【作者】 黄衍章；

【导师】 杨长举；

【作者基本信息】 华中农业大学 ， 动物学， 2007， 博士

【摘要】 粮食仓储工作是我国粮食工作的重点，而如何有效地防治储粮害虫又是粮食仓储工作的关键。植物源农药属绿色无公害农药，对害虫不易产生抗药性，符合可持续农业发展的客观要求。玉米象、谷蠹、赤拟谷盗是我国的三种主要储粮害虫，其中玉米象已被国内粮食部门列为储粮头号害虫，谷蠹则为南方储藏谷物的重要害虫，在我国已对磷化氢等化学杀虫剂产生了严重的抗药性。至此，本研究得到了国家十五科技重点攻关项目“植物杀虫剂在储粮害虫防治中的应用与示范”（2004BA523803-2）的有力资助。1石菖蒲根茎提取物有效杀虫成分的分离与鉴定1．1石菖蒲根茎提取物有效杀虫成分的分离采用甲醇冷浸法和有机溶剂萃取法，对石菖蒲新鲜根茎干粉进行了提取和初步分离。结果表明：甲醇对石菖蒲根茎干粉具有较高的提取率，浓缩后提取物得率为19.2％。石油醚萃取物浓缩后得率为23.5％。采用硅胶柱层析法，以碘蒸气为显色剂，对石油醚萃取物有效杀虫成分逐步进行初分和细分，浓缩后得到三种活性组分。选取活性最好的两种组分用硅胶柱层析法进行纯化，得到两种浅黄色油状活性单体。1．2石菖蒲根茎提取物各分离组分对试虫的初步毒效采用药膜法，以玉米象、谷蠹、赤拟谷盗为试虫，研究了石菖蒲根茎提取物各分离组分对试虫的初步毒效，结果表明：石菖蒲根茎提取物四种溶剂萃取物以石油醚萃取层的生物活性最高。药膜法314.40μg/cm²密闭处理72h后，石油醚萃取物对玉米象、谷蠹和赤拟谷盗的校正死亡率均为100.00％。水萃取物也具有一定的毒杀活性，对谷蠹和赤拟谷盗以314.40μg／cm²密闭处理72 h后的校正死亡率分别为88.51％和56.98％。将初分馏分用157.20μg／cm²剂量进行生物测定，结果表明活性馏分为极性较低的化合物。8种洗脱馏分以61～72号洗脱馏分的毒杀活性最高，处理72h后对三种试虫的校正死亡率均高于80.00％，显著高于其余7种洗脱馏分。将细分馏分用157.20μg/cm²剂量进行活性测定，结果表明组分Ⅱ、Ⅳ和Ⅵ对试虫具有较高的毒杀活性，处理72h后对玉米象的校正死亡率依次为81.11％、97.78％、100.00％，对谷蠹的校正死亡率分别为57.78％、97.78％、97.78％。组分Ⅲ、Ⅵ用157.20μg／cm²剂量处理72h后对赤拟谷盗的校正死亡率均为100.00％。采用药膜法对三种优选组分进一步进行初步毒效测定，结果表明组分Ⅲ、Ⅵ对试虫具有较好的毒杀效果。组分Ⅲ对试虫的毒杀效果比较缓慢，其毒效与处理时间关系密切。组分Ⅲ对谷蠹和赤拟谷盗具有较好的生物活性，用39．30μg／cm²剂量处理72h后的校正死亡率分别为64.44％和90.00％。组分Ⅵ对玉米象和谷蠹具有较高的毒效，19.65μg／cm²剂量处理72h后的校正死亡率分别为54.44％和87.78％。1.3石菖蒲根茎提取优选组分有效成分初步分析采用气相色谱-质谱联用法（GC-MS），对石菖蒲根茎提取三种优选活性组分的有效成分进行了初步分析。在组分Ⅱ中，化合物1～2的分子量均约为205，且互为同分异构体。化合物3～5的分子量均约为220，也互为同分异构体。初步推断组分Ⅱ主要由5种倍半萜类化合物组成。在组分Ⅲ中，化合物7为主要活性成分，分子量为218。在组分Ⅵ中，初步推断化合物8为榄香素（1，2，3-trimethoxy-5-（2-propenyl）benzene），化合物9为β-细辛醚（cis-1，2，4-trimethoxy-5-（1-propenyl）benzene），化合物9为组分Ⅵ的主要活性成分。1．4石菖蒲根茎提取活性单体结构鉴定1．4．1化合物7结构鉴定质谱鉴定结果表明：单体7为愈创烷型倍半萜，分子量为218，可能含有CH₃-，＞CHCH₂-，＞C=O片段。紫外吸收光谱鉴定结果表明：单体7中含有共轭双键（-C=C-C=C-）或类似键结构。红外吸收光谱鉴定结果表明，单体7中含有＞C=O和不饱和的共轭酰基碳结构（-C=C-C=O）。¹HNMR鉴定结果表明单体7中含有2个CH₃-CH＜片段，2个烯甲基（CH₃-C=C），2个烯丙基结构（-C=C-CH₂-C=C-），且存在-CH₂-CO-片段，不含-CH=CH-结构。¹³CNMR鉴定结果显示单体7中含有1个＞C=O，2个＞C=C＜。综合分析各光谱特征，确定单体7为一种已知的倍半萜类化合物菖蒲烯酮（3，8-二甲基-5-（1-甲基乙叉基）-1，2，3，4，5，6，7，8-八氢奠-6-酮）。1．4．2化合物9结构鉴定质谱鉴定结果表明，单体9与β-asarone有很大的匹配度，含有CH₃-和CH₃CH=片段，分子量为208。紫外吸收光谱鉴定结果表明，单体9与标准品β-asarone均含有1个双键与苯环共轭。红外吸收光谱鉴定结果表明，单体9含有-CH=CH-，CH₃O-及苯环。¹HNMR鉴定结果显示单体9含有1个CH₃-CH=结构，3个CH₃O-，CH₃CH=CH-基团，-CH=CH-C=基团且苯环上共有2个H。¹³CNMR鉴定结果表明单体9苯环上有3个H被甲氧基所取代，且含有CH₃-与-CH=CH-相连。综合分析各光谱特征，确定化合物9为一种已知的苯丙素类化合物β-细辛醚（1，2，4-三甲氧基-5-（1-Z-丙烯基）苯）。2石菖蒲根茎浸膏石油醚萃取物，β-细辛醚和菖蒲烯酮对试虫的生物活性2．1石菖蒲根茎浸膏石油醚萃取物对试虫的生物活性药膜法测定结果表明：石油醚萃取物对谷蠹和玉米象具有较高的触杀活性。用39.30μg／cm²剂量处理72h后对谷蠹的校正死亡率为88.37％。石油醚萃取物对谷蠹、玉米象、赤拟谷盗的LD₅₀（72h）依次为8.85、40.04、64.88μg／cm²。三角瓶药纸熏蒸试验结果表明：石油醚萃取物对谷蠹具有较高的熏蒸活性，随着处理时间的延长，对谷蠹的熏杀效果显著加强，25μL／L与100μL／L熏蒸处理120h后对谷蠹的校正死亡率无显著差异。石油醚萃取物对玉米象具有一定的熏蒸活性，100μL／L熏蒸处理120h后对玉米象的校正死亡率为83.30％。滤纸药膜选择法测试结果表明：石油醚萃取物对赤拟谷盗具有很高的驱避效应，各剂量处理72h三次调查的平均驱避率均为Ⅴ级水平。石油醚萃取物对玉米象和谷蠹具有一定的驱避活性，用157.20μg／cm²剂量处理72h的平均驱避率均为Ⅳ级水平。粮食拌药法试验结果表明：石油醚萃取物对谷蠹和玉米象具有较好的种群抑制活性和防治效果，对谷蠹兼具较高的毒杀作用。用250mg／Kg剂量处理3d后对谷蠹的毒杀率为97.73％，显著高于5mg／Kg马拉硫磷处理，对小麦的防效达100.00％，与5mg／Kg马拉硫磷处理相当；125mg／Kg剂量处理对谷蠹子代种群的抑制率为96.47％，与5mg／Kg马拉硫磷处理没有显著差异。2．