1329001 九、發明說明： 【發明所屬之技術領域】 本發明乃有關用以預防及消除有害生物之組成物，更 具體而言，係含有3,5_二氯]_(3,3_二氣：丙烯氧 基）-4-[3/(5-二氟曱基D比咬-2-基氧基）丙氧基]苯以及蘇雲金 芽孢桿菌（Bacillus thuringiensis(同 thuringiesis))之殺蟲性 蛋白質而用以預防及消除有害生物之組成物。 【先前技術】 己知3,5-二氯-1-(3,3-二氯丙烯氧基）_4_[3_(5_三氟 甲基吡啶_2_基氧基）丙氧基]苯具備殺蟲活性（參考 WO96/11909 號公報）。 因此，利用3,5_二氯-1·(3,3-二氣_2_丙烯氧基）·4·[3_(5_ :氟甲基吡啶-2-基氧基）丙氧基]笨，可期待研究開發對於 辰、園蟄上之有害生物，以更低劑量即可顯示優異防除效 果之有害生物防除劑。 【發明内容】 ^本發明研究者在上述情況下銳意探討之結果，發現可 獲得較單純混合時所得相加效果更高之相乘效果，且能減 少所用劑量之用以預防及消除有害生物之組成物中之調配 組合而完成了本發明。 即’本發明提供下列項目： 1·含有 3,5-二氯-1_(3,3-二氯 _2_ 丙烯氧基）_4_[3_(5_三 氟甲基吡啶-2-基氧基）丙氧基]苯（下文中，或簡稱本 化合物）及蘇雲金芽孢桿菌（Bac⑴us thuringiensis)1329001 IX. Description of the invention: [Technical field to which the invention pertains] The present invention relates to a composition for preventing and eliminating harmful organisms, and more particularly, 3,5-dichloro]-(3,3_digas : propyleneoxy)-4-[3/(5-difluorodecyl D-biti-2-yloxy)propoxy]benzene and insecticidal protein of Bacillus thuringiensis (with thuringiesis) It is used to prevent and eliminate the constituents of harmful organisms. [Prior Art] It is known that 3,5-dichloro-1-(3,3-dichloropropenyloxy)_4_[3_(5-trifluoromethylpyridine-2-yloxy)propoxy]benzene has Insecticidal activity (refer to WO96/11909). Therefore, using 3,5-dichloro-1·(3,3-dioxa-2-propenyloxy)·4·[3_(5_:fluoromethylpyridin-2-yloxy)propoxy] It is expected to research and develop harmful insecticides that exhibit excellent control effects at lower doses for pests on the ground and garden. SUMMARY OF THE INVENTION The inventors of the present invention have been keenly discussing the results under the above circumstances, and found that a multiplication effect obtained by a simple mixing effect can be obtained, and the dose used can be reduced to prevent and eliminate harmful organisms. The present invention has been completed by a combination of the compositions. That is, the present invention provides the following items: 1. Containing 3,5-dichloro-1_(3,3-dichloro-2-propenyloxy)_4_[3_(5-trifluoromethylpyridin-2-yloxy) Propoxy]benzene (hereinafter, or simply referred to as the present compound) and Bacillus thuringiensis (Bac(1)us thuringiensis)
3J640J 5 1329001 之殺蟲性蛋白質（下文中，或簡稱本蛋白質） 之用以預防及消除有害生物之組成物（下文中，或三 稱本發明組成物）； 2·含有相乘效果量（a synergistic effective 〇f) 之3,5-二氯-l-(3,3_二氣_2_丙烯氧基）牡[3三一 甲基吡啶-2·基氧基）丙氧基]苯及蘇雲金芽孢桿= (Bacillus thuringiensis)之殺蟲性蛋白質之用以預 及消除有害生物之組成物； 防 3. 如上述第1項或第2項之用以預防及消除有害生物 之組成物，其中含有以重量比計為1 : 1〇〇〇至1〇: 1 比例之 3,5-二氯 _ι_(；3,3_ 二氣 _2_ 丙烯氧基 三氟甲基吡啶-2-基氧基）丙氧基]苯及蘇雲金芽孢桿 菌（Bacillus thuringiensis)之殺蟲性蛋白質者； 4. 如上述第丨項或第2項之用以預防及消除有害生物 之組成物，其中殺蟲性蛋白質係蘇雲金芽孢桿菌 (Bacillus thuringiensis)之培養物、活菌體，或芽胞， 或其經熱處理或化學方法死菌化處理過之菌體，或 其菌體之破碎物中所含之殺蟲性蛋白質者； 5·—種用以預防及消除有害生物之方法，其特徵為將 有放里之3,5- 一氣-l-(3,3-二氯-2-丙焊氧 基）-4-[3-(5-三氟甲基吡啶_2_基氧基）丙氧基]苯及蘇 4孟芽抱桿囷（Bacillus thuringiensis)之殺蟲性蛋白 質兩者，施用於須保護植物，或有害生物或有害生 物之棲息場所者； 316401 6 1329001 6.—種用以帛防及消除有害生物之方其特徵為將 相乘效果量（a synergistic effective am〇unt 〇f)之 3,5·二氯-丨-^夂二氯_2•丙烯氧基）_4_[3_(5_三氟甲 基D比疋-2-基氧基）丙氧基]苯及蘇雲金芽孢桿菌 (BacUlus thuringiensis)之殺蟲性蛋白質施用於須保 護植物，有害生物或有害生物之棲息場所者； 7,為預防及消除有害生物而調配組合-二氯 二氯-2-丙烯氧基）_4_[3_(5_三氟甲基吡啶_2_基氧基） 丙氧基]笨及蘇雲金芽孢桿菌（Bacil丨us thuringiensis) 之殺蟲性蛋白質以供使用；以及 8·為預防及消除有害生物而調配組合相乘效果量 synergistic effective amount of)之 3,:5-二氣-1-(3,3- 二氣-2-丙烯氧基）_4_[3_(5_三氟甲基吡啶_2_基氧基） 丙氧基]苯及蘇雲金芽孢桿菌（Bacillus thuringiensis) 之殺蟲性蛋白質以供使用等。 【實施方式】 (實施本發明之最佳形態） 本發明組成物中所使用作為有效成分之一的本化合物 乃係下式（1)所示具有殺蟲活性之化合物。3J640J 5 1329001 insecticidal protein (hereinafter, or simply referred to as the protein) for preventing and eliminating the composition of pests (hereinafter, or three is called the composition of the present invention); 2·containing the amount of multiplication effect (a Synergistic effective 〇f) 3,5-dichloro-l-(3,3_dioxa-2-propenyloxy)mu [3 trimethylpyridin-2-yloxy)propoxy]benzene and The insecticidal protein of Bacillus thuringiensis is used to pre-treat the composition of the pest; 3. The composition for preventing and eliminating harmful organisms according to item 1 or 2 above, wherein Contains a ratio of 1:1〇〇〇 to 1〇: 1 ratio of 3,5-dichloro_ι_(;3,3_diox_2_ propyleneoxytrifluoromethylpyridin-2-yloxy a propionyl protein of phenyloxy]benzene and Bacillus thuringiensis; 4. A composition for preventing and eliminating harmful organisms according to the above item or item 2, wherein the insecticidal protein system Cultures, live cells, or spores of Bacillus thuringiensis, or heat treated or chemically treated a method for preventing and eliminating pests, which is a method for preventing and eliminating harmful organisms, which is a method for preventing and eliminating harmful organisms; 5-mono-l-(3,3-dichloro-2-propeneoxy)-4-[3-(5-trifluoromethylpyridin-2-yloxy)propoxy]benzene and su 4 Both insecticidal proteins of Bacillus thuringiensis are applied to those who need to protect plants, or habitats of harmful organisms or