JP2015503548A - Composition for controlling phytopathogenic fungi containing Bacillus subtilis KBC1010 strain and its culture as an active ingredient - Google Patents
Composition for controlling phytopathogenic fungi containing Bacillus subtilis KBC1010 strain and its culture as an active ingredient Download PDFInfo
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Abstract
本発明は、バチルスサブチリスＫＢＣ１０１０菌株及びその培養液を有効成分として含有する植物病原菌防除用組成物に関するもので、より詳細には、土壌から分離したバチルスサブチリスＫＢＣ１０１０菌株が芝生の病害を誘発する病原体であるリゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］を含む様々な植物病原菌に対して優れた抗真菌活性を示し、従来の有機合成農薬と比較して毒性が少なく、生態系と人畜に害を与えず、芝生や他の植物病原菌を防除することができる環境に優しい組成物として有用に用いることができる。【選択図】図１The present invention relates to a Bacillus subtilis KBC1010 strain and a composition for controlling phytopathogenic fungi containing the culture solution thereof as an active ingredient. More specifically, the Bacillus subtilis KBC1010 strain isolated from soil induces lawn diseases. Has excellent antifungal activity against various phytopathogenic fungi including Rhizoctonia solani AG-2-2 (IV), the pathogen Rhizoctonia solani AG-2-2 (IV), compared with conventional organic synthetic pesticides Therefore, it can be usefully used as an environmentally friendly composition that can control turf and other phytopathogenic fungi without causing harm to ecosystems and human animals. [Selection] Figure 1
Description
本発明は、松に生息する蝉の幼虫の腸内から分離したバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株及びその培養液を有効成分として含有する芝生およびその他植物病原菌防除用組成物に関するものである。 The present invention relates to a Bacillus subtilis KBC1010 (Bacillus subtilis KBC1010) strain isolated from the intestines of moth larvae that inhabit pine and a composition for controlling turf and other phytopathogenic fungi containing a culture solution thereof as an active ingredient. .
芝生は、元来野性が強く、病気に対する強い抵抗性を持っており、正常な発育状態の下では病気が発生しない。しかし、ゴルフ場など、特定の目的のために栽培管理され商業的に利用される場合、集約管理による栽培環境の変化、競技者によって踏まれると傷などのストレスを受けて発症環境が造成されると、正常の生育条件下では問題にならない病原体にも容易に発症する傾向がある。 Lawn is naturally wild and has a strong resistance to disease, and disease does not occur under normal developmental conditions. However, when it is cultivated and managed for specific purposes, such as golf courses, changes in the cultivated environment due to intensive management, and when it is stepped on by competitors, it creates stress-onset environments. And there is a tendency to easily develop pathogens that are not a problem under normal growth conditions.
韓国のゴルフ場で植栽されている芝生類に発生する病気は約１４種であり、関連病原体は、１７余種が知られている（韓国植物病理学会、２００９年。韓国植物病名目録）。主な病害としては、リゾクトニアソラニＡＧ−１（ＩＡ）［Rhizoctonia solani AG-1(IA)］によって誘発されるブラウンパッチ、スクレロチニアホモエオカルパ（Sclerotinia homoeocarpa）によって誘発されるダラースポット、ピシウム属（Pythium spp.）により誘発されるピシウム立枯病、コレトトリカムアキテイタム（Colletotrichum acutatum）により誘発される炭疽病、リゾクトニアケレアリス（Rhizoctonia cerealis）により誘発されるイエローパッチ、リゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］により誘発されるリゾクトニア葉腐病（ラージパッチ）などがある。 There are about 14 diseases that occur in lawns planted at golf courses in Korea, and more than 17 related pathogens are known (Korea Society for Plant Pathology, 2009. Korean Plant Disease List). Major diseases include brown patches induced by Rhizoctonia solani AG-1 (IA), dollar spots induced by Sclerotinia homoeocarpa, Pythium blight caused by Pythium spp., Anthracnose caused by Colletotrichum acutatum, Yellow patch induced by Rhizoctonia cerealis, Rhizoctonia solani AG-2-2 (IV) Rhizoctonia leaf rot (large patch) induced by [Rhizoctonia solani AG-2-2 (IV)].
これら芝生の病害の防除のために様々な種類の有機合成農薬を使用しますが、ゴルフ場周辺の住民だけでなく、競技者から農薬の土壌残留による生態系破壊などの問題として否定的な認識を受けている。したがって、環境にやさしい防除の一環として、無毒性の殺菌剤の開発が切実に求められている。国内外では、農業に適用していた微生物を用いて、様々な芝生病害用組成物が開発されているが、芝生の病害に対して優れた抗真菌活性を有する組成物は、不十分なのが実情である。 Various kinds of organic synthetic pesticides are used to control these turf diseases. However, not only the residents around the golf course but also athletes have negative recognition as problems such as the destruction of the ecosystem due to the residue of pesticides on the soil. Is receiving. Therefore, there is an urgent need for the development of non-toxic fungicides as part of environmentally friendly control. In Japan and overseas, various lawn disease compositions have been developed using microorganisms that have been applied to agriculture, but compositions having excellent antifungal activity against lawn diseases are insufficient. It is a fact.
