JP4737671B2 - Agricultural / horticultural fungicide composition - Google Patents
Agricultural / horticultural fungicide composition Download PDFInfo
- Publication number
- JP4737671B2 JP4737671B2 JP2005171785A JP2005171785A JP4737671B2 JP 4737671 B2 JP4737671 B2 JP 4737671B2 JP 2005171785 A JP2005171785 A JP 2005171785A JP 2005171785 A JP2005171785 A JP 2005171785A JP 4737671 B2 JP4737671 B2 JP 4737671B2
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- JP
- Japan
- Prior art keywords
- agricultural
- bacillus
- control
- fungicide composition
- horticultural
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 238000000034 method Methods 0.000 claims description 21
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- IQGKIPDJXCAMSM-UHFFFAOYSA-N triazoxide Chemical compound N=1C2=CC=C(Cl)C=C2[N+]([O-])=NC=1N1C=CN=C1 IQGKIPDJXCAMSM-UHFFFAOYSA-N 0.000 description 1
- HSMVPDGQOIQYSR-KGENOOAVSA-N triflumizole Chemical compound C1=CN=CN1C(/COCCC)=N/C1=CC=C(Cl)C=C1C(F)(F)F HSMVPDGQOIQYSR-KGENOOAVSA-N 0.000 description 1
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- FJBGIXKIXPUXBY-UHFFFAOYSA-N {2-[3-(4-chlorophenyl)propyl]-2,4,4-trimethyl-1,3-oxazolidin-3-yl}(imidazol-1-yl)methanone Chemical compound C1=CN=CN1C(=O)N1C(C)(C)COC1(C)CCCC1=CC=C(Cl)C=C1 FJBGIXKIXPUXBY-UHFFFAOYSA-N 0.000 description 1
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Description
本発明は、農園芸用殺菌剤に関し、更に詳しくはバシルス属に属する細菌の芽胞と特定の化学合成殺菌剤を含有する、病害防除作用に優れた農園芸用殺菌剤組成物に関する。 The present invention relates to an agricultural and horticultural fungicide, and more particularly to an agricultural and horticultural fungicide composition excellent in disease control action, which contains bacterial spores belonging to the genus Bacillus and a specific chemically synthesized fungicide.
植物病害防除法としては、輪作や太陽熱を利用した耕種的あるいは物理的防除、化学農薬を用いる化学的防除、病害抵抗性品種の利用による防除、弱毒ウィルスや病原菌に対する拮抗微生物を用いた生物的防除等が挙げられる。これらのうち、化学農薬、特に有機合成殺菌剤の開発研究は近年に至るまで目覚ましく発達し、効力が高く様々な作用を有する多数の薬剤が次々と開発され、更には様々な施用法も開発された。これらを用いた化学的防除法は植物の病害防除並びに防除作業の省力化等に大きく貢献し、広く普及している。しかしながら、近年いわゆる薬剤耐性菌の出現により、化学的防除法による防除効果が低下するという現象が一部作物、病害で認められ、問題となってきている。また作物の指定産地化が進むにつれて連作を余儀なくされる結果、化学農薬では難防除とされる土壌伝染性病害の発生も各地で深刻な問題となっている。さらに、化学農薬を大量に何度も繰り返して用いる方法は、自然界に存在しない化学物質を環境中に放出するため、動植物に直接毒性を有する薬剤のみならず、そうでない薬剤であっても、環境へ悪影響を引き起こすことが懸念されている。 Plant disease control methods include cultivating or physical control using crop rotation and solar heat, chemical control using chemical pesticides, control using disease resistant varieties, and biological control using antagonistic microorganisms against attenuated viruses and pathogens. Etc. Among these, research and development of chemical pesticides, especially organic synthetic fungicides, has developed remarkably until recently, many drugs having high efficacy and various actions have been developed one after another, and various application methods have also been developed. It was. Chemical control methods using these have greatly contributed to the control of plant diseases and the labor saving of control work, and are widely spread. However, due to the emergence of so-called drug-resistant bacteria in recent years, the phenomenon that the control effect by the chemical control method is reduced has been recognized in some crops and diseases and has become a problem. In addition, as crops are designated as local production areas, continuous cropping is unavoidable. As a result, the occurrence of soil-borne diseases that are difficult to control with chemical pesticides has become a serious problem. Furthermore, the method of repeatedly using a large number of chemical pesticides repeatedly releases chemical substances that do not exist in nature into the environment, so that not only drugs that are directly toxic to animals and plants, There are concerns that it will cause adverse effects.
以上のように化学農薬による病害防除は耐性菌の出現によって防除効果が低下する可能性が高く、その場合新たなる殺菌剤の開発が必要となってしまう。また化学農薬では難防除とされる病害防除に対しては、代替手段あるいは他の方法を併用する手段を講じなくてはならない。さらに、環境に対してより安全性の高い防除技術の確立も望まれている。 As described above, disease control with chemical pesticides is likely to reduce the control effect due to the emergence of resistant bacteria, and in this case, development of a new fungicide is required. In addition, for the control of diseases that are difficult to control with chemical pesticides, alternative means or other means must be taken together. Furthermore, establishment of control technology with higher safety for the environment is also desired.
近年このような背景のもと、化学農薬の使用に偏った防除方法を見直し、化学農薬からより環境への安全性が高いと想定される微生物を利用した生物防除（いわゆる生物農薬）方法が提案され、その一部は実用化されている。このような農園芸植物の病害防除に用いる微生物として、トリコデルマ属、グリオクラディウム属、タラロマイゼス属に属する糸状菌、アグロバクテリウム属、シュードモナス属、バシルス属に属する細菌等が挙げられる。 In recent years, based on this background, we reviewed the control methods that are biased toward the use of chemical pesticides, and proposed biocontrol methods using so-called microorganisms (so-called biological pesticides) that are expected to be safer from the environment. Some of them have been put into practical use. Examples of microorganisms used for controlling diseases of such agricultural and horticultural plants include filamentous fungi belonging to the genus Trichoderma, genus Gliocladium, genus Talaromyces, bacteria belonging to the genus Agrobacterium, Pseudomonas and Bacillus.
この中でバシルス属に属する細菌については、例えば、バシルス・ズブチリス（Ｂａｃｉｌｌｕｓ ｓｕｂｔｉｌｉｓ）ＪＢ３菌株（ＮＣＩＢ１２３７５）、バシルス・ズブチリスＪＢ３．６菌株（ＮＣＩＢ１２３７６）、バシルス・ズブチリスＲ１菌株（ＮＣＩＢ１２６１６）あるいはこれらの変異株等から得られる抗菌物質が植物の病害、動物およびヒトの微生物感染を抑制し、更に一般的な微生物汚染を抑制するとし、上記各菌株の培養物を用いて各種農園芸用植物の病害を防除する試みがなされている（特許文献１）。 Among the bacteria belonging to the genus Bacillus, for example, Bacillus subtilis JB3 strain (NCIB12375), Bacillus subtilis JB3.6 strain (NCIB12376), Bacillus subtilis R1 strain (NCIB12616) or mutants thereof. Antibacterial substances obtained from the above control plant diseases, microbial infections of animals and humans, and further suppress general microbial contamination, and control diseases of various agricultural and horticultural plants using cultures of the above strains Attempts have been made (Patent Document 1).
このような現状で、化学合成殺菌剤と拮抗微生物との併用による病害防除が考えられる。しかし、化学合成殺菌剤は拮抗微生物の生育に悪影響を与える場合が多く、相加もしくは相乗的な防除効果を得ることが難しい。このような状況下で、病原菌と拮抗する微生物と化学合成殺菌剤を併用することで、相乗的な防除効果を得ようとする試みが行われている（例えば、特許文献２）。また、病原菌と拮抗する微生物と化学合成殺菌剤を併用することで、耐性菌の抑制効果を得ようとする試みも行われている（例えば、特許文献３）。 Under such circumstances, it is conceivable to control diseases by the combined use of chemically synthesized fungicides and antagonistic microorganisms. However, chemical synthetic fungicides often adversely affect the growth of antagonistic microorganisms, and it is difficult to obtain an additive or synergistic control effect. Under such circumstances, attempts have been made to obtain a synergistic control effect by using a microorganism that antagonizes the pathogen and a chemically synthesized fungicide (for example, Patent Document 2). Attempts have also been made to obtain an inhibitory effect on resistant bacteria by using a microorganism that antagonizes pathogenic bacteria and a chemically synthesized fungicide (for example, Patent Document 3).
