JP5001518B2 - Plant disease control agent using Fusarium strain and control method using the same - Google Patents
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Description
本発明は、植物病害の防除剤、及びそれを利用した植物病害の防除方法に関するものである。 The present invention relates to a plant disease control agent and a plant disease control method using the same.
これまで、農作物の生産における植物病害の防除には化学農薬が多く利用されている。しかし、化学農薬による病害防除では、耐性菌の出現により防除効果が低下する場合が多いため、新たな化学農薬の開発が必要となる場合がある。また、近年では、食物の安全性や環境保護などの面から、化学農薬に代わるべき、または化学農薬と併用すべき手段として、年々環境に影響の少ない微生物資材を利用した病害防除（いわゆる生物農薬）に期待が高まっている。 Until now, many chemical pesticides have been used to control plant diseases in the production of agricultural products. However, in disease control with chemical pesticides, since the control effect often decreases due to the emergence of resistant bacteria, it may be necessary to develop new chemical pesticides. In recent years, as a means to replace chemical chemical pesticides or to be used in combination with chemical pesticides in terms of food safety and environmental protection, disease control using microbial materials that have little environmental impact every year (so-called biological pesticides). ) Expectations are rising.
農園芸作物の病害防除に用いられてきた微生物、特に灰色かび病防除に用いられてきた微生物としては、バチルス属に属する細菌、トリコデルマ属、ペニシリウム属、グリオクラディウム属に属する糸状菌が挙げられる（例えば、特許文献１、特許文献２参照）。しかし、灰色かび病防除にフザリウム属菌が使用された例はない。 Microorganisms that have been used to control diseases of agricultural and horticultural crops, especially those that have been used to control gray mold, include bacteria belonging to the genus Bacillus, fungi belonging to the genus Trichoderma, Penicillium, and Glyocladium. (For example, see Patent Document 1 and Patent Document 2). However, there is no example of Fusarium genus used for gray mold control.
また、イネばか苗病防除に用いられてきた細菌としては、シュードモナス・グラディオリ（Ｐｓｅｕｄｏｍｏｎａｓ ｇｌａｄｉｏｌｉ）、シュードモナス属の一種（Ｐｓｅｕｄｏｍｏｎａｓ ｓｐ.）が挙げられる。イネばか苗病防除に用いられてきた糸状菌としては、トリコデルマ属、非病原性フザリウム属に属する微生物が挙げられ、これまでにこれらの微生物を含有する農園芸用殺菌剤も数多く研究されてきている（例えば、特許文献３、特許文献４参照）。 Examples of bacteria that have been used for controlling rice sapling seedling diseases include Pseudomonas gladioli and a kind of Pseudomonas genus (Pseudomonas sp.). Examples of the filamentous fungi that have been used for the control of rice seedling diseases include microorganisms belonging to the genus Trichoderma and non-pathogenic Fusarium, and many agricultural and horticultural fungicides containing these microorganisms have been studied. (For example, see Patent Document 3 and Patent Document 4).
しかしながら、斑点細菌病防除や褐斑病防除のための、微生物を使用した農園芸用殺菌剤についての例はこれまでに報告されていない。 However, no examples of agricultural and horticultural fungicides using microorganisms for controlling bacterial spot disease and brown spot disease have been reported so far.
一方、フザリウム属菌を用いた植物病害防除については、非病原性フザリウム属菌を利用したサツマイモつる割病、トマト萎凋病など、これらの病害に対する防除法の報告はいくつか知られている。 On the other hand, regarding plant disease control using Fusarium spp., Several reports on control methods for these diseases such as sweet potato vine split disease and tomato wilt disease using non-pathogenic Fusarium spp. Are known.
フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）のうち、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１（ＦＥＲＭ Ｐ−１８２６６）はシンビジウム病害の発病抑制に有効性があり、植物に弱病原性であるとして同定された（特許文献５参照）。この菌株は、シンビジウム黄斑病、腐敗病などのシンビジウム病害全般に効果があることは確認されているが、他の植物病害に効果があるかどうかは不明であった。 Among Fusarium subglutinans, Fusarium subglutinans HPF-1 (FERM P-18266) was identified as being effective in suppressing the pathogenesis of cymbidium diseases and weakly pathogenic to plants ( (See Patent Document 5). Although it has been confirmed that this strain is effective for all cymbidium diseases such as cymbidium macular disease and rot disease, it has been unclear whether it is effective for other plant diseases.