2石菖蒲根茎提取β-细辛醚对试虫的生物活性β-细辛醚对玉米象和谷蠹具有较高的触杀活性，药膜法处理72h后的LD₅₀分别为51.29μg／cm²和30.27μg／cm²。β-细辛醚对谷蠹具有很高的熏蒸活性，25μL／L熏蒸处理120h后的校正死亡率为97.93％，与100μL／L处理无显著差异。β-细辛醚对谷蠹的熏杀效果比较缓慢，延长熏蒸时间可显著提高熏杀效果。β-细辛醚对玉米象具有较高的熏蒸活性，25μL／L熏蒸处理120h后的校正死亡率为89．81％。β-细辛醚对赤拟谷盗具有很高的驱避活性，各剂量处理72h三次调查的平均驱避率均≥Ⅳ级水平。β-细辛醚对玉米象和谷蠹具有较好的毒杀活性、种群抑制作用和防治效果。200mg／Kg剂量处理3d后对谷蠹的毒杀率为99.00％，显著高于5mg／Kg马拉硫磷处理，100mg／Kg处理对谷蠹子代种群的抑制率为85.25％，对小麦的防效可达90.35％，与5mg／Kg马拉硫磷处理没有显著差异。250mg／Kg剂量处理后对玉米象子代种群的抑制率和小麦的防效分别为98.82％和88.08％，与5mg／Kg马拉硫磷处理效果相当。2．3石菖蒲根茎提取菖蒲烯酮对试虫的生物活性菖蒲烯酮对玉米象和谷蠹具有一定的触杀活性。药膜法处理72h时的LD₅₀分别为61.84μg／cm²和71.34μg／cm²。菖蒲烯酮对玉米象和谷蠹具有较好的熏蒸活性，166.67μL／浓度处理120h后的校正死亡率分别为72.52％和86.43％。菖蒲烯酮对谷蠹的熏蒸效果比较缓慢，随着熏蒸时间的延长，对谷蠹的熏杀作用显著加强。菖蒲烯酮对赤拟谷盗具有很高的驱避效应，19.65μg／cm²剂量处理72h三次调查的平均驱避率为Ⅴ级水平。菖蒲烯酮对谷蠹具有较好的毒杀活性、种群抑制作用和防治效果。用500mg／Kg剂量处理3d后对谷蠹的毒杀率为94.47％，与10mg／Kg马拉硫磷处理无显著差异。250mg／Kg剂量处理后对谷蠹子代种群的抑制率为98.70％，与10mg／Kg马拉硫磷处理效果相当，125mg／Kg剂量处理后对小麦的防效为100.00％，与10mg／Kg马拉硫磷处理无显著差异。2．4β-细辛醚和菖蒲烯酮对试虫的联合毒力药膜法测试结果表明：β-细辛醚和菖蒲烯酮混用对玉米象和谷蠹成虫具有一定的增效作用。随着β-细辛醚含量的增加，对谷蠹的增效作用逐步增强。当β-细辛醚和菖蒲烯酮按4：1（W／W）混用时，对谷蠹的共毒系数为127.53（＞120），表现为显著的增效作用。β-细辛醚和菖蒲烯酮按1：1（W／W）混用时，对玉米象表现为显著的拮抗作用，其共毒系数为69.42（＜80），当β-细辛醚和菖蒲烯酮按2：1（W／W）混用时，其共毒系数为142.33，对玉米象具有显著的增效作用。3β-细辛醚对玉米象和谷蠹的杀虫机理3．1对玉米象成虫的杀虫机理用β-细辛醚LD₅₀（485.00mg／Kg）剂量拌粮处理试虫后，对乙酰胆碱酯酶、谷胱甘肽S-转移酶的活性整体表现为抑制作用，并且在处理24h时的酶活力最低。随着处理时间的延长，β-细辛醚对羧酸酯酶的诱导作用有所加强。β-细辛醚不同处理时间对试虫的酯酶同工酶具有一定的影响，处理12h时酯酶E4、E5的表达量有所降低，36h后酯酶E1、E2、E3的表达量则有所增加。用不同剂量的β-细辛醚处理试虫，结果表明低剂量（≤250.00mg／Kg）的β-细辛醚处理对乙酰胆碱酯酶具有诱导作用，高剂量（≥500.00 mg／Kg）处理对乙酰胆碱酯酶则表现为抑制作用。β-细辛醚对谷胱甘肽S-转移酶具有抑制作用，但抑制效果与处理剂量关系并不密切。随着β-细辛醚处理剂量的增高，羧酸酯酶的活力显著升高。提高β-细辛醚的处理剂量对酯酶E5的抑制作用有所增强。3．2β-细辛醚对谷蠹成虫的杀虫机理用β-细辛醚LD₅₀（94.49mg／Kg）剂量处理试虫，12h时乙酰胆碱酯酶活性被诱导，24h后主要表现为抑制作用。β-细辛醚对谷胱甘肽S-转移酶的时间效应并不明显，对酯酶同工酶具有一定的诱导作用，诱导效果与处理时间有一定的关系，处理12h后酯酶E4被明显诱导。低剂量（67.5mg／Kg）的β-细辛醚对乙酰胆碱酯酶具有显著的诱导作用，随着处理剂量的升高，对乙酰胆碱酯酶的活力多数表现为抑制作用。低剂量（≤100.0mg／Kg）的β-细辛醚对谷胱甘肽S-转移酶具有诱导作用，而高剂量（≥133.3mg／Kg）的β-细辛醚对谷胱甘肽S-转移酶具有抑制作用。β-细辛醚对羧酸酯酶的活性多数表现为诱导作用，提高β-细辛醚的处理剂量可提高羧酸酯酶的活力。不同剂量的β-细辛醚处理对谷蠹酯酶同工酶均具有显著的诱导作用，但诱导效果与处理剂量关系并不密切。更多还原

【Abstract】 The storage work was the emphasis of foodstuff management in China, and how to effectively control the stored product pest was the key task for food storage. The botanical pesticide belongs to the bio-rational pesticide, and it is hard for pests to develop resistance to the botanical pesticide. As a result, the botanical pesticide answered for the impersonal demand of sustainable agriculture. The maize weevil Sitophilus zeamais Motsch., grain borer Rhizopertha dominica （Fab.） and red flur beetle Tribolium castaneum （Herbst） were the three key stored product pests in China, of which the maize weevil had been named as the first key stored product pest by national foodstuff department, and the grain borer was the primary pest of stored grain in south China. At present, the grain borer had developed serious resistance to some chemical pesticides, such as phosphine, etc. Thus, this research was effectively financed by the Chinese 15th key task project through the