JP3907218B2 - Matsunoi Sentiyu Control Agent - Google Patents

Description

【０００９】
式中、Ｒ¹はヒドロキシル基であり且つＲ²は水素原子であるか、或いはＲ¹とＲ²は一緒になつて＝Ｎ−ＯＣＨ₃を表わす、
で示される化合物を有効成分として含有することを特徴とするマツノザイセンチユウ防除剤を提供するものである。
【００１０】
Ｒ¹がヒドロキシル基であり且つＲ²が水素原子である上記式（Ｉ）の化合物は、ストレプトマイセス・シアネオグリセウス・ノンシアノゲヌス（Streptomyces cyaneogriseus noncyanogenus）ＬＬ−Ｆ２８２４９（ＮＲＲＬ １５７７３）を好気的に発酵させることによつて生産される下記の理化学的性質：
（１） 分子量：６１２（ＦＡＢ−ＭＳ）
（２） 分子式：Ｃ₃₆Ｈ₅₂Ｏ₈
（３） 比旋光度：［α］_D ²⁶＝＋１３３±３°（ｃ０.３、アセトン）
（４） 紫外線吸収スペクトル：ＵＶ_max ^CH ₃ ^OH＝２４４ｎｍ(ε２８,０００)
（５） 赤外線吸収スペクトル（ＫＢｒ ｄｉｓｃ）：３４３９、２９６０、２９２５、１７１４、１４５４、１３７４、１３３８、１１７１、１１２０、９９６、９６７ｃｍ^-1
（６） Ｃ¹³核磁気共鳴スペクトル（ＣＤＣｌ₃）：

(1) ＴＭＳから低磁場；ＣＤＣｌ₃溶液
(2) ｑ＝４級炭素
(3)、(4) ２つの未解像シグナル
（７） 電子衝撃質量スペクトル：
m ／ Z 元素組成
612.3705 C₃₆H₅₂O₈
594.3543 C₃₆H₅₀O₇
576.3472 C₃₆H₄₈O₆
484.3211 C₃₀H₄₄O₅
482.2648 C₂₉H₃₈O₆
466.3097 C₃₀H₄₂O₄
448.2987 C₃₀H₄₀O₃
442.2375 C₂₆H₃₄O₆
425.2327 C₂₆H₃₃O₅
354.2181 C₂₃H₃₀O₃
314.1877 C₂₀H₂₆O₃
278.1144 C₁₅H₁₈O₅
265.1786 C₁₆H₂₅O₃
248.1405 C₁₅H₂₀O₃
247.1705 C₁₆H₂₃O₂
237.1838 C₁₅H₂₅O₂
219.1740 C₁₅H₂₃O
151.0753 C₉H₁₁O₂
を有する物質であり、その製造方法の詳細は特開昭６１−１０５８９号公報に記載されている（同公報ではＲ¹がヒドロキシル基であり且つＲ²が水素原子である上記式（Ｉ）の化合物は“ＬＬ−Ｆ２８２４９α”化合物と称されており、本明細書においてもそのように称することとする）ので、ここではその引用を以つて詳細な記述に代える。
【００１１】
ストレプトマイセス・シアネオグリセウス・ノンシアノゲヌスの好気的発酵によれば、上記特開昭６１−１０５８９号公報に記載されているとおり、ＬＬ−Ｆ２８２４９α化合物のみならず、外部及び内部寄生虫撲滅性抗生物質活性を有する以下に示すＬＬ−Ｆ２８２４９β、γ、δ、ε、ζ、η、θ、ι、κ、λ、μ、ν及びωと指称される化合物のいずれか１種又はそれ以上も同時に生成する。
【００１２】
【化３】