前者の試みにおいては、本発明と同様にバシルス属に属する細菌と化学合成殺菌剤とからなる殺菌剤組成物が使用されているが、実施例については種子処理の場合にとどまり、例えば、直接植物に散布する方法、土壌に散布する方法、植物や土壌に添加する水や肥料に添加する方法などによる相乗的な防除効果を利用した植物病害防除についての記載はない。 In the former attempt, a fungicide composition comprising a bacterium belonging to the genus Bacillus and a chemically synthesized fungicide as in the present invention is used. However, the examples are limited to seed treatment, for example, a direct plant. There is no description of plant disease control using a synergistic control effect such as a method of spraying on a soil, a method of spraying on soil, a method of adding to water or fertilizer added to plants or soil.
後者の試みにおいても、本発明と同様にバシルス属に属する細菌と化学合成殺菌剤を併用することで、耐性菌の抑制効果を得ようとする技術が開示されているが、化学合成殺菌剤との併用による相乗的な防除効果を利用した植物病害防除についての記載はない。 Even in the latter attempt, a technique for obtaining an inhibitory effect on resistant bacteria by using a bacterium belonging to the genus Bacillus and a chemically synthesized fungicide in the same manner as in the present invention has been disclosed. There is no description about plant disease control using the synergistic control effect by the combined use.
また、本発明に用いられるバシルス属に属する細菌の芽胞と農園芸用殺菌化合物の混合剤については知られていない。
本発明は、優れた病害防除作用を求めながら、同時に安全性や環境への十分な配慮といった、当業界のニーズに応えるため、化学合成殺菌剤の使用量を減らしても、なおかつ高い防除効果を有し、環境に対してより安全性が高く病害防除作用にも優れた農園芸用殺菌剤組成物を提供することを目的とする。 In order to meet the needs of the industry such as safety and sufficient consideration for the environment while seeking an excellent disease control effect, the present invention has a high control effect even if the amount of chemically synthesized fungicides is reduced. An object of the present invention is to provide an agricultural and horticultural fungicide composition that is safer for the environment and superior in disease control.
本発明者らは、上記課題を解決するため鋭意研究を重ねた結果、バシルス属に属する細菌の芽胞と化学合成殺菌剤との組合せにより、上記の課題が解決されることを見出した。本発明はかかる知見に基づいて完成させたものである。 As a result of intensive studies to solve the above problems, the present inventors have found that the above problems can be solved by a combination of bacterial spores belonging to the genus Bacillus and a chemically synthesized fungicide. The present invention has been completed based on this finding.
本発明の要旨は以下のとおりである。農園芸用殺菌化合物は市販されているか、または農園芸用殺菌剤として知られた化合物であり、これらの化合物は日本植物防疫協会発行の農薬ハンドブック（２００２年）、全国農業協同組合連合会発行のクミアイ農薬総覧（２００５年）及び同連合会発行のＳＨＩＢＵＹＡ ＩＮＤＥＸ（２００５年）などで知られる。 The gist of the present invention is as follows. Agricultural and horticultural bactericidal compounds are commercially available or known as agricultural and horticultural bactericides. These compounds are published by the Japanese Agricultural Handbook (2002), published by the National Federation of Agricultural Cooperatives. Kumiai Agricultural Chemicals Directory (2005) and SHIBUYA INDEX (2005) published by the association are known.
（１）化学合成殺菌化合物、すなわち無機銅化合物、例えば塩基性硫酸銅、無水硫酸銅、水酸化第二銅、塩基性塩化銅等、有機銅化合物、例えば有機銅、ノニルフェノールスルホン酸銅等、無機硫黄化合物、例えば硫黄、全硫化態硫黄等、有機硫黄化合物、例えばジネブ、マンネブ、プロピネブ、チアジアジン、チウラム、ポリカーバメート等、アニリノピリミジン系化合物、例えばシプロジニル、ピリメタニル、メパニピリム等、フェニルピロール系化合物、例えばフルジオキソニル等、有機塩素系化合物、例えばクロロタロニル、キャプタン、トリアジン、フルアジナム、スルフェン酸、フサライド等、炭酸水素塩剤、例えば炭酸水素ナトリウム、炭酸水素カリウム等、有機リン系化合物、例えばＥＤＤＰ、ホセチル、トルクロホスメチル、ＩＢＰ等、ベンズイミダゾール系化合物、例えばカルベンダジム、チオファネートメチル、チアベンダゾール、ベノミル、フベリダゾール等、ジカルボキシイミド系化合物、例えばイプロジオン、プロシミドン、ビンクロゾリン等、アゾール系化合物、例えばフェンブコナゾール、シメコナゾール、ジクロブトラゾール、トリチコナゾール、イプコナゾール、フルコナゾール、ミクロブタニル、ペンコナゾール、ビテルタノール、ブロムコナゾール、オキスポコナゾール、シプロコナゾール、ジフェノコナゾール、ジニコナゾール、エポキシコナゾール、フェンブコナゾール、フルキンコナゾール、フルシラゾール、フルトリアホール、ヘキサコナゾール、イミベンコナゾール、メトコナゾール、プロピコナゾール、シプコナゾール、テブコナゾール、テトラコナゾール、トリアジメホン、トリアジメノール等、イミダゾール系化合物、例えばトリフルミゾール、プロクロラズ、イマザリル、ペフラゾエート等、ピペラジン系化合物、例えばトリホリン等、モルホリン系化合物、例えばフェンプロピモルフ、トリデモルフ、フェンプロピジン等、ヒドロキシピリミジン系化合物、例えばエチリモル、ジメチリモル等、グアニジン化合物、例えばイミノクタジン酢酸塩、イミノクタジンアルベシル酸塩、グアザチン等、酸アミド系化合物、例えばオキシカルボキシン等、ベンゾアニリド系化合物、例えばメプロニル、ジクロメジン、フルトラニル、ペンシクロン、フラメトピル、チフルザミド等、アシルアラニン系化合物、例えば、オキサジキシル、メタラキシル、メトキシアクリレート系化合物、例えばアゾキシストロビン、クレソキシムメチル、メトミノストロビン、トリフロキシストロビン、ピコキシストロビン、ピラクロストロビン、オリサストロビン等、キノキサリン系化合物、例えばキノメチオネート等、ヒドロキシアニリド系化合物、例えばフェンヘキサミド等、シアノアセトアミド系化合物、例えばシモキサニル等、シアノイミダゾール系化合物、例えばシアゾファミド等、その他ファモキサドン、スピロキサミン、トリアゾキシド、ピラゾホス、フルオルイミド、ジメトモルフ、イプロバリカルブ、フェナミドン、エタボキサム、シフルフェナミド、ジチアノン、カルプロパミド、プロベナゾール、メタスルホカルブ、ピロキロン、ヒドロキシイソキサゾール、トリシクラゾール、ジフルメトリム、フェナジンキシド、イソプロチオラン、オキソリニック酸、アシベンゾラル−Ｓ−メチル、キノキシフェン、ベンチアバリカルブイソプロピル、チアジニルから選択される１種または２種以上の化学殺菌剤、及びバシルス属に属する細菌の芽胞を含有することを特徴とする農園芸用殺菌剤組成物。 (1) Chemically synthetic bactericidal compounds, that is, inorganic copper compounds such as basic copper sulfate, anhydrous copper sulfate, cupric hydroxide, basic copper chloride, etc., organic copper compounds such as organic copper, copper nonylphenol sulfonate, etc., inorganic Sulfur compounds such as sulfur, total sulfide sulfur, etc.Organic sulfur compounds such as dineb, manneb, propineb, thiadiazine, thiuram, polycarbamate, anilinopyrimidine compounds such as cyprodinil, pyrimethanil, mepanipyrim, phenylpyrrole compounds, Organic chlorine compounds such as fludioxonil, such as chlorothalonil, captan, triazine, fluazinam, sulfenic acid, fusalide, etc., hydrogen carbonate agents such as sodium hydrogen carbonate, potassium hydrogen carbonate, etc.Organic phosphorus compounds such as EDDP, fosetyl, torquelophos Mechi , IBP, etc., benzimidazole compounds such as carbendazim, thiophanate methyl, thiabendazole, benomyl, fuberidazole, etc., dicarboximide compounds such as iprodione, procymidone, vinclozoline, etc., azole compounds such as fenbuconazole, cimeconazole, diclobutra Sol, triticonazole, ipconazole, fluconazole, microbutanyl, penconazole, viteltanol, bromconazole, oxpoconazole, cyproconazole, difenoconazole, diniconazole, epoxiconazole, fenbuconazole, fluquinconazole, flusilazole, flutriahol , Hexaconazole, imibenconazole, metconazole, propiconazole, cypconazole, Conazole, tetraconazole, triadimethone, triadimenol, etc., imidazole compounds such as triflumizole, prochloraz, imazalyl, pefrazoate, piperazine compounds such as triphorin, morpholine compounds such as fenpropimorph, tridemorph, fenpro Pyridine, hydroxypyrimidine compounds such as ethylimyl, dimethylimole, etc., guanidine compounds such as iminotadine acetate, iminoctadine arbesylate, guazatine, acid amide compounds such as oxycarboxyl, benzoanilide compounds such as mepronil, Acylalanine compounds such as dichromedin, flutolanil, pencyclon, furametopil, tifluzamide, etc., such as oxadixyl, metalaxyl, methoxyacrylic Rate compounds such as azoxystrobin, cresoxime methyl, metminostrobin, trifloxystrobin, picoxystrobin, pyraclostrobin, orissastrobin, quinoxaline compounds such as quinomethionate, hydroxyanilide compounds such as fenhexamide Cyanoacetamide compounds such as simoxanil, cyanoimidazole compounds such as cyazofamide, etc.Famoxadone, spiroxamine, triazoxide, pyrazophos, fluorimido, dimethomorph, iprovaricarb, fenamidone, ethaboxam, cyflufenamide, dithianone, carpropamide, probenazole, metasulfocarbamide , Pyroxylone, hydroxyisoxazole, tricyclazole, diflumetrim, Containing one or more chemical fungicides selected from nazinoxide, isoprothiolane, oxolinic acid, acibenzoral-S-methyl, quinoxyphene, benchavaricarb isopropyl, thiazinyl, and bacterial spores belonging to the genus Bacillus A feature of a fungicidal composition for agriculture and horticulture.