science and technology ministry of state （Application and Demonstration of Botanical Insecticide in Controlling Stored Product Pest, serial number: 2004BA523B03-2）.1 Isolation and identification of effective insecticidal ingredients in Acorus gramineus Soland rhizome extract1.1 Isolation of effective insecticidal ingredients in A. gramineus rhizome extractThe dry powder of A. gramineus rhizome was firstly soaked using methanol in cool condition, and then the extract was pretreated by extraction method using organic solvent correspondingly. The results indicated that the methanol had better extraction effect to dry powder of A. gramineus rhizome as 19.2% extract yield after concentration, and the yield of petroleum ether fraction was up to 23.5% after concentration.Adopting the silica gel Column Chromatography （CC）, the effective insecticidal ingredients of petroleum ether extract were separated as I₂ vapor acted as display reagent step by step. Three active ingredients were obtained after concentration, and two ingredients with better activity were selected for purification in silica gel CC. At last, two active monomers were obtained, and the monomers were kind of yellowish oily liquid.1.2 Primary insecticidal effects of ingredients isolated from A. gramineus rhizome extractThe primary toxicities of insecticidal ingredients isolated from A. gramineus rhizome extract to S. zeamais, R. dominica and T. castaneum adults were tested using drug-film method. The results were listed as follows: As for four solvents extracts of A. gramineus rhizome extract, the petroleum ether extract had highest bioactivities to tested pests. At 314.40μg/cm² dosage, the corrected mortalities of petroleum ether extract against three tested pests were 100.00% at 72h post-treatment. Furthermore, the water extract had some toxicities to R. dominica and T. castaneum adults with 88.51% and 56.98% corrected mortalities accordingly at 72h post-treatment.The results of bioassay for primary fractions at 157.20μg/cm² dosage indicated the active ingredients belonged to the compound with low polarity.The C （number 61～72） fraction of 8 eluted fractions had highest toxicities against three tested pests as the corrected mortalities at 72 h post-treatment were more than 80.00%, these mortalities were significantly higher than that of other 7 eluted fractions. The bioassay results for sub-separated fractions at 157.20μg/cm² dosage indicated ingredientsⅡ,ⅢandⅥhad better toxicities as the corrected mortalities to S. zeamais at 72 h post-treatment were 81.11%, 97.78% and 100.00% correspondingly, and the corrected mortalities to R. dominica at 72 h post-treatment were 57.78%, 97.78% and 97.78% correspondingly. At 157.20μg/cm² dosage, the corrected mortalities of ingredientⅢandⅥto T. castaneum at 72 h post-treatment were 100.00%.Further primary bioassay for three selected ingredients demonstrated