【００１３】
【表１】

【００１４】
【化４】

【００１５】
従つて、本発明の防除剤における有効成分であるＬＬ−Ｆ２８２４９α化合物は、発酵培養液から抽出、単離された実質的に純粋な形態で使用することができ、或いはまた、上記のＬＬ−Ｆ２８２４９β、γ、δ、ε、ζ、η、θ、ι、κ、λ、μ、ν及びω化合物の１種もしくはそれ以上との混合物の形態、又はこれらを含有する発酵培養液もしくはマツシユ（mash）の形態で使用することもできる。通常、ＬＬ−Ｆ２８２４９α化合物と、ＬＬ−Ｆ２８２４９β、γ、δ、ε、ζ、η、θ、ι、κ、λ、μ、ν及びω化合物の１種もしくはそれ以上とを含むマツシユの形態のものが好適に使用される。
【００１６】
他方、Ｒ¹とＲ²が一緒になつて＝Ｎ−ＯＣＨ₃を表わす前記式（Ｉ）の化合物は、「モキシデクチン（moxidectin）」なる一般名で呼ばれているそれ自体既知の化合物であり（特開昭６３−５４３７５号公報）、例えば、前記のＬＬ−Ｆ２８２４９α化合物を酸化した後、メトキシアミン又はその塩と反応させることにより製造することができる。
【００１７】
式（Ｉ）の化合物をマツノザイセンチユウの防除に使用する場合、該化合物は、松の樹幹に注入しやすく且つ樹体内移行速度の大きい剤型に製剤化することができる。例えば、式（Ｉ）の化合物は、適当な溶媒と共に液体とすることができ、特に水をベースとする水性液剤とすることが好ましい。該水性液剤のための溶媒としては、水単独又は好ましくは、水と水混和性有機溶媒、例えばメタノール、エタノール、アセトニトリル、ブトキシエタノール等との混合溶媒が挙げられる。特に水混和性有機溶媒としてアセトニトリルを使用すると、液剤の樹体内への浸透移行速度が著るしく向上するので好適である。
【００１８】
該混合溶媒中の水混和性有機溶媒の割合は、水／水混和性有機溶媒の重量比で、一般に１／０.１〜１／２、特に１／１〜１／１.５の範囲内が適当である。
【００１９】
また、上記水性液剤における式（Ｉ）の有効成分化合物の濃度は、厳密に制限されるものではなく、該化合物の種類、溶媒の種類等に応じて変えることができるが、通常、０.１〜２０重量％、好ましくは１.８〜５.４重量％の範囲内とすることができる。
【００２０】
また、式（Ｉ）の化合物を含む液剤には、さらに必要に応じて、適当な補助剤、例えば、界面活性剤［例：ポリオキシエチレンアルキルフエニルエーテル（エチレンオキサイド付加モル数８〜２０）、ポリオキシエチレンアリルフエニルエーテル（エチレンオキシド付加モル数８〜２０）、ポリオキシエチレンスチリルフエニルエーテル及びその縮合物（エチレンオキシド付加モル数８〜２０）、ポリオキシエチレングリコール型界面活性剤、アルキルベンゼンスルホネート型界面活性剤、ジオクチルサクシネート］等を含ませることもできる。
【００２１】
本発明の防除剤の松への適用時期及び適用方法は、現在市販されているものと同様とすることができ、例えば、適用時期は媒介昆虫であるマツノマダラカミキリの成虫発生約３ケ月前までが好ましく、また、適用方法としては、例えば、樹幹の地際から高さ約０.５〜約１ｍまでの間に、ドリル等でやや斜め下方に向けて深さ約５〜約９ｃｍ、直径６〜８ｍｍの注入孔をあけ、液剤を注入器を用いて注入する方法が挙げられる。１樹あたりの注入孔数は樹幹の太さや高さ等に応じて変えることができるが、通常胸高直径１０〜２０ｃｍの松で１〜２孔／樹程度とすることができる。
【００２２】
また、本発明の防除剤の使用量は、対象とする松の樹幹部の胸高直径等に応じて変えることができ、例えば、式（Ｉ）の化合物を３.６重量％含有する水性液剤の場合には、下記に示す量を標準的な使用量とすることができる。
【００２３】

【００２４】
注入は通常大気圧下に行なわれるが、場合によつては加圧注入してもよい。加圧注入の場合の圧力は２ｋｇ／ｃｍ²以下とすることが望ましい。
【００２５】
以上述べた本発明のマツノザイセンチユウ防除剤は、マツノザイセンチユウに対して、現在市販されている防除剤に比べてはるかに高い活性を有しており、従つて、低薬量で高い殺マツノザイセンチユウ効果を期待することができる。
【００２６】
また、本発明の水性液剤は樹体内移行速度が大きく、松に対する薬害もなく、マツノザイセンチユウ防除剤として実用的に極めて有用である。
【００２７】
以下、製剤例及び試験例によつて本発明をさらに具体的に説明する。
【００２８】

以上の各成分を充分に混合し、有効成分濃度３.６重量％の水性液剤を得た。
【００２９】

以上の各成分を充分に混合し有効濃度１.８重量％の水性液剤を得た。
【００３０】

以上の各成分を充分に混合し、有効濃度３.６重量％の水性液剤を得た。
【００３１】

以上の各成分を充分に混合し、有効濃度３.６重量％の水性液剤を得た。
【００３２】

以上の各成分を充分に混合し、有効濃度３.６重量％の水性液剤を得た。
【００３３】

以上の各成分を充分に混合し、有効濃度３.６重量％の水性液剤を得た。
【００３４】

以上の各成分を充分に混合し、有効濃度３.６重量％の水性液剤を得た。
【００３５】
試験例１：マツノザイセンチユウに対する増殖抑制効果の in vitro テスト
（１） 試験方法
供試動物：マツノザイセンチユウＳ−１０系統
区制 ：各区２反復
処理方法：ＰＳＡ培地に灰色カビの菌系（マツノザイセンチユウの栄養分）を繁殖させたシヤーレ（直径９０ｍｍ）に所定の濃度の薬液を注入し、１分間放置した後、薬液を捨てた。薬剤処理を行なつたシヤーレにマツノザイセンチユウの懸濁液０.２ｃｃを滴下し、２５℃の温度条件下においた。懸濁液０.６ｃｃ中のセンチユウ数を顕微鏡下で計測し０.２ｃｃ中の平均センチユウ数を求め、これを滴下センチユウ数（初期個体数）とした。接種７日後にシヤーレに水２０ｃｃを注入し１５秒間撹拌した後、その水を回収した。回収した水のうち各０.２ｃｃをとり３回顕微鏡下で観察し、生存センチユウ数を測定しシヤーレ内の生存個体数（７日後個体数）を推定し、下記式により増殖率を算出した。
【００３６】
【数１】