（２）バシルス属に属する細菌が、バシルス・ズブチリスである前記記載の農園芸用殺菌剤組成物。 (2) The agricultural and horticultural fungicide composition as described above, wherein the bacterium belonging to the genus Bacillus is Bacillus subtilis.
（３）バシルス属に属する細菌が、バシルス ｓｐ．Ｄ７４７（受託番号ＦＥＲＭ ＢＰ−８２３４）である前記記載の農園芸用殺菌剤組成物。 (3) Bacteria belonging to the genus Bacillus are Bacillus sp. The agricultural and horticultural fungicide composition as described above, which is D747 (Accession No. FERM BP-8234).
（１）〜（３）の何れかに記載の農園芸用殺菌剤組成物を用いた植物病害防除方法。 (1) A plant disease control method using the agricultural and horticultural fungicide composition according to any one of (1) to (3).
以下、本発明について詳細に説明する。 Hereinafter, the present invention will be described in detail.
＜１＞バシルス属に属する細菌の芽胞
本発明の農園芸用殺菌剤組成物は、バシルス属に属する細菌の培養物より得られる芽胞を有効成分として含有する。
<1> Spores of bacteria belonging to the genus Bacillus The agricultural and horticultural fungicide composition of the present invention contains spores obtained from a culture of bacteria belonging to the genus Bacillus as an active ingredient.
本発明に用いるバシルス属に属する細菌としては、バシルス属に属する細菌であれば特に制限されないが、好ましくは植物病原菌と拮抗するバシルス属に属する細菌が挙げられ、その中でもより好ましくはバシルス・ズブチリスやバシルス（Ｂａｃｉｌｌｕｓ） ｓｐ．Ｄ７４７が挙げられる。 Bacteria belonging to the genus Bacillus used in the present invention are not particularly limited as long as they belong to the genus Bacillus, but preferably include bacteria belonging to the genus Bacillus that antagonize phytopathogenic fungi, more preferably Bacillus subtilis and Bacillus sp. D747.
このＤ７４７菌株は、静岡県菊川市の空気中から単離された菌株であって、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おいて、Ｂａｃｉｌｌｕｓ ｓｐ．Ｄ７４７（受託番号ＦＥＲＭ ＢＰ−８２３４）として国際寄託されている。 This strain D747 is a strain isolated from the air in Kikukawa City, Shizuoka Prefecture, and is currently being used by the National Institute of Advanced Industrial Science and Technology, the Patent Biological Deposit Center as Bacillus sp. It is deposited internationally as D747 (accession number FERM BP-8234).
このバシルス ｓｐ．Ｄ７４７株の細菌的性質は以下に示す通りである。なお、菌学的性質の試験はＢｅｒｇｅｙ’ｓ Ｍａｎｕａｌ ｏｆ Ｓｙｓｔｅｍａｔｉｃ Ｂａｃｔｅｒｉｏｌｏｇｙ、ｖｏｌｕｍｅ １（１９８４）に基づいて行った。 This Bacillus sp. The bacterial properties of strain D747 are as shown below. The bacteriological properties were tested based on Bergey's Manual of Systematic Bacteriology, volume 1 (1984).
（Ａ）形態学的性質
形態： 桿菌
大きさ： 幅１．０〜１．２μｍ、長さ３〜５μｍ
運動性： ＋
鞭毛の着生状態： 周鞭毛
内生胞子： ＋
胞子の位置： 中央
胞子の膨張： −
(A) Morphological properties Form: Neisseria gonorrhoeae Size: Width 1.0-1.2 μm, Length 3-5 μm
Mobility: +
Flagellar state: Periflagellate Endospore: +
Spore position: center Spore expansion: −
（Ｂ）培養的性質
コロニーの色： 白色〜薄い茶色
肉汁寒天平板培養：白色〜クリーム色のコロニーを形成し、表面はしわ状
(B) Culture properties Colony color: White to light brown Meat broth agar plate culture: White to cream-colored colonies, wrinkled surface
（ｃ）生理学的性質
グラム染色性： ＋
硝酸塩の還元： ＋
ＭＲ試験： −
ＶＰ試験： ＋
インドールの生成： −
澱粉の加水分解： ＋
クエン酸の資化性： ＋
無機窒素源： ＋
オキシダーゼ： −
カタラーゼ： ＋
生育ｐＨ
６．８、肉エキス培地： ＋
５．７、肉エキス培地： ＋
生育温度
３０℃： ＋
５０℃： −
生育ＮａＣｌ濃度
２％： ＋
５％： ＋
７％： ＋
好気的生育： ＋
嫌気的生育： −
Ｏ−Ｆテスト： Ｏ
卵黄反応： −
グルコースからの酸生成： ＋
マンニトールからの酸生成： −
Ｌ−アラビノースからの酸生成： −
Ｄ−キシロースからの酸生成： −
グルコースからのガス生成： −
β−ガラクトシダーゼ： −
ＮａＣｌ及びＫＣｌ要求性： −
(C) Physiological properties Gram stainability: +
Reduction of nitrate: +
MR test: −
VP test: +
Indole production: −
Hydrolysis of starch: +
Utilization of citric acid: +
Inorganic nitrogen source: +
Oxidase: −
Catalase: +
Growth pH
6.8, meat extract medium: +
5.7, meat extract medium: +
Growth temperature 30 ° C: +
50 ° C .: −
Growth NaCl concentration 2%: +
5%: +
7%: +
Aerobic growth: +
Anaerobic growth: −
OF test: O
Egg yolk reaction: −
Acid production from glucose: +
Acid production from mannitol: −
Acid production from L-arabinose: −
Acid production from D-xylose: −
Gas production from glucose: −
β-galactosidase: −
NaCl and KCl requirements: −
本発明の農園芸用殺菌剤組成物は、上記のように植物病害を防除できるバシルス ｓｐ．Ｄ７４７を含有するものである。本発明の植物病害防除剤においては、Ｄ７４７菌株を単体で使用することもできるが、Ｄ７４７菌株の変異体も、単体でもしくはＤ７４７菌株とともに使用することができる。変異体は、上記Ｄ７４７菌株の細菌学的特性を有し、植物病害防除作用を有するものであり、自然突然変異株、紫外線や化学変異剤を用いての突然変異株、また細胞融合株および遺伝子組み換え株も利用が可能である。本発明の植物防除剤に含有されるＤ７４７菌株には、Ｄ７４７菌株の変異体が含まれる。 The agricultural and horticultural fungicide composition of the present invention comprises Bacillus sp. Which can control plant diseases as described above. D747 is contained. In the plant disease control agent of the present invention, the D747 strain can be used alone, but a mutant of the D747 strain can also be used alone or together with the D747 strain. The mutant has the bacteriological characteristics of the above-mentioned D747 strain and has a plant disease control action, and is a natural mutant, a mutant using ultraviolet rays or chemical mutants, a cell fusion strain and a gene. Recombinant strains can also be used. The D747 strain contained in the plant control agent of the present invention includes mutants of the D747 strain.