the ingredientⅢandⅥhad better insecticidal effects against pests. The insecticidal effect of ingredientⅢwas slow relatively, and the toxicity effect had close relationship with exposure time. IngredientⅢhad better bioactivities to R. dominica and T. castaneum adults with 64.44% and 90.00% corrected mortalities （72 h） accordingly at 39.30μg/cm² dosage, and ingredientⅥhad better bioactivities to S. zeamais and R. dominica with 54.44% and 87.78% corrected mortalities （72 h） accordingly at 19.65μg/cm² dosage.1.3 Primary analysis of effective insecticidal components for selected ingredients isolated from A. gramineus rhizomeActive components for three selected ingredients isolated from A. gramineus rhizome were primarily analyzed using Gas Chromatography-Mass Spectrometry （GC-MS）.In ingredientⅡ, the molecular weight of compound 1～2 were 205, which were isomeric compounds. The molecular weight of compound 3～5 were 220, and were isomeric compounds as well. It was primarily concluded that ingredientⅡwas composed of 5 sesquiterpenoids. As for ingredientⅢ, compound 7 was the leading active component with 218 molecular weight. In ingredientⅥ, it was primarily concluded that compound 8 was elemine [1,2,3-trimethoxy-5-（2-propenyl） benzene], and compound 9 wasβ-asarone [cis-1,2,4-trimethoxy-5-（1-propenyl） benzene] acted as a leading active component in ingredientⅥ.1.4 Structure identification of active monomer isolated from A. gramineus rhizome1.4.1 Structure identification for compound 7Mass Spectrum （MS） data indicated the monomer 7 was a kind of guaiane-type sesquiterpene, and its molecular weight was 218. The monomer might contain methyl （CH₃^-）, secondary carbon linked with tertiary carbon （＞CHCH₂^-） and ketonic bond （-C=O）. Ultraviolet Spectrum （UV） showed monomer 7 contained conjugated double bond （-C=C-C=C-） or similar bond. Infrared Spectrum （IR） indicated the monomer 7 contained ketonic bond and conjugated unsaturated carbonyl unit （-C=C-C=O）. ¹H Nuclear Magnetic Resonance （¹HNMR） spectrum indicated monomer 7 contained two methyl linked with tertiary carbon （CH₃-CH＜）, two methyl linked with double bond （CH₃^-C=C）, a doubly allylic methylene group appearing as a tylical AB doublet （-C=C-CH₂-C=C-）, and a secondary carbon linked with ketonic bond （-CH₂-CO-）, without hydrogen linked with unsaturated carbon （-CH=CH-）. ¹³C Nuclear Magnetic Resonance (¹³CNMR) spectrum reconfirmed monomer 7 had one ketonic bond, and two double bonds without hydrogen （＞C=C＜）.Integrated with above spectrum characters, this monomer could be confirmed as a known sequiterpenoid, calamusenone named as 3,8-dimethyl-5-（1-methylethylidene）-1,2,3,4,5,6,7,8-octahydro-6-azulenone.1.4.2 Structure identification for compound 9MS data indicated the monomer 9 has much matching degree withβ-asarone, and its molecular weight was 208. UV spectrum showed monomer 9 and standard sampleβ-asarone