摘要
八角尺蠖(Dilophodes elegans sinica Prout)是危害八角树的主要食叶害虫,大发生时造成不可估量的经济损失,严重阻碍了八角产业的发展。为实现对该害虫的有效管理与利用,进一步提高八角产业的附加值,作者对八角尺蠖幼虫粪便及其寄主植物提取物的生物活性进行了初步研究。结果如下:
1、采用固相微萃取技术和气相色谱-质谱联用(GC-MS)技术检测出八角尺蠖幼虫虫粪挥发物有6种主要成分,分别为反式茴香醚(83.24%)、α-合金欢烯(2.95%)、2,6-二甲基-6-(4-甲基-3-戊烯基)-庚二烯(4.20%)、十一酸甲酯(0.71%)、十九酸乙酯(1.37%)、11-十八酸甲酯(2.40%),占总峰面积的94.87%。
2、八角尺蠖雌、雄成虫对幼虫粪便挥发物具有强烈的EAG反应。其中,雌、雄成虫对反式茴香醚的EAG反应较强,对十九酸乙酯的EAG反应最弱;反式茴香醚表现出性别差异,其余成分未表现出性别差异。剂量反应曲线表明,在测定的浓度范围内,雌、雄成虫对十一酸甲酯、反式茴香醚、11-十八酸甲酯等3种化合物的EAG反应与浓度的变化呈正相关;对a-合金欢烯的EAG反应呈“U”型;对十九酸乙酯的触角电位反应值在0.01(v/v)浓度下达到最大。雌、雄成虫对不同成分混合物的EAG反应表明,对于雌虫,各成分对反式茴香醚没有增效作用;对于雄虫,a-合金欢烯和十一酸甲酯对反式茴香醚存在增效作用。
3、幼虫虫粪不同溶剂提取物对不同菌种的抑制作用不同,其中丙酮提取物对细菌(如金黄色葡萄球菌、大肠杆菌)的抑菌作用较强,蒸馏水提取物对真菌(如枯青霉、根霉菌)的抑菌作用较强。
4、幼虫虫粪不同溶剂的萃取物对三种自由基均有一定的清除能力。其中,幼虫虫粪萃取物对DPPH自由基的清除能力较小,明显小于同浓度下八角叶片萃取物的清除能力;氯仿相和乙酸乙酯相对·OH自由基的清除能力较强,石油醚相、正丁醇相和水相对·OH自由基的清除能力均小于八角叶片萃取物的清除能力;幼虫虫粪萃取物对O2-自由基的清除能力均小于相同条件下八角叶片萃取物的清除能力。
5、八角叶的乙醇、丙酮、乙酸乙酯和石油醚4种溶剂提取物对榕管蓟马成虫均有较强的驱避作用和熏蒸作用,其中丙酮提取物的驱避作用和熏蒸作用最强,石油醚最弱。在丙酮提取物的各种萃取物中,石油醚萃取物的熏蒸作用最强。石油醚萃取层主要包括烯烃类、醇类、酸类、酮类等21种化学成分,其中,含量最多的成分为大茴香醛、反式茴香脑。
6、八角叶片乙醇和丙酮两种溶剂提取物对榕母管蓟马成虫体内3种保护酶(CAT、POD和SOD)活性均表现为处理初期有促进作用,而在处理后期有抑制作用。两种提取物处理榕母管蓟马2h以后,成虫体内酯酶活性显著升高。丙酮提取物处理6h后,试虫酯酶活性迅速下降到低于对照的水平;而乙醇提取物处理的试虫酯酶活性在6h后继续上升,在较长时间内维持较高的水平。两种提取物处理6h以前对GSTs的活力起促进作用,而处理6h以后则对GSTs的活力起抑制作用。
Dilophodes elegans sinica Prout was the main defoliator in Illicium verum. The development of Illicium verum industry was seriously hindered, and the major economic was immeasurably lost when this insect attack severely occurring. In order to realize the effective management and make use of the pests, and improve value-added of Illicium verum industry, the author preliminary studied on the biological activity of Dilophodes elegans sinica Prout frass and its host plant extracts. The results were summarized as follows:
1. The main volatile matters of Dilophodes elegans sinica Prout frass were detected by means of Solid Phase microextraction and Gas Chromatography-Mass Spectrometry (GC-MS), they were anisole p-propenyl(83.24%),α-farnesene, (2.95%), bicycle[3.1.1]hept-2-ene,2,6-dimethyl-6-(4-methyl-3-pentenyl)(4.20%), undecanoic acid, methyl ester (0.71%), nonadecanoic acid, ethyl ester(1.37%),11-octadecenoic acid methyl ester(2.40%) respectively. The six main volatile matters occupied 94.87% of the total in the peak area.