一方、バチルスサブチリスを用いた植物病原菌防除用組成物として、特許文献１では、新規なバチルスサブチリスＳ３７−２菌株およびそれを用いた微生物肥料がレタスの菌核菌に対して抗真菌活性を示すことが開示されており、特許文献２では、新規なバチルスサブチリスＤＢＢ１５０１菌株がキュウリうどん粉病、イチゴうどん粉病、真桑瓜うどん粉病、桃の穿孔細菌病と高麗人参斑点病に対する抗真菌活性を示すことが開示されており、特許文献３には、新規なバチルスサブチリスＫＫＧ−１およびそれを用いた微生物農薬がトウガラシ疫病に対する抗真菌活性を示すことが開示されている。また、特許文献４では、新規な高温性バチルスサブチリスＤＯＮＫ０３２７菌株がレタス菌核病に対する抗真菌活性を示すことが開示されているが、芝生の病害を誘発する病原体であるリゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］により誘発されるリゾクトニア葉腐病（ラージパッチ）に対する強力な抗真菌活性を示す微生物を用いた病原菌防除用組成物は、公知されていない。 On the other hand, as a composition for controlling phytopathogenic fungi using Bacillus subtilis, in Patent Document 1, a novel Bacillus subtilis S37-2 strain and a microbial fertilizer using the same have antifungal activity against lettuce sclerotia. Patent Document 2 discloses that the novel Bacillus subtilis DBB1501 strain exhibits antifungal activity against cucumber powdery mildew, strawberry powdery mildew, mulberry powdery mildew, peach perforated bacterial disease and ginseng spot disease Patent Document 3 discloses that a novel Bacillus subtilis KKG-1 and a microbial pesticide using the same exhibit antifungal activity against a cabbage epidemic. Further, Patent Document 4 discloses that a novel thermophilic Bacillus subtilis DONK0327 strain exhibits antifungal activity against lettuce nuclear disease, but Rhizoctonia solani AG-2-2, which is a pathogen that induces lawn disease 2 (IV) [Rhizoctonia solani AG-2-2 (IV)] A composition for controlling pathogens using a microorganism exhibiting a strong antifungal activity against Rhizoctonia leaf rot (large patch) is known. Absent.
そこで、本発明者らは様々な植物病原体に対する優れた抗真菌活性を有する組成物を開発するために努力した結果、受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株及びその培養液が、芝生病害を誘発する病原菌であるリゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］を含む様々な植物病原菌に対して、優れた抗真菌活性を示すことを確認することにより、本発明を完成した。 Accordingly, as a result of efforts to develop a composition having excellent antifungal activity against various plant pathogens, the present inventors, as a result, Bacillus subtilis KBC1010 strain deposited under the deposit number KCTC10304BP and culture thereof. The liquid exhibits excellent antifungal activity against a variety of plant pathogens including Rhizoctonia solani AG-2-2 (IV), a pathogen that induces lawn diseases By confirming this, the present invention was completed.
本発明の目的は、受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株及びその培養液を有効成分として含有する芝生およびその他植物病原菌防除用組成物およびその製造方法を提供することである。 An object of the present invention is to provide a composition for controlling turf and other phytopathogenic fungi containing a Bacillus subtilis KBC1010 strain deposited as accession number KCTC10304BP, a culture solution thereof as an active ingredient, and a method for producing the same. It is.
また、本発明の他の目的は、受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０菌株及びその培養液を有効成分として含有する芝生リゾクトニア葉腐病（ラージパッチ）防除用組成物を提供することである。 Another object of the present invention is to provide a Bacillus subtilis KBC1010 strain deposited under the accession number KCTC10304BP and a composition for controlling lawn lysoctonia leaf rot (large patch) containing the culture solution as an active ingredient. is there.
前記目的を達成するために、本発明は、受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株及びその培養液を有効成分として含有する植物病原菌防除用組成物を提供する。 To achieve the above object, the present invention provides a Bacillus subtilis KBC1010 strain deposited under the accession number KCTC10304BP and a composition for controlling phytopathogenic fungi containing a culture solution thereof as an active ingredient.
また、本発明は、受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株及びその培養液を有効成分として含有する芝生リゾクトニア葉腐病（ラージパッチ）防除用組成物を提供する。 The present invention also provides a composition for controlling turf rhizobonia leaf rot (large patch) containing the Bacillus subtilis KBC1010 strain deposited under the deposit number KCTC10304BP and a culture solution thereof as active ingredients.