contained a double bond conjugated with benzene. IR spectrum indicated the monomer 9 contained hydrogen linked with unsaturated carbon （-CH=CH-）, methoxyl （CH₃O-） and benzenoid form. ¹HNMR indicated monomer 9 contained one methyl linked with unsaturated carbon （CH₃CH=）, three methoxyl groups, CH₃CH=CH-group,-CH=CH-C= group and two hydrogen located in benzenoid form. ¹³CNMR reconfirmed monomer 9 had three methoxyl groups linked with benzenoid form as well as one methyl linked with unsaturated carbon. Integrated with above spectrum characters, monomer 9 could be confirmed as a known phenylpropanoid named as cis-1,2,4-trimethoxy-5-（1-propenyl） benzene.2 Bioactivities of petroleum ether extract,β-asarone and calamusenone isolated from A. gramineus rhizome extract to tested pests2.1 Bioactivities of petroleum ether extract isolated from A. gramineus rhizome extract to tested pestsThe bioassay results of drug-film method showed petroleum ether extract had better contact activities against R. dominica and S. zeamais adults. The corrected mortality of petroleum ether extract at 39.30μg/cm² to R. dominica was 88.37% at 72 h post-treatment. LD₅₀ （72 h） of petroleum ether extract to R. dominica, S. zeamais and T. castaneum were 8.85, 40.04, 64.88μg/cm² correspondingly. Drug-paper fumigant experiment showed the petroleum ether extract had better fumigant activities against R. dominica, and the effect increased markedly with increasing of exposure time. After 120 h treatment, the corrected mortality to R. dominica at 25μL/L concentration hadn’t significant （P＞0.05） difference to 100μL/L treatment. Petroleum ether extract had moderate fumigant activities against S. zeamais, and the corrected mortality at 100μL/L was 83.30% after 120 h treatment. The petroleum ether extract had excellent repellency action on T. castaneum by selection test on filter paper held with drug. As for three dosages, all the mean repellecy rates based on 24, 48, 72 h were V grade level. Petroleum ether extract had moderate repellency effect on R. dominica and 5. zeamais, and both of the mean repellency rates wereⅣlevel at 157.20μg/cm² dosage.The mixture experiment of foodstuff and drug showed that petroleum ether extract had better population inhibition activity and control effect against R. dominica and S. zeamais, and better poisonous effect against R. dominica as well. At 250 mg/Kg dosage, the mortality of this extract to R. dominica was 97.73% at 3 d post-treatment, which was markedly higher than that of malathion treatment at 5 mg/Kg dosage. At 250 mg/Kg dosage, the control effect of petroleum ether extract against R. dominica was up to 100.00%, which was similar to the malathion treatment at 5 mg/Kg dosage. When mixed with wheat at 125 mg/Kg dosage, the population inhibition rate of petroleum ether extract to R. dominica was 96.47%, and there hadn’t markedly difference with malathion treatment at 5 mg/Kg dosage. 