【作者】 程彬；

【导师】 高长启；

【作者基本信息】 东北林业大学 ， 森林保护， 2010， 博士

【摘要】 本文首先通过顶空取样法对来源于寄主植物的信息化学物质进行了分析鉴定,包括健康完整的1年生榆苗、受不同虫口密度的榆紫叶甲成虫危害的榆苗和受不同虫口密度的榆紫叶甲幼虫危害的榆苗的挥发物进行分析,同野外寄主和非寄主的挥发物、家榆韧皮部萃取液的成分进行比较,并分析了挥发物的释放节律、影响挥发物释放的主要因素等。然后通过萃取法提取来源于榆紫叶甲虫体的信息化学物质,包括榆紫叶甲各龄幼虫体表、成虫和幼虫粪便以及取食前后雌雄二性榆紫叶甲成虫后肠的化学物质的鉴定,结合来源于植物的化学信息物质,分析二者的联系和交流。最后通过室内外生物测定,筛选出对榆紫叶甲有引诱作用的信息化学物质。受虫害诱导的榆苗的挥发物相比健康完整的榆苗不仅种类增加,而且释放量也增加。10头榆紫叶甲成虫危害后挥发物增加了benzyl benzoate、tmtt、(+)-longifolene、benzoic acid, butyl ester、methyl salicylate和2-heptanol 6种组分,40头榆紫叶甲成虫危害后挥发物增加了anisole、cis-3-hexenyl acetate、1-undecene、undecanal、methyl salicylate、linalool、caryophyllene、naphthalene,1,2-dimethyl-和tmtt 10种组分；20头榆紫叶甲幼虫危害后挥发物增加了2,4,6-tris(1,1-dimethylethyl) phenol、tmtt、cis-nerolidol、(+)-longifolene、caryophyllene、naphthalene, 1,2,3-trimethyl-4-propenyl-, (e)-、benzoic acid butyl ester、butanedioic acid dibutyl ester、methyl salicylate、linalool、2-heptanol和benzaldehyde 12种组分,40头榆紫叶甲幼虫危害后挥发物增加了anisole、benzaldehyde、cis-3-hexenyl acetate、1-undecene、methyl salicylate、linalool、undecanal、butylated hydroxytoluene、tmtt、heptadecane、和benzyl benzoate11种组分。虫害诱导之后的挥发物有一些是立即释放,如：绿叶气味cix-β-hexenyl acetate在40头成虫和40头幼虫危害后1h内就立即释放。而多数的挥发物组分则是在危害后8-9h的释放量达到最大,少部分是在危害后第三天释放量达到最大。野外家榆主要挥发物的释放基本遵循上午释放量逐渐增加,到中午时达到最大,在下午逐渐减少；在日夜交替的晚19：00-20：00这一时间段有一个爆发的规律。相比室内寄主挥发物的组成,野外寄主未检测到2-heptanol、anisole、benzaldehyde、β-caryophyllene、cis-3-hexenyl acetate和cis-β-ocimene这几种物质。韧皮部的成分增加了pentanal-2-methy、cis-2-peten-1-ol、hexanal、1,3-dimethyl benzene、methyl-pentylketo、myrcene、γ-terpinene、naphthalene, decahydro-2,6-dimethyl-、naphthalene, decahydro-2,3-dimethyl-、α-humulene、globulo、octadecane、nonadecane、n-eicosane和n-heneicosane 15种物质,其余13种物质和家榆的挥发物一致。但家榆挥发物中的β-caryophyllene、cis-3-hexenyl acetate、linalool、methyl salicylate、1-tridecene、cis-nerolidol、tmtt等是家榆韧皮部中没有的。从榆紫叶甲4个龄期幼虫的表皮中共检测到benzyl butyl ether、cis-2-peten-l-ol、n-octane、hexanal、benzene,1,2-dimethyl-、myrcene、benzeneacetaldehyde、phenylacetate、methyl benzoate、naphthalene, decahydro-2,6-dimethyl-、naphthalene, decahydro-2,3-dimethyl-、1-methyl naphthalene、tetradecane、hexadecen-1-ol, trans-9-、heptadecane、1,2-benzenedicarboxylic acid, diundecyl ester、nonadecane、tetracontane, 3,5,24-trimethyl-、dodecane,2,6,10-trimethyl-、-2-tetradecen-1-ol、1,2-benzenedicarboxylic acid, diundecyl ester、citronellylacetone、tetradecane,2,6,10-trimethyl-、dotriacontane、1-hexadecanol,2-methyl-、13-methylheptacosane 26种物质,其中1龄幼虫表皮的物质的量较少,4龄幼虫表皮一些化合物的量较高,与其他3龄相比有显著差异。榆紫叶甲后肠化学物质在取食前后雌雄二性都发生了变化,雌性在取食之后增加了cis-2-peten-l-ol、cyclohexane,1,4-dimethyl-, cis-、(-)-β-pinene、limonene、naphthalene、naphthalene, decahydro-1,5-dimethyl-、tetradecane,2,6,10-trimethyl-、1,2-benzenedicarboxylic acid, butyl 8-methylnonyl ester、dotriacontane、vitamin E、13-methylheptacosane 11种化合物,雄性取食之后增加了n-octane、styrene、α-thujene、camphene、3-carene、benzene, 1-ethynyl-4-methyl-、naphthalene、acenaphthylene、9,10-ethanoanthracene,9,10-dihydro-11,12-diacetyl-、vitamin E、dotriacontane 11种化合物。其中一些物质是寄主植物所具有的,如：cis-2-peten-1-ol、(-)-β-pinene、limonene、dotriacontane、tetradecane,2,6,10-trimethyl。榆紫叶甲成虫的粪便相比幼虫的粪便来说,含有更多的化合物。粪便中的limonene、cis-β-ocimene、α-pinene、phenyl acetate、linalool、methyl salicylate、α-farnesene、δ-cadinene、tmtt、nerolidol、n-hexadecane、heptadecane也是寄主植物所具有的；而benzene,1,2-dimethyl-、α-pinene、limonene、1-heptatriacotanol、dotriacontane、vitamin E是榆紫叶甲后肠的化合物。本文选取了13种化合物进行室内外生物测定。y形管的行为测定表明榆紫叶甲更喜欢受害24h之内的榆树挥发物,而在受害后第二天的挥发物和健康完整榆苗的挥发物比较时,则没有显著差异。室内触角电位、触角电位浓度实验和y形管实验都表明雌性榆紫叶甲对linalool、α-farnesene、dibutyl phthalate、indole和cis-3-hexen-1-ol的反应超过了其他几种物质,雄性榆紫叶甲对γ-terpinene、α-farnesene和β-caryophyllene的反应较强。雌性对0.1、10和100μg浓度的linalool的反应显著高于雄性。室外生物测定的结果也基本同室内结果一致,但多种因素影响使诱捕到的数量较少。混合化合物作为榆紫叶甲的引诱剂在野外还需要进一步进行研究。更多还原