【００３７】
使用薬剤：ＬＬ−Ｆ２８２４９αの４重量％メタノール溶液
比較のための薬剤として
保土谷化学工業（株）の「センチユリー注入剤」（塩酸レバミゾール４％含有）
及び
フアイザー製薬（株）の「グリンガード・エイト」（酒石酸モランテル８％含有）
を使用。
【００３８】
なお、使用薬剤には、薬液の拡散をよくするため、界面活性剤（ベタリンＡ、５０００倍希釈）を添加した。
【００３９】
（２） 試験結果
【００４０】
【表２】

【００４１】
試験例２：薬害試験
（１） 試験方法
供試薬剤：ＬＬ−Ｆ２８２４９α
注入日時：１月３０日
供試植物：クロマツ３年生
区制 ：１区１樹３反復
処理方法：各試験木に１ケ所ドリルで直径２.５ｍｍの孔を開け、薬液を加圧注入し、２５℃の人工気象室にいれた。
【００４２】
試験に用いた薬剤と注入量は第２表に示す。
【００４３】

試験例３：フイールドテスト（その１）
１．試験方法
（１） 試験実施場所
試験はアカマツ１８〜３０年生林で行つた。試験地周辺は５、６年前からマツノザイセンチユウ被害が発生し、現在では１０％程度の枯損率である。
【００４４】
（２） 供試薬剤
供試薬剤としてＬＬ−Ｆ２８２４９αを用い、前記製剤例１で調製した水性液剤（３.６％ＡＳ）
（３） 処理方法および処理月日
薬剤処理方法は、地上高５０〜８０ｃｍ部にストレートドリルで直径６ｍｍ、深さ５ｃｍ程度の注入孔をあけ、ここに容器を挿入して自然圧注入を行つた。施用量はｍ³当り１２５ｍｌと２５０ｍｌの２処理で、各供試木の処理量は第４表に示すとおりである。
【００４５】
薬剤処理は２月２２日に行つた。
【００４６】
なお、いずれの薬剤とも注入孔周辺の腐朽を防止するため、トツプジンＭペースト剤を注入孔に注入した。
【００４７】
（４） 供試木
供試木は胸高直径１０〜１４ｃｍ、樹高１０〜１２ｍのアカマツ小径木である。各処理の供試本数はいずれも１０本で、これ以外に無処理区１０本と薬害区５本を使用した。
【００４８】
（５） 線虫接種
線虫接種は７月２日に行つた。接種方法は供試木の地上高４ｍ部の樹幹部にストレートドリルで直径９ｍｍ、深さ６ｃｍ程度の地上孔を１ケ所あけ、ここに１ｃｃ当り１.５万頭に調整したマツノザイセンチユウの懸濁液を２ｃｃ（３万頭）注入した。なお、供試した線虫は森林総研で培養したものを使用した。
【００４９】
（６） 調査方法
▲１▼ 薬剤効果
線虫接種後、樹脂量調査（８月、１１月）を行い、供試木の異常状況を調べるとともに、１１月中旬に枯損調査を行い、これらの結果から薬剤の処理効果を判定した。なお、樹脂流出量は５段階にわけ「＋＋＋、＋＋」は正常木、「＋、−、０」は異常木とした。ただ、この場合外見上健全でも樹脂流出量が少ないものは異常木とし、逆に一部変色部分が見られても樹脂流出量が正常なものは正常木として扱つた。
【００５０】
▲２▼ 薬害調査
薬害区は供試木１本につきｍ³当り５００ｍｌを注入し、その後の枯損状態にいて調査した。
【００５１】
▲３▼ 注入孔周辺の形成層障害
薬害区の処理木を枯損調査時に注入孔部を剥皮し、形成層部の変色状況を調査した。
【００５２】
２．結果と考察
（１） 薬剤効果
試験結果は第４表、第５表に示すとおりである。
【００５３】
まず、無処理区の枯損率は線虫接種後３２日目の８月５日が９０％、さらに１００日経過後の１１月１５日には１００％となつた。
【００５４】
これに対し式（Ｉ）の化合物による処理は、１２５ｍｌ及び２５０ｍｌ処理ともいずれも樹脂量が＋＋＋で、枯損率が０％、それに全く線虫が検出されなかつたことから、この薬剤処理の著しい効果が認められた。また、１２５ｍｌと２５０ｍｌ処理間に効果の違いが見られなかつたので、実用化に当つては１２５ｍｌ／ｍ³処理で十分と思われる。
【００５５】
それにこの薬剤は施用量が少ないので運搬、注入処理等の現場での取り扱いが容易であることと、注入時間が極めて速い（この試験の場合はすべて１時間以内に完了）ので、薬剤効果も含めれば実用性の高い防除技術と考えられる。
【００５６】
（２） 薬害調査
調査結果は第６表に示すとおりである。供試木５本とも、葉の変色、異常落葉等の症状は見られなかつた。
【００５７】
（３） 注入孔周辺の形成層障害
いずれの供試木とも注入孔の周りがわずかに変色していたが、明らかに形成層障害と思われるような変色部（１ｃｍ以上）は見られなかつた。
【００５８】
３．摘要
（１） マツノザイセンチユウに対するＬＬ−Ｆ２８２４９αの著しい効果が認められた。
【００５９】
（２） 本薬剤は施用量が少なく、注入時間が極めて速いので実用性の高い防除技術と考えられる。
【００６０】
（３） 薬量はｍ³当り１２５ｍｌでよいものと考えられる。
【００６１】
（４） 本薬剤の薬害は認められなかつた。
【００６２】
（５） 注入孔周辺における大きな形成層障害は認められなかつた。
【００６３】
【表３】