また、フザリウム属菌を用いてイネ種子に対する伝染性病害を防除する方法は、いくつか知られている（特許文献６〜８参照）が、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）を用いた例はない。
上記の通り、環境負荷の少ない、微生物を用いた生物農薬に対する需要は高まっているが、斑点細菌病防除や褐斑病防除のための、微生物を使用した農園芸用殺菌剤についての例はこれまでに報告されていない。また、上記の病害を含む、広い病害防除スペクトラムを有し、植物に対して弱病原性のフザリウム属菌は未だ見いだされていない。
そこで、本発明は、灰色かび病、斑点細菌病、褐斑病及びイネばか苗病を含む植物病害の防除方法を提供することを目的とする。
As mentioned above, there is an increasing demand for biopesticides using microorganisms that have a low environmental impact, but this is an example of agricultural and horticultural fungicides using microorganisms for controlling bacterial spot disease and brown spot disease. It has not been reported until. In addition, no Fusarium spp. That has a broad disease control spectrum including the above-mentioned diseases and is weakly pathogenic to plants has not yet been found.
Then, an object of this invention is to provide the control method of the plant disease containing a gray mold disease, a spot bacterial disease, a brown spot disease, and a rice seedling disease.
上記の目的を達成するため、本発明者らは、弱病原性フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）の植物の茎葉部病害への有用性を検討した。その結果、当該菌株は、灰色かび病、褐斑病および斑点細菌病に活性を示すことを見出した。また、イネ種子伝染性病害への有用性を検討したところ、糸状菌性病害であるイネばか苗病に活性を示すことを見出し、本発明を完成した。 In order to achieve the above-mentioned object, the present inventors examined the usefulness of a weakly pathogenic Fusarium subglutinans (Fusarium subglutinans) for plant foliage diseases. As a result, it was found that the strain was active against gray mold disease, brown spot disease and spotted bacterial disease. Moreover, when the usefulness to a rice seed infectious disease was examined, it discovered that it showed activity in the rice seedling disease which is a filamentous fungal disease, and completed this invention.
すなわち、本発明は、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）を有効成分として含有する、植物の灰色かび病、斑点細菌病又は褐斑病の防除剤を提供する。ここで、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）は、植物に対して弱病原性であることが確認されていることから、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１（ＦＥＲＭ Ｐ−１８２６６）を用いるのが望ましい。また、本発明の防除剤を使用する植物として、果菜を含む野菜を挙げることができる。 That is, the present invention provides a plant gray mold disease, spot bacterial disease or brown spot disease control agent containing Fusarium subglutinans as an active ingredient. Here, since it has been confirmed that Fusarium subglutinans are weakly pathogenic to plants, it is necessary to use Fusarium subglutinans HPF-1 (FERM P-18266). desirable. Moreover, vegetables containing fruit vegetables can be mentioned as plants using the control agent of the present invention.
また、本発明は、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）を有効成分として含有する、イネばか苗病の防除剤を提供する。ここで、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）は、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１（ＦＥＲＭ Ｐ−１８２６６）が望ましい。 Moreover, this invention provides the control agent of the rice seedling disease which contains Fusarium subglutinans (Fusarium subglutatinans) as an active ingredient. Here, the Fusarium subglutinans is preferably Fusarium subglutinans HPF-1 (FERM P-18266).
さらに、本発明は、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）を含有する溶液により植物体を処理することによる、植物の灰色かび病、斑点細菌病又は褐斑病の防除方法を提供する。ここで、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）は、植物に対して弱病原性であることが確認されていることから、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１（ＦＥＲＭ Ｐ−１８２６６）を用いるのが望ましい。本発明の防除方法は、果菜を含む野菜に適用可能である。 Furthermore, the present invention provides a method for controlling plant gray mold disease, spot bacterial disease or brown spot disease by treating a plant with a solution containing Fusarium subglutinans. Here, since it has been confirmed that Fusarium subglutinans are weakly pathogenic to plants, it is necessary to use Fusarium subglutinans HPF-1 (FERM P-18266). desirable. The control method of the present invention is applicable to vegetables including fruit vegetables.
さらに、本発明は、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）を含有する溶液により種子を処理することによる、イネばか苗病の防除方法を提供する。ここで、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）は、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１（ＦＥＲＭ Ｐ−１８２６６）が望ましい。 Furthermore, the present invention provides a method for controlling rice seedling disease by treating seeds with a solution containing Fusarium subglutinans. Here, the Fusarium subglutinans is preferably Fusarium subglutinans HPF-1 (FERM P-18266).