2. Dilophodes elegans sinica Prout adult showed the strong EAG responses to the volatile matters of Dilophodes elegans sinica Prout frass. Among the five compounds tested, EAG responses of Dilophodes elegans sinica Prout adult to Anisole, p-propenyl were stronger than that of other compounds tested, but the weakest to nonadecanoic acid, ethyl ester, all of the compounds no representation sex difference except anisole, p-propenyl. Dosage response curves of EAG of Dilophodes elegans sinica Prout adult showed that the male and the female adult showed positive correlation EAG response to concentration's varying of undecanoic acid, methyl ester and anisole, p-propenyl and 11-octadecenoic acid, methyl ester, showed "U" model EAG response to the a-farnesene, showed the maximum value of EAG response to nonadecanoic acid, ethyl ester in 0.01 (v/v). The EAG responses of female and male adults to different compositions mixture showed that there were no synergy effects between anisole, p-propenyl and other compositions to female. But the males showed synergy effects between anisole, p-propenyl and a-farnesene, undecanoic acid, methyl ester.
3. The bacteriostasis effect of different extracts of larvae frass to different strains existed some difference. Acetone extracts had a stronger effect to bacterium which including S. aureus and E. coli, and distilled water extracts had a stronger effect to fungus which including P. curinum and R. stolonifer.
4. All of the different extracts from the larvae frass in this paper had scavenging activity to the free radicals including DPPH free radical,-OH free radical,and O2- free radical. Concretely speaking, the extracts from the larvae frass had a weak scavenging activity to DPPH free radical, which was significantly weaker than the extract from Illicium verum leaves under the same concentration. The extracts of chloroform and ethyl acetate had a much stronger scavenging activity to the·OH free radical, while the scavenging activity of extracts of Petroleum ether, butanol and distilled water to·OH free radical was weaker than that of extracts from Illicium verum leaves. The scavenging activity from larvae frass to the O2- free radical was weaker than that of extracts from I. verum leaves under the same conditions.
5. The ethanol, acetone, ethyl acetate and petroleum ether, etc. four extracts all had a strong repellent and fumigate effect, and the repellent and fumigate effect of the acetone extract was strongest while that of petroleum ether extract was weakest among them. In various extracts of acetone extract, the petroleum ether extracts had the best fumigate action, which included 21 kinds of chemical composition of olefine, alcohols, acids, ketones and so on, and the most content were p-anisaldehyde and anethole among all kinds of chemical composition.
6. The ethanol and acetone extracts of I, verum leaves showed a stimulative effect to three protective enzymes (CAI、POD and SOD) of adults of G. ficorum in the early treatment and a inhibition effect in the late treatment. The carboxylesterase activity in adults of G. ficorum significantly increased when the two solvent extracts treated on G. ficorum after two hours. The CAT activity of the treated G. ficorum rapidly decreased down to lower than untreated group when the acetone extract treated after six hours. On the contrary, the CAT activity kept to rise during a long time to maintain a high level when the ethanol extract treated after six hours. The two solvent extracts above showed a stimulative effect to GSTs six hours before treatment and showed a inhibition effect six hours after treatment.