harmful organisms; 316401 6 1329001 6. - Used to prevent and eliminate harmful organisms It is characterized by the amount of synergistic effect (a synergistic effective am〇unt 〇f) of 3,5·dichloro-indole-^夂dichloro-2-propenyloxy)_4_[3_(5_trifluoromethyl) The insecticidal protein of D is more than 疋-2-yloxy)propoxy]benzene and BacUlus thuringiensis is applied to the habitats of plants, pests or pests; 7, for prevention and elimination Combination of pests-dichlorodichloro-2-propenyloxy)_4_[3_(5-trifluoromethylpyridine-2-yloxy)propoxy] stupid and Bacillus thuringiensis (Bacil丨us thur Ingiensis) insecticidal protein for use; and 8) synergistic effective amount of 3,5-diqi-1-(3,3-diox) for the prevention and elimination of pests -2-Propoxyoxy)_4_[3_(5-trifluoromethylpyridin-2-yloxy)propoxy]benzene and an insecticidal protein of Bacillus thuringiensis for use or the like. [Embodiment] (Best Mode for Carrying Out the Invention) The present compound which is one of the active ingredients used in the composition of the present invention is a compound having insecticidal activity represented by the following formula (1).
本化合物記載於WO96/11909號公報中，可依照該公 316401 7 1329001 報中所記載方法而製造。 本發明組成物中所使用作為有效成分之一的本蛋白質 祇要是蘇雲金芽孢桿菌（Bacillus thuringiensis)之殺蟲性蛋 白質即可，別無限制，具體而言，為例如CrylA(a)、 CrylA(b)、CrylA(c)、CrylB、CrylC、CrylD、CrylF、CryllA、 CryllB、CrylllB、CrylllC、CrylllD、CryV、CryIV 等。 這些本蛋白質係天然由蘇雲金芽孢桿菌（Bacillus thuringiensis)所屬細菌所生產，其中以由卻氏卡奇亞種 (kurstaki)、亞莎華亞種（aizawai)、尼布甲亞種 (tenebrionis)、曰本亞種（japonensis Buibui)等亞種所屬之上 述細菌所生產者做為本發明組成物使用較佳。 又，本蛋白質為天然存在之蘇雲金芽抱桿菌（Bacillus thuringiensis)所屬細菌所生產者，但是亦可為一般基因工 程學方法所製成之轉形體（例如大腸菌、枯草桿菌、植物等） 所生產者。又，製作上述轉形體之際所使用，具有編碼本 蛋白質胺基酸序列之鹼基序列之基因（下文中，或簡稱為本 基因）可為：（1)由天然存在之基因中經選殖之基因；（2)由 天然存在之基因中經選殖之基因之鹼基序列中，其一部分 驗基之缺失、取代或加成係以人為方式導入而得之基因 [即，天然存在之基因經變異處理（施以局部變異導入處 理、突然變異處理等）而得之基因];(3)人為合成之基因等 皆可行。 上述微生物可自天然中分離，或由菌種保存機構等購 入，又可從上述微生物藉一般微生物學方法或基因工學方 8 316401 1329001 法等進行人工合成。 從自然界分離時，例如首先自野外採取土壤，懸濁所 採取土壤於滅菌水中之後，所得懸濁液塗布於例如枯草桿 菌等微生物分離用固體培養基上，在251下培養之。數曰 後切取得增殖之獨立菌叢，例如移植於枯草桿菌等微生物 分離用固體培養基上，再於25°C下培養之。就所生長之細 菌按照 SNEATH，（P.H.A.)，MAIR，（N.S·) SHARPE, (M.E.) and HOLT, (J.G.) :Bergey’s manual of Systematic Bacteriology,第 2 卷，1984 年，Williams and Wilkins, Baltimore等所記載之方法等，鑑定是否屬於蘇雲金芽孢桿 菌（Bacillus thuringiensis)之細菌而選拔屬於蘇雲金芽孢桿 菌（Bacillus thuringiensis)之細菌即可。 其次，就所選拔之屬於蘇雲金芽孢桿菌（Bacillus thuringiensis)之細菌，例如依據生物活性評估法或顯微鏡 觀察法等確認該細菌中有無殺蟲性蛋白質，而選拔屬於蘇 雲金芽孢桿菌（Bacillus thuringiensis)之細菌即可。 本發明組成物中所使用之本蛋白質，可直接為該蛋白 質本身，但是也可以是例如蘇雲金芽孢桿菌（Bacillus thuringiensis)之培養物、活菌體、芽孢，或其經熱處理或 化學方法經死菌化處理之菌體，或該等菌體之破碎物中所 含之殺蟲性蛋白質。 又，培養能生產殺蟲性蛋白質之蘇雲金芽孢桿菌 (Bacillus thuringiensis)所屬細菌之培養基，例如可使用一 般細菌用培養基，一般肉汁液體培養基等，祇要是該細菌 9 31640] 可利用。如上述將含有使用上述培養基 及复生產物蛛跑桿菌（Baciuus thuHngiensis)之細菌 或乾烤而養物，連同培養基—起施與離心分理及/ it而做為本發明組成物中之另—種有效成分之本蛋白 虫。&，按照周知方法，從上述培養物分離而得之殺 :、蛋白質本身也可以直接作為本蛋白質使用。本蛋白質 刀子量通常為數萬至十數萬道爾吞(―)。本蛋白質在 =複分裂-定次數之屬於蘇雲金芽孢桿菌（Baciiius ^nnglensis)之、細菌崩解之際，可藉結晶性物質之形態和 芽孢一起釋放出來。 又，本發明組成物中所使用之本蛋白質，也可使用市 販之BT製劑品’其中以使用台波爾⑼ρ〇ι)(商標名，住友 化學工業公司製品）、愛司馬克（商標名，住友化學工業公 司製品）、浮羅派克（商標名，住友化學工業公司製品）、仙 達利（商標名，住化武田農藥公司製品）、諾佛都耳 (Novodor)(商標名，Valent Bio Sciences公司製品）、布衣漢 特（商標名，住友化學工業公司製品：由久保田生技公司在 2003 年 12 月獲得轉讓）、Design(商標名，Thermo Trilogy 公司製品）等。 本發明組成物可利用在廣範圍之農園藝中以預防或消 除有害之生物（例如昆蟲類或有害蟎類等）。其典型之有害 生物種類如下述： 半翅目害蟲：稻灰飛虱（Laodelphax striatellus)、稻褐 飛虱（Nilaparvata lugens)、白背飛虱（Sogatella furcifera)等 316401 1329001 飛il類、黑尾葉輝（Nephotettix cincticeps)、微葉蟬 (Empoasca onukii)等葉蟬類、棉蚜（Aphis gossypii)、赤蚜 (Myzus persicae)、油菜财蟲（Lipaphis pseudobrassicae)等管 虫牙類、蝽類、溫室白粉風（Trialeurodes vaporariorum)、棉 粉風（Bemisia tabaci)、銀葉粉風（Bemisia argentifolli)等粉 虱類、介殼蟲類、軍配蟲類、木虱類等。 鱗翅目害蟲：二化镇（Chilo suppressalis)、稻縱捲煩 (Cnapha locrocis medinalis)、歐州玉米螟（Ostrinia nubilalis)、镇蛾科（Parapediasiateterrella)等填蛾類、斜紋 夜蛾（Spodoptera litura)、甜菜葉蛾（Spodoptera exigua)、東 方黏蟲（Pseudaletia separata)、甘藍夜蛾（Mamestra brasicae)、球菜葉蛾（Agrotis ipsilon)、擬尺礎（Trichoplusia ni)、紋葉蛾（Autographa nigrisigna)、夜蛾屬（Heliothis spp)、煙葉蛾種（Helicoverpa spp)、鑽夜蛾種（Earias spp) 等夜蛾類、日本紋白蝶（Pieris rapae crucivora)等粉蝶類、 茶姬捲葉蛾（Adoxophyes honmai)、茶小卷夜蛾（Adoxophyes orana fasciata)、梨小食心蟲（Grapholita molesta)、蘋果蠹 蛾（Cydia pomonella)等葉捲蛾類、桃小食心蟲（Carposina niponensis)等食心蟲、桃潛葉蛾（Lyonetia clerkella)等潛葉 蛾類、麗金龜（Phyllonorycter ringoniella)等細蛾類、柑橘 潛夜蛾（Phyllocnistis citrella)等小潛葉蛾類、小菜蛾 (Plutella xylostella)等菜蛾類、台灣黃毒蛾（Euproctis taiwana)、舞毒蛾（Lymantria dispar)、黃尾毒蛾（Euproctis similis)等毒蛾類、背刺蛾（Scopelodes contracus)等刺蛾 316401 1329001 類、赤松毛蟲（Dendrolimus spectabilis)等枯葉蛾類、紅鈴 蟲（Pectinophora gossypiella)等紅铃蟲類、燈蛾類、廣巢小 峨類等。 雙翅目害蟲：非州菊斑潛繩（Liriomyza trifolii)、番另S 斑潛繩（Liriomyza bryoniae)、美州斑潛繩（Liriomyza sativae)、昭和草斑潛蠅（Liriomyza asterivora)、婉豆彩潛 蠅（Chromatomyia horticola)等潛葉蛾類、家蚊類、藪蚊類、 葉斑蚊類、搖蚊類、家蠅類、墨蠅類、肉蠅類、花蠅類、 玉繩類、實绳類、果繩類、蝶蠅類、虹：蚊類、蚋類、刺蜗 類等。 鞘翅目害蟲：葉蟲類、金龜子類、象蟲類、捲葉象蟲 類、瓢蟲類、天牛類、偽步行蟲類等。 薊馬目害蟲：黃紋薊馬（Thrips palmi)等薊馬屬、首稽 薊馬（Frankliniella occidentalis)等花莉馬屬 茶黃薊馬 (Sciltothrips dorsalis)等茶黃薊馬屬等薊馬類、管莉馬〶 等。 