また、本発明は、
１）受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株を液体培地で培養し、培養物を製造する工程と、
２）前記工程１）で製造された培養物を乾燥し、粉末化して原剤を製造する工程を含む植物病原菌防除用組成物の製造方法を提供する。
The present invention also provides:
1) culturing a Bacillus subtilis KBC1010 strain deposited under accession number KCTC10304BP in a liquid medium to produce a culture;
2) A method for producing a composition for controlling phytopathogenic fungi comprising a step of drying and pulverizing the culture produced in step 1) to produce an active ingredient.
本発明のバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株またはその培養液は、芝生の病害を誘発する病原体であるリゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］を含む様々な植物病原菌に対して優れた抗真菌活性を示すので、環境にやさしい植物病原菌防除用組成物として有用に用いることができる。 The Bacillus subtilis KBC1010 strain or the culture solution thereof of the present invention can be obtained by using Rhizoctonia solani AG-2-2 (IV), which is a pathogen that causes lawn diseases. Since it exhibits excellent antifungal activity against various phytopathogenic fungi, it can be usefully used as an environment-friendly composition for controlling phytopathogenic fungi.
以下、本発明を詳細に説明する。 Hereinafter, the present invention will be described in detail.
本発明は、受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株またはその培養液を有効成分として含有する植物病原菌防除用組成物を提供する。 The present invention provides a composition for controlling phytopathogenic fungi containing, as an active ingredient, a Bacillus subtilis KBC1010 strain deposited under the deposit number KCTC10304BP or a culture solution thereof.
また、本発明は、バチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010；受託番号ＫＣＴＣ１０３０４ＢＰ）菌株またはその培養液を植物や土壌に処理する工程を含む植物病原菌の防除方法を提供する。 The present invention also provides a method for controlling phytopathogenic fungi comprising a step of treating a Bacillus subtilis KBC1010 (accession number KCTC10304BP) strain or a culture solution thereof with plants or soil.
また、本発明は、バチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010；受託番号ＫＣＴＣ１０３０４ＢＰ）菌株またはその培養液を植物病原菌防除用組成物に用いる用途を提供する。 Moreover, this invention provides the use which uses the Bacillus subtilis KBC1010 (Bacillus subtilis KBC1010; accession number KCTC10304BP) strain or its culture solution for the composition for plant pathogen control.
また、本発明は、バチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010；受託番号ＫＣＴＣ１０３０４ＢＰ）菌株をデキストロース（dextrose）０．１〜１０．０重量部およびペプトン（peptone）０．１〜１０．０重量部を含む培地で２５〜３５℃およびｐＨ５．５〜８．５で培養して得られた培養物を乾燥し、粉末化した原剤５．０〜５０．０％（ｗ／ｗ）、および増量剤を含む組成物を水に２００倍〜１０００倍に希釈し、１Ｌ／ｍ２を土壌灌注する工程を含む植物病原体の防除方法を提供する。 The present invention also includes 0.1 to 10.0 parts by weight of dextrose and 0.1 to 10.0 parts by weight of peptone of Bacillus subtilis KBC1010 (accession number KCTC10304BP) strain. A culture obtained by culturing at 25 to 35 ° C. and pH 5.5 to 8.5 in a medium was dried, and powdered drug substance 5.0 to 50.0% (w / w), and a bulking agent were added. Provided is a method for controlling plant pathogens, comprising a step of diluting a composition comprising 200 to 1000 times in water and irrigating 1 L / m 2 with soil.
また、本発明は、バチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010；受託番号ＫＣＴＣ１０３０４ＢＰ）菌株をデキストロース０．１〜１０．０重量部およびペプトン０．１〜１０．０重量部を含む培地で２５〜３５℃およびｐＨ５．５〜８．５で培養して得られた培養物を乾燥し、粉末化した原剤５．０〜５０．０％（ｗ／ｗ）、および増量剤を含む組成物を植物病原体の防除のための水和剤組成物に用いる用途を提供する。 The present invention also relates to a medium containing 0.1 to 10.0 parts by weight of dextrose and 0.1 to 10.0 parts by weight of peptone of a Bacillus subtilis KBC1010 (accession number KCTC10304BP) strain at 25 to 35 ° C. And a composition obtained by drying and pulverizing a culture obtained by culturing at pH 5.5 to 8.5 and a bulking agent 5.0 to 50.0% (w / w), and a bulking agent. Provided is a use for a wettable powder composition for the control of water.
前記バチルスサブチリスＫＢＣ１０１０菌株は、本発明者によって２００２年０７月１８日、韓国生命工学研究院遺伝子銀行に受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されている。 The Bacillus subtilis KBC1010 strain was deposited by the present inventor on July 18, 2002 at the Korea Institute of Biotechnology, Bank of Japan under the accession number KCTC10304BP.