【Abstract】 In this paper, infochemicals from the plants were first analyzed and identified by the headspace sampling method. Volatiles from a year-old elm seedlings including intact and induced by adult and larvae Ambrostoma quadriimpressum of different population was identified. Comparing with volatiles from host and non-host in the field, and extract from elm’s phloem, the release rhythm of volatile and the main factors of affecting the release of volatile were analysis. And then infochemicals from A. quadriimpressum by extraction, including the cuticular of larvae, the hindgut pre-and post feed and the feces produced by adults and larvaes. Combined with compounds from plants, chemical communication and link between the two were analysis. At last the bioassay in the lab and field was finished, so as to choosing suitable chemicals that lured adult A. quadriimpressum.Herbivore-induced volatiles increased both quantitative and qualitative compared to the undamaged intact elm, the volatiles induced by 10 adult A. quadriimpressum increased 6 chemicals which were benzyl benzoate, tmtt, (+)-longifolene, benzoic acid, butyl ester, methyl salicylate and 2-heptanol. The volatiles induced by 40 adults A. quadriimpressum increased 10 chemicals which were anisole, cis-3-hexenyl acetate,1-undecene, undecanal, methyl salicylate, linalool, caryophyllene, naphthalene,1,2-dimethyl-and tmtt. The volatiles induced by 20 larvaes A. quadriimpressum increased 12 chemicals which were 2,4,6-tris(1,1-dimethylethyl) phenol, tmtt, cis-Nerolidol, (+)-longifolene, caryophyllene, naphthalene, 1,2,3-trimethyl-4-propenyl-, (E)-, benzoic acid butyl ester, butanedioic acid dibutyl ester, methyl salicylate, linalool,2-heptanol and benzaldehyde. The volatiles induced by 40 larvaes A. quadriimpressum increased 11 chemicals which were anisole, benzaldehyde, cis-3-hexenyl acetate,1-undecene, methyl salicylate, linalool, undecanal, butylated hydroxytoluene, tmtt, heptadecane and benzyl benzoate. Some herbivore-induced volatile components were released immediately, such as green leaf odor, cis-3-hexenyl acetate released within 1h after 40 adults and 40 larvae induced. Release quantity reached the max when elm was damaged after 8-9h for most volatiles, whereas minority was reached the max when elm was damaged in the third day. Release of volatile for Ulmus pumila in the field followed the regulations that release quantity increased gradually in the morning, and reach the maximum at noon, and gradually reduced in the afternoon, there is an outbreak in day and night that is the time 19:00-20:00. Compared with volatiles in the lab, volatiles in the field increased 2-heptanol, anisole, benzaldehyde, (3-caryophyllene, cis-3-hexenyl acetate and cis-β-ocimene. Chemicals in the phloem inceased pentanal-2-methy, cis-2-peten-l-ol, hexanal 1,3-dimethyl benzene, methyl-pentylketo, myrcene, y-terpinene, naphthalene, decahydro-2,6-dimethyl-, naphthalene, decahydro-2,3- dimethyl-,α-humulene, globule, octadecane, nonadecane, n-eicosane and n-heneicosane, butβ-caryophyllene, cis-3-hexenyl acetate, linalool, methyl salicylate,1-tridecene, cis-nerolidol and tmtt from volatiles were not found in the phloem. There were 13 chemicals were in accordance with volatiles from elm.There are 26 