本発明に用いる芽胞は、上記バシルス属に属する細菌の培養物から得られる。バシルス属に属する細菌の培養は、例えば往復式振とう培養、ジャーファーメンター培養、培養タンク等の液体培養や固体培養等、バシルス属に属する細菌の通常の培養方法に準じて行うことができる。例えば肉エキス培地など一般的な培地の他、グルコース、ペプトン、イーストエキスを含む培地などが挙げられる。また液体培地以外に寒天入りの斜面培地および平板培地等の固体培地を用いてもよい。培養によってＤ７４７菌株を増殖させて、所望の菌体量を得ることができる。 The spore used in the present invention is obtained from a culture of bacteria belonging to the genus Bacillus. Bacteria belonging to the genus Bacillus can be cultured according to the usual culture method for bacteria belonging to the genus Bacillus, such as reciprocal shaking culture, jar fermenter culture, liquid culture such as a culture tank, and solid culture. For example, in addition to a common medium such as a meat extract medium, a medium containing glucose, peptone, yeast extract, and the like can be mentioned. In addition to the liquid medium, a solid medium such as a slope medium containing agar and a plate medium may be used. The D747 strain can be grown by culturing to obtain the desired cell mass.
培地の炭素源としては、上記菌株が同化しうるあらゆるものが利用可能である。例えばグルコース、ガラクトース、ラクトース、スクロース、マルトース、麦芽エキス澱粉加水分解物などの糖の外に、Ｄ７４７菌株が利用し得る各種の合成または天然炭素源をあげることができる。培地の窒素源として、同様に、ペプトン、肉エキス、酵母エキスなどの有機窒素含有物をはじめ、該菌株が利用し得る各種の合成又は天然物も利用可能である。微生物培養の常法に従って、食塩、リン酸塩などの無機塩類、カルシウム、マグネシウム、鉄などの金属の塩類、ビタミン、アミノ酸などの微量栄養源も必要に応じて添加することができる。 Any carbon source that can be assimilated by the above strain can be used as the carbon source of the medium. For example, in addition to sugars such as glucose, galactose, lactose, sucrose, maltose, and malt extract starch hydrolysate, various synthetic or natural carbon sources that can be used by the strain D747 can be mentioned. Similarly, as a nitrogen source of the medium, various synthetic or natural products that can be used by the strain, including organic nitrogen-containing materials such as peptone, meat extract, and yeast extract, can be used. In accordance with a conventional method for culturing microorganisms, inorganic salts such as sodium chloride and phosphate, metal salts such as calcium, magnesium and iron, trace nutrients such as vitamins and amino acids can be added as necessary.
培養は、振盪培養、通気培養などの好気的条件下で行なうことができる。培養温度は２０〜３０℃、好ましくは２５〜３０℃、ｐＨは５〜８、好ましくは６〜７、培養期間は１〜４日、好ましくは２〜３日が適当である。 The culture can be performed under aerobic conditions such as shaking culture and aeration culture. The culture temperature is 20-30 ° C., preferably 25-30 ° C., pH 5-8, preferably 6-7, and the culture period is 1-4 days, preferably 2-3 days.
上記のようにして得られたバシルス属に属する細菌の培養物より芽胞を分離する方法としては、膜分離、遠心分離、濾過分離等の方法を用いて行うことができる。得られた芽胞画分は、そのままある程度の水分を含んだ状態で本発明の農園芸用殺菌剤組成物に用いることも、また、必要に応じて凍結乾燥、スプレードライ等の乾燥法を用いて乾燥物として本発明の農園芸用殺菌剤組成物に用いることも可能である。 As a method for separating spores from a culture of bacteria belonging to the genus Bacillus obtained as described above, a method such as membrane separation, centrifugation, or filtration separation can be used. The obtained spore fraction can be used as it is for the agricultural and horticultural fungicide composition of the present invention in a state containing a certain amount of water, or by using a drying method such as freeze drying or spray drying, if necessary. It can also be used in the agricultural and horticultural fungicide composition of the present invention as a dried product.
＜２＞化学合成殺菌剤
本発明において用いる化学合成殺菌剤は、バシルス属に属する細菌の芽胞との併用で、相乗的な防除効果が得られる。
<2> Chemically synthetic fungicide The chemically synthesized fungicide used in the present invention provides a synergistic control effect when used in combination with bacterial spores belonging to the genus Bacillus.
＜３＞農園芸用殺菌剤組成物
本発明の農園芸用殺菌剤組成物は、上記バシルス属に属する細菌の芽胞及び上記化学合成殺菌剤をそれぞれ１種または２種以上含有するものである。
<3> Agricultural and horticultural fungicide composition The agricultural and horticultural fungicide composition of the present invention contains one or more spore of bacteria belonging to the genus Bacillus and one or more of the above chemically synthesized fungicides.
本発明の農園芸用殺菌剤組成物は、通常の微生物製剤の製造方法に従って、上記バシルス属に属する細菌の芽胞及び上記化学合成殺菌剤を必要に応じて各種任意成分と共に、粉剤、粒剤、水和剤、乳剤、液剤、フロアブル剤、塗布剤等に製剤して使用することができる。 The agricultural and horticultural fungicide composition of the present invention, according to a normal method for producing a microbial preparation, the spore of the bacterium belonging to the genus Bacillus and the chemical synthetic fungicide together with various optional components, if necessary, powder, granule, It can be formulated and used in wettable powders, emulsions, liquids, flowables, coatings and the like.
さらに本発明による植物病害防除剤においてＤ７４７菌株は、菌体または培養物を単独で用いるほか、不活性な液体または固体の担体で希釈し、必要に応じて界面活性剤、その他の補助剤を加えた薬剤として用いてもよい。具体的な製剤例としては、粒剤、粉剤、水和剤、懸濁製剤、乳剤等の剤型等があげられる。好ましい担体の例としては、タルク、ベントナイト、クレー、カオリン、珪藻土、ホワイトカーボン、バーミキュライト、消石灰、珪砂、硫安、尿素、多孔質などの固体担体、水、イソプロピルアルコール、キシレン、シクロヘキサノン、メチルナフタレン、アルキレングリコール、などの液体担体等があげられる。界面活性剤および分散剤としては、例えばジナフチルメタンスルホン酸塩、アルコール硫酸エステル塩、アルキルアリールスルホン酸塩、リグニンスルホン酸塩、ポリオキシエチレングリコールエーテル、ポリオキシエチレンアルキルアリールエーテル、ポリオキシエチレンソルビタンモノアルキレート等があげられる。補助剤としては、カルボキシメチルセルロース、ポリエチレングリコール、プロピレングリコール、アラビアゴム、キサンタンガム等、保護剤としてはスキムミルク、ｐＨ緩衝剤等があげられる。この場合、Ｄ７４７菌株および／またはその培養物の量、さらには適用時期および適用量は上記生菌の場合に準じて適宜決定することができる。 Furthermore, in the plant disease control agent according to the present invention, the D747 strain is used as a cell or culture alone, diluted with an inert liquid or solid carrier, and a surfactant or other auxiliary agent is added as necessary. It may be used as a drug. Specific examples of preparations include dosage forms such as granules, powders, wettable powders, suspension preparations, and emulsions. Examples of preferred carriers include solid carriers such as talc, bentonite, clay, kaolin, diatomaceous earth, white carbon, vermiculite, slaked lime, silica sand, ammonium sulfate, urea, porous, water, isopropyl alcohol, xylene, cyclohexanone, methylnaphthalene, alkylene Examples thereof include liquid carriers such as glycol. Examples of the surfactant and dispersant include dinaphthyl methane sulfonate, alcohol sulfate ester salt, alkyl aryl sulfonate, lignin sulfonate, polyoxyethylene glycol ether, polyoxyethylene alkyl aryl ether, and polyoxyethylene sorbitan. Examples include monoalkylates. Examples of the auxiliary agent include carboxymethyl cellulose, polyethylene glycol, propylene glycol, gum arabic, and xanthan gum, and examples of the protective agent include skim milk and pH buffering agent. In this case, the amount of the D747 strain and / or its culture, as well as the application time and application amount, can be appropriately determined according to the case of the above live bacteria.
このようにして得られる本発明の農園芸用殺菌剤組成物は、製剤の態様によらず長期間の保存が可能で、例えば、水和剤として４８ヶ月間室温にて保存した後にも、化学合成殺菌剤無添加のものと同等の防除価が得られる。 The agricultural and horticultural fungicide composition of the present invention thus obtained can be stored for a long period of time regardless of the form of the preparation. For example, even after being stored as a wettable powder at room temperature for 48 months, A control value equivalent to that without a synthetic fungicide is obtained.