本発明のフザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）を含む植物病害の防除剤は、灰色かび病、斑点細菌病、褐斑病を含む病害に対して発病を強く抑制する作用がある。また、イネの育苗時に発生する糸状菌性病害であるイネばか苗病に対しても、発病抑制効果が強い。本発明の防除剤は、これらの病害に対して現在使用されている化学農薬と同等またはそれ以上の防除効果が期待でき、化学農薬の代替として、または化学農薬と併用して使用できるものである。また、自然界に存在する微生物を使用するため、環境に対する負荷も少なく、化学農薬を使用した場合に比べて耐性菌が出現しにくいと考えられる点でも有効である。 The plant disease control agent containing Fusarium subglutinans of the present invention has an action of strongly suppressing the onset of diseases including gray mold disease, spot bacterial disease, and brown leaf disease. Moreover, it has a strong disease-suppressing effect against rice sapling disease, which is a filamentous fungal disease that occurs during rice seedling raising. The control agent of the present invention can be expected to have a control effect equivalent to or higher than that of currently used chemical pesticides against these diseases, and can be used as an alternative to chemical pesticides or in combination with chemical pesticides. . Moreover, since microorganisms that exist in nature are used, there is little impact on the environment, and it is also effective in that resistant bacteria are less likely to appear than when chemical pesticides are used.
本発明では、フザリウム・サブグルチナンスの任意の菌株を使用できるが、植物に対する弱病原性が確認されていることから、菌株ＨＰＦ−１（ＦＥＲＭ Ｐ−１８２６６）を用いるのが望ましい。本菌株は、産業技術総合研究所生命工業技術研究所（現在、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）に平成１３年３月１９日付けで寄託されたものである。
表１にフザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の形態的特徴を示す。
In the present invention, any strain of Fusarium subglutinance can be used. However, since weak pathogenicity against plants has been confirmed, it is desirable to use strain HPF-1 (FERM P-18266). This strain was deposited at the National Institute of Advanced Industrial Science and Technology (Now, the National Institute of Advanced Industrial Science and Technology (AIST)) on March 19, 2001.
Table 1 shows the morphological characteristics of Fusarium subglutinans HPF-1.
コロニー形状は、ポテトデキストロース寒天平板培地で２５℃、２週間培養後に観察した。形態的特徴は、ＢＬＢ照射下においてＳＮＡ培地で２５℃、５８日で培養したものを光学顕微鏡下で観察した。その結果、本菌は、コロニー形態および菌の形態的特徴からＦｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ（Ｎｅｌｓｏｎら、１９８３）と同定されている。 The colony shape was observed after culturing at 25 ° C. for 2 weeks on a potato dextrose agar plate medium. Morphological characteristics were observed under an optical microscope after culturing in SNA medium at 25 ° C. for 58 days under BLB irradiation. As a result, the present bacterium has been identified as Fusarium subglutinans (Nelson et al., 1983) from the colony morphology and the morphological characteristics of the bacterium.
本発明の植物病害防除剤組成物の培養には、特別な方法を用いる必要はなく、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ-１を液体培養あるいは固体培養等の公知の手段で増殖させた菌体を用いることができる。培養に用いる培地としては、微生物が生育可能な炭素源、窒素源および無機物および必要な生育促進物質を適当に含有する培地であれば、天然培地、合成培地いずれも利用できる。具体的な培地を例示すると、液体培養の場合、ポテトデキストロース液体培地、Ｃｚａｐｅｋ Ｄｏｘ液体培地などを、固体培養の場合、ポテトデキストロース寒天培地、Ｃｚａｐｅｋ Ｄｏｘ寒天培地、フスマ培地などを挙げることができる。なお、液体培養では、胞子又は芽胞状菌体が生産され、固体培養では小型分生子又は大型分生子が生産される。
培養に際しては、培養温度を１５〜３５℃、好ましくは２５〜３０℃、培養日数は１〜３０日、液体培養では好ましくは３〜７日、固体培養では７〜２０日間培養することが望ましい。
The cultivation of the plant disease control agent composition of the present invention does not require any special method, and the fungus body obtained by growing Fusarium subglutinans HPF-1 by a known means such as liquid culture or solid culture. Can be used. As a medium used for the culture, any of a natural medium and a synthetic medium can be used as long as the medium appropriately contains a carbon source, a nitrogen source, an inorganic substance, and a necessary growth promoting substance capable of growing microorganisms. Specific examples of the medium include potato dextrose liquid medium and Czapek Dox liquid medium in the case of liquid culture, and potato dextrose agar medium, Czapek Dox agar medium and bran medium in the case of solid culture. In the liquid culture, spores or spores are produced, and in the solid culture, small conidia or large conidia are produced.
In culturing, the culture temperature is 15 to 35 ° C., preferably 25 to 30 ° C., the culture period is 1 to 30 days, preferably 3 to 7 days for liquid culture, and 7 to 20 days for solid culture.