chemicals from the cuticular of larvae A. quadriimpressum which were benzyl butyl ether, cis-2-peten-l-ol, n-octane, hexanal, benzene,1,2-dimethyl-, myrcene, benzeneacetaldehyde, phenyl acetate, methyl benzoate, naphthalene, decahydro-2,6-dimethyl-, naphthalene, decahydro-2,3-dimethyl-,1-methyl naphthalene, tetradecane, hexadecen-1-ol, trans-9-, heptadecane,1,2-benzenedicarboxylic acid, diundecyl ester, nonadecane, tetracontane, 3,5,24-trimethyl-, dodecane,2,6,10-trimethyl-, E-2-tetradecen-l-ol,1,2-benzenedicarboxylic acid, diundecyl ester, citronellylacetone, tetradecane,2,6,10-trimethyl-, dotriacontane,1-hexadecanol,2-methyl-,13-methyl heptacosane. Quality and quantity of infochemicals from 1st instar larvae is low, meanwhile quality and quantity of infochemicals from 4th instar larvae was the highest among each star, and had significant difference. The chemicals from the hindgut of female and male A. quadriimpressum changed pre-and post feed. For females cis-2-peten-1-ol, cyclohexane,1,4-dimethyl-, cis-, (-)-β-pinene, limonene, naphthalene, naphthalene, decahydro-1,5-dimethyl-, tetradecane,2,6,10-trimethyl-,1,2-benzenedicarboxylic acid, butyl 8-methylnonyl ester, dotriacontane, vitamin E,13-methylheptacosane were increased after feed, meanwhile for males, n-octane, styrene, a-thujene, camphene,3-carene, benzene, 1-ethynyl-4-methyl-, naphthalene, acenaphthylene,9,10-ethanoanthracene,9,10-dihydro-11,12-diacetyl-, vitamin E and dotriacontane were increased after feed. Some of these compounds were according with volatiles from elm, such as cis-2-peten-l-ol, (-)-β-pinene, limonene, dotriacontane and tetradecane,2,6,10-trimethyl. The compounds in the feces produced by adults were much than by larvaes,12 compounds which were limonene, cis-β-ocimene,α-pinene, phenyl acetate, linalool, methyl salicylate,α-farnesene,δ-cadinene, tmtt, nerolidol, n-hexadecane, heptadecane were according with volatiles from elm, moreover benzene,1,2-dimethyl-,α-pinene, limonene,1-heptatriacotanol, dotriacontane, vitamin E also were founding the hindgut of A. quadriimpressum.In this paper,13 compounds were chosen to conduct the bioassay in the lab and field. Experiments using the Y-tube olfactometer indicated that beetle-damaged elm seedlings were more attractive to additional beetles during the first 24 h of damage as compared with intact elm seedlings. Volatiles from feeding-damaged trees play important roles in attracting A. quadriimpressum. Electrophysiological and behavioral responses of male and female beetles to 13 semiochemical compounds from elm were investigated to determine their potential ecological functions prior to more detailed field studies. Electroantennograms (EAGs), EAG dose-response curves and Y-tube olfactometer bioassays using synthetic compounds revealed that females had a stronger response to linalool, a-farnesene, dibutyl phthalate, indole and cis-3-hexen-l-ol than to other compounds; males had a stronger response toγ-terpinene, a-farnesene and P-caryophyllene than to other compounds. EAG responses to linalool at doses of 0.1,10 and 100μg were significantly different between females and males. The results in the field were generally consistent with the results in the lab. But the quantity of A. quadriimpressum in the trap was less due to many factors. These factors could provide important information when synthetic attractants are evaluated as baits for controlling adult A. quadriimpressum.更多还原