【００６４】
【表４】

【００６５】
【表５】

【００６６】
試験例４：フイールドテスト（その２）
１．試験地の概況
（１） 立地条件
沿岸部に植栽された防潮林で、標高５ｍ前後、一部に５％未満の傾斜があることを除けばほぼ平坦地、土質は表層は砂、その下は砂礫土で深部は礫がほとんどである。
【００６７】
（２） 林況
クロマツ２５〜２７年生、立木本数はｈａ当たり３,０００本程度、樹高８〜１５ｍ、胸高直径６〜１８ｃｍ。
【００６８】
２．処理条件
（１） 処理日時
２月２７日
（２） 処理方法・薬剤名・処理量等
薬剤処理は樹脂流出量を調査し、正常と認められた供試木に樹幹注入法により、地上約５０ｃｍの樹幹にφ９ｍｍ、深さ３ｃｍの穴をあけアンプルにより自然圧により注入した。供試薬剤は試験例３で用いたと同じもの、処理区分および供試本数は次のとおりである。
【００６９】
５００ｍｌ／ｍ³区 （薬害区） ５本
２５０ 〃 区 １０本
１２５ 〃 区 １０本
対 照 区 １０本
３．線虫接種
６月３０日、供試木の樹脂流出量を調査し、正常と認められたものについて線虫接種を行つた。線虫は森林総合研究所から送付されたものを培養・増殖し、１ｍｌ当たり３０,０００頭に調整した懸濁液を１供試木あたり１ｍｌ接種した。接種部位は地上高４〜５ｍの樹幹部とし、可能なものはその付近の枝を切断してその切り口にφ９ｍｍ、深さ３ｃｍの穴をあけ接種した。
【００７０】
４．調査方法および調査結果
効果調査は小田式判定法により、樹脂流出量をもつて行つた。調査日は線虫接種後３ケ月の９月２８日と、同５ケ月の１１月１９日の２回とし、最終調査時点で枯死木あるいは異常木については、樹幹胸高部からドリルで材を採取し、線虫の再検出を行つた。
【００７１】
調査結果は第７表のとおりであつた。
【００７２】
５．結果の考察
対照区の枯損率は９０％、異常木率は１００％であり、今回使用した線虫の病原性は高かつたものと考えられる。
【００７３】
これに対して薬剤処理区は全てが健全であり、当薬剤のマツノザイセンチユウに対する防除効果は非常に高いものと考えられた。なお、薬害区も全て健全であり、外観上の異常も見られなかつた。
【００７４】
【表６】

【００７５】
【表７】

【００７６】
試験例５：マツノザイセンチユウに対する増殖抑制効果の in vitro テスト
（１） 試験方法
供試動物：マツノザイセンチユウＳ−１０系統
区制 ：各区、２反復
処理方法：前記試験例１と同じ
使用薬剤：モキシデクチンの１重量％メタノール溶液
比較のための薬剤として
保土谷化学工業（株）の「センチユリー注入剤（塩酸レバミゾール４％含有）
及び
フアイザー製薬（株）の「グリンガード・エイト（酒石酸モランテル８％含有）
を使用。
【００７７】
なお、使用薬剤には、薬剤の拡散をよくするため、界面活性剤（ベタリンＡ、５０００倍希釈）を添加した。
【００７８】
（２） 試験結果
【００７９】
【表８】