戈 膜翅目害蟲：葉蜂類、螞蟻類、虎頭蜂類等， 網翅目害蟲：蟑螂類、茶翅蟑螂類等， 直翅目害蟲：飛蝗類、螻蛄類， 隱翅目害蟲：跳蚤類， 蝨目害蟲：人蝨等， 白蟻目害蟲：曰本白蟻（Reticulitermes speratus)、臺灣 家白蟻（coptotermes formosanus)等之白議類， 碑目害蟲：蜘蛛種（Tetranychus spp.)、紅蜘蛛種 316401 12 1329001 番茄刺皮瘿蟎（Aculops 真蜱類等。 (Panonychus spp.)等葉蜱類 lycopersici)等節蜱類、粉蜱類 本發明組成物之用途並無特別限制，例如可使 監、白菜、青梗菜、蘿萄、青花菜、葛苣、渡二、 庙子、青椒、辣椒、胡瓜、西瓜、香瓜、蘆筍、：、 香焦、稻米、裸麥、小麥、大麥、燕麥、粟、㈣ 善、玉米、菜豆、田豆（field bean)、黃豆、紅豆甜^ 甘庶、茶、料、飼料作物類、食用甜菜、 1 、雛豆、柑桔類、可可椰子、咖啡、棉花二 化果、_、頻果、櫻桃、梨、桃、梅、柿、番石 : 黃秋葵、觀賞植物、木瓜、花生、胡椒、樹豆 ） =物或果實中，多種農園藝方面，用以預防或 生物。 。 另外，例如草皮、康乃馨、櫻花、薔薇、非洲菊、菊、 盆栽卓化及仙人㈣栽培花卉也能利用本發明加以保護。 又’栽培植物之例舉應包括所有觀賞用或開花植物類。 本發明組成物中，本化合物及本蛋白f間之混 例：祇要是可獲得相乘效果之混合比例即可，例如以重‘比 计為1 : 10000至10000:丨之比例，而以丨：1〇〇〇至⑺Μ 之比例較佳’以1 : 1_至5 : i之比例更佳，而以^ 4〇 至1 : 1之比例又更優，尤以i : 2〇至i :】之比例為最佳。 本發明組成物在使用時，也可以不加 僅混合本化合物及本蛋白質而使用，-般在二t;: 可再添加固體載劑、液體載劑、氣體載劑等，必要時再加 316401 13 界面活化劑、其他製劑用 合劑、顆粒性水合劑、水“/丨“以油劑、乳劑、水 動劑、粉劑、粒劑、氣=蜀劑、水中乳濁劑等之可流 毒刪作為各種製宜微熱燻蒸劑、 有效成分之本發日月/，、 上述衣悧中，所含做為 95%。本么明組成物通常以重量比計宜為約0.01至 土 之際，所使用固體載劑之例舉如黏土類（高嶺 人/，潤土、夫巴沙密黏土、酸性白土等)、合成 ;Μ類、陶磁類' 其他無機礦物（絹雲母、石 二：Γ碳、碳叫水合氧化")、化學肥料(硫 =㈣叙、硝㈣、尿素、氣化銨等）等之微粉末或粒 .液體載劑可例舉如水、醇類（甲醇、乙醇等）'酮類（丙 嗣甲基乙基酮等）、芳族烴類（甲苯、二甲苯、乙基苯、 甲基蔡等）、非芳族煙類（己烧、環己烧、燈油、輕油等）、 酿類（乙酸乙醋、乙酸丁酉旨等）、腈類（乙腈、異丁膳等）、喊 類（”丙醚一噁烧等）、醯胺類（Ν，Ν-二甲基甲醯胺、ν，Ν-二：基乙醯胺類）、齒化烴類(二氣曱⑥、三氯乙烷、四氯 化碳等）、一曱亞碉、植物性油（黃豆油、棉籽油等）等。 又，氣體載劑可例舉例如夫隆氣體、丁烷氣體、液化 煤氣、二曱醚、二氧化碳等噴射劑。 界面活化劑可例舉如烷基硫酸酯鹽、烷基磺酸鹽、烷 基芳基磺酸鹽、烷基芳基醚類及其聚氧化乙烯化合物、聚 乙二醇醚類、多元醇酯類、糖醇衍生物等。 316401 1329001 -他衣劑用佐劑可例舉如路朊、明膠、_類（殿粉、阿 拉伯膠一纖維素衍生物、澡酸等）、木質素衍生物、膨潤土、 口成水冷性尚分子化合物(聚乙烯醇、聚乙烯吡咯烷酮、聚 丙烯m)' PAP(酸性磷酸異两自旨）、ΒΗτ(2,6_二第三丁 基基㈣、ΒΗΑ(2_第三丁基_4·甲氧㈣及]第三丁基 4曱氧基酚）、植物性油、礦物性油、界面活化劑、脂肪 酸、脂肪酸酯等。 毋名耳月j之基材可例舉如穀粉、植物性油、冑、結晶纖 維素等稱料成分、二丁基經基甲苯' 降二氯愈瘡木酸等抗 軋化^…脫氣乙酸等保存料、辣椒末等防止誤食劑、乾路 香料、洋蔥香料等誘引性香料等。 水中曰懸濁劑、水中乳濁劑等可流動劑，通常係將約工 至^重量％之有效成分化合物微量分散於含有約0.5至15 懸濁補助劑(例如保護膠體或可賦與觸變性之物 二）’約^至10重量％補助劑（例如消泡劑、防錄劑、安定 展著d滲透助劑、防凍劑、防腐劑、防霉劑等）之水 中即可獲得。 流本發明組成物可將各種有效成分依照上述製劑方法製 ::後：將此等製劑混合而製成。即，本發明組成物隨其 :劑形態’可將預先將本化合物製劑化而得之組成物及預 將本蛋白貝製劑化而得之組成物加以混合而製成。另 外，也可以在施用時將二者混合或併用之。 按照上述操作製劑化之本發明組成物可 以水等稀釋而使用。另外，也可以和其他之殺蟲劑、殺： 316401 15 y、殺線蟲劑、殺菌劑、 劑、肥料、土壤改良劑、動又：、植物生長調整劑、協力 本發明相杰―動物用飼料等混用或併用。 所，須從有害生物保護之上生::有害生物之棲息場 係製劑成為乳劑、水合劑、：：：•，當本發明組成物 通常以水等稀釋處理。㈣水合劑、可流動劑等時， 本發明組成物對於農園藝 本發明組成物之有㈠U 有#生物如處理時’ Λ ，里或施用量，通常以每1000m2之施 用里為約0.1至1000 ^ 有效成分直接以粒劑"或消除有害生物用之 丨叔劑、油劑等施用時’通常，不經 ° >稀釋直接按照上述有效量或施用量施用即可。 又’將預防或消除有害生物用之有效成分以乳劑、水 顆粒劑水合劑、可流動劑等用水稀釋而施用時，可 用水稀釋成為上述有效量或施用量而使用。具體而言，例 乂水稀釋礼劑、水合劑、顆粒水合劑、可流動劑等而使 用時，本發明組成物之施用濃度通常為約1至iooooppm。 上述之有效量或施用量皆隨製劑種類、施用時期、施 用％所、施用方法、害蟲種類、為害程度等狀況而不同， 並不限制在上述範圍而可適度增減使用。 實施例 本發明藉製劑例及試驗例詳細說明如下。但是本發明 不侷限在其範圍。 又’下述「BT菌培養物」之意義如下： 將屬於蘇雲金牙抱桿菌（Bacillus thuringiensis)細菌由 16 316401 1329001 來之活芽跑’每lml中含有1Χ ι〇3個孢子之懸濁液 1’接種在裝有經殺菌過之L-肉汁液體培養基（1〇〇mi)之容 量為500ml之燒瓶中，在25。(：下振盪培養（150rpm)2至5 曰左右，然後藉離心法從所得培養液回收屬於蘇雲金芽孢 桿菌（Bacillus thuringiensis)細菌由來之芽孢以及殺蟲性蛋 . 白質二者，並將二者乾燥所得之培養物（即，本蛋白質之形. 態之一）。 製劑例1乳劑 _ 溶解1份之本化合物及10份之BT菌培養物於含有 36·5份之二甲苯及36·5份之二曱基曱醯胺中，再加入1〇 伤之t氧化乙烯笨乙基苯基鱗及6份之十二烧基苯續酸 赶’經充分攪拌混合而製得乳劑。 製劑例2 水合劑 、添加5份之本化合物及45份之BT菌培養物於含有4馨 份之月桂基硫酸鈉，2份之木質素磺酸鈣，22份之合成含 虱氧化矽微粉末及22份之矽藻土之混合物中，充分混合 拌而製得水合劑。 見 製劑例3粒劑 末添，5份之BT菌培養物，5份之合成含氫氧化矽微粉 伤之十一烧基苯續酸鈉’ 30份之膨潤土以及54份之 妯土於1份之本化合物中，充分攪拌混合，繼之，加入適 31640] 17 量之水於上述混合物中， ， 通風乾燥而得粒劑。 充分攪拌，藉造粒機製成粒狀， 製劑例4 粉劑 將0.5份之本化合物 成含氫氧切微粉末 固培養物，1份之合 品）及7份之黏土在^之凝聚劑多利士6(三共公司製 拌’所得混合物中加：86=:分混合後，藉果汁機混合攪 得粉劑。 .”之黏土月，充分攪拌混合而 製劑例5可流動劑 添加1份之本化合物，1 之山犁插^ ^ 10伤之BT函培養物及1.5份 二：㈣酸醋於含有2份之聚乙稀醇之水溶液％ 充刀攪拌混纟’然後，藉砂磨冑將 :二為3…下)之後，於其中加入含有。。= # .1伤之矽酸鋁鎂之水溶液48份，再加入15份之 醇充分授拌混合而得可流動劑。 製劑例7 微膠囊劑 316401 1 _ 6油劑 之一 ’奋解0.1份之本化合物及〇 8份之BT菌培養物於5份 本及5伤之二氣乙院中，再混合以891份之脫臭燈 油而得油劑。 、 1329001 將2份之本化合物，8份之BT菌培養物，10份之苯 基二甲苯基乙烷及0.5份之士米珠L-75(住友貝爾脲烷公司 製品之曱苯撐二異氰酸酯）混合之後，加入20份之10%阿 拉伯膠水溶液中，藉均質機攪拌而得平均粒徑為20 // m之 乳化液。然後加入2份之乙二醇，再於60°C之溫浴中反應 24小時而得微膠囊淤漿液。另外，將0.2份之咕噸膠及1.0 份之蜂膠R(三洋化成公司製品之矽酸鋁鎂）分散於56.3份 之離子交換水中而得增黏劑溶液。混合42.5份之上述微膠 囊淤漿液及57.5份之增黏劑溶液而得微膠囊劑。 以下藉試驗例示本發明組成物所具有優異之有害生物 防除效果。又，混合兩種有效成分而處理之際可預測之殺 蟲率之理論值係由E=X+Y — (X+Y/100)(上式為一般被稱 為Colby之計算式，乃計算藥劑組合之相乘效果及拮抗效 果之判定基準而使用者）而求得。該式中， X :將有效成分A以Mppm處理之際之殺蟲率（％) Y :將有效成分B以Nppm處理之際之殺蟲率（％) E :當有效成分A以Mppm，有效成分B以Nppm處 理之際所預測之殺蟲率之理論值（％) 試驗例1 對於夜蛾（Heliothis virescens)之人工試料試驗 將10%之本化合物乳劑和BT菌製劑品：Design(商標 名，蘇雲金芽孢桿菌變種亞莎草株（Bacillus thuringiensis var. aizawai strain)GC91 接合體（conjugant)，可濕性顆粒製 劑品，Thermo Trilogy公司製品）之稀釋水溶液混合而調製 19 316401 1329001 成各個預定濃度之稀釋試驗液。於容量為30ml之塑膠杯 内注入人工飼料（約2g)於杯底之後，以0.2ml之試驗液處 理之。處理後約風乾2小時後，放飼供試夜蛾（Heliothis virescens)之2齡幼蟲一隻（20個重複）。處理5日後求得死 蟲率，該死蟲率以下式校正而算出殺蟲率（％)，其結果示於 表1。 