前記の植物病原体は、リゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］、リゾクトニアケレアリス（Rhizoctonia cerealis）、リゾクトニアソラニＡＧ−１（ＩＡ）［Rhizoctonia solani AG-1(IA)］、ボトリティスシネリア（Botrytis cinerea）、コレトトリカムアキティタム（Colletotrichum acutatum）とスクレロチウムロルフシイ（Sclerotium rolfsii）からなる群から選択されるいずれか一つ以上であることが好ましく、リゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］であることがより好ましい。 The plant pathogens are Rhizoctonia solani AG-2-2 (IV) [Rhizoctonia solani AG-2-2 (IV)], Rhizoctonia cerealis, Rhizoctonia solani AG-1 (IA) [Rhizoctonia solani AG- 1 (IA)], preferably one or more selected from the group consisting of Botrytis cinerea, Colletotrichum acutatum and Sclerotium rolfsii Rhizoctonia solani AG-2-2 (IV) [Rhizoctonia solani AG-2-2 (IV)] is more preferable.
本発明の具体的な実施形態において、本発明者らはバチルスサブチリスＫＢＣ１０１０菌株の植物病原菌に対する生育阻止効果を確認した結果、本発明のバチルスサブチリスＫＢＣ１０１０菌株は、６種の植物病原体に対して有意な抗真菌効果を示すことを確認し、特に芝生リゾクトニア葉腐病（ラージパッチ）を誘発する病原体であるリゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］には、顕著な抗真菌効果を示すことを確認した（表１および図１を参照）。 In a specific embodiment of the present invention, as a result of confirming the growth inhibitory effect of the Bacillus subtilis KBC1010 strain on phytopathogenic fungi, the present inventors found that the Bacillus subtilis KBC1010 strain of the present invention is against Rhizoctonia solani AG-2-2 (IV) [Rhizoctonia solani AG-2-2 (IV)], a pathogen that induces lawn rhizobonia leaf rot (large patch) Was confirmed to show a remarkable antifungal effect (see Table 1 and FIG. 1).
また、本発明は、
１）受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株を液体培地で培養し、培養物を製造する工程と
２）前記工程１）で製造された培養物を乾燥し、粉末化して原剤を製造する工程を含む植物病原菌防除用組成物の製造方法を提供する。
The present invention also provides:
1) a step of culturing a Bacillus subtilis KBC1010 strain deposited under the accession number KCTC10304BP in a liquid medium and producing a culture; and 2) drying the culture produced in the above step 1) to obtain a powder. Provided is a method for producing a composition for controlling plant pathogens, which comprises a step of producing an active ingredient by converting into an agent.
前記の工程１）の培地は、デキストロース０．１〜１０．０重量部およびペプトン０．１〜１０．０重量部を含むことが好ましいが、これに限定されない。 The medium of the above step 1) preferably contains 0.1 to 10.0 parts by weight of dextrose and 0.1 to 10.0 parts by weight of peptone, but is not limited thereto.
前記の工程１）の培養は、２５〜３５℃およびｐＨ５．５〜８．５で培養することが好ましいが、これに限定されない。 The culture in step 1) is preferably performed at 25 to 35 ° C. and pH 5.5 to 8.5, but is not limited thereto.
前記の工程２）の乾燥粉末化は、噴霧乾燥または凍結乾燥機を用いて培養液を粉末化することが好ましいが、これに限定されない。 The dry pulverization in step 2) is preferably performed by pulverizing the culture solution using a spray dryer or a freeze dryer, but is not limited thereto.
前記組成物は、前記の菌株またはその培養液を有効成分として含むことができる。 The said composition can contain the said strain or its culture solution as an active ingredient.
本発明による組成物は、通常の方法で植物病原菌の防除用に製剤化することができ、乾燥粉末の形態または液状形態で製造することができる。 The composition according to the present invention can be formulated for the control of phytopathogenic fungi by conventional methods and can be produced in the form of a dry powder or in a liquid form.
具体的には、本発明による組成物は、液状生物農薬の形態で製造することができ、増量剤を添加して粉末の形態で利用したり、またはそれを剤形化し、顆粒化することもできる。しかし、その剤形には特に限定されない。 Specifically, the composition according to the present invention can be manufactured in the form of a liquid biological pesticide, and can be used in the form of a powder by adding a bulking agent, or it can be formulated and granulated. it can. However, the dosage form is not particularly limited.