引文
[1]陈长琨.昆虫生理生化实验.北京:农业出版社.1993.61-68
[2]陈尚文.马尾松毛虫过氧化氢酶及过氧化物酶与耐药性的关系.昆虫学报,2001,44(1):9-14
[3]陈俊,韦立秀,韦娜,等.肉桂、八角提取物抑菌活性研究.林业科技开发,2008,22(2):38-39
[4]程红,严善春,徐波,等.青杨脊虎天牛触角主要感器的超微结构及其分布.昆虫知识,2008,45(2);223-232
[5]丁保福.光肩星天牛取食寄主后虫粪挥发性半挥发性物质的比较.内蒙古农业大大学硕士学位论文.2009
[6]杜芝兰.中华蜜蜂工蜂触角感受器的扫描电镜观察.昆虫学报,1989,32:166-169
[7]高泽正.中华微刺盲蝽触角环境扫描电镜观察.华南农业大学学报,2006,27(4):18-20
[8]郭线茹,原国辉,蒋金,等.植物挥发性次生物质对昆虫触角电位反应的影响.河南农业大学学报,2003,37(1):18-22
[9]郭莹.山植提取物抗氧化活性的研究.天津科技大学硕士学位论文.2009
[10]郭志永.东亚飞蝗粪便挥发物活性成分鉴定.中国农业大学硕士学位论文.2004
[11]韩丽,符健.秋茄提取物的体外抗菌活性研究.海南大学学报,2009,27(1):30-33
[12]何冬梅,刘红星,黄初升,等.广西八角茴油的提取及其成分研究.食品研究与开发,2009,30(6):3-6
[13]何荣,中丽.超简述八角尺蠖的发生和防治.农家之友,2008,(8):35-36
[14]胡裕进,杨伟秋,杨长举,等.水菖蒲超临界CO2萃取物对德国小蠊的触杀和驱避活性.昆虫知识,2009,46(3):437-441
[15]金鑫,张善干,张龙.东亚飞蝗四种类型的触角感受器超微结构(昆虫纲:直翅目).农业生物技术学报,2004,12(3):300-305
[16]赖荣泉,赖成连,钟秀金,等.大蒜乙醇抽提物对烟蚜忌避及斜纹夜蛾拒食作用.福建农业学报,2009,24(1):85-88
[17]李明,杨振德,张增强.八角尺蠖对八角叶及性信息素腺体浸提液的触角电位反应.基因组学与应用生物学,2010a,29(3):490-494
[18]李明.八角尺蠖与寄主植物气味互作研究.广西大学硕十学位论文.2010b
[19]李水清,张钟宁.松墨天牛成虫对幼虫虫粪挥发性物质的触角电位反应及林间驱避试验.昆虫 学报,2008,51(3):284-289
[20]李周直,沈惠娟,蒋巧根,等.几种昆虫体内保护酶系统活力的研究.昆虫学报,1994,37(4):399-403
[21]凌冰,向亚林,董易之,等.葫芦素B对美洲斑潜蝇成虫产卵和取食的抑制作用研究.天然产物研究与开发,2006,18:29-32
[22]刘惠霞,董育新,吴文君.苦皮藤素V对东方粘虫中肠细胞及其消化酶活性的影响.昆学报,1998,41(3):258-262
[23]刘明霞,陈小丹,王春龙.我国植物源杀虫剂研究进展.辽宁农业科技,2004,2:29-30
[24]刘源,周光宏,王锡昌,等.顶空固相微萃取气质联用检测八角挥发性风味成分.中国调味品,2007,(1):67-69
[25]刘昭明,黄翠姬,田玉红,等.八角挥发油成分分析与抑菌活性研究.中国调料品,2009,34(10):52-55
[26]卢传兵,薛明,刘雨晴,等.黄荆精油对玉米象的杀虫活性成分、毒力及作用机制.昆虫学报,2009,52(2):159-167
[27]鲁玉茶,张孝羲.棉铃虫雄虫对人工合成性信息素的触角电位反应.河南农业大学学报,2004,38(1):49-53
[28]马瑞燕.杜家纬.昆虫的触角感器.昆虫知识,2000,37(3):179-183
[29]马晓乾,魏霞,周琦,等.2种趋避剂对樟子松球果象甲的趋避作用.林业科技,2008,33(5):28-29
[30]马志卿,张兴.植物源杀虫物质的作用特点.植物保护,2000,26(2):37-39
[31]孟琳琳,王进忠,张志勇,等.蛇床子素对小菜蛾幼虫的胃毒作用及对虫体内两种酶活性的影响.北京农学院学报,2010,25(3):24-27
[32]慕立义.植物化学保护研究方法.北京:中国农业出版社.1994.55-56
[33]那杰,于维熙,李玉萍,等.昆虫触角感器的种类及其生理生态学意义.沈阳师范大学学报,2008,26(2):213-216
[34]宁玲.小叶榕榕管蓟马的发生及防治.热带农业科技,2005,28(4):39-40
[35]乔海霞,甄攀,李秀娟.吴茱萸提提取物体外抗菌作用初步研究.河北北方学院学报,2009,26(1):27-29
[36]钦俊德.昆虫与植物的关系—论昆虫与植物的相互作用及演化.北京:科学出版社.1987.38-61
[37]孙红霞,占志雄,吴珍泉,等.5种植物精油对小菜蛾幼虫的触杀作用及两种酶活性的影响.福建农业学报,2008,23(3):274-276
[38]王有年,李青,李照会.篇蓄提取物对朱砂叶螨的触杀活性及相关酶活性的影响.林业科学,2010,46(2):103-107
[39]王占娣,葛长荣,徐志强,等.五种芳香类挥发物质对酪蝇的引诱效果.昆虫知识,2009,46(3):423-426
[40]吴小锋,徐俊良,崔为正,等.家蚕血液过氧化氢酶活力及其与蚕体抗逆性的关系.昆虫学报,1998,41(2):124-129
[41]贤振华,孙晋,邹啸文.紫苏提取物对黄曲条跳甲成虫的拒食活性研究.广东农业科学,2009(8):126-128