2.2 Bioactivities ofβ-asarone isolated from A. gramineus rhizome extract to tested pestsβ-asarone had better contact activities against S. zeamais and R. dominica adults, and the LD₅₀ （72 h） were 51.29μg/cm² and 30.27μg/cm² respectively.β-asarone had excellent fumigant activities against R. dominica. At 120 h post-treatment, the corrected mortality to R. dominica at 25μL/L concentration was 97.93%, which hadn’t significant （P＞0.05） difference to 100μL/L treatment. The fumigant effect ofβ-asarone was relatively slow, which increased markedly with prolonging of exposure time.β-asarone had better fumigant activities against S. zeamais, and the corrected mortality at 25μL/L concentration was 89.81% after 120 h treatment.β-asarone had excellent repellency action on T. castaneum, all the mean repellecy rates based on 24 h, 48 h and 72 h were no less thanⅣgrade level.Theβ-asarone had better poisonous activity, population inhibition action and control effect against R. dominica and S. zeamais. At 200 mg/Kg dosage, the mortality ofβ-asarone to R. dominica was 99.00% at 3 d post-treatment, which was markedly higher than that of malathion treatment at 5 mg/Kg dosage. At 100 mg/Kg dosage, the population inhibition rate and control effect ofβ-asarone against R. dominica was 85.25% and 90.35% accordingly, which was similar to the malathion treatment at 5 mg/Kg dosage. When mixed with wheat at 250 mg/Kg dosage, the population inhibition rate and control effect ofβ-asarone against 5. zeamais was 98.82% and 88.08% accordingly, which hadn’t significant （P＞0.05） difference to malathion treatment at 5 mg/Kg dosage.2.3 Bioactivities of calamusenone isolated from A. gramineus rhizome extract to tested pestsCalamusenone had moderate contact activities against S. zeamais and R. dominica adults, and the LD₅₀ （72 h） were 61.84μg/cm² and 71.34μg/cm² respectively. Calamusenone had better fumigant activities against S. zeamais and R. dominica adults. At 120 h post-treatment, the corrected mortalities to S. zeamais and R. dominica at 166.67μL/L concentration were 72.52% and 86.43% correspondingly. The fumigant effect of calamusenone against R. dominica increased markedly with prolonging of exposure time. Furthermore, calamusenone had excellent repellency action on T. castaneum, the mean repellecy rate during 72 h at 19.65μg/cm² dosage wasⅤgrade level.The calamusenone had better poisonous activity, population inhibition action and control effect against R. dominica. At 500 mg/Kg dosage, the mortality of calamusenone to R. dominica was 94.47% at 3 d post-treatment. The population inhibition rate （250 mg/Kg） and control effect （125 mg/Kg） of calamusenone against R. dominica were 98.70% and 100.00% accordingly, which hadn’t significant （P＞0.05） difference to the malathion treatment at 10 mg/Kg dosage.2.4 Combined toxicities ofβ-asarone and calamusenone