[0001]
The present invention relates to a novel pinewood nematode control agent, and more particularly to a pinewood nematode control agent that exhibits an excellent control effect at a low dosage and a method for controlling pinewood nematode.
[0002]
The pine wilt phenomenon is caused by the pine wood nematode (Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle) invading a healthy pine and causing it to die and cause it to die. This is the so-called “pine wilt disease”.
[0003]
Although the emphasis was placed on extinguishing the pine pine beetle, a conventional carrier, as a countermeasure against this disease, insecticides must be sprayed extensively to eliminate vector insects, and environmental health and safety, etc. There is a big restriction from this aspect.
[0004]
On the other hand, for pine trees that are important for environmental conservation, such as golf courses, historic sites, parks, shrines and temples, a method of removing the pine trees that have penetrated into the pine trees by single-tree processing such as trunk injection. Has been proposed, and control agents for this purpose have already been marketed. For example, Morrantel tartrate is marketed under the trade name “Green Guard Eight” from Faiser Pharmaceutical Co., Ltd., and levamisole hydrochloride under the trade name “Sentily Injection” from Hodogaya Chemical Industries, Ltd. Solutions are commercially available.
[0005]
Although these commercially available pinewood nematode control agents (solutions) exert a considerable effect on the control of pinewood nematode invading the pine tree, the control effect is not sufficient, and the trunk is large and long-lasting. There is a disadvantage that it must be injected.
[0006]
Accordingly, the present inventors have intensively studied for an active ingredient compound that exhibits a high effect with a relatively small amount of injection, and as a result, have produced certain species produced by fermentation of Streptomyces microorganisms. The present inventors have found that the macrolide compound and its derivative exhibit extremely high activity against pinewood nematode and have completed the present invention.
[0007]
Thus, the present invention provides the following formula:
[Chemical 2]

[0009]
In which R ¹ is a hydroxyl group and R ² is a hydrogen atom, or R ¹ and R ² together represent ═N—OCH ₃ ,
The present invention provides a pinewood nematode control agent characterized by containing a compound represented by formula (1) as an active ingredient.
[0010]
The compound of the above formula (I) in which R ¹ is a hydroxyl group and R ² is a hydrogen atom is aerobically Streptomyces cyaneogriseus noncyanogenus LL-F28249 (NRRL 15773). The following physicochemical properties produced by fermenting to:
(1) Molecular weight: 612 (FAB-MS)
(2) Molecular formula: C ₃₆ H ₅₂ O ₈
(3) Specific rotation: [α] _D ²⁶ = + 133 ± 3 ° (c0.3, acetone)
(4) Ultraviolet absorption spectrum: UV _max ^CH ₃ ^OH = 244 nm (ε28,000)
(5) Infrared absorption spectrum (KBr disc): 3439, 2960, 2925, 1714, 1454, 1374, 1338, 1171, 1120, 996, 967 cm ⁻¹
(6) C ¹³ nuclear magnetic resonance spectrum (CDCl ₃ ):

(1) Low magnetic field from TMS; CDCl ₃ solution
(2) q = quaternary carbon
(3), (4) Two unresolved signals (7) Electron impact mass spectrum:
m / Z Elemental composition
612.3705 C ₃₆ H ₅₂ O ₈
594.3543 C ₃₆ H ₅₀ O ₇
576.3472 C ₃₆ H ₄₈ O ₆
484.3211 C ₃₀ H ₄₄ O ₅
482.2648 C ₂₉ H ₃₈ O ₆
466.3097 C ₃₀ H ₄₂ O ₄
448.2987 C ₃₀ H ₄₀ O ₃
442.2375 C ₂₆ H ₃₄ O ₆
425.2327 C ₂₆ H ₃₃ O ₅
354.2181 C ₂₃ H ₃₀ O ₃
314.1877 C ₂₀ H ₂₆ O ₃
278.1144 C ₁₅ H ₁₈ O ₅
265.1786 C ₁₆ H ₂₅ O ₃
248.1405 C ₁₅ H ₂₀ O ₃
247.1705 C ₁₆ H ₂₃ O ₂
237.1838 C ₁₅ H ₂₅ O ₂
219.1740 C ₁₅ H ₂₃ O
151.0753 C ₉ H ₁₁ O ₂
The details of the production method are described in JP-A No. 61-10589 (in the same publication, R ¹ is a hydroxyl group and R ² is a hydrogen atom. The compound is referred to as the “LL-F28249α” compound, and will be referred to as such in this specification), and therefore, the detailed description will be replaced with the reference herein.
[0011]
According to the aerobic fermentation of Streptomyces cyaneoglyceus non-cyanogenus, as described in the above-mentioned JP-A 61-10589, not only the LL-F28249α compound, but also eradication properties of external and internal parasites Any one or more of the following compounds having antibiotic activity, designated as LL-F28249β, γ, δ, ε, ζ, η, θ, ι, κ, λ, μ, ν, and ω, shown below: Generate.
[0012]
[Chemical 3]

[0013]
[Table 1]

[0014]
[Formula 4]