殺蟲率（％)=100x (Mt—Mc)/(100—Me)This compound is described in WO96/11909, and can be produced according to the method described in the publication No. 316401 7 1329001. The present protein used as one of the active ingredients in the composition of the present invention may be any insecticidal protein of Bacillus thuringiensis, and is not limited, specifically, for example, CrylA(a), CrylA (b) ), CrylA (c), CrylB, CrylC, CrylD, CrylF, CryllA, CryllB, CrylllB, CrylllC, CrylllD, CryV, CryIV, and the like. These proteins are naturally produced by bacteria belonging to Bacillus thuringiensis, including kurstaki, aizawai, tenebrionis, and alfalfa. The producer of the above-mentioned bacteria to which the subspecies such as the subspecies (Japonensis Buibui) belong is preferably used as the composition of the present invention. Further, the protein is produced by a naturally occurring bacterium belonging to Bacillus thuringiensis, but can also be produced by a general genetic engineering method (for example, coliform, Bacillus subtilis, plant, etc.). . Further, when the above-described transformant is produced, a gene having a base sequence encoding the amino acid sequence of the present protein (hereinafter, or simply referred to as a gene) may be: (1) selected from a naturally occurring gene. a gene; (2) a gene derived from a naturally-occurring gene in a base sequence, wherein a part of the base is deleted, substituted or added is a human-derived gene [ie, a naturally occurring gene) The gene obtained by the mutation treatment (the local mutation introduction treatment, the sudden mutation treatment, etc.); (3) the artificially synthesized gene is feasible. The above microorganisms may be isolated from nature, purchased by a strain storage mechanism, or artificially synthesized from the above microorganisms by a general microbiological method or a genetic engineering method. When separating from the natural world, for example, the soil is first taken from the field, and the soil is suspended in sterilized water, and the resulting suspension is applied to a solid medium for microbial separation such as Bacillus subtilis, and cultured at 251. After several times, an independent colony which has been proliferated is obtained, for example, transplanted on a solid medium for microbial isolation such as Bacillus subtilis, and cultured at 25 °C. The bacteria grown are according to SNEATH, (PHA), MAIR, (NS·) SHARPE, (ME) and HOLT, (JG): Bergey's manual of Systematic Bacteriology, Vol. 2, 1984, Williams and Wilkins, Baltimore et al. The method of recording, etc., identifies whether the bacteria belonging to Bacillus thuringiensis and the bacteria belonging to Bacillus thuringiensis. Secondly, the bacteria belonging to Bacillus thuringiensis are selected, for example, according to the biological activity evaluation method or the microscopic observation method, and the bacteria belonging to Bacillus thuringiensis are selected, and the bacteria belonging to Bacillus thuringiensis are selected. Just fine. The present protein used in the composition of the present invention may be directly the protein itself, but may also be, for example, a culture of Bacillus thuringiensis, a living cell, a spore, or a heat-treated or chemically-treated fungus. The bacterial cells treated, or the insecticidal proteins contained in the broken bodies of the cells. Further, a medium for cultivating a bacterium belonging to Bacillus thuringiensis capable of producing an insecticidal protein can be used, for example, a general bacterium medium, a general broth liquid medium or the like, as long as the bacterium is available. As described above, the above-mentioned culture medium and the compound Bacchuus thuHngiensis bacteria or dry-baked nutrients, together with the culture medium, centrifuged and/or it will be used as another species in the composition of the present invention. The active ingredient of the protein worm. &, according to a known method, the above culture can be isolated and killed: the protein itself can also be directly used as the present protein. The amount of this protein knife is usually tens of thousands to tens of thousands of dolphins (-). The protein can be released by the form of a crystalline substance and the spore when the protein is disintegrated and the number of times belongs to Baciiius nnglensis. Further, as the present protein used in the composition of the present invention, a commercially available BT preparation product can also be used, in which Tai Bo (9) ρ〇ι (trade name, product of Sumitomo Chemical Industries Co., Ltd.), Aisike (trade name, Sumitomo Chemical Industries Co., Ltd.), Float Parker (trade name, Sumitomo Chemical Industries Co., Ltd.), Xiandali (trade name, Shouhua Takeda Pesticide Company), Novodor (trade name, Valent Bio Sciences) Products), Cloth Hante (trade name, Sumitomo Chemical Industries Co., Ltd.: transferred by Kubota Biotech Co., Ltd. in December 2003), Design (trade name, products of Thermo Trilogy), etc. The composition of the present invention can be utilized in a wide range of agricultural and horticultural plants to prevent or eliminate harmful organisms (e.g., insects or harmful mites, etc.). Typical pest species are as follows: Hemiptera pests: Laodelphax