to tested pestsTest of drug-film method showed mixted application forβ-asarone and calamusenone had moderate synergism to tested pests, and the synergism was gradually enhanced with increasing ofβ-asarone content. When the ratio （W/W） ofβ-asarone and calamusenone was 4:1, the co-toxicity coefficient （CTC） was 127.53 （＞120）, it denoted there had marked synergism to R. dominica. As for S. zeamais, there had marked antagonism （CTC=69.42＜80） when the ratio （W/W） ofβ-asarone and calamusenone was 1:1, whereas marked synergism （CTC=142.33） was given when the ratio （W/W） was 2:1.3 Insecticidal mechanism ofβ-asarone to S. zeamais and R. dominica adults3.1 Insecticidal mechanism ofβ-asarone to S. zeamais adultWhen S. zeamais was treated withβ-asarone at LD₅₀ （485.00 mg/Kg） by mixture test for foodstuff and drug, the activities of acetylcholinesterase （AChE） and glutathione S-transferase （GST） were restrained at most times, and their activities were at lowest level at 24 h post-treatment. With the prolonging of exposure time, the induced action ofβ-asarone on Carboxylesterases （CarE） intro-pest was strengthened accordingly. The exposure time ofβ-asarone had some effects on the activity of est isozyme. At 12 h post-treatment, expression of isozyme E4 and E5 were restrained in some extent, whereas expression of isozyme E1, E2 and E3 were enhanced somewhat at 36 h post-treatment.After treated in five dosages, theβ-asarone at low dosage （=250.00 mg/Kg） showed induced action to AChE of intro-pest, whereas theβ-asarone at high dosage （=500.00 mg/Kg） gave inhibiting action to AChE.β-asarone had some inhibiting action to GST, but the inhibiting effect hadn’t close relationship with dosage. With the enhancing of dosage forβ-asarone, the activity of CarE was significantly promoted. On the contrary, activity of Est E5 was somewhat restrained as the dosage was enhanced.3.2 Insecticidal mechanism ofβ-asarone to R. dominica adult When treated withβ-asarone at LD₅₀ （94.49 mg/Kg）, the activity of AChE to R. dominica was induced at 12 h, yet it was mostly restrained at （after） 24 h post-treatment. The exposure time ofβ-asarone to R. dominica hadn’t distinct effect on GST. Whereas, the inducement ofβ-asarone on est isozyme was somewhat related with exposure time, and the isozyme E4 was induced distinctly at 12 h post-treatment.Theβ-asarone at low dosage （67.5 mg/Kg） showed marked induced action to AChE of intro-pest. With the increasing of dosage, the activity ofβ-asarone on AChE intro-pest was mostly restrained.β-asarone showed induced action to GST at low dosage （=100.0 mg/Kg）, but inhibiting action to GST at high dosage （=133.3 mg/Kg）. With the enhancing of dosage forβ-asarone, the activity of CarE was somewhat promoted.β-asarone had induced action to Est isozyme intro-pest at different dosages, whereas the induced effect hadn’t close relationship with treated dosage.更多还原