[0015]
Therefore, the LL-F28249α compound which is an active ingredient in the control agent of the present invention can be used in a substantially pure form extracted and isolated from the fermentation broth, or alternatively, the LL-F28249β described above. , Γ, δ, ε, ζ, η, θ, ι, κ, λ, μ, ν and ω in the form of a mixture with one or more compounds, or a fermentation broth or mash containing them It can also be used in the form of Usually in the form of a mash comprising LL-F28249α compound and one or more of LL-F28249β, γ, δ, ε, ζ, η, θ, ι, κ, λ, μ, ν and ω compounds Are preferably used.
[0016]
On the other hand, the compound of the above formula (I) in which R ¹ and R ² together represent ═N—OCH ₃ is a compound known per se called by the general name “moxidectin” ( JP, 63-54375, A), for example, it can manufacture by making the above-mentioned LL-F28249 alpha compound oxidize, and making it react with methoxyamine or its salt.
[0017]
When the compound of the formula (I) is used for controlling pinewood nematodes, the compound can be formulated into a dosage form that can be easily injected into a pine tree trunk and has a high rate of migration into the tree. For example, the compound of the formula (I) can be made into a liquid together with an appropriate solvent, and particularly preferably an aqueous liquid based on water. Examples of the solvent for the aqueous liquid preparation include water alone or, preferably, a mixed solvent of water and a water-miscible organic solvent such as methanol, ethanol, acetonitrile, butoxyethanol and the like. In particular, the use of acetonitrile as the water-miscible organic solvent is preferable because the permeation transfer rate of the liquid into the tree is significantly improved.
[0018]
The ratio of the water-miscible organic solvent in the mixed solvent is generally within a range of 1 / 0.1 to 1/2, particularly 1/1 to 1 / 1.5, by weight ratio of water / water-miscible organic solvent. Is appropriate.
[0019]
The concentration of the active ingredient compound of formula (I) in the aqueous liquid is not strictly limited and can be changed according to the type of the compound, the type of the solvent, etc., but is usually 0.1. It can be in the range of -20% by weight, preferably 1.8-5.4% by weight.
[0020]
In addition, the liquid preparation containing the compound of formula (I) may further include a suitable auxiliary agent, for example, a surfactant [Example: polyoxyethylene alkylphenyl ether (8 to 20 ethylene oxide addition moles), if necessary. , Polyoxyethylene allyl phenyl ether (8 to 20 ethylene oxide addition mole), polyoxyethylene styryl phenyl ether and its condensate (8 to 20 ethylene oxide addition mole), polyoxyethylene glycol surfactant, alkylbenzene sulfonate Type surfactant, dioctyl succinate] and the like.
[0021]
The application time and application method of the control agent of the present invention to the pine can be the same as those currently on the market. For example, the application time is up to about 3 months before the occurrence of the adult insect of the pine beetle In addition, as an application method, for example, between about 0.5 to about 1 m in height from the base of the trunk, a depth of about 5 to about 9 cm and a diameter of 6 are slightly inclined downward with a drill or the like. A method of forming an injection hole of ˜8 mm and injecting the liquid agent using an injector is mentioned. The number of injection holes per tree can be changed according to the thickness, height, etc. of the trunk of the tree, but can usually be about 1-2 holes / tree with a pine tree having a breast height of 10 to 20 cm.
[0022]
The amount of the control agent of the present invention can be changed according to the breast height diameter of the target pine tree trunk, for example, an aqueous solution containing 3.6% by weight of the compound of formula (I). In some cases, the following amounts can be used as standard amounts.
[0023]

[0024]
Injection is usually performed under atmospheric pressure, but in some cases, pressurized injection may be performed. The pressure in the case of pressure injection is desirably ² kg / cm ² or less.
[0025]
As described above, the pinewood nematode control agent of the present invention has a much higher activity against pinewood nematode than the currently marketed control agents, and is therefore low and high in dosage. It can be expected to have a killing effect.
[0026]
In addition, the aqueous liquid preparation of the present invention has a high rate of migration into the tree, no phytotoxicity to pine, and is extremely useful practically as a pinewood nematode control agent.
[0027]
Hereinafter, the present invention will be described in more detail with reference to formulation examples and test examples.
[0028]

The above components were thoroughly mixed to obtain an aqueous liquid having an active ingredient concentration of 3.6% by weight.
[0029]

The above components were thoroughly mixed to obtain an aqueous liquid having an effective concentration of 1.8% by weight.
[0030]

The above components were sufficiently mixed to obtain an aqueous liquid having an effective concentration of 3.6% by weight.
[0031]

The above components were sufficiently mixed to obtain an aqueous liquid having an effective concentration of 3.6% by weight.
[0032]

The above components were sufficiently mixed to obtain an aqueous liquid having an effective concentration of 3.6% by weight.
[0033]

The above components were sufficiently mixed to obtain an aqueous liquid having an effective concentration of 3.6% by weight.
[0034]

