrow angle with each other, forming a large, green, ball-shaped asexual spore mass.
Asexual spores are short elliptical 4.5-6 × 3.5-4 μm, and the surface is smooth.
Since these morphological features are consistent with those described in the literature, the strain in the present invention is classified as Gliocladium virens. However, the nutrient medium produces a high concentration of gliotoxin and autolyzes it. The property that the amount of spore formation is extremely small, and the solid medium using a natural product as a substrate, the amount of mycelium is extremely large, and the properties such as slow asexual spore formation are not observed in the same type of bacteria. For this reason, the strain in the present invention is entrusted to the National Institute of Advanced Industrial Science and Technology as the accession number FERM P-14123.
Among the mutant strains of the present invention, there are strains that produce a high concentration of gliotoxin when the strain is naturally mutated or mutated by radiation or chemicals.
[0011]
As described above, the soil disease suppressing material of the present invention includes a soil improvement material type and a fertilizer type. Among these, the soil improvement material used for the preparation of the soil improvement material type of the present invention includes peat, peat moss, vermiculite, zeolite, charcoal, and the like, and the substrate includes rice bran, rice bran, rapeseed oil lees, and the like. Usually, 7 to 9 parts by weight of soil amendment material and 3 to 1 part by weight of the substrate are mixed to prepare a solid medium, inoculated with Gliocladium bilens NT-10, and 7 to 10 at 20 to 30 ° C. Incubate for days. The culture is used as a soil disease control material as it is or by mixing it with a raw soil improving material to form a granule or a tablet, or by converting the culture into a dry powder at a low temperature.
The culture used for fertilizer type preparation is cultivated in the same manner as the soil improvement material type, and the resulting culture is prepared by blending organic fertilizer raw materials such as vegetable oil cakes, rice bran, fish cake, bone meal, and skin powder. To do. The mixing ratio of the culture and the organic fertilizer raw material is not particularly limited, but in order for the strain of the present invention to exert soil disease deterrence in the soil, it is desirable that the bacterial density in the material is high. Ideally, the upper limit should be 100 times the weight of the object. A granulation aid is added to the blended product to form granules or tablets, or dried at a low temperature to obtain a powdery soil disease control material.
[0012]
The soil disease-suppressing material of the present invention is used for preventing or suppressing the occurrence of soft rot of Chinese cabbage, seedling blight of cucumber, and large patch disease of turf. As for the amount used, 10 to 100 kg of soil disease control material per 10 g of field or turf (containing 10 4 to 10 6 colony forming units of live cells or spores of gliocladium virens per gram of material), powder, granule It is desirable to spray as an agent or liquid. The time of spraying may be before seedling planting, when the above symptoms appear, or when normal growth is observed.
[0013]
The effects of the present invention will be specifically described below with reference to examples.
[Example 1]
Regarding the antibacterial activity of the metabolite of Gliocladium bilens NT-10 strain (FERM P-14123) contained in the material of the present invention, beetle soft rot fungi and major soil-borne pathogenic fungi that are considered to be agricultural problems The target was investigated.
The material of the present invention is the cultivation of Glyocladium bilens NT-10 (FERM P-14123) for 5 days in a solid medium containing 1 part by weight of cocoon to 9 parts by weight of a carrier composed of a mixture of peat moss and peat. Manufactured. The metabolite was extracted by adding 100 ml of ethyl acetate to 100 g of the culture, then concentrated and developed in a thin layer of silica gel (Merck).
For soft rot fungi, use a bioautography technique to embed the developed TLC plate in an agar medium, and for filamentous fungi, develop a suspension of pathogenic bacteria asexual spores or vegetative mycelia on a chromatographic spray. The antibacterial activity of each fraction was examined using the bioautography method applied to the TLC plate of No. 1 using the growth inhibition band formation of the fungus as an index.
The results are as shown in Table 1. The reduced gliotoxin fraction having an Rf value of 0.35 and the gliotoxin fraction having an Rf value of 0.54 showed strong antibacterial activity against all the bacteria tested.
[0014]
[Table 1]
[0015]
[Example 2]
In the Chinese cabbage continuous cropping field of Nippon Fertilizer Co., Ltd. (location: Fujioka City, Gunma Prefecture), the effect of the present invention material on soft rot disease was demonstrated.
The field was carried out in two places, a soil disinfection field with chlorpicrin and a non-disinfection field. The material of the present invention comprises 9 parts by weight of a culture in which Gliocladium bilens NT-10 strain (FERM P-14123) is grown in a medium mainly composed of peat moss and 1 part by weight of rice bran, and is formed into granules. I tried the thing that I did.