本発明の農園芸用殺菌剤組成物は、施用形態により、藻菌類（Ｏｏｍｙｃｅｔｅｓ）、子嚢菌類（Ａｓｃｏｍｙｃｅｔｅｓ）、担子菌類（Ｂａｓｉｄｉｏｍｙｃｅｔｅｓ）、接合菌類（Ｚｙｇｏｍｙｃｅｔｅｓ）、及び不完全菌類（Ｄｅｕｔｅｒｏｍｙｃｅｔｅｓ）に属する菌類、および細菌類に起因する植物の病害を防除することができる。 The agricultural and horticultural fungicide composition of the present invention belongs to algal fungi (Omycetes), ascomycetes (Ascomycetes), basidiomycetes, Zygomycetes, and incomplete fungi (Deuteromycetes), depending on the application form. Plant diseases caused by fungi and bacteria can be controlled.
本発明の農園芸用殺菌剤組成物が防除することのできる植物の病原菌として、具体的にはシュードペロノスポラ（Ｐｓｅｕｄｏｐｅｒｏｎｏｓｐｏｒａ）属菌、例えばキュウリべと病菌（Ｐｓｅｕｄｏｐｅｒｏｎｏｓｐｏｒａ ｃｕｂｅｎｓｉｓ）、ベンチュリア（Ｖｅｎｔｕｒｉａ）属菌、例えばリンゴ黒星病菌（Ｖｅｎｔｕｒｉａ ｉｎａｅｑｕａｌｉｓ）、エリシフェ（Ｅｒｙｓｉｐｈｅ）属菌、例えばコムギうどんこ病菌（Ｅｒｙｓｉｐｈｅ ｇｒａｍｉｎｉｓ）、ピリキュラリア（Ｐｙｒｉｃｕｌａｒｉａ）属菌、例えばイネいもち病菌（Ｐｙｒｉｃｕｌａｒｉａ ｏｒｙｚａｅ）、ボトリチス（Ｂｏｔｒｙｔｉｓ）属菌、例えばキュウリ灰色かび病菌（Ｂｏｔｒｙｔｉｓ ｃｉｎｅｒｅａ）、リゾクトニア（Ｒｈｉｚｏｃｔｏｎｉａ）属菌、例えばイネ紋枯病菌（Ｒｈｉｚｏｃｔｏｎｉａ ｓｏｌａｎｉ）、クラドスポリウム（Ｃｌａｄｏｓｐｏｒｉｕｍ）属菌、例えばトマト葉かび病菌（Ｃｌａｄｏｓｐｏｒｉｕｍ ｆｕｌｖｕｍ）、コレトトリカム（Ｃｏｌｌｅｔｏｔｒｉｃｈｕｍ）属菌、例えばイチゴ炭そ病菌（Ｃｏｌｌｅｔｏｔｒｉｃｈｕｍ ｆｒａｇａｒｉａｅ）、パクシニア（Ｐｕｃｃｉｎｉａ）属菌、例えばコムギ赤さび病菌（Ｐｕｃｃｉｎｉａ ｒｅｃｏｎｄｉｔａ）、セプトリア（Ｓｅｐｔｏｒｉａ）属菌、例えばコムギふ枯病菌（Ｓｅｐｔｏｒｉａ ｎｏｄｏｒｕｍ）、スクレロティニア（Ｓｃｌｅｒｏｔｉｎｉａ）属菌、例えばキュウリ菌核病菌（Ｓｃｌｅｒｏｔｉｎｉａ ｓｃｌｅｒｏｔｉｏｒｕｍ）、ピシウム（Ｐｙｔｈｉｕｍ）属菌、例えばキュウリ苗立枯病菌（Ｐｙｔｈｉｕｍ ｄｅｂａｒｙａｎｕｍ Ｈｅｓｓｅ）、ゲウマノマイセス（Ｇａｅｕｍａｎｎｏｍｙｃｅｓ）属菌、例えばコムギ立枯病菌（Ｇａｅｕｍａｎｎｏｍｙｃｅｓ ｇｒａｍｉｎｉｓ）、また細菌として、バークホルデリア（Ｂｕｒｋｈｏｌｄｅｒｉａ）、例えばイネ苗立枯細菌病（Ｂｕｒｋｈｏｌｄｅｒｉａ ｐｌａｎｔａｒｉｉ）などをあげることができるが、本発明はこれらの例により限定されるものではない。 The plant pathogens that can be controlled by the agricultural and horticultural fungicide composition of the present invention are specifically Pseudoperonospora, such as Pseudoperonospora cubensis and Venturia. Bacteria such as Venturia inaequalis, Erysiphe genus such as wheat powdery mildew, Pyricularia genus, such as rice blast fungus For example, Botrytis cinerea, Rhizoctonia ) Genus fungi such as Rhizoctonia solani, Cladosporium genus such as tomato leaf mold fungus (Cladosporum fulvum), Collototricum genus fungus such as strawberry charcot rot Puccinia genus bacteria, such as wheat red rust fungus (Puccinia recondita), Septoria genus bacteria such as Septoria nodorum, Sclerotinia nuclei, sclerotiorum), Pythium spp. Phythium debaryanum Hesse, genus Gaeumanomyces, for example, wheat stem blight (Gaeumanomyces graminis), and bacteria such as Burkolderia, for example, rice seedling However, the present invention is not limited to these examples.
＜４＞本発明の農園芸用殺菌剤組成物の施用方法
本発明の農園芸用殺菌剤組成物は、そのまま直接施用するか、あるいは水などで希釈して施用することができる。植物病害防除剤としての施用方法は、特に限定されず、例えば、直接植物に散布する方法、土壌に散布する方法、植物や土壌に添加する水や肥料に添加する方法などがあげられる。その他、製剤の施用量は、対象病害、対象作物、施用方法、発生傾向、被害の程度、環境条件、使用する剤型などによって変動するので、適宜調整されることが好ましい。
<4> Method for applying agricultural and horticultural fungicide composition of the present invention The agricultural and horticultural fungicide composition of the present invention can be applied directly or diluted with water or the like. The application method as a plant disease control agent is not specifically limited, For example, the method of spraying directly to a plant, the method of spraying to soil, the method of adding to the water and fertilizer added to a plant or soil, etc. are mention | raise | lifted. In addition, since the dosage of the preparation varies depending on the target disease, target crop, application method, occurrence tendency, degree of damage, environmental conditions, dosage form used, etc., it is preferably adjusted as appropriate.
また、栽培植物への農園芸用殺菌剤組成物の施用に際して、殺虫剤、殺線虫剤、除草剤、植物生長調節剤、肥料、土壌改良資材等を混合施用、交互施用、または同時施用することも可能である。 In addition, when applying the agricultural and horticultural fungicide composition to the cultivated plant, the insecticide, nematicide, herbicide, plant growth regulator, fertilizer, soil improvement material, etc. are mixed, applied alternately or simultaneously. It is also possible.
本発明の農園芸用殺菌剤組成物の施用量は、病害の種類、適用植物の種類、殺菌剤組成物の剤型等によって異なるため、一概に規定できないが、例えば液剤の農園芸用殺菌剤組成物を散布する場合には、芽胞濃度として１０５〜１０１０ｃｆｕ（コロニー形成単位）／ｍｌであり、好ましくは１０７〜１０９ｃｆｕ／ｍｌである。また施用量は１０アールあたり液剤の農園芸用殺菌剤組成物を０．５〜１０００リットル施用するのが好ましい。 The application amount of the agricultural and horticultural fungicide composition of the present invention varies depending on the type of disease, the type of plant to be applied, the dosage form of the fungicide composition, etc. When the composition is sprayed, the spore concentration is 10 5 to 10 10 cfu (colony forming unit) / ml, preferably 10 7 to 10 9 cfu / ml. Moreover, it is preferable to apply 0.5-1000 liters of the agricultural and horticultural fungicide composition as a liquid agent per 10 ares.
本発明の農園芸用殺菌剤組成物は、キュウリ灰色かび病、べと病、コムギうどんこ病、イネいもち病等の各種病原菌に対して優れた防除効果を示し、化学合成殺菌剤の使用量を減らすことができる。また有用作物に対する安全性が極めて高い。 The agricultural and horticultural fungicide composition of the present invention exhibits excellent control effects against various pathogens such as cucumber gray mold, downy mildew, wheat powdery mildew, rice blast, etc. Can be reduced. Also, the safety for useful crops is extremely high.
本発明を以下の実施例によって具体的に説明するが、本発明はこれらの実施例に限定されるものではない。 The present invention will be specifically described by the following examples, but the present invention is not limited to these examples.