本発明では、灰色かび病害、斑点細菌病害および褐斑病害防除法は、フザリウム・サブグルチナンスを有効成分とする防除剤により処理することにより行う。この処理は、通常、茎葉散布することによる。また、イネばか苗病害防除法は、本菌株菌体を有効成分とする防除剤でイネ種子を消毒することにより行う。 In the present invention, the method for controlling gray mold disease, spotted bacterial disease and brown spot disease is carried out by treating with a control agent containing Fusarium subglutinance as an active ingredient. This treatment is usually by spraying foliage. In addition, the rice shoot seedling disease control method is carried out by disinfecting rice seeds with a control agent comprising the present bacterial strain as an active ingredient.
本発明における防除剤として用いる場合には、本菌株の菌体または培養液を単独で用いてもよいが、微生物の保存法（根井外喜男編、東京大学出版会、１９７７）で示されているような凍結菌体を用いてもよい。すなわち、遠心分離等の操作で濃縮した菌体を、サッカロースなどの保護剤に懸濁し、予備凍結した後、凍結乾燥機により乾燥菌体を作成する。さらに乾燥菌体を粉砕し、粉砕菌体を担体または補助剤等と配合して常法により、例えば、粉剤、粒剤、水和剤などの形態にして使用すると更に好ましい。好適な担体としては、例えば、鉱物質粉末（カオリン、ベントナイト、クレー、タルク、ゼオライト、ケイソウ土、バーミキュライト、炭酸カルシウム、ホワイトカーボン、消石灰など）、植物質粉末(大豆粉、小麦粉、でんぷんなど)等の固体担体が挙げられる。また、補助剤としては、例えば、カルボキシメチルセルロース（ＣＭＣ）、ゼラチン、アルギン酸ナトリウム等が挙げられる。 When used as a control agent in the present invention, the cell or culture solution of this strain may be used alone, but it is shown in a method for preserving microorganisms (edited by Yoshio Nei, University of Tokyo Press, 1977). Such frozen cells may be used. That is, microbial cells concentrated by an operation such as centrifugation are suspended in a protective agent such as saccharose, pre-frozen, and then dried microbial cells are prepared by a freeze dryer. Further, it is more preferable to pulverize the dried microbial cells, mix the pulverized microbial cells with a carrier or an auxiliary agent, etc., and use them in the form of, for example, powders, granules, wettable powders, etc. Suitable carriers include, for example, mineral powder (kaolin, bentonite, clay, talc, zeolite, diatomaceous earth, vermiculite, calcium carbonate, white carbon, slaked lime, etc.), vegetable powder (soy flour, wheat flour, starch, etc.), etc. Solid support. Examples of the auxiliary agent include carboxymethyl cellulose (CMC), gelatin, sodium alginate and the like.
使用量としては、製剤の剤型、適用方法、適用場所、適用すべき病害の種類などに応じて適宜選択可能である。水和剤の場合、フザリウム・サブグルチナンスの菌体濃度は、１０4〜１０9細胞／ｍｌ程度、好ましくは１０6〜１０8細胞／ｍｌ程度の範囲で使用することができる。菌体は、芽胞状菌体、小型分生子、大型分生子またはそれら混合物のいずれをも使用可能である。 The amount used can be appropriately selected according to the dosage form of the preparation, the application method, the application location, the type of disease to be applied, and the like. In the case of a wettable powder, the cell concentration of Fusarium subglutinance can be used in the range of about 10 4 to 10 9 cells / ml, preferably about 10 6 to 10 8 cells / ml. As the fungus body, any of spore-like fungus bodies, small conidia, large conidia or a mixture thereof can be used.
本発明の防除剤及び防除方法は、任意の植物に適用でき、果菜を含む野菜のような農業用植物、果樹、園芸植物等に使用できる。 The control agent and control method of the present invention can be applied to any plant, and can be used for agricultural plants such as vegetables including fruit vegetables, fruit trees, garden plants, and the like.
以下に実施例を挙げて、本発明をさらに詳細に説明するが、本実施例により本発明を制限または限定することを意図するものではない。
〔実施例〕
EXAMPLES The present invention will be described in more detail with reference to the following examples. However, the present invention is not intended to be limited or limited by the examples.
〔Example〕
＜調製例１：フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１芽胞状菌体の調製＞
２００ｍｌのポテトデキストロース液体培地を含む、５本の５００ｍｌコルベン（計１０００ｍｌ）を、１２１℃で２０分間オートクレーブ滅菌した。滅菌後の培地に、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の菌体を接種し、２７℃で４日間振とう培養した。培養終了後、培養物を３重のガーゼで濾過し、遠心分離することにより、１×１０9〜３×１０9細胞／ｍｌ程度に濃縮された芽胞状菌体を得た。
<Preparation Example 1: Preparation of Fusarium subglutinans HPF-1 spores>
Five 500 ml Kolben (total 1000 ml) containing 200 ml of potato dextrose liquid medium were autoclaved at 121 ° C. for 20 minutes. The sterilized medium was inoculated with Fusarium subglutinans HPF-1 and cultured with shaking at 27 ° C. for 4 days. After completion of the culture, the culture was filtered through triple gauze and centrifuged to obtain spore-like cells concentrated to about 1 × 10 9 to 3 × 10 9 cells / ml.