本発明の具体的な実施形態において、本発明者らは、本発明の菌株を液体培養した後、粉末原剤を製造した後、前記培養液と粉末原剤の温度およびｐＨによる生胞子数を調査した結果、１．０×１０９ｃｆｕ／ｍｌおよび１．０×１０９ｃｆｕ／ｇ以上の含有量を示し、一般的なバチルス菌株と同様の生産性を示すことを確認した（表２および表３を参照）。 In a specific embodiment of the present invention, after liquid culture of the strain of the present invention, the present inventors produced a powder base, and then determined the number of viable spores according to the temperature and pH of the culture medium and the powder base. As a result of the investigation, it was confirmed that the contents of 1.0 × 10 9 cfu / ml and 1.0 × 10 9 cfu / g or more were exhibited, and the productivity was similar to that of a general Bacillus strain (Table 2 and (See Table 3).
また、本発明者らは、本発明のバチルスサブチリスＫＢＣ１０１０菌株原剤および増量剤などを混合して水和剤として製剤し、理化学的特性と胞子数の含有量を調査した結果、本発明の水和剤は、粉末度および水和性が良好で有意な胞子数含有量を有することを確認した（表４参照）。 In addition, the present inventors mixed the Bacillus subtilis KBC1010 strain base material and the bulking agent of the present invention and formulated as a wettable powder, and as a result of investigating the physicochemical properties and the content of the spore count, It was confirmed that the wettable powder had a fine powder content and hydration property and a significant spore number content (see Table 4).
前記組成物に添加される増量剤は、微生物のような有効成分組成において適切な割合で含有することができるように、生物農薬組成物の全体量を調節する役割をする。本発明で使用することができる増量剤には、ナトリウムアルギネート、ゼラチン化デンプン、トウモロコシデンプン、大豆粕、ふすま、粒状繊維質、硫安、珪藻土、ゼオライト、ベントナイト、タルク、カオリン、パイロフイライト、ホワイトカーボン、糖類などを使用することができる。 The bulking agent added to the composition serves to adjust the total amount of the biopesticidal composition so that it can be contained in an appropriate ratio in the active ingredient composition such as a microorganism. Bulking agents that can be used in the present invention include sodium alginate, gelatinized starch, corn starch, soybean meal, bran, granular fiber, ammonium sulfate, diatomaceous earth, zeolite, bentonite, talc, kaolin, pyrophyllite, white carbon. Sugars can be used.
前記したように、本発明のＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株及びその培養液を有効成分として含有する組成物は、様々な剤形に製造することができ、前記の様々な剤形には、有意な生胞子数を含有しているので、芝生の病害を誘発する病原体であるリゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］を含む様々な植物病原菌に対して、環境にやさしい植物病原菌防除用組成物として有用に用いることができる。 As described above, the composition containing the Bacillus subtilis KBC1010 strain deposited as KCTC10304BP of the present invention and its culture solution as active ingredients can be produced in various dosage forms. Rhizoctonia solani AG-2-2 (IV) is a pathogen that induces turf disease because various dosage forms contain significant viable spores. Can be usefully used as an environmentally friendly composition for controlling plant pathogens.
前記の植物病原菌防除用組成物を用いた防除方法は、通常、一般的に行なわれている方法、すなわち、散布（例えば、噴霧、ミスティング、アトマイジング、粉末スプレー、顆粒散布、水面施用、床施用など）、土壌施用（例えば、混入、灌注など）、表面施用（例えば、塗布、塗抹法、被覆等）、浸漬、燻煙施用等により行うことができる。その使用量は、被害状況、適用方法、適用場所等に応じて適切に決定することができる。 The control method using the above-mentioned composition for controlling plant pathogens is usually a commonly performed method, that is, spraying (for example, spraying, misting, atomizing, powder spraying, granule spraying, water surface application, floor application, flooring). Application), soil application (for example, mixing, irrigation, etc.), surface application (for example, application, smearing, coating, etc.), immersion, smoke application, etc. The amount used can be appropriately determined according to the damage situation, application method, application location, and the like.
また、本発明は、ＫＣＴＣ１０３０４ＢＰで寄託されたバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株またはその培養液を有効成分として含有する芝生リゾクトニア葉腐病（ラージパッチ）防除用組成物を提供する。 The present invention also provides a composition for controlling lawn rhizobonia leaf rot (large patch) containing, as an active ingredient, the Bacillus subtilis KBC1010 strain deposited with KCTC10304BP or a culture solution thereof.
また、本発明は、バチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010；受託番号ＫＣＴＣ１０３０４ＢＰ）菌株またはその培養液を植物や土壌に処理する工程を含む、リゾクトニアソラニＡＧ−２−２（ＩＶ）によって誘発された芝生のリゾクトニア葉腐病（ラージパッチ）防除方法を提供する。 The present invention also relates to lawn induced by Rhizoctonia solani AG-2-2 (IV), which comprises a step of treating a plant or soil with a Bacillus subtilis KBC1010 (accession number KCTC10304BP) strain or a culture solution thereof. Of Rhizoctonia leaf rot (large patch).