（製造例１：湿菌体の製造）
静岡県内の空気中より分離されたバシルス ｓｐ．Ｄ７４７菌株（ＦＥＲＭ ＢＰ−８２３４）を平板培地上で培養し、分離したコロニーをフラスコに植菌し、２０ｍｌのブイヨン培地（肉エキス１％、ペプトン１％、塩化ナトリウム０．５％）で、２７℃、１２０ｒｐｍ、１日間振盪培養後、得られた培養液をグルコース１％、可溶性澱粉２％、ポリペプトン０．５％、乾燥酵母１％、脱脂大豆１％、ＫＨ２ＰＯ４ ０．２％、ＮａＣｌ ０．２％、炭酸カルシウム０．３％を含む、ｐＨ ６．０の培地２０Ｌに植菌し２７℃、１２０ｒｐｍ、３日間振盪培養した後、遠心集菌（１０，０００×ｇ，１５分間）し、滅菌水中に懸濁し、培地成分を洗浄した。この操作を２回行い、湿重量約１ｋｇの湿菌体（芽胞画分）を得た。この芽胞画分はバシルス ｓｐ．Ｄ７４７の芽胞を乾燥重量で５０重量％含有するものである。
(Production Example 1: Production of wet cells)
Bacillus sp. Isolated from the air in Shizuoka Prefecture D747 strain (FERM BP-8234) was cultured on a plate medium, and the isolated colonies were inoculated into a flask. In a 20 ml broth medium (meat extract 1%, peptone 1%, sodium chloride 0.5%), 27 ° C., 120 rpm, after 1 day shaking culture, resulting culture with 1% glucose, soluble starch 2%, polypeptone 0.5%, dried yeast 1%, defatted soybean 1% KH 2 PO 4 0.2% After inoculating 20 L of a medium of pH 6.0 containing 0.2% NaCl and 0.3% calcium carbonate and shaking culture at 27 ° C., 120 rpm for 3 days, centrifugation (10,000 × g, 15 minutes) And suspended in sterile water to wash the medium components. This operation was performed twice to obtain wet cells (spore fraction) having a wet weight of about 1 kg. This spore fraction is a Bacillus sp. It contains 50% by weight of D747 spores by dry weight.
（製造例２：粉末芽胞の製造）
上記製造例１によって得られたバシルス ｓｐ．Ｄ７４７の芽胞画分１ｋｇを蒸留水５Ｌに懸濁後、スプレードライヤー（ニロジャパン社）にて処理（入口温度１５０℃、出口温度１００℃）。スプレードライヤーにて得られた乾燥物を破砕することにより、約１００ｇの乾燥粉末芽胞を得た。この乾燥粉末はバシルス ｓｐ．Ｄ７４７の芽胞を５×１０１１ｃｆｕ／ｇ以上を含むものである。
(Production Example 2: Production of powder spore)
Bacillus sp. Obtained in Production Example 1 above. 1 kg of the D747 spore fraction was suspended in 5 L of distilled water and then treated with a spray dryer (Niro Japan) (inlet temperature 150 ° C., outlet temperature 100 ° C.). By crushing the dried product obtained by the spray dryer, about 100 g of dry powder spores were obtained. This dry powder is Bacillus sp. D747 spores containing 5 × 10 11 cfu / g or more.
次に本発明の農園芸用殺菌剤組成物の代表的な製剤例をあげて製剤方法を具体的に説明する。以下の説明において「％」は重量百分率を示す。 Next, the preparation method will be specifically described with reference to typical preparation examples of the agricultural and horticultural fungicide composition of the present invention. In the following description, “%” indicates a weight percentage.
（製剤例１：水和剤）
上記のようにして得られたバシルス ｓｐ．Ｄ７４７菌株の粉末芽胞を２０％、農園芸用殺菌化合物メパニピリムを１．８％、ナフタレンスルホン酸ホルマリン縮合物ナトリウム塩１．５％、ポリオキシエチレンアルキルアリール１．５％、珪藻土２６％、クレー４９．２％を均一に混合粉砕して、水和剤とした。
(Formulation example 1: wettable powder)
Bacillus sp. Obtained as described above. 20% powder spores of D747 strain, 1.8% of agricultural and horticultural fungicide mepanipyrim, 1.5% naphthalenesulfonic acid formalin condensate sodium salt, 1.5% polyoxyethylene alkylaryl, 26% diatomaceous earth, clay 49 .2% was mixed and ground uniformly to obtain a wettable powder.
（製剤例２：水和剤）
上記のようにして得られたバシルス ｓｐ．Ｄ７４７菌株の粉末芽胞を３０％、農園芸用殺菌化合物フルジオキソニルを０．９％、ナフタレンスルホン酸ホルマリン縮合物ナトリウム塩１．５％、ポリオキシエチレンアルキルアリール１．５％、珪藻土２６％、クレー４０．１％を均一に混合粉砕して、水和剤とした。
(Formulation example 2: wettable powder)
Bacillus sp. Obtained as described above. D747 strain powder spore 30%, agricultural and horticultural bactericidal compound fludioxonil 0.9%, naphthalenesulfonic acid formalin condensate sodium salt 1.5%, polyoxyethylene alkylaryl 1.5%, diatomaceous earth 26%, clay 40 .1% was mixed and ground uniformly to obtain a wettable powder.
（製剤例３：水和剤）
上記のようにして得られたバシルス ｓｐ．Ｄ７４７菌株の粉末芽胞を１０％、農園芸用殺菌化合物炭酸水素ナトリウムを３０％、ナフタレンスルホン酸ホルマリン縮合物ナトリウム塩１．５％、ポリオキシエチレンアルキルアリール１．５％、珪藻土２６％、クレー３１％を均一に混合粉砕して、水和剤とした。
(Formulation example 3: wettable powder)
Bacillus sp. Obtained as described above. 10% powder spore of D747 strain, 30% sodium bicarbonate of agricultural and horticultural compound, 1.5% naphthalenesulfonic acid formalin condensate sodium salt, 1.5% polyoxyethylene alkylaryl, 26% diatomaceous earth, clay 31 % Was uniformly mixed and ground to obtain a wettable powder.
（製剤例４：水和剤）
上記のようにして得られたバシルス ｓｐ．Ｄ７４７菌株の粉末芽胞を１０％、農園芸用殺菌化合物アゾキシストロビンを９％、ナフタレンスルホン酸ホルマリン縮合物ナトリウム塩１．５％、ポリオキシエチレンアルキルアリール１．５％、珪藻土２６％、クレー５２％を均一に混合粉砕して、水和剤とした。
(Formulation example 4: wettable powder)
Bacillus sp. Obtained as described above. 10% powder spore of D747 strain, 9% azoxystrobin, agricultural and horticultural bactericidal compound, 1.5% naphthalenesulfonic acid formalin condensate sodium salt, 1.5% polyoxyethylene alkylaryl, 26% diatomaceous earth, clay 52% was uniformly mixed and ground to obtain a wettable powder.
（製剤例５：水和剤）
上記のようにして得られたバシルス ｓｐ．Ｄ７４７菌株の粉末芽胞を６０％、農園芸用殺菌化合物トリシクラゾールを０．０５％、ナフタレンスルホン酸ホルマリン縮合物ナトリウム塩１．５％、ポリオキシエチレンアルキルアリール１．５％、珪藻土２６％、クレー１０．９５％を均一に混合粉砕して、水和剤とした。
(Formulation example 5: wettable powder)
Bacillus sp. Obtained as described above. D747 strain powder spore 60%, agricultural and horticultural bactericidal compound tricyclazole 0.05%, naphthalenesulfonic acid formalin condensate sodium salt 1.5%, polyoxyethylene alkylaryl 1.5%, diatomaceous earth 26%, clay 10 .95% was mixed and ground uniformly to obtain a wettable powder.
（製剤例６：粉剤）
上記のようにして得られたバシルス ｓｐ．Ｄ７４７菌株の粉末芽胞を１０％、農園芸用殺菌化合物フサライドを１％、珪藻土６％及びクレー８３％を均一に混合粉砕して粉剤とした。
(Formulation Example 6: Powder)
Bacillus sp. Obtained as described above. The powder spore of D747 strain was uniformly mixed and pulverized with 10% of the powder spore, 1% of the agricultural and horticultural bactericidal compound fusaride, 6% of diatomaceous earth and 83% of clay to obtain a powder.