The above components were sufficiently mixed to obtain an aqueous liquid having an effective concentration of 3.6% by weight.
[0035]
Test Example 1: In vitro test of growth inhibitory effect against pinewood nematode (1) Test method Test animals: pinewood sentiyu S-10 strain system: Each group 2 repeated treatment method: Gray mold fungus system on PSA medium A chemical solution having a predetermined concentration was poured into a shyare (diameter 90 mm) in which (the nutrient of pinewood nematode) was bred, and left for 1 minute, and then the chemical solution was discarded. 0.2 cc of a suspension of pinewood nematode was added dropwise to a shearle that had been subjected to chemical treatment and placed under a temperature condition of 25 ° C. The number of centimeters in 0.6 cc of the suspension was measured under a microscope to determine the average number of centimeters in 0.2 cc, and this was taken as the number of dropped centimeters (initial number of individuals). Seven days after the inoculation, 20 cc of water was poured into the shear and stirred for 15 seconds, and then the water was recovered. Each 0.2 cc of the collected water was taken and observed three times under a microscope, the number of surviving centimeters was measured to estimate the number of surviving individuals (number of individuals after 7 days), and the growth rate was calculated by the following formula.
[0036]
[Expression 1]

[0037]
Agent used: “Sentily Injection” by Hodogaya Chemical Co., Ltd. (contains 4% levamisole hydrochloride) as a drug for comparison of 4% by weight methanol solution of LL-F28249α
And "Glingard Eight" from Faiser Pharmaceutical Co., Ltd. (containing 8% Morantel tartrate)
use.
[0038]
In addition, in order to improve the diffusion of the chemical solution, a surfactant (betalin A, diluted 5000 times) was added to the drug used.
[0039]
(2) Test results [0040]
[Table 2]

[0041]
Test Example 2: Chemical hazard test (1) Test method Reagent: LL-F28249α
Injection date / time: January 30 Test plant: Japanese black pine 3rd year ward system: 1 ward 1 tree 3 repetitive treatment method: A hole of 2.5 mm in diameter is drilled in each test tree with a drill in one place, and a chemical solution is injected under pressure. I was in an artificial weather room at 25 ° C.
[0042]
The drugs used for the test and the injection amount are shown in Table 2.
[0043]

Test Example 3: Field Test (Part 1)
1. Test method (1) The test site test was conducted in a red pine 18-30 year old forest. The area around the test site has been damaged by pinewood nematode from 5 to 6 years ago, and now it has a depletion rate of about 10%.
[0044]
(2) Aqueous solution (3.6% AS) prepared in Formulation Example 1 using LL-F28249α as a reagent agent
(3) In the treatment method and the treatment date drug treatment method, an injection hole having a diameter of about 6 mm and a depth of about 5 cm was made in a 50-80 cm portion above the ground with a straight drill, and a natural pressure was injected by inserting a container therein. . The application rate is 2 treatments of 125 ml and 250 ml per m ^3, and the treatment amount of each sample tree is as shown in Table 4.
[0045]
The drug treatment took place on February 22.
[0046]
In any case, in order to prevent decay around the injection hole, Toppsin M paste was injected into the injection hole.
[0047]
(4) Test tree The test tree is a red pine small-diameter tree having a chest height of 10 to 14 cm and a tree height of 10 to 12 m. The number of test specimens for each treatment was 10, and in addition, 10 untreated areas and 5 phytotoxic areas were used.
[0048]
(5) Nematode Inoculation Nematode inoculation took place on July 2. The inoculation method is a pine tree with a diameter of about 9 mm and a depth of about 6 cm in the trunk of a 4 m high above the test tree. 2 cc (30,000 heads) of the suspension was injected. The nematodes used were those cultured at Forest Research Institute.
[0049]
(6) Survey method (1) After inoculating the drug effect nematode, survey the amount of resin (August, November), investigate the abnormal condition of the sample tree, and conduct the withering survey in mid-November. From this, the treatment effect of the drug was determined. In addition, the resin outflow amount was divided into five stages, and “++, ++” was a normal tree and “+, −, 0” was an abnormal tree. However, in this case, the trees that were healthy but had a small resin spillage were treated as abnormal trees, and conversely, even if some discolored parts were seen, those with normal resin spillage were treated as normal trees.
[0050]
(2) Investigation of phytotoxicity In the phytotoxicity area, 500 ml per m ³ was infused per one sample tree, and then it was investigated in the state of withering.
[0051]
(3) The treated hole in the formation layer obstacle chemical damage zone around the injection hole was peeled off at the time of investigating withering, and the discoloration state of the formation layer part was investigated.
[0052]
2. Results and Discussion (1) The drug effect test results are as shown in Tables 4 and 5.
[0053]
First, the withering rate in the untreated area was 90% on August 5th, 32 days after nematode inoculation, and 100% on November 15th after 100 days had passed.
[0054]
On the other hand, the treatment with the compound of formula (I) had a remarkable effect of this chemical treatment because both the 125 ml treatment and the 250 ml treatment had a resin amount of ++, the withering rate was 0%, and no nematode was detected. Was recognized. In addition, since there was no difference in effect between the 125 ml treatment and the 250 ml treatment, 125 ml / m ³ treatment seems to be sufficient for practical use.
[0055]
In addition, since this drug is applied in a small amount, it can be easily handled in the field such as transportation and injection processing, and the injection time is extremely fast (in this case, all are completed within one hour), so the drug effect is included. This is considered to be a highly practical control technique.
[0056]
(2) The results of the survey on phytotoxicity are as shown in Table 6. None of the five test trees showed signs of leaf discoloration or abnormal fallen leaves.
[0057]
(3) Formation layer hindrance around the injection hole Although all the sample trees were slightly discolored around the injection hole, no discoloration (1 cm or more) apparently appears to be a formation layer hindrance. .
[0058]
3. Summary (1) A remarkable effect of LL-F28249α on pinewood nematode was observed.
[0059]
(2) This drug is considered to be a highly practical control technique because of its small application amount and extremely fast injection time.
[0060]
(3) It is considered that the dose may be 125 ml per m ³ .
[0061]
(4) No phytotoxicity was observed with this drug.
[0062]
(5) There was no significant formation layer failure around the injection hole.
[0063]
[Table 3]