＜調製例２：フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ-１芽胞状菌体の調製＞
２００ｍｌの半合成培地（脱イオン水１リットルあたり、２０.０ｇのＳｏｌｕｂｌｅ Ｓｔａｒｃｈ、５．０ｇのグルコース、２.０ｇのポリペプトン、３.０ｇの乾燥酵母抽出物、１．０ｇのＫＨ2ＰＯ4、０．５ｇのＭｇＳＯ4・７Ｈ2Ｏ、０.５ｇのＫＣｌを含む）を含む、５本の５００ｍｌコルベン（計１０００ｍｌ）を、１２１℃で２０分間オートクレーブ滅菌した。滅菌後の培地に、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の菌体を接種し、２７℃で４日間振とう培養した。培養終了後、培養物を３重のガーゼで濾過し、遠心分離することにより、１×１０9〜３×１０9細胞／ｍｌ程度に濃縮された芽胞状菌体を得た。
<Preparation Example 2: Preparation of Fusarium subglutinans HPF-1 spores>
200 ml semi-synthetic medium (20.0 g Soluble Starch, 5.0 g glucose, 2.0 g polypeptone, 3.0 g dry yeast extract, 1.0 g KH 2 PO 4 per liter of deionized water, Five 500 ml Kolbens (1000 ml total) containing 0.5 g MgSO 4 .7H 2 O, 0.5 g KCl) were autoclaved at 121 ° C. for 20 minutes. The sterilized medium was inoculated with Fusarium subglutinans HPF-1 and cultured with shaking at 27 ° C. for 4 days. After completion of the culture, the culture was filtered through triple gauze and centrifuged to obtain spore-like cells concentrated to about 1 × 10 9 to 3 × 10 9 cells / ml.
＜調製例３：フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ-１小型分生子の調製＞
１５ｍｌのＳＮ培地（脱イオン水１リットルあたり、１.０ｇのＫＨ2ＰＯ4、１.０ｇのＫＮＯ3、０．５ｇのＭｇＳＯ4・７Ｈ2Ｏ、０．５ｇのＫＣｌ、０．２ｇのグルコース、０．２ｇのスクロースを含む）と２０ｇのフスマとを含む２００ｍｌコルベンを、１２１℃で２０分間オートクレーブ滅菌した。滅菌後の培地に、フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の菌体を接種し、２５℃で９日間静置培養した。培養物を３重のガーゼで濾過し、小型分生子を調製した。
<Preparation Example 3: Preparation of Fusarium subglutinans HPF-1 small conidia>
15 ml of SN medium (1.0 g KH 2 PO 4 , 1.0 g KNO 3 , 0.5 g MgSO 4 .7H 2 O, 0.5 g KCl, 0.2 g glucose per liter of deionized water 200 ml Kolben containing 0.2 g of sucrose and 20 g of bran was autoclaved at 121 ° C. for 20 minutes. The sterilized medium was inoculated with Fusarium subglutinans HPF-1 and statically cultured at 25 ° C. for 9 days. The culture was filtered through triple gauze to prepare small conidia.
＜調製例４：フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ-１芽胞状菌体の水和剤調製＞
調製例２で得られた芽胞状菌体を遠心分離することにより、２．５×１０10胞子／ｍｌに濃縮した芽胞状菌体２０部を、ゼオライト８０部と均一に混合することにより水和剤を調製した。
〔実施例１〕
<Preparation Example 4: Preparation of wettable powder of Fusarium subglutinans HPF-1 spores>
By centrifuging the spore-like cells obtained in Preparation Example 2, 20 parts of spore-like cells concentrated to 2.5 × 10 10 spores / ml are hydrated by uniformly mixing with 80 parts of zeolite. An agent was prepared.