[42]徐海燕.源于棉铃虫幼虫粪便的产卵信息素的研究.南京农业大学硕士学位论文.2006
[43]徐汉虹,赵善欢,周俊,等.八角茴香精油的杀虫活性与化学成分研究.植物保护学报,1996,23(4):338-342
[44]许鹏翔,贾卫民,毕良武,等.芳香植物精油气相色谱分析进展.分析科学学报,2004,20(3):312-316.
[45]阎凤鸣.化学生态学.北京:科学出版社,2003.85-87
[46]于立梅,赵谋明,崔春.山竹壳提取物不同溶剂相的体外抗氧化作用与成分分析.天然产物研究与开发,2007,19:1-5
[47]杨彬.植物精油熏蒸杀虫活性研究.华中农业大学硕士学位论文.2008
[48]杨玲.蓝靛果提取物抗氧化及抗癌作用的研究.东北林业大学硕士学位论文.2009
[49]杨元,张风娟,等.桑枝和桑天牛虫粪挥发物对桑天牛长尾啮小蜂的引诱作用.蚕业科学,2007,33(3):360-366
[50]杨振德,常明山,孙艳娟,等.八角尺蠖生物学特性研究.中国森林病虫,2008,27(5):7-817
[51]杨振德.柳蓝叶甲和分月扇舟蛾的生物学特性及其对苦豆草生物碱的行为反应.南京林业大学博士学位论文.2004
[52]张国洲,吴恭谦.植物性杀虫剂研究进展.安徽农业大学学报,2002,29(1):25-28
[53]张红兵,李小鹰,戴华国,等.台湾乳白蚁和黄胸散白蚁对三种饵剂的嗅觉行为反应.昆虫知识,2005,42(3):298-301
[54]张红英,赤国彤,张金林.昆虫解毒酶系与抗药性研究进展.河北农业大学学报,2005,25(5):193-195
[55]张军,宋敦伦,陈建新.家蚕感染蛹虫草后的生理生化变化.昆虫学报,2003,46(6):674-678
[56]张丽霞, 管志斌,管艳红,等.榕母管蓟马危害榕树盆景.植物保护,2004,30(1):89-90
[57]赵国富,陈伟洋.榕母管蓟马生物学特性及防治措施.广西农业科学,2008,39(4):493-496
[58]赵新成,阎云花,王睿,等.昆虫神经生物学研究技术:触角电位图记录.昆虫知识,2004,41(3):270-274
[59]甄昭世,廖超林.检测掺假香叶油技术方法的探讨.香料香精化妆品,2001,(1):9
[60]钟国华,胡美英,徐伟松,等.闹羊花素-Ⅲ对菜粉蝶幼虫血淋巴和中肠酷酶的影响.华中农业大学学报,2001,20(1):15-19
[61]Anderson P, Hilker M, Hanssen BS, et al. Sehildknecht H.Oviposition deterring components in larval frass of Spodoptera littoralis (Lepidoptera:Noctuidae):behavioural and electrophy siolosical evaluation. Insect Physical,1993,39:129-137
[62]Anne B L. Benefits and costs to pollinating, seed-eating insects:the effect of flower size and fruit abortion on larval performance. Oecologia,2009,161:87-98
[63]Barbehenn R V. Digestion of uncrushed leaf tissues by leaf-snipping larval Lepidotera. Department of Entomological Sciences,1992,89:229-235
[64]Birch MC, Tilden PE, Wood DL, et al. Biological activity of compounds isolated from air condensates and frass of the bark beetle. Insect Physiology,1977,23:1373-1376
[65]Chuche J, Xuereb A, Thiery D. Attraction of dibrachys cavus (Hymenoptera:Pteromalidae)to its host frass volatiles. Chem Ecol,2006,32:2721-2731
[66]David K. W, Mcfarlane J E, and Alli I. Aggergation in yellow mealoworms, Tenebrio molitor L. larvae I. individual and group attraction to frass and isolation of an aggregant. Journal of Chemical Ecology,1989,15(5):1605-1615
[67]FettkOther R, Reddy G VP, et al. Effect of host and larval frass volatiles on behavioural response of the old house borer, Hylotrupes bajulas (L.) (Coleoptera:Cerambycidae), in a wind tunnel bioassay. Chemiecology,2000,10:1-10.