The whole amount of granular solid fertilizer was fertilized with the original fertilizer, and the material was cultivated by applying 100 kg per 10a before planting seedlings. The test field was a Chinese cabbage continuous cropping field, and soft rot fungi were left to occur naturally without artificial inoculation. Cultivation management was followed, but no pesticide spray was applied to control soft rot.
To determine the disease control effect, the cross section of the Chinese cabbage base stem was examined during the harvest period, the degree of browning and rot was evaluated in four stages, and the disease severity was calculated by the following equation.
Disease severity (%) = (3 ・ n 3 ＋2 ・ n 2 +1 ・ n 1 ＋0 ・ n 0 ) ÷ (3N) × 100
n 3 : Number of severe strains n 2 : Number of moderate strains n 1 : Number of mild strains n 0 : Number of healthy strains is as shown in Table 2, and the incidence of soft rot in the district where the material of the present invention is applied regardless of the presence or absence of soil disinfection Was significantly reduced.
[0016]
[Table 2]
[0017]
[Reference example]
In the spring of 1991, a recovery test by applying the material of the present invention was carried out at a golf course located in Okabe-cho, Osato-gun, Saitama, where large patch disease due to Rhizoctonia bacterium occurred on the Korai Shiba fairway.
In other words, in the fall of 1991, 200 g / m 2 of the material of the present invention was applied to the severe part of large patch symptoms, and in the spring of 1992, a thick and active runner extended, and large patch disease was completely recovered.
On the other hand, generation of a new large patch was observed in a portion where the material of the present invention was not applied.
Furthermore, from 1992 to 1994, a test for confirming the effect was conducted on the large patch generated at the golf course. After spraying 100 g of the present invention material per 1 m 2 on the surface in spring or autumn, half a year later Has been demonstrated to recover all large patch diseases.
The material of the present invention used for the test was a medium composed of 9 parts by weight of a carrier consisting of a mixture of peat moss and peat and 1 part by weight of 麩, and Gliocladium bilens NT-10 strain (FERM P-14123). Was cultivated for 10 days, formed into granules having a particle size of 2 mm or less, dried, and contained 10 6 colony forming units of gliocladium virens spores per gram of material.
[0018]
[Example 3 ]
The effect of the present invention material application on the suppression of seedling blight was demonstrated.
Apply the present material (1) (soil improvement material type) or the present material (2) (organic fertilizer type) to artificial soil contaminated with Rhizoctonia solani (AG-4) 3% of the soil weight. Was cultivated to investigate the onset of seedling blight.
The soil, pathogenic bacteria, and materials are as follows.
Soil: Using a commercially available horticultural cultivated soil mainly composed of black soil, the product was treated as non-sterilized soil, steam sterilized, and used as sterilized soil.
Pathogen: Rhizoctonia solani (AG-4) was cultured in a mixed medium of peat moss and straw for 7 days and inoculated with 0.05% of soil weight to make artificially contaminated soil.
Material of the present invention (1) : Glyocladium bilens NT-10 strain (RERM P-) was added to 8 parts by weight of peat moss and peat carrier and 1 part by weight of water and 1 part by weight of water and pasteurized with high pressure steam. 14123) was cultured for 10 days.
Invention material (2) : Invention material (1) It was manufactured by adding 1 part by weight of 9% by weight.
The material application rate was 3% of the soil weight.
The morbidity rate of cucumber seedling blight was as shown in Table 3, and the seedling blight was significantly suppressed by application of the material of the present invention in both sterilized soil and non-sterilized soil.
[0019]
[Table 3]
Claims (8)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP04333294A JP3641492B2 (en) | 1994-02-18 | 1994-02-18 | Soil disease control materials |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP04333294A JP3641492B2 (en) | 1994-02-18 | 1994-02-18 | Soil disease control materials |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH07233019A JPH07233019A (en) | 1995-09-05 |
| JP3641492B2 true JP3641492B2 (en) | 2005-04-20 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP04333294A Expired - Lifetime JP3641492B2 (en) | 1994-02-18 | 1994-02-18 | Soil disease control materials |
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| Country | Link |
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| JP (1) | JP3641492B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2000236536A1 (en) * | 2000-03-31 | 2001-10-08 | Yuen Foong Yu Paper Mfg Co., Ltd. | A method for preventing plant disease |
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| JPH07233019A (en) | 1995-09-05 |
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