[Example 1]
＜インゲン灰色かび病に対する発病抑制効果１＞
調製例１で調製したフザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１芽胞状菌液を、インゲン（品種：姫手芒）の初生葉に散布した。散布濃度は、１×１０6細胞／ｍｌ、１×１０7細胞／ｍｌ又は１×１０8細胞／ｍｌとした。その後、２４℃下で管理し、３日後に灰色かび病の病原菌であるボトリシス・シネレア（Ｂｏｔｒｙｔｉｓ ｃｉｎｅｒｅａ）の分生子懸濁液（２．０×１０6胞子／ｍｌ）を散布した。散布処理後、２０℃で５日間インキュベートした後、初生葉の病斑面積率を調査し、式１により防除価を算出した。また、灰色かび病に対する化学農薬であるセイビアーフロアブル２０を１０００倍希釈したものを散布し、本発明の防除剤と比較した。なお、試験は３反復で行った。
<Onset inhibitory effect 1 against gray bean mildew>
The Fusarium subglutinans HPF-1 spore-like fungus solution prepared in Preparation Example 1 was sprayed on the primary leaves of green beans (variety: Himete-don). The application concentration was 1 × 10 6 cells / ml, 1 × 10 7 cells / ml or 1 × 10 8 cells / ml. Thereafter, the mixture was controlled at 24 ° C., and after 3 days, a conidial suspension (2.0 × 10 6 spores / ml) of Botrytis cinerea, a pathogen of gray mold disease, was sprayed. After spraying treatment, after incubation at 20 ° C. for 5 days, the lesion area ratio of the primary leaves was investigated, and the control value was calculated by Formula 1. Moreover, what diluted 1000 times the Savior flowable 20 which is a chemical pesticide with respect to a gray mold disease was sprayed, and it compared with the control agent of this invention. Note that the test was repeated three times.
防除価＝（１−処理区の平均病斑面積率÷無処理区の平均病斑面積率）×１００…式１ Control value = (average lesion area ratio of the average lesion area ratio ÷ non-treated group of 1-treated group) × 100 ... formula 1
結果を表２に示す。フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の芽胞状菌液を処理した区は、インゲン灰色かび病に対して高い発病抑制効果を示した。 The results are shown in Table 2. The group treated with the spore-like fungal solution of Fusarium subglutinans HPF-1 showed a high disease-inhibiting effect against green bean mold.
＜インゲン灰色かび病に対する発病抑制効果２＞
調製例３で調製したフザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の小型分生子の菌液を、インゲン（品種：姫手芒）の初生葉に散布した。散布濃度は、１×１０7細胞／ｍｌ又は１×１０8細胞／ｍｌとした。その後、２４℃下で管理し、３日後灰色かび病の病原菌であるボトリシス・シネレア（Ｂｏｔｒｙｔｉｓ ｃｉｎｅｒｅａ）の分生子懸濁液（２．０×１０6胞子／ｍｌ）を散布した。散布処理後、２０℃で５日間インキュベートした後、初生葉の病斑面積率を調査し、式１により防除価を算出した。また、灰色かび病に対する化学農薬であるセイビアーフロアブル２０を１０００倍希釈したものを散布し、本発明の防除剤と比較した。なお、試験は３反復で行った。
<Onset inhibitory effect 2 against green beans mildew>
The liquid solution of Fusarium subglutinans HPF-1 small conidia prepared in Preparation Example 3 was sprayed on the primary leaves of green beans (variety: Himete-don). The application concentration was 1 × 10 7 cells / ml or 1 × 10 8 cells / ml. Thereafter, the mixture was controlled at 24 ° C., and after 3 days, a conidial suspension (2.0 × 10 6 spores / ml) of Botrytis cinerea, a pathogen of gray mold disease, was sprayed. After spraying treatment, after incubation at 20 ° C. for 5 days, the lesion area ratio of the primary leaves was investigated, and the control value was calculated by Formula 1. Moreover, what diluted 1000 times the Savior flowable 20 which is a chemical pesticide with respect to a gray mold disease was sprayed, and it compared with the control agent of this invention. Note that the test was repeated three times.
結果を表３に示す。フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の小型分生子を処理した区は、インゲン灰色かび病に対して高い発病抑制効果を示した。 The results are shown in Table 3. The group treated with small conidia of Fusarium subglutinans HPF-1 showed a high disease-inhibiting effect on green bean mold.
＜インゲン灰色かび病に対する発病抑制効果３＞
調製例２及び調製例４で調製したフザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の芽胞状菌液又はその水和剤を、インゲン（品種：姫手芒）の初生葉に散布した。散布濃度は、調製例２で調製した菌液は１×１０7細胞／ｍｌ及び１×１０8細胞／ｍｌ、調製例４で調製した水和剤は５００倍希釈とした。その後、２４℃下で管理し、３日後灰色かび病の病原菌であるボトリシス・シネレア（Ｂｏｔｒｙｔｉｓ ｃｉｎｅｒｅａ）の分生子懸濁液（２．０×１０6胞子／ｍｌ）を散布した。散布処理後、２０℃で５日間インキュベートした後、初生葉の病斑面積率を調査し、式１により防除価を算出した。また、灰色かび病に対する化学農薬であるセイビアーフロアブル２０を１０００倍希釈したものを散布し、本発明の防除剤と比較した。なお、試験は３反復で行った。
<Disease inhibitory effect 3 against green beans fungus>
The spore-like fungus solution of Fusarium subglutinans HPF-1 prepared in Preparation Example 2 and Preparation Example 4 or a wettable powder thereof was sprayed on the primary leaves of green beans (variety: Himete-an). The spray concentration was 1 × 10 7 cells / ml and 1 × 10 8 cells / ml for the bacterial solution prepared in Preparation Example 2, and the wettable powder prepared in Preparation Example 4 was diluted 500 times. Thereafter, the mixture was controlled at 24 ° C., and after 3 days, a conidial suspension (2.0 × 10 6 spores / ml) of Botrytis cinerea, a pathogen of gray mold disease, was sprayed. After spraying treatment, after incubation at 20 ° C. for 5 days, the lesion area ratio of the primary leaves was investigated, and the control value was calculated by Formula 1. Moreover, what diluted 1000 times the Savior flowable 20 which is a chemical pesticide with respect to a gray mold disease was sprayed, and it compared with the control agent of this invention. Note that the test was repeated three times.