[68]Hagus T H, Qi R D. Types of frass produced by the ambrosia beetle Platypus quercivorus during gallery construction, and host suitability of five tree species for the bettle. J For Res,2011, 16:68-75
[69]Hilker M, Klien B. Investigation of oviposition deterrent in larval frass of Spodopter alittoralis (Boisd). Chem. Ecol,1989,15:929-938
[70]Jeremy D.A, John H.B, Steven J.S A review of the chemical ecology of the Cerambycidea. Chenical,2004,14:123-150
[71]Johanna S D. Consequences of mixed species infestation on the searching behavior and parasitism success of a larval parasitoid. Biocontral,2008,53:725-736
[72]Jones A S, Hodges R.J, Hall. D. R et al. Responses of teretrius nigrescens toward the dust and frass of its prey Journal of Chemical Ecology 2004,30(8):1269-1644
[73]Letizia Mattiacci and Marcel Dicke. The parasitoid Cotesia glomerata (Hymenoptera:Braconidae) discriminates between first and fifth larval instars of its host Pieris Brassicae on the basis of contact cues from frass, silk, and herbivore-damaged leaf tissue. Journal of Insect Behavior,1995, 8(4):485-498
[74]Li GQ, Ishikawa. Y.Oviposition deterents in larval frass of four Ostrinia species fed on an artificial diet. Chem Ecol,2004,30:1445-1455.
[75]Li S.Q, Zhang Z.N. Influence of larval frass eatracts on the oviposition behavior of Monochamus alternatus (Col., Cerambycidae) J. Appl. Entomol.,1996,130 (3):177-182
[76]Malik M.M., Muiutaba N.S.H. Screening of some indigenous plants as repellents of antifeedants for stored grain insect. Stored Products Research,1984,20:45-48
[77]Mitchell E.R, heath, R.R. Influence of Amaranthus hybridus L. allelochemics on oviposition behavior of Spodoptera exigue and Spodoptera eridania (Lepidoptera:Noctuidae).J.chem.Ecol. 1985,11:609-618
[78]Poirier L M and Borden J H. Oral exudate as a mediator of behavior in larval eastern and western spruce budworms. Journal of Insect Behavior,1995,8(6):801-811
[79]Sandrine G, Ted C J, et al. Induction of volatile emissions in maize by different larval instars of Spodotera Littoralis. Journal of Chemical Ecology,2003,29(1):145-160
[80]Suiter D R, Carlson D A, Patterson R S, et al. Host location kairomone from Periplaneta Americana (L.) for parasitoid aprostocetus hagenowii (Ratzeburg). Chem Ecol,1996,22:637-651