striatellus, Nilaparvata lugens, Sogatella furcifera, etc. 316401 1329001 Fly il, black tail Nephotettix cincticeps, Empoasca onukii, Aphis gossypii, Myzus persicae, Lipaphis pseudobrassicae, tubeworms, mites, greenhouse white wind (Trialeurodes vaporariorum), cotton powder wind (Bemisia tabaci), silver leaf powder wind (Bemisia argentifolli) and other whitefly, scale insects, military insects, hibiscus and so on. Lepidoptera pests: Chilo suppressalis, Cnapha locrocis medinalis, Ostrinia nubilalis, Parapedia siateterrella, Moth, Spodoptera litura, beet Spodoptera exigua, Pseudaletia separata, Mamestra brasicae, Agrotis ipsilon, Trichoplusia ni, Autographa nigrisigna, Noctuidae Genus (Heliothis spp), tobacco leaf moth (Helicoverpa spp), E. sinensis (Earias spp), etc., Japanese butterfly (Pieris rapae crucivora) and other pink butterflies, Adoxophyes honmai, tea small roll night Moth (Adoxophyes orana fasciata), Gapholita molesta, Cydia pomonella, etc., such as leaf moths, Carposina niponensis, and other leaf miners such as Lyyonetia clerkella Small leaf moths such as the fine moth, such as Phyllonycter ringoniella, and Phyllocnistis citrella. Plutella xylostella (Plutella xylostella), such as the diamond moth, Euproctis taiwana, Lymantria dispar, Euproctis similis, and other species such as the Scopelodes contracus, 316401 1329001 , such as the red leaf moth, Dendrolimus spectabilis, red bellworm (Pectinophora gossypiella) and other red bollworms, lamp moths, and broad-necked mites. Diptera pests: Liriomyza trifolii, Liriomyza bryoniae, Liriomyza sativae, Liriomyza asterivora, and peas Leafy moths such as Chromatomyia horticola, house mosquitoes, cockroaches, leaf spotted mosquitoes, chironomids, house flies, ink flies, flies, flower flies, jade ropes, real Ropes, fruit ropes, butterfly flies, rainbow: mosquitoes, mites, thorns and so on. Coleoptera pests: leaf worms, chafers, worms, leafy worms, ladybugs, beetles, pseudo-walking worms, etc.蓟马目虫虫: Thrips palmi, such as Thrimas genus, Frankliniella occidentalis, etc., such as Scultothrips dorsalis, etc. Guan Li Ma Wei and so on. Hymenoptera pests: leaf bees, ants, tiger head bees, etc., nettle-like pests: mites, tea-winged mites, etc., Orthoptera pests: locusts, mites, cryptidae pests: fleas Class, 虱目虫虫: human mites, etc., termite worms: Reticulitermes speratus, coptotermes formosanus, etc., monumental insects: spider species (Tetranychus spp.), red spider species 316401 12 1329001 Tomato thorny scorpion (Aculops genus, etc. (Panonychus spp.), such as spider lycopersici), etc., and the use of the composition of the present invention is not particularly limited, and for example, it can be used for supervision. Cabbage, green stalk, radish, broccoli, chicory, Duji, temple, green pepper, pepper, courgette, watermelon, cantaloupe, asparagus,:, fragrant, rice, rye, wheat, barley, oats, millet, (4) Good, corn, kidney bean, field bean, soybean, red bean sweet ^ Ganzi, tea, material, feed crops, edible beet, 1, young beans, citrus, coconut palm, coffee, cotton Fruit, _, frequency fruit, cherry , Pear, peach, plum, persimmon, Fan Stone: okra, ornamental plants, papaya, peanut, pepper, bean tree) = or fruit, a variety of agricultural and horticultural aspects to prevent or biological. . In addition, for example, turf, carnation, cherry, rose, gerbera, chrysanthemum, potted Zhuohua, and Xianren (4) cultivated flowers can also be protected by the present invention. Further, examples of cultivated plants should include all ornamental or flowering plants. In the composition of the present invention, a mixture of the present compound and the present protein f may be used as long as it is a mixing ratio at which a multiplication effect can be obtained, for example, a ratio of 1:10000 to 10000:丨 in terms of a weight ratio, and The ratio of 1〇〇〇 to (7)Μ is better. The ratio of 1: 1 to 5: i is better, and the ratio of ^ 4〇 to 1:1 is better, especially i : 2〇 to i : The ratio is the best. When the composition of the present invention is used, it may be used without mixing only the present compound and the present protein, and is generally used in two t;: a solid carrier, a liquid carrier, a gas carrier, etc. may be further added, and if necessary, 316401 may be added. 13 Interfacial activators, other preparations, granular hydrating agents, water “/丨” can be used as an oil, emulsion, hydrodynamic agent, powder, granule, gas = sputum, water emulsion, etc. The various preparations of micro-heat fumigants, the active ingredients of the hair, the sun, the month, /, the above-mentioned clothes, 95%. The composition of the present invention is usually about 0.01 to soil by weight, and the solid carrier used is exemplified by clay (Gaolin//, Runcheng, Fuabasha, clay, acid clay, etc.), synthesis. ; bismuth, ceramic magnetic class 'Other inorganic minerals (sericulture, stone two: strontium carbon, carbon called hydration oxidation "), chemical fertilizer (sulfur = (four) Syrian, nitrate (four), urea, vaporized ammonium, etc.) Or the granule. The liquid carrier may, for example, be water, alcohol (methanol, ethanol, etc.) 