（製剤例７：粒剤）
上記のようにして得られたバシルス ｓｐ．Ｄ７４７菌株の粉末芽胞を１０％、農園芸用殺菌化合物フルトラニルを１０％、ラウリルアルコール硫酸エステルのナトリウム塩２％、リグニンスルホン酸ナトリウム５％、カルボキシメチルセルロース２％及びクレー７１％を均一に混合粉砕した。この混合物１００重量部に水２０重量部を加えて練合し、押出式造粒機を用いて１４〜３２メッシュの粒状に加工した後、乾燥して粒剤とした。
(Formulation Example 7: Granules)
Bacillus sp. Obtained as described above. 10% powder spores of D747 strain, 10% agricultural and horticultural bactericidal compound flutolanil, 2% sodium lauryl alcohol sulfate, 5% sodium lignin sulfonate, 2% carboxymethylcellulose and 71% clay were uniformly mixed and ground. . 20 parts by weight of water was added to 100 parts by weight of the mixture, kneaded, processed into granules of 14 to 32 mesh using an extrusion granulator, and dried to form granules.
（製剤例８：水和剤）
上記のようにして得られたバシルス ｓｐ．Ｄ７４７菌株の粉末芽胞を１０％、ナフタレンスルホン酸ホルマリン縮合物ナトリウム塩１．５％、ポリオキシエチレンアルキルアリール１．５％、珪藻土２６％、クレー６１％を均一に混合粉砕して、水和剤とした。
(Formulation example 8: wettable powder)
Bacillus sp. Obtained as described above. 10% powder spore of D747 strain, 1.5% naphthalenesulfonic acid formalin condensate sodium salt, 1.5% polyoxyethylene alkylaryl, 26% diatomaceous earth, 61% clay, and uniformly pulverized, wettable powder It was.
（製剤例９：水和剤）
農園芸用殺菌化合物メパニピリムを１．８％、ナフタレンスルホン酸ホルマリン縮合物ナトリウム塩１．５％、ポリオキシエチレンアルキルアリール１．５％、珪藻土２６％、クレー６９．２％を均一に混合粉砕して、水和剤とした。
(Formulation example 9: wettable powder)
Agricultural and horticultural bactericidal compound mepanipyrim 1.8%, sodium naphthalene sulfonic acid formalin condensate 1.5%, polyoxyethylene alkylaryl 1.5%, diatomaceous earth 26%, clay 69.2% uniformly mixed and ground. To obtain a wettable powder.
次に本発明の農園芸用殺菌剤組成物が植物病害防除剤として有用であることを試験例で示す。 Next, test examples show that the agricultural and horticultural fungicide composition of the present invention is useful as a plant disease control agent.
（試験例１：キュウリ灰色かび病防除効果試験）
９ｃｍ×９ｃｍの塩ビ製鉢にキュウリ種子（品種：相模半白）を１０粒づつ播種し、温室内で７日間育成し、子葉が展開したキュウリ幼苗を供試植物として用いた。製剤例１〜５に準じながら供試濃度になるように適宜配合量を変えて調製した本発明の農園芸用殺菌剤組成物、製剤例８のバシルス ｓｐ．Ｄ７４７、及び製剤例９に準じて調製した化学合成殺菌剤を、有効成分が所定の濃度になるように水で希釈し、１ポット当たり１０ｍｌ散布した。風乾後、キュウリ灰色かび病菌（Ｂｏｔｒｙｔｉｓ ｃｉｎｅｒｅａ）の胞子懸濁液を浸したぺーパーディスクをキュウリ子葉表面に置床接種し、２０℃の湿室で管理した。接種３日後に子葉の病斑直径を求め、得られた数値をもとに、数１により防除価（％）を求めた。
(Test Example 1: Cucumber gray mold control effect test)
Ten cucumber seeds (variety: Sagamihanjiro) were sown in 9 cm x 9 cm PVC pots, grown in a greenhouse for 7 days, and cucumber seedlings with cotyledons were used as test plants. The agricultural and horticultural fungicide composition of the present invention prepared by appropriately changing the blending amount so as to obtain the test concentration according to Formulation Examples 1 to 5, Bacillus sp. D747 and a chemically synthesized fungicide prepared according to Formulation Example 9 were diluted with water so that the active ingredient had a predetermined concentration, and 10 ml per one pot was sprayed. After air-drying, a paper disk soaked with a spore suspension of Botrytis cinerea was placed on the surface of the cucumber cotyledon and managed in a wet chamber at 20 ° C. Three days after the inoculation, the lesion diameter of the cotyledon was obtained, and the control value (%) was obtained from Equation 1 based on the obtained numerical value.
２種類の活性化合物や微生物を混合して処理した際に期待される防除効果は、コルビー（Ｃｏｌｂｙ）の計算式より求めることができる（除草剤の組み合わせの相乗的及び拮抗的反応の計算：Ｃａｌｃｕｌａｔｉｎｇ Ｓｙｎｅｒｇｉｓｔｉｃ ａｎｄ Ａｎｔａｇｏｎｉｓｔｉｃ Ｒｅｓｐｏｎｓｅｓ ｏｆ Ｈｅｒｂｉｃｉｄｅ Ｃｏｍｂｉｎａｔｉｏｎ、 Ｗｅｅｄ １５、 ２０〜２２ページ、 １９６７）。コルビー（Ｃｏｌｂｙ）の計算式を数２に示す。 The control effect expected when two types of active compounds and microorganisms are mixed and processed can be obtained from the calculation formula of Colby (calculation of synergistic and antagonistic responses of herbicide combinations: Calculating) Synergistic and Antagonistic Responses of Herbide Combination, Weed 15, pages 20-22, 1967). The calculation formula of Colby is shown in Formula 2.
数２で、Ｘは一方の活性化合物や微生物をｍ（ｐｐｍもしくはｃｆｕ／ｍｌ）の濃度で処理した場合の防除効果（防除価）を表し、Ｙは他方の活性化合物や微生物をｎ（ｐｐｍもしくはｃｆｕ／ｍｌ）の濃度で処理した場合の防除効果（防除価）を表わす。その場合、Ｅは前者の活性化合物や微生物をｍ（ｐｐｍもしくはｃｆｕ／ｍｌ）、後者の活性化合物や微生物をｎ（ｐｐｍもしくはｃｆｕ／ｍｌ）の濃度になるように混合して処理した場合の防除効果（防除価）の相加作用による期待値を表している。 In Equation 2, X represents the control effect (control value) when one active compound or microorganism is treated at a concentration of m (ppm or cfu / ml), and Y represents the other active compound or microorganism n (ppm or The control effect (control value) when treated at a concentration of cfu / ml). In that case, E is the control when the former active compound or microorganism is mixed and treated so that the concentration of the latter active compound or microorganism is n (ppm or cfu / ml) and n (ppm or cfu / ml). It represents the expected value due to the additive effect (control value).
本発明の農園芸用殺菌剤組成物の防除効果が、Ｘとしてバシルス ｓｐ．Ｄ７４７のみを処理した防除効果、Ｙとして化学合成殺菌剤のみを処理した防除効果を用いてコルビー（Ｃｏｌｂｙ）の計算式より求められた計算値（Ｅ）より大きくなれば、この組み合わせによる防除効果が相乗効果により発揮されていることを示すこととなる。試験によって得られた実験値の防除価（％）、及びコルビーの計算式より求められた計算値の防除価（％）を表１に示した。 The control effect of the agricultural and horticultural fungicide composition of the present invention is X as Bacillus sp. If the control effect obtained by treating only D747 and the control effect obtained by treating only the chemically synthesized fungicide as Y is larger than the calculated value (E) obtained from the Colby formula, the control effect by this combination is obtained. It shows that it is being demonstrated by a synergistic effect. Table 1 shows the control value (%) of the experimental value obtained by the test and the control value (%) of the calculated value obtained from the calculation formula of Colby.
（試験例２：コムギうどんこ病防除効果試験）
９ｃｍ×９ｃｍの塩ビ製鉢に小麦種子（品種：農林６１号）を９粒づつ播種し、温室内で８日間育成し供試植物として用いた。製剤例１〜５に準じながら供試濃度になるように適宜配合量を変えて調製した本発明の農園芸用殺菌剤組成物、製剤例８のバシルス ｓｐ．Ｄ７４７、及び製剤例９に準じて調製した化学合成殺菌剤を、有効成分が所定の濃度になるように水で希釈し、１鉢当たり１０ｍｌ散布した。風乾後、コムギうどんこ病菌（Ｅｒｙｓｉｐｈｅ ｇｒａｍｉｎｉｓ）の胞子を接種し、２０〜２５℃の温室内に入れた。接種１０日後に各々の第１葉の発病面積を下記の基準に従って指数調査し、得られた指数値をもとに、数３により被害度を求め、さらに数４により防除価（％）を求めた。試験によって得られた実験値の防除価（％）、及びコルビーの計算式より求められた計算値の防除価（％）を表２に示した。
(Test Example 2: Wheat powdery mildew control effect test)
Nine wheat seeds (variety: Norin 61) were sown in a 9 cm × 9 cm PVC pot, grown in a greenhouse for 8 days, and used as a test plant. The agricultural and horticultural fungicide composition of the present invention prepared by appropriately changing the blending amount so as to obtain the test concentration according to Formulation Examples 1 to 5, Bacillus sp. D747 and a chemically synthesized fungicide prepared according to Formulation Example 9 were diluted with water so that the active ingredient had a predetermined concentration, and 10 ml was sprayed per pot. After air drying, spores of wheat powdery mildew (Erysiphe graminis) were inoculated and placed in a 20-25 ° C. greenhouse. 10 days after the inoculation, each diseased area of the first leaf was indexed according to the following criteria. Based on the obtained index value, the degree of damage was obtained by Equation 3, and the control value (%) was obtained by Equation 4. It was. Table 2 shows the control value (%) of the experimental value obtained by the test and the control value (%) of the calculated value obtained from the calculation formula of Colby.