[0064]
[Table 4]

[0065]
[Table 5]

[0066]
Test example 4: Field test (2)
1. Outline of the test site (1) Location conditions A tideland forest planted on the coastal area, except for an altitude of around 5m and a slope of less than 5% in some areas, and the soil is sand and the surface is below it. Is sand and gravel, and the deep part is mostly gravel.
[0067]
(2) Forest condition Black pine 25-27 years old, the number of standing trees is about 3,000 per ha, tree height 8-15m, chest height 6-18cm.
[0068]
2. Treatment conditions (1) Treatment date and time February 27 (2) Treatment method, drug name, treatment amount, etc. For chemical treatment, the amount of resin spillage was investigated. A hole with a diameter of 9 mm and a depth of 3 cm was made in the tree trunk and injected with an ampoule by natural pressure. The same reagent as used in Test Example 3, the processing category and the number of samples used are as follows.
[0069]
500ml / m ³ ward (phytotoxic zone) 5 250 250 〃 10 １２５ 125 区 10 Nematode inoculation On June 30th, the amount of resin spilled on the test tree was investigated, and those found to be normal were inoculated with nematodes. Nematodes were cultivated and propagated from the National Forest Research Institute and inoculated 1 ml of a suspension adjusted to 30,000 per ml of test tree. The inoculation site was a tree trunk with a height of 4 to 5 m above ground, and what was possible was cut off a branch nearby and inoculated a hole with a diameter of 9 mm and a depth of 3 cm.
[0070]
4). The investigation method and the investigation result effect investigation were carried out with the resin outflow amount by the Oda method judgment method. The survey date is two times, September 28, 3 months after nematode inoculation, and November 19, 5 months after the nematode inoculation, and at the time of the final survey, dead trees or abnormal trees were collected by drilling from the trunk chest height. The nematode was rediscovered.
[0071]
The survey results are shown in Table 7.
[0072]
5. Consideration of the results The control area has a withering rate of 90% and an abnormal tree rate of 100%, and the nematode used this time has a high pathogenicity.
[0073]
On the other hand, all of the chemical treatment areas were healthy, and it was considered that the control effect of this drug against pinewood nematode was very high. The phytotoxic areas were all healthy and there were no abnormalities in appearance.
[0074]
[Table 6]

[0075]
[Table 7]

[0076]
Test Example 5: In vitro test of growth inhibitory effect against pinewood nematode (1) Test method Test animals: pinewood nematode S-10 strain system: Each group, 2 iterations: Same use as in test example 1 Drug: “Sentily Injection (containing 4% levamisole hydrochloride) from Hodogaya Chemical Co., Ltd.”
And "Glingard Eight" (containing 8% Morantel tartrate)
use.
[0077]
In addition, in order to improve the diffusion of the drug, a surfactant (betalin A, diluted 5000 times) was added to the drug used.
[0078]
(2) Test results [0079]
[Table 8]

Claims (5)

formula

A pine wilt control agent in the form of an aqueous solution prepared by dispersing the compound represented by formula (1) in a mixed solvent of water and acetonitrile .   The control agent according to claim 1, comprising 0.1 to 20% by weight of the compound of formula (I). The control agent according to claim 1, wherein the mixing ratio of water and acetonitrile is in the range of 1/1 to 1 / 1.5 by weight ratio of water / acetonitrile . The active ingredient contains a compound of formula (I) according to claim 1, obtained by aerobic fermentation of Streptomyces cyaneogriseus noncyanogenus LL-28249 (NRRL 15773). The control agent according to claim 1, which is a fermentation broth. An effective amount of the control agent according to claim 1 is injected into a pine tree trunk.