'ketones (propionyl methyl ethyl ketone, etc.), aromatic hydrocarbons (toluene, xylene, ethylbenzene, methyl cyano Etc.), non-aromatic cigarettes (hexilated, cyclohexane, kerosene, light oil, etc.), brewing (ethyl acetate, butyl acetate, etc.), nitriles (acetonitrile, isobutylate, etc.), shouting ( "propyl ether-oxygen, etc.", guanamines (Ν, Ν-dimethylformamide, ν, Ν-di: acetamide), dentate hydrocarbons (dioxane 6, trichloroethane) An alkane, carbon tetrachloride, etc., a scorpion, a vegetable oil (such as soybean oil, cottonseed oil, etc.), etc. Further, the gas carrier may, for example, be a fulong gas, a butane gas or a liquefied gas. a propellant such as diterpene ether or carbon dioxide. The interface activator may, for example, be an alkyl sulfate salt, an alkyl sulfonate, an alkyl aryl sulfonate, an alkyl aryl ether or a polyoxyethylene compound thereof, Polyglycol ethers, polyhydric alcohol esters, sugar alcohol derivatives, etc. 316401 1329001 - Adjuvants for other coating agents can be exemplified by roller, gelatin, _ class (Dian powder, gum arabic-cellulose derivative, Bath acid, etc.), lignin derivatives, bentonite, oral water-cooling molecular compounds (polyvinyl alcohol, polyvinylpyrrolidone, polypropylene m) 'PAP (acidic phosphate different from the purpose), ΒΗτ (2,6_ two Third butyl group (tetra), hydrazine (2_t-butyl-4 methoxy (tetra) and] tert-butyl 4-methoxy phenol), vegetable oils, mineral oils, interfacial activators, fatty acids, fatty acids Ester, etc. The base of the 耳名耳月j can be exemplified by ingredients such as cereal flour, vegetable oil, strontium, crystalline cellulose, etc., and anti-rolling of dibutyl perylene toluene-dichloro-hypoxic acid ... decontamination of acetic acid and other preservation materials, pepper powder and other anti-feeding agents, dry-line spices, onion spices and other attractive spices. A flowable agent such as a turbid agent or an aqueous emulsion, usually in an amount of about 0.5% by weight of the active ingredient compound, is contained in a suspension agent containing about 0.5 to 15 (for example, a protective colloid or a thixotropic substance) It can be obtained from about 0 to 10% by weight of a supplementary agent (for example, an antifoaming agent, an anti-recording agent, a d-permeation aid, an antifreeze agent, a preservative, an antifungal agent, etc.). The various active ingredients can be prepared according to the above-mentioned preparation method:: after: these preparations are mixed and prepared. That is, the composition of the present invention can be prepared by preliminarily formulating the compound according to the form of the agent: The composition obtained by formulating the protein shell is mixed and prepared. Alternatively, the two may be mixed or used at the time of application. The composition of the present invention which has been formulated according to the above operation may be used by dilution with water or the like. In addition, it can also be used with other insecticides, killing: 316401 15 y, nematicides, fungicides, agents, fertilizers, soil amendments, kinetics: plant growth regulators, synergistic inventions Mix or use. It must be born from the protection of pests: the habitat of the pests is an emulsion, a hydrating agent, and:::•, when the composition of the present invention is usually diluted with water or the like. (4) When a hydrating agent, a flowable agent, or the like, the composition of the present invention is used for agricultural and horticultural compositions of the present invention. (1) U has a biological treatment such as Λ, 里, or application amount, usually about 0.1 to 1000 per 1000 m2 of application. ^ When the active ingredient is applied directly to the granules " or the herbicide for removing harmful agents, oils, etc., it is usually applied directly without the > dilution, in accordance with the above effective amount or application amount. Further, when the active ingredient for preventing or eliminating harmful organisms is applied by dilution with an emulsion, a water granule hydrating agent, a flowable agent or the like, it can be used by diluting with water to the above effective amount or application amount. Specifically, when used in the form of a hydrophobic diluent, a hydrating agent, a particulate hydrating agent, a flowable agent or the like, the composition of the present invention is usually