また、本発明は、バチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010；受託番号ＫＣＴＣ１０３０４ＢＰ）菌株またはその培養液をリゾクトニアソラニＡＧ−２−２（ＩＶ）によって誘発される芝生リゾクトニア葉腐病（ラージパッチ）防除用組成物に用いる用途を提供する。 In addition, the present invention is for controlling Bacillus subtilis KBC1010 (Accession No. KCTC10304BP) strain or its culture solution against lawn Rhizoctonia leaf rot (large patch) induced by Rhizoctonia solani AG-2-2 (IV). Applications for use in the composition are provided.
前記芝生リゾクトニア葉腐病（ラージパッチ）は、国内のゴルフ場の韓国芝で最も多く発生する病気であり、リゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］の病原体により誘発されるものであり、本発明の組成物は、顕著な芝生リゾクトニア葉腐病（ラージパッチ）の防除効果を示すことを確認した（図２および表５を参照）。 The lawn Rhizoctonia leaf rot (large patch) is the most commonly occurring disease in Korean turf at golf courses in Japan. Rhizoctonia solani AG-2-2 (IV) [Rhizoctonia solani AG-2-2 (IV)] It was confirmed that the composition of the present invention exhibited a remarkable lawn rhizobonia leaf rot (large patch) control effect (see FIG. 2 and Table 5).
したがって、本発明の受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０（Bacillus subtilis KBC1010）菌株及びその培養液は、リゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］、病原体により誘発される芝生リゾクトニア葉腐病（ラージパッチ）に対して優れた抗真菌活性を示すので、芝生リゾクトニア葉腐病（ラージパッチ）防除用組成物の有効成分として有用に用いることができる。 Accordingly, the Bacillus subtilis KBC1010 strain deposited under the accession number KCTC10304BP of the present invention and its culture solution are Rhizoctonia solani AG-2-2 (IV) [Rhizoctonia solani AG-2-2 (IV)]. Since it exhibits excellent antifungal activity against lawn lysoctonia leaf rot (large patch) induced by pathogens, it can be usefully used as an active ingredient in a composition for controlling lawn lysoctonia leaf rot (large patch). .
以下、本発明を実施例により詳細に説明する。 Hereinafter, the present invention will be described in detail with reference to examples.
但し、下記の実施例は本発明を具体的に例示するものであり、本発明の内容が実施例により限定されるものではない。 However, the following examples specifically illustrate the present invention, and the contents of the present invention are not limited to the examples.
＜実施例１＞植物病原体に対する生育阻止効果の確認
本発明者らは、受託番号ＫＣＴＣ１０３０４ＢＰとして寄託されたバチルスサブチリスＫＢＣ１０１０の植物病原体に対する生育阻止効果を確認するために、殺菌したジャガイモ寒天培地上に様々な植物病原体とバチルスサブチリスＫＢＣ１０１０を培養した。ここで、植物病原体は、ジャガイモ寒天培地で継代培養した個体群を直径５ｍｍのサイズの円形切片にしたものを使用し、ＬＢ培地で培養したバチルスサブチリスＫＢＣ１０１０を用いて接種した。次に、様々な病原菌の生育に良い温度である２８℃で１０日間培養した後、阻止帯(Inhibition zone)の直径を測定した。
<Example 1> Confirmation of growth inhibitory effect on plant pathogens In order to confirm the growth inhibitory effect of Bacillus subtilis KBC1010 deposited under the deposit number KCTC10304BP on a sterilized potato agar medium, Various plant pathogens and Bacillus subtilis KBC1010 were cultured. Here, the plant pathogen was inoculated with Bacillus subtilis KBC1010 cultured in an LB medium using a population obtained by subcultured in a potato agar medium in a circular section having a diameter of 5 mm. Next, after culturing at 28 ° C., which is a good temperature for growth of various pathogenic bacteria, for 10 days, the diameter of the inhibition zone was measured.
前記の植物病原菌には、作物に大きな被害を与える病原体である芝生植物病原菌としてリゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］、リゾクトニアケレアリス（Rhizoctonia cerealis）、リゾクトニアソラニＡＧ−１（ＩＡ）［Rhizoctonia solani AG-1(IA)］、その他一般的な植物病原菌としてボトリティスシネリア（Botrytis cinerea）、コレトトリカムアキティタム（Colletotrichum acutatum）およびスクレロチウムロルフシイ（Sclerotium rolfsii）の６種を使用した。 The plant pathogenic fungi include Rhizoctonia solani AG-2-2 (IV), Rhizoctonia cerealis, a lawn plant pathogen that is a pathogen that causes great damage to crops. , Rhizoctonia solani AG-1 (IA), and other common plant pathogens such as Botrytis cinerea, Colletotrichum acutatum and Sclerotium rolfushii Six species (Sclerotium rolfsii) were used.
