【作者】 江志利；

【导师】 张兴；

【作者基本信息】 西北农林科技大学 ， 农药学， 2013， 博士

【摘要】 植物精油是一类存在于植物不同组织中的重要次生代谢物质,为易挥发的具有强烈香味的油状液体。其在医学、保健、保鲜等方面具有极其重要的作用和功能。研究表明,植物精油具有杀虫、杀菌、抗病毒等多种生物活性,并对多种农药具增效作用。由于其具有挥发性,因而尤适于家居环境卫生害虫的控制。尽管许多学者在植物精油的杀虫活性研究方面取得了不少可喜的成绩,但鲜见在生产实践中应用。基于此,本文在前期研究基础上,对所筛选出的高活性植物精油,主要从精油活性成分的互作、制剂及精油对拟除虫菊酯类农药的增效作用等方面进行了较为系统的研究。结果如下：1.以白纹伊蚊(Aedes albopictus)为试虫,对37种植物精油的驱避活性和熏杀活性进行筛选,发现：(1)黄樟精油、柠檬桉油、木姜子油、五倍橙油、山苍子油、玳玳花油等6种精油2-6h的驱避效果优于或相当于隆力奇花露水,其中木姜子油和山苍子油效果更为突出,该2种精油4-6h的趋避率均在50%以上,显著高于隆力奇花露水的效果(<30%),值得进一步研究；(2)供试植物精油对白纹伊蚊成虫均有一定的熏杀活性,但不同品种对白纹伊蚊的毒力存在明显差异。6h熏杀LC50在5μL·L-1以内的植物精油有3种,分别为冬青油、柠檬草油和肉豆蔻油,其熏杀LC5o分别为2.2365μL·L-1、3.8554μL-L-1和4.8797μL-L-1,表现出较强的熏蒸活性；6h熏杀LC5o在5-10μL·L-1以内的植物精油有10种,表现出较好的熏蒸活性；(3)茶树油、肉豆蔻油、缬草油、丁香罗勒油等4种植物精油对白纹伊蚊表现出一定的引诱作用,有必要进一步研究。2.通过溶剂和乳化剂的系统筛选,研制出符合商品农药要求的10%天然冬青油乳油和10%柠檬草油乳油制剂配方：(1)10%天然冬青油乳油配方：天然冬青油10%、乳化剂02045%,95%乙醇定容至100%；(2)10%柠檬草油乳油制剂配方：柠檬草油10%,专用乳化剂N1：0204(6：4)的乳化剂组合5%,乙酸乙酯：95%乙醇(4：6)的混合溶剂定容至100%。活性测试表明,10%天然冬青油乳油和10%柠檬草油乳油对白纹伊蚊的熏杀LC5o分别为0.4951μL·L-1和0.2384μL-L-1；在LC95剂量下,10%冬青油乳油和10%柠檬草油乳油的LTso分别为13.85min和10.06min。该2种精油乳油制剂环保,杀虫效果高,值得进一步产业化开发。3.以代替蚊香市场中已被禁止使用的增效剂S2为目标,在对增效剂所用溶剂筛选的基础上,对植物精油增效剂配方进行了筛选。结果表明：乙酸丁酯、甲酸乙酯、碳酸二乙酯等4种溶剂在脱臭煤油中溶解性较好,可作为配制植物精油增效剂的溶剂；通过对黄樟油、八角叶油、天然冬青油和柠檬草油及其混用组合增效作用的系统测试,筛选出11种对烯丙菊酯具增效作用的植物精油增效剂,该11种增效剂基础配方为40.0%-80.0%植物精油+13.5%～53.5%溶剂+6.5%乳化剂；以淡色库蚊为试虫,采用密闭圆筒法测试分别添加11种增效剂蚊香的杀蚊活性,结果表明11种蚊香样品的KTso均小于2min,达到国家A级标准,其中14#、16#、25#、26#、28#等5个配方样品对蚊虫的SR值分别为1.33、1.34、1.29、1.36和1.30,均高于或相当于添加对照增效剂S2蚊香的SR值1.28,具有良好的增效效果。4.以市售“榄菊”牌气雾剂药液配方为基础,采用5.8m3小屋法及28m3模拟现场药效试验法进行基于植物精油的气雾剂增效剂研究。从供试的6种植物精油筛选出冬青油和p-蒎烯表现出较好的增效活性,添加二者的气雾剂样品对淡色库蚊的KT50分别比添加化学增效剂S-1的样品缩短了2.3927min和1.3819min;以冬青油为主要增效成分,研制出S-101、S-102和S-103等3个增效剂配方；5.8m3小屋法活性测试发现,S-102的增效效果最好,其样品KTso比对照(化学增效剂S1)缩短4.1875min,增效系数达1.38。S-103和S-101也表现出较好的增效效果,增效系数分别为1.23和1.16；基于28m3模拟现场的测试结果,据农药卫生用杀虫剂药效评价标准,气雾剂B和气雾剂C达到A级标准,气雾剂A则达到B级标准。5.采用GC-MS对木姜子油和山苍子油的成分进行了分析检测,并以粉纹夜蛾(Trichoplusia ni)三龄幼虫为试虫,探讨了该两种植物精油及其各组分的杀虫活性。结果表明：(1)从木姜子油中共检出11种成分,占全部精油的92.7%。其中,1,8-桉叶素(1,8-Cineole)和香芹酮(Carvone)为其主要成分,分别占总精油的52.4%和10.3%；(2)从山苍子油中共检出6种成分,占全部精油的89.9%。其中,丫-松油烯(Gamma-Terpinene)和柠檬烯(Limonene)为山苍子油的主要成分,分别占总精油的43.6%和27.2%；(3)以粉纹夜蛾三龄幼虫为试虫的毒力测试表明,木姜子油的触杀致死中量(LD50)为87.1μg/头,山苍子油的LDso为112.4μg/头；(4)精油成分间的相互作用研究表明,1,8-桉叶素和香芹酮为木姜子油的主效成分；γ-松油烯和柠檬烯为山苍子油的主效成分,对伞花烃也是其活性成分之一：两种植物精油中的其它单体成分或其混合物对粉纹夜蛾三龄幼虫均无明显的触杀活性,但这些单体成分或其混合物对精油的活性成分均有明显的增效作用。这些结果表明,植物精油中无论是主效成分,还是非活性成分,均是植物精油表现杀虫活性不可或缺的组成。6.在研究确定木姜子油超临界CO2萃取工艺基础上,测试、比较了水蒸气蒸馏法和超临界CO2萃取技术对木姜子植物挥发油的提取效果,以及两种方法提取挥发油的杀虫活性。结果表明：(1)在上样量为1g时,木姜子油超临界C02萃取的较佳工艺为：采用先静态、后动态的萃取方式。静态萃取压力4000psi,时间10min；动态萃取压力4000psi,萃取温度55℃,C02体积为40mL,不使用夹带剂。在此条件下,计算挥发油最佳得率为7.85%；(2)超临界CO2萃取木姜子挥发油的得油率明显高于水蒸气蒸馏提取法；(3)以白纹伊蚊成虫为试虫的活性测定表明,木姜子水蒸气提取物熏杀效果明显优于超临界C02萃取物,二者熏杀致死中浓(LC50)分别为68.35μL/L和135.94μL/L。植物精油可谓一复杂、深奥,又充满希望的研究领域。王年前的多部古书中都记有“驱虫辟邪”,一语道破了其神奇的功效。本研究结果进一步证实了其在害虫防治,特别是在家庭卫生害虫防控中的潜能和实用价值,对该研究领域的深入探讨将会在人类疾病控制、改善生活质量方面做出大的贡献。更多还原