applied at a concentration of about 1 to ioooo ppm. The above-mentioned effective amount or application amount varies depending on the type of the preparation, the application period, the application rate, the application method, the type of pest, the degree of damage, and the like, and is not limited to the above range, and can be appropriately increased or decreased. EXAMPLES The present invention will be described in detail by way of formulation examples and test examples. However, the invention is not limited in scope. The meaning of the following "BT culture" is as follows: The live bud belonging to Bacillus thuringiensis bacteria from 16 316401 1329001 runs 1 suspension of 1 Χ 3 spores per lml 1 'Inoculated in a 500 ml flask containing sterilized L-broth liquid medium (1 〇〇mi) at 25. (: 2 to 5 Torr under shaking culture (150 rpm), and then recover the spores belonging to Bacillus thuringiensis bacteria and the insecticidal egg white matter from the obtained culture solution by centrifugation, and dry the two The obtained culture (i.e., one of the forms of the protein). Formulation Example 1 Emulsion _ Dissolve 1 part of the present compound and 10 parts of the BT bacteria culture containing 36. 5 parts of xylene and 36.5 parts In the bis-indolylamine, an additional 1 〇 之 氧化 氧化 氧化 氧化 氧化 氧化 及 及 及 及 及 及 及 及 及 及 及 及 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 。 Mixture, add 5 parts of the compound and 45 parts of BT culture in 4 parts of sodium lauryl sulfate, 2 parts of calcium lignin sulfonate, 22 parts of synthetic cerium oxide containing fine powder and 22 parts In the mixture of diatomaceous earth, mix well and mix to obtain hydrating agent. See Formulation Example 3 granules at the end, 5 parts of BT bacteria culture, and 5 parts of synthetic cesium hydroxide containing micro-powder. Sodium '30 parts of bentonite and 54 parts of bauxite in 1 part of this compound, fully Stirring and mixing, then adding water of appropriate amount of 31640] 17 to the above mixture, and drying by air to obtain granules. Stir well, granulate into granules, preparation example 4 powder, 0.5 parts of the compound is formed into Hydrogen-oxygen cut micro-powder solid culture, 1 part of the mixture) and 7 parts of the clay in the coagulant Dolly 6 (the mixture produced by the Sankyo Company): 86 =: after mixing, by the juice machine Mix and stir the powder. "Clay month, stir well and mix and add 5 parts of this compound to the flowable agent of Formulation 5, 1 shan plucking ^ ^ 10 wound BT letter culture and 1.5 parts 2: (4) sour vinegar In a solution containing 2 parts of polyethylene glycol, the mixture is stirred and mixed with a knife. Then, by sanding the mash, the second is 3 (under), and then the content is added. . = # .1 injured aluminum magnesium citrate 48 parts of the aqueous solution, and then added 15 parts of the alcohol to mix and mix to obtain a flowable agent. Formulation Example 7 Microcapsule 316401 1 _ 6 oil agent 'strives 0.1 part of the compound and 8 parts of BT bacteria The culture was mixed with 891 parts of deodorized lamp oil in 5 parts of this and 5 wounds of the second gas, and the oil was obtained. 1329001 2 parts of the compound, 8 parts of BT bacteria culture, 10 parts of phenyl xylyl ethane and 0.5 parts of miltam beads L-75 (Sumitomo urethane diphenyl phthalate) After mixing, add 20 parts of 10% aqueous gum arabic solution and stir by homogenizer to obtain an emulsion with an average particle size of 20 // m. Then add 2 parts of ethylene glycol and then in a warm bath at 60 ° C. The reaction was carried out for 24 hours to obtain a microcapsule slurry. In addition, 0.2 parts of xanthan gum and 1.0 part of propolis R (aluminum magnesium silicate of Sanyo Chemical Co., Ltd. product) were dispersed in 56.3 parts of ion-exchanged water to obtain a tackifier solution. . 42.5 parts of the above microcapsule slurry and 57.5 parts of a tackifier solution were mixed to obtain a microcapsule. The following examples demonstrate the excellent pest control effect of the composition of the present invention. Moreover, the theoretical value of the predictable insecticidal rate when the two active ingredients are mixed and processed is E=X+Y — (X+Y/100) (the above formula is generally calculated as the calculation formula of Colby, which is calculated It is obtained by the user based on the multiplication effect of the drug combination and the criterion for determining the antagonistic effect. In the formula, X: insecticidal rate (%) when the active ingredient A is treated at Mppm Y: insecticidal rate (%) when the active ingredient B is treated at N ppm E: when the active ingredient A is Mppm, effective Theoretical value (%) of the insecticidal rate predicted