発病指数 ０ ： 発病をみとめず
１ ： 葉の面積の５％未満の発病面積
２ ： 葉の面積の５％以上１０％未満の発病面積
３ ： 葉の面積の１０％以上２５％未満の発病面積
４ ： 葉の面積の２５％以上の発病面積
Disease index 0: Does not detect disease
1: Disease area less than 5% of leaf area
2: Disease area of 5% or more and less than 10% of leaf area
3: Disease area of 10% or more and less than 25% of leaf area
4: Onset area of 25% or more of leaf area
（試験例３：キュウリべと病防除効果試験）
９ｃｍ×９ｃｍの塩ビ製鉢にキュウリ種子（品種：相模半白）を１０粒づつ播種し、温室内で７日間育成し、子葉が展開したキュウリ幼苗を供試植物として用いた。製剤例１〜５に準じながら供試濃度になるように適宜配合量を変えて調製した本発明の農園芸用殺菌剤組成物、製剤例８のバシルス ｓｐ．Ｄ７４７、及び製剤例９に準じて調製した化学合成殺菌剤を、有効成分が所定の濃度になるように水で希釈し、１鉢当たり１０ｍｌを噴霧散布した。風乾後、キュウリべと病菌（Ｐｓｅｕｄｏｐｅｒｏｎｏｓｐｏｒａ ｃｕｂｅｎｓｉｓ）の分生胞子懸濁液を噴霧接種し、直ちに２０℃の湿室内に２４時間入れた。その後温室内に移し、７日後に各子葉の発病程度を下記の発病指数の基準に従って調査し、得られた指数値をもとに、数３により被害度を求め、さらに数４により防除価（％）を求めた。試験によって得られた実験値の防除価（％）、及びコルビーの計算式より求められた計算値の防除価（％）を表３に示した。
(Test Example 3: Cucumber downy mildew control effect test)
Ten cucumber seeds (variety: Sagamihanjiro) were sown in 9 cm x 9 cm PVC pots, grown in a greenhouse for 7 days, and cucumber seedlings with cotyledons were used as test plants. The agricultural and horticultural fungicide composition of the present invention prepared by changing the blending amount appropriately so as to obtain the test concentration according to Formulation Examples 1 to 5, and Bacillus sp. D747 and a chemically synthesized fungicide prepared according to Formulation Example 9 were diluted with water so that the active ingredient had a predetermined concentration, and sprayed at 10 ml per pot. After air drying, a conidial spore suspension of cucumber downy mildew (Pseudoperonospora cubensis) was spray-inoculated and immediately placed in a 20 ° C. humidity chamber for 24 hours. After that, after moving into the greenhouse, after 7 days, the degree of disease of each cotyledon was investigated according to the following criteria of disease index, and based on the obtained index value, the degree of damage was calculated by Equation 3, and the control value by Equation 4 ( %). Table 3 shows the control value (%) of the experimental value obtained by the test and the control value (%) of the calculated value obtained from the calculation formula of Colby.
発病指数 ０ ： 発病をみとめず
１ ： 葉の面積の５％未満の発病面積
２ ： 葉の面積の５％以上３３．３％未満の発病面積
３ ： 葉の面積の３３．３％以上６６．６％未満の発病面積
４ ： 葉の面積の６６．６％以上の発病面積、または落葉
Disease index 0: Does not detect disease
1: Disease area less than 5% of leaf area
2: Onset area of 5% or more and less than 33.3% of leaf area
3: Onset area of 33.3% to less than 66.6% of leaf area
4: Disease area of 66.6% or more of leaf area, or fallen leaves
（試験例４：イネいもち病防除効果試験）
直径７ｃｍの素焼鉢に水稲種子（品種：愛知旭）を約１５粒ずつ播種し、温室内で２〜３週間育成し、第４葉が完全に展開したイネ苗を供試植物として用いた。製剤例１〜５に準じながら供試濃度になるように適宜配合量を変えて調製した本発明の農園芸用殺菌剤組成物、製剤例８のバシルス ｓｐ．Ｄ７４７、及び製剤例９に準じて調製した化学合成殺菌剤を、有効成分が所定の濃度になるように水で希釈し、１鉢当たり１０ｍｌを噴霧散布した。風乾後、イネいもち病菌（Ｐｙｒｉｃｕｌａｒｉａ ｏｒｙｚａｅ）の分生胞子懸濁液を噴霧接種し、直ちに２５℃の湿室内に２４時間入れた。その後温室内に移し、接種５日後に第４葉の病斑数を調査し、数５により防除価を求めた。試験によって得られた実験値の防除価（％）、及びコルビーの計算式より求められた計算値の防除価（％）を表４に示した。
(Test Example 4: Rice Blast Control Effect Test)
About 15 seeds of rice seed (variety: Aichi Asahi) were sown in 7 cm diameter clay pots, grown in a greenhouse for 2 to 3 weeks, and rice seedlings in which the fourth leaf was completely developed were used as test plants. The agricultural and horticultural fungicide composition of the present invention prepared by changing the blending amount appropriately so as to obtain the test concentration according to Formulation Examples 1 to 5, and Bacillus sp. D747 and a chemically synthesized fungicide prepared according to Formulation Example 9 were diluted with water so that the active ingredient had a predetermined concentration, and sprayed at 10 ml per pot. After air-drying, a conidial spore suspension of Pyricularia oryzae was spray-inoculated and immediately placed in a humid chamber at 25 ° C. for 24 hours. Then, it moved to the greenhouse, and the number of lesions on the 4th leaf was investigated 5 days after the inoculation. Table 4 shows the control value (%) of the experimental value obtained by the test and the control value (%) of the calculated value obtained from the calculation formula of Colby.
上記の試験例から明らかなように、個々の拮抗微生物や化学合成殺菌剤のみでは不十分な防除効果しか示さない濃度でも、本発明の農園芸用殺菌剤組成物は相乗効果による優れた防除効果を示すので、化学合成殺菌剤の使用量を減らしても高い防除効果を得ることができる。
As is clear from the above test examples, the agricultural and horticultural fungicide composition of the present invention has an excellent control effect due to a synergistic effect even at a concentration at which only individual antagonistic microorganisms and chemically synthesized fungicides alone show an insufficient control effect. Therefore, even if the amount of the chemically synthesized fungicide is reduced, a high control effect can be obtained.
Claims (2)
以上 A plant disease control method using the agricultural and horticultural fungicide composition according to claim 1.
more than
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| EP2192841A2 (en) | 2007-09-20 | 2010-06-09 | Basf Se | Combinations comprising a fungicidal strain and at least one additional fungicide |
| WO2009124707A2 (en) * | 2008-04-07 | 2009-10-15 | Bayer Cropscience Ag | Combinations of biological control agents and insecticides or fungicides |
| AR076150A1 (en) * | 2009-03-26 | 2011-05-18 | Basf Se | USE OF SYNTHETIC AND BIOLOGICAL FUNGICIDES IN COMBINATION TO CONTROL DANGINE FUNGES |
| CN103430962A (en) * | 2013-08-14 | 2013-12-11 | 南京农业大学 | Bacillus subtilis and kresoxim-methyl composition |
| CA2972579A1 (en) * | 2014-12-29 | 2016-07-07 | Timothy M. Martin | Compositions and methods for use of insecticide with bacillus sp. d747 |
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| WO2017018464A1 (en) | 2015-07-27 | 2017-02-02 | 住友化学株式会社 | Plant disease control composition and plant disease control method |
| US10375965B2 (en) * | 2015-07-27 | 2019-08-13 | Sumitomo Chemical Company, Limited | Plant disease control composition and plant disease control method |
| US10694750B2 (en) | 2015-07-27 | 2020-06-30 | Sumitomo Chemical Company, Limited | Plant disease control composition and plant disease control method |
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