【Abstract】 Plant essential oils, volatile and highly fragrant liquids, are important secondary metabolites present in different tissues of the plant. Essential oils are used in the field of medicine, healthcare, and fruit industry (keeping the fruit fresh). Several studies have shown that these oils possess different kinds of bioactivities (pesticidal, funficidal and antiviral activities and may own synergistic effects. Because of the strong volatile nature, these oils may be suitable for controlling house and sanitary insect pests. Based on that, this study was aimed at investigating the interaction of constituents of essential oil and the synergistic effects on pyrethroids and making preparations. The results are as follows:1. Using Aedes albopictus as the target insect,37plant essential oils were screened for the repellent action and fumigation action.(1) sassafras oil, citriodora oil, Litsea cubeba oil, Citrus aurantium oil, L., Five times of orange oil and dai-dai flower oil showed comparable strong repellent effects than Longlife floral water after2-4h treatment, among which citrus aurantium oil and litsea cubeba oil showed50%repellent ratio after4-6h treatment worthy further study;(2) Though essential oils all showed fumigation action to adult aedes albopictus, different cultivars had different effects. Wintergreen oil, lemon grass oil and myristica oil, had LC50level lower than5μL·L-1after6h treatment and had LC50level of2.2μL·L-1,3.8μL·L-1and4.9μL·L-1, respectively. Ten oils showed some fumigation activity with less than5-10μL/L-1LC50level.(3) Tea tree oil, myristica oil, valerian oil and Ocimum gratissimum oil showed unexpectedly attractant effect and need to be considered for future research.2. For the screening of solvent and emulsifier,10EC wintergreen oil and10EC lemon grass oil were selected fitting the commodity commands:(1) recipe of10EC wintergreen oil: wintergreen oil10%, emulsifier02045%and95%ethanol for the rest;(2)10EC lemon grass oil recipe:lemon grass oil10%, exclusive used emulsifier N1:0204(6:4)5%and ethyl acetate:95%ethanol (4:6) for the rest. Fumigation active tests showed that10EC natural wintergreen oil and10EC lemongrass oil’s LC50of fumigation active against Aedes albopictus were0.4951μL·L-1and0.2384μL·L-1respectively; under the dose of LC95, LT50of10EC natural wintergreen oil and10EC lemongrass oil were13.85min and10.06min respectively. The two kinds of essential oils emulsifiable solution (EC) were environment-friendly, and had high insecticidal effects, needed further industrialization development.3. In terms of mosquito coils synergistic agent, S2, had been banned from the market. Plant essential oil solvents were screened on the basis of mosquito-repellent incense synergist as synergist formula. The results show that four kinds of solvent, butyl acetate, ethyl formate and diethyl carbonate etc., had good solubility in the deodorization of kerosene oil and could be used as a solvent in the preparation of plant essential oil synergist. To study the synergistic effect of single compound and mixtures using sassafras oil, anise leaf oil, natural wintergreen oil and lemon grass oil, we screened11kinds of plant essential oil synergist effects for allyl chrysanthemum ester, the11kinds of synergistic agent recipe were40.0%～80.0%plant essential oil+13.5%～53.5%solvent+6.5%emulsifier OP-10. Sealed cylinder method was used to test11kinds of synergist mosquito-repellent incense for fumigation activity against Culex pipiens. The results showed that KT50of11kinds of mosquito-repellent incense samples were less than2min, reaching the national A-level standard; The SRs of five mosquito-repellent incenses (14#、16#、25#、26#、28#) were1.33,1.34,1.29,1.36and1.30respectively, showing good synergistic effects comparing with control synergist S2’s SR (1.28).4. Based on the liquid formula of the commercially available "lanju" aerosol, we studied the plant essential oil of aerosol synergist by using5.8m3cottage method and the28m3simulated field efficacy method. The wintergreen oil and β-pinene screened from the selected six plant essential oil showed good synergistic activity, KT50of the aerosol by mixing these two compounds was reduced by2.3927min and1.3819min respectively compared with the sample by adding chemical synergistic agent; using wintergreen oil as the main synergistic ingredient and YW, Y001, Y002, and YN as surfactant or penetrating agent, we developed three synergistic agent formulas, S-101, S-102and S-103. Activity test through5.8m3cottage method showed that S-102had the best synergistic effect reducing the level of KT50(chemical synergistic agent S-1) by4.1875min with SR1.38. S-103and S-101also showed strong synergistic actions with SRs1.23and1.16respectively. According to the test results of28m3simulated field method and the sanitary insecticide efficacy evaluation standard, aerosol B and C reached A-level standard, and aerosol A reached B-level standard.5. GC-MS analysis of Listsea pungent and L. cubeba oils showed that:(1)11compounds of L. pungent fruit oil comprised92.7%of the total oil, with1,8-cineole (43.6%) and limonene (27.2%) being the main constituents;(2) Six components from L. cubeba oil comprised89.9%of the total with gamma-Terpinene (43.6%) and limonene (27.2%) being the main constituents;(3) Bioassay showed that the LD50values of L. pungent fruit oil and L. cubeba oil against third instar larvae of cabbage loopers were87.1μ.g/larva and112.4μg/larva respectively;(4) Though the single compounds or the mixture of the main constituents of L. pungent fruit oil, L. cubeba oil,1,8-cineole, limonene, gamma-Terpinene and limonene, didn’t show any contact activity, their synergistic effects were obvious. Our results showed that all constituents whether active or inactive, were required for the total activity of the oil.6. Comparing the different extraction methods (steam distillation method versus supercritical CO2extraction) using Qinling Mountains aromatic plant, L. pungent fruit oil as material, we got the following results:(1) The efficiency of supercritical CO2extraction was far more superior than steam distillation method;(2) After1g was loaded, the best craft for supercritical CO2extraction was:static extraction for10min, holding pressure at4000psi at55℃and40mL CO2. The production under this optimal condition was7.85%;(3) Pesticidal assay against housefly showed reversed results for these two methods and the LC50of steam distillation method and supercritical CO2extraction were170.54μL/L and566.79μL/L respectively.Studying bioactivity of essential oils is a very promising field of research, although it is also complicated, and esoteric. Many ancient books on essential oils claim that essential oils not only repel the insects but the evil too, pointing towards the magic function of essential oils. The results of our study further confirm their potential as botanical insecticides against pests, especially the hygienic insects.更多还原