その結果、図１および表１に示したように、バチルスサブチリスＫＢＣ１０１０菌株は、前記の植物病原体すべてに対して有意な抗真菌効果を示すことを確認し、特に、芝生病害を誘発する病原体であるリゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］に対しては、顕著な抗真菌活性を示すことを確認した（表１、図１）。 As a result, as shown in FIG. 1 and Table 1, it was confirmed that the Bacillus subtilis KBC1010 strain exhibits a significant antifungal effect against all the above-mentioned plant pathogens, particularly in the pathogens that induce lawn diseases. It was confirmed that a certain Rhizoctonia solani AG-2-2 (IV) [Rhizoctonia solani AG-2-2 (IV)] showed remarkable antifungal activity (Table 1, FIG. 1).
＋：０＜Ｘ≦２ｍｍ
＋＋：２ｍｍ＜Ｘ≦５ｍｍ
＋＋＋：５ｍｍ＜Ｘ≦１０ｍｍ
＋＋＋＋：１０ｍｍ＜Ｘ≦１５ｍｍ
＋＋＋＋＋：１５ｍｍ＜Ｘ
+: 0 <X ≦ 2 mm
++: 2 mm <X ≦ 5 mm
+++: 5 mm <X ≦ 10 mm
+++++: 10 mm <X ≦ 15 mm
++++++: 15 mm <X
＜実施例２＞バチルスサブチリスＫＢＣ１０１０菌株の液体培養および粉末原剤製造
本発明のバチルスサブチリスＫＢＣ１０１０菌株を１．２ＫＬインキュベーターを用いて液体培養を行った。
<Example 2> Liquid culture of Bacillus subtilis KBC1010 strain and production of powder bulk agent The Bacillus subtilis KBC1010 strain of the present invention was subjected to liquid culture using a 1.2 KL incubator.
具体的には、培地成分（ｗ／ｖ）にデキストロース０．１〜１０．０％、ペプトン０．１〜１０．０％を含む液体培地にバチルスサブチリスＫＢＣ１０１０菌株を、温度２５〜３５℃、ｐＨ５．５〜８．５に調節して１．２ＫＬ発酵器を用いて培養した。 Specifically, a Bacillus subtilis KBC1010 strain is added to a liquid medium containing dextrose 0.1 to 10.0% and peptone 0.1 to 10.0% in a medium component (w / v) at a temperature of 25 to 35 ° C. The pH was adjusted to 5.5 to 8.5 and cultured using a 1.2 KL fermenter.
また、全培養時間は４８時間にし、培養終了後、前記培養液、および噴霧乾燥や凍結乾燥をした粉末原剤を、温度およびｐＨによる生胞子数の含量を調査した結果、いずれも１．０×１０９ｃｆｕ／ｍｌおよび１．０×１０９ｃｆｕ／ｇ以上の含有量を示し、一般的なバチルス菌株と類似の生産性を示すことを確認した（表２および表３）。 In addition, the total culture time was 48 hours, and after completion of the culture, as a result of investigating the content of viable spores according to temperature and pH, the culture broth and the powdered base material spray-dried or freeze-dried were both 1.0. The contents of × 10 9 cfu / ml and 1.0 × 10 9 cfu / g or more were shown, and it was confirmed to show productivity similar to general Bacillus strains (Tables 2 and 3).
＜実施例３＞バチルスサブチリスＫＢＣ１０１０を含有する組成物の製造
前記＜実施例２＞で製造されたバチルスサブチリスＫＢＣ１０１０菌株原剤５．０〜５０．０％（ｗ／ｗ）と増量剤５０．０〜９５．０％（ｗ／ｗ）を混合した後、水和剤を製造した。前記のように水和剤として製剤化した結果、本発明の水和剤の製剤は、農村振興庁告示「農薬の検査方法」に開示された粉末度および水和性が良好な結果を示し、水和剤内のバチルスサブチリスＫＢＣ１０１０胞子数の含量は、５．１×１０７ｃｆｕ／ｇ以上を示し、適切な有効量を有することを確認した（表４）。
<Example 3> Manufacture of a composition containing Bacillus subtilis KBC1010 5.0 to 50.0% (w / w) of bulk agent of Bacillus subtilis KBC1010 strain prepared in the above <Example 2> After mixing 0.0 to 95.0% (w / w), a wettable powder was prepared. As a result of the formulation as a wettable powder as described above, the wettable powder formulation of the present invention showed good results in fineness and hydration disclosed in the Rural Development Agency announcement "Agricultural Chemicals Inspection Method" The content of the number of Bacillus subtilis KBC1010 spores in the wettable powder was 5.1 × 10 7 cfu / g or more, and it was confirmed to have an appropriate effective amount (Table 4).