結果を表４に示す。フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の芽胞状菌体及びその水和剤を処理した区は、インゲン灰色かび病に対して高い発病抑制効果を示した。 The results are shown in Table 4. The group treated with Fusarium subglutinans HPF-1 spores and its hydrating agent showed a high disease-inhibiting effect on bean gray mold.
＜キュウリ斑点細菌病に対する発病抑制効果＞
調製例１で調製したフザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１芽胞状菌液を、キュウリ（品種：四葉）のポットに散布した。散布濃度は、１×１０7細胞／ｍｌ又は１×１０8細胞／ｍｌとした。その後、２４℃下で管理し、３日後に斑点細菌病の病原菌であるシュードモナス・シリンゲ（Ｐｓｅｕｄｏｍｏｎａｓ ｓｙｒｉｎｇａｅ）の懸濁液（１．０×１０8ＣＦＵ／ｍｌ）を散布した。散布処理後、２４℃で１０日間インキュベートした後、第２本葉および第３本葉の病斑数を調査し、式２により防除価を算出した。また、斑点細菌病に対する化学農薬であるカスミンボルドーを１０００倍希釈したものを散布し、本発明の防除剤と比較した。なお、試験は３反復で行った。
<Suppression effect against cucumber spotted bacterial disease>
The Fusarium subglutinans HPF-1 spore-like fungus solution prepared in Preparation Example 1 was sprayed onto a cucumber (variety: Yotsuba) pot. The application concentration was 1 × 10 7 cells / ml or 1 × 10 8 cells / ml. Thereafter, the suspension was controlled at 24 ° C., and after 3 days, a suspension (1.0 × 10 8 CFU / ml) of Pseudomonas syringae, a pathogenic bacterium of spotted bacterial disease, was sprayed. After spraying treatment, after incubation at 24 ° C. for 10 days, the number of lesions on the second true leaf and the third true leaf was examined, and the control value was calculated according to Formula 2. Moreover, what diluted 1000 times Casmin Bordeaux which is a chemical pesticide with respect to a spot bacterial disease was sprayed, and it compared with the control agent of this invention. Note that the test was repeated three times.
防除価＝（１−処理区の平均病斑数÷無処理区の平均病斑数）×１００…式２ Control value = (average disease the number of plaques of average lesion number ÷ non-treated area of the 1-treated group) × 100 ... Equation 2
結果を表５に示す。フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１芽胞状菌液を処理した区は、キュウリ斑点細菌病に対して高い発病抑制効果を示した。 The results are shown in Table 5. The group treated with Fusarium subglutinans HPF-1 spore-like bacterial solution showed a high disease-inhibiting effect against cucumber spotted bacterial disease.
＜キュウリ褐斑病に対する発病抑制効果＞
調製例１で調製したフザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の芽胞状菌液を、キュウリ（品種：相模半白）のポットに散布した。散布濃度は、１×１０7細胞／ｍｌ又は１×１０8細胞／ｍｌとした。その後、２４℃下で管理し、３日後に褐斑病の病原菌であるコリネスポーラ・キャシコーラ（Ｃｏｒｙｎｅｓｐｏｒａ ｃａｓｓｉｉｃｏｌａ）の分生子懸濁液（２．０×１０5胞子／ｍｌ）を散布した。散布処理後、２４℃で８日間インキュベートした後、第１本葉の病斑数を調査し、式２により防除価を算出した。また、褐斑病等に対する化学農薬であるダコニール１０００を１０００倍希釈したものを散布し、本発明の防除剤と比較した。なお、試験は３反復で行った。
<Onset inhibitory effect on cucumber brown spot disease>
The spore-like fungal solution of Fusarium subglutinans HPF-1 prepared in Preparation Example 1 was sprayed on a pot of cucumber (variety: Sagamihanjiro). The application concentration was 1 × 10 7 cells / ml or 1 × 10 8 cells / ml. Thereafter, the mixture was controlled at 24 ° C., and after 3 days, a conidial suspension (2.0 × 10 5 spores / ml) of Corynespora casicola, which is a pathogen of brown spot disease, was sprayed. After spraying treatment, after incubation at 24 ° C. for 8 days, the number of lesions on the first true leaf was examined, and the control value was calculated by Equation 2. In addition, a 1000-fold diluted product of Daconil 1000, which is a chemical pesticide against brown spot disease, was sprayed and compared with the control agent of the present invention. Note that the test was repeated three times.