when component B was treated at N ppm Test Example 1 For the artificial sample test of Heliothis virescens, 10% of the compound emulsion and BT bacteria preparation: Design (trade name) , a Bacillus thuringiensis var. aizawai strain GC91 conjugant, a wettable granule preparation, a diluted aqueous solution of a Thermo Trilogy product, mixed to prepare 19 316401 1329001 to each predetermined concentration Dilute the test solution. Inject a manual feed (about 2 g) into a plastic cup having a capacity of 30 ml after the bottom of the cup, and treat it with 0.2 ml of the test solution. After about 2 hours of air drying, one of the 2nd instar larvae of Heliothis virescens (20 replicates) was fed. After 5 days from the treatment, the mortality rate was determined, and the insecticidal rate was corrected by the following formula to calculate the insecticidal rate (%). The results are shown in Table 1. Insecticide rate (%) = 100x (Mt - Mc) / (100 - Me)
Mt示供試化合物處理區之死蟲率（％)Mt shows the mortality rate of the test compound treatment area (%)
Me示供試化合物無處理區之死蟲率（％) 表1 供試藥劑 施用濃度 殺蟲率理論值 殺蟲率實測值 有無相乘效果 (ppm) (〇/〇) (%) 判定結果 本化合物 1.56 0.0 — 本蛋白質 15.6 11.1 — 本發明組成物 1.56+15.6 11.1 88.3 有 本化合物 6.25 27.8 — 本蛋白質 62.5 50.0 — 本發明組成物 6.25+62.5 63.9 100.0 有 試驗例2 對斜紋夜蛾（Spodoptera litura)之食葉浸潰試驗 將10%本化合物乳劑和BT菌製劑：夫羅派克（商標 名，蘇雲金芽胞桿菌變種亞莎草（Bacillus thuringiensis var. aizawai)之乾燥可流動性製劑品，住友化學工業公司製品） 之稀釋水溶液混合成為各個預定濃度之稀釋液，以該稀釋 液之1/5000量添加展著劑（特製利諾，曰本農藥公司製品） 而調製成試驗用藥液。將盆植之5至8葉期之甘藍葉片剪 20 316401 1329001 下並浸潰於上述試驗用藥液中60秒，然後於500ml容量之 塑膠杯内舖以直徑90mm之濾紙，上面放置浸潰後風乾之 甘藍葉片。該甘藍葉上飼放供試斜紋夜蛾（Spodoptera litura) 之3齡幼蟲10隻（3重複）。處理6曰後求得死蟲率，該死 蟲率依據下式校正而算出殺蟲率（％)，其結果示於表2。 殺蟲率（％)=100x (Mt—Mc)/(100 —Me)Me shows the mortality rate of the test compound in the untreated area (%). Table 1 Test agent application concentration insecticidal rate theoretical value insecticidal rate measured value with or without multiplicative effect (ppm) (〇 / 〇) (%) Compound 1.56 0.0 - Present protein 15.6 11.1 - Composition of the invention 1.56+15.6 11.1 88.3 Present compound 6.25 27.8 - Present protein 62.5 50.0 - Composition of the invention 6.25+62.5 63.9 100.0 Test Example 2 Spodoptera litura The food leaf impregnation test will be 10% of the compound emulsion and BT bacteria preparation: Froppak (trade name, Bacillus thuringiensis var. aizawai) dry flowable preparation, Sumitomo Chemical Industry Co., Ltd. The dilute aqueous solution of the product is mixed into a predetermined concentration of the diluted solution, and a spreading agent (a special product manufactured by Rinocco, Sakamoto Agrochemical Co., Ltd.) is added in an amount of 1/5000 of the diluted solution to prepare a test chemical solution. The cabbage leaves of the 5 to 8 leaf stage of the potted plants were cut and immersed in the test liquid for 60 seconds, and then coated with a filter paper of 90 mm in diameter in a plastic cup of 500 ml capacity. Cabbage leaves. Ten (3 repeats) 3rd instar larvae of Spodoptera litura were tested on the cabbage leaves. The insecticidal rate was determined after 6 weeks of treatment, and the insecticidal rate was calculated according to the following formula to calculate the insecticidal rate (%). The results are shown in Table 2. Insecticide rate (%) = 100x (Mt - Mc) / (100 - Me)
Mt :供試化合物處理區之死蟲率（％)Mt : mortality rate of the test compound treatment zone (%)
Me :供試化合物無處理區之死蟲率（％) 表2 供試藥劑 施用濃度 殺蟲率理論值 殺蟲率實測值 有無相乘效果 (ppm) (%) (%) 判定結果 本化合物 0.78 30.0 本蛋白質 25.0 20.0 — 本發明組成物 0.78+25.0 44.0 56.7 有 本化合物 1.56 30.0 _ 本蛋白質 25.0 20.0 — 本發明組成物 1.56+25.0 44.0 76.7 有 本化合物 1.56 30.0 _ 本蛋白質 12.5 10.0 — 本發明組成物 1.56+12.5 37.0 90.0 有 本化合物 1.56 30.0 — 本蛋白質 6.25 10.0 _ 本發明組成物 1.56+6.25 37.0 86.0 有 (產業上之應用可行性） 本發明可提供僅單純混合時即獲得相加效果以上之相 乘效果，而可降低所用藥量之用以預防及消除有害生物之 組成物。 21 316401Me: mortality rate of the test compound in the untreated area (%) Table 2 Test agent application concentration insecticidal rate theoretical value insecticidal rate measured value with or without multiplication effect (ppm) (%) (%) judgment result this compound 0.78 30.0 The present protein 25.0 20.0 - The composition of the present invention 0.78 + 25.0 44.0 56.7 The present compound 1.56 30.0 _ The present protein 25.0 20.0 - The composition of the present invention 1.56 + 25.0 44.0 76.7 The present compound 1.56 30.0 _ The present protein 12.5 10.0 - The composition of the present invention 1.56+12.5 37.0 90.0 The present compound is 1.56 30.0 — the present protein is 6.25 10.0 _ The composition of the present invention is 1.56+6.25 37.0 86.0 (industrial application feasibility) The present invention can provide a phase in which the addition effect is obtained only by simple mixing. By multiplying the effect, the amount of the drug used can be reduced to prevent and eliminate the composition of the pest. 21 316401