＜実施例４＞バチルスサブチリスＫＢＣ１０１０の植物病原体の防除効果
慶尚南道宣寧で芝生（品種：韓国芝）を対象に、芝生のリゾクトニア葉腐病（ラージパッチ）発生初期に、前記＜実施例３＞で製造した本発明の水和剤を水に２００倍〜１０００倍に希釈して１Ｌ／ｍ２で、土壌灌注処理を１回実施した（２０１１年８月３０日）。最終的な薬剤処理の９、２１日と一ヶ月後の芝生リゾクトニア葉腐病（ラージパッチ）試験群の写真を撮影し、一ヶ月後の被害面積率を調査し、防除効果を評価した。
<Example 4> Control effect of plant pathogens of Bacillus subtilis KBC1010 In lawn (variety: Korean turf) in Namyeong, Gyeongsangnam-do, at the initial stage of occurrence of rhizobonia leaf rot (large patch) The wettable powder of the present invention produced in 3> was diluted 200 to 1000 times in water and subjected to soil irrigation once at 1 L / m 2 (August 30, 2011). Photographs were taken of the lawn lysoctonia leaf rot (large patch) test group on the 9th and 21st days and one month after the final drug treatment, and the damage area rate after one month was investigated to evaluate the control effect.
その結果、図２および表５に示したように、本発明のＫＢＣ１０１０水和剤は、最大８９．４％の高い防除価を示し、生物農薬としての実用性が高いことを確認した（図２および表５）。 As a result, as shown in FIG. 2 and Table 5, the KBC1010 wettable powder of the present invention showed a high control value of 89.4% at the maximum, and was confirmed to be highly practical as a biopesticide (FIG. 2). And Table 5).
本発明のバチルスサブチリスＫＢＣ１０１０菌株及びその培養液を有効成分として含有する植物病原菌防除用組成物は、芝生の病害を誘発する病原体であるリゾクトニアソラニＡＧ−２−２（ＩＶ）［Rhizoctonia solani AG-2-2(IV)］を含む様々な植物病原菌に対して優れた抗真菌活性を示すことにより、芝生のリゾクトニア葉腐病（ラージパッチ）を含む様々な植物の病気のための環境にやさしい植物病原菌防除用組成物の開発に有用に用いることができる。したがって、化学農薬による生態系の破壊と人畜毒性の問題を解消することができるので、農薬産業、環境産業上非常に有用に用いることができる。 A composition for controlling a phytopathogenic fungus containing the Bacillus subtilis KBC1010 strain of the present invention and a culture solution thereof as an active ingredient is Rhizoctonia solani AG-2-2 (IV) [Rhizoctonia solani AG- 2-2 (IV)] is an environmentally friendly plant for various plant diseases, including lawn rhizobonia leaf rot (large patch), by exhibiting excellent antifungal activity against various plant pathogens including It can be usefully used for developing a composition for controlling pathogens. Therefore, the problem of ecosystem destruction and human and animal toxicity caused by chemical pesticides can be solved, so that it can be very useful in the agricultural chemical industry and the environmental industry.
寄託機関名：韓国生命工学研究院
受託番号：ＫＣＴＣ１０３０４ＢＰ
受託日：２００２．０７．２０
Depositary name: Korea Biotechnology Institute Accession number: KCTC10304BP
Date of entrustment: 2002.7.20
Claims (17)
２）前記工程１）で製造された培養物を乾燥し、粉末化して原剤を製造する工程を含む植物病原菌防除用組成物の製造方法。 1) culturing a Bacillus subtilis KBC1010 strain deposited under accession number KCTC10304BP in a liquid medium to produce a culture;
2) A method for producing a composition for controlling phytopathogenic fungi, comprising a step of drying and pulverizing the culture produced in the step 1) to produce an active ingredient.
Bacillus subtilis KBC1010 (Accession No. KCTC10304BP) strain in a medium containing 0.1 to 10.0 parts by weight of dextrose and 0.1 to 10.0 parts by weight of peptone at 25 to 35 ° C. and pH 5.5 to 5.5 The composition obtained by culturing at 8.5 was dried and pulverized 5.0 to 50.0% (w / w), and a bulking agent for controlling a plant pathogen. Use for wettable powder compositions.
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| PCT/KR2012/010162 WO2013100405A1 (en) | 2011-12-28 | 2012-11-28 | Bacillus subtilis kbc1010 strain, and composition for controlling plant pathogens containing culture broth of same as active ingredient |
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| JP2018525420A (en) * | 2015-08-28 | 2018-09-06 | アグバイオーム， インコーポレイテッド | Bacterial strains and their use for controlling plant diseases |
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