結果を表６に示す。フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の芽胞状菌液を処理した区は、キュウリ褐斑病に対して高い発病抑制効果を示した。 The results are shown in Table 6. The group treated with the spore-like fungus solution of Fusarium subglutinans HPF-1 showed a high disease-inhibiting effect against cucumber brown spot disease.
＜イネばか苗病に対する発病抑制効果（自然感染籾）＞
調製例１で調製したフザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の芽胞状菌液（１×１０8細胞／ｍｌ）内に、イネばか苗病罹病籾（品種：新潟早生、自然感染籾）を、１５℃、浸種前２４時間浸漬処理（浸漬液量比＝１：１）した後、１５℃で５日間浸種（浸種液量比＝１：２）した。なお、浸種３日後に水交換を１回行った。その後、水を捨て３２℃の催芽器内に１７時間保って催芽させた。育苗培土（くみあい合成培土３号）を充填した育苗箱に催芽種子を播種し、播種後２日間、３２℃の育苗庫内に静置し出芽処理を行った。次に、出芽処理した苗を温室内で２１日間管理した後、全苗について発病の有無（徒長苗および枯死苗）を調査し、式３により発病苗率（％）を、また式４により防除価を算出した。また、イネばか苗病に対する化学農薬であるモミガードＣ水和剤を２００倍希釈したもので１５℃、浸種前２４時間浸漬処理し、本発明の防除剤と比較した。なお、試験は３反復で行った。
<Disease control effect against rice sapling seedling disease (natural infection)>
In a spore-like fungus solution (1 × 10 8 cells / ml) of Fusarium subglutinans HPF-1 prepared in Preparation Example 1 After immersion at 15 ° C. for 24 hours before soaking (immersion liquid volume ratio = 1: 1), seeding was performed at 15 ° C. for 5 days (immersion liquid volume ratio = 1: 2). In addition, water exchange was performed once 3 days after immersion. Thereafter, the water was discarded and germination was carried out in a germinator at 32 ° C. for 17 hours. Germination seeds were sown in a seedling box filled with seedling culture soil (Kumiai Synthetic Culture soil No. 3), and the seedlings were allowed to stand in a seedling storage room at 32 ° C. for 2 days after sowing and budding treatment was performed. Next, after the germinated seedlings were managed in the greenhouse for 21 days, all seedlings were examined for the presence or absence of disease (length seedlings and dead seedlings), and the percentage of diseased seedlings (%) was controlled by Formula 3 and controlled by Formula 4 The value was calculated. Further, 200-fold diluted Momigard C wettable powder, which is a chemical pesticide against rice sapling seedling disease, was immersed at 15 ° C. for 24 hours before soaking, and compared with the control agent of the present invention. Note that the test was repeated three times.
発病苗率（％）＝（徒長苗数＋枯死苗数）／全調査苗数×１００…式３ Onset seedling ratio (%) = (spindly seedlings number + withering number of seedlings) / total survey seedlings number × 100 ... Equation 3
防除価＝（１−処理区の平均発病苗率÷無処理区の平均発病苗率）×１００…式４
Control value = (average disease seedling rate of the average disease seedling rate ÷ non-treated area of the 1-treated group) × 100 ... Equation 4
結果を表７に示す。フザリウム・サブグルチナンス（Ｆｕｓａｒｉｕｍ ｓｕｂｇｌｕｔｉｎａｎｓ）ＨＰＦ−１の芽胞状菌液を処理した区は、イネばか苗病に対して高い発病抑制効果を示した。 The results are shown in Table 7. The group treated with the spore-like fungus solution of Fusarium subglutinans HPF-1 showed a high disease-inhibiting effect against rice sapling seedling disease.
Claims (6)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2005062176A JP5001518B2 (en) | 2005-03-07 | 2005-03-07 | Plant disease control agent using Fusarium strain and control method using the same |
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| JP3807243B2 (en) * | 2001-03-30 | 2006-08-09 | 山梨県 | Efficient inoculation method for weakly pathogenic Fusarium fungus to cymbidium |
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