【作者】 王广利；

【导师】 迟德富； 周志强；

【作者基本信息】 东北林业大学 ， 森林生物工程， 2009， 博士

【摘要】 青杨脊虎天牛(Xylotrechus rusticus L.)是东北地区近年来新爆发成灾的重要蛀干害虫。由于其钻蛀危害的特点给防治带来了极大困难。杨树是东北地区的主栽树种,现今局部地区遭受青杨脊虎天的危害,对当地林业生产造成严重威胁。为探索青杨脊虎天牛的生态控制技术途径,本论文重点研究了青杨脊虎天牛自身及其与植物之间通讯联系所涉及的信息化学物质,分析探索这些信息化学物质的功能,力求找到对青杨脊虎天牛具有生物活性的信息化合物单体或者不同比例的混合物,为该虫的生态控制提供理论依据。本研究采用水蒸气蒸馏(Steam distillation SD)、超声波提取(Ultrasonicextraction)方法分别提取了抗虫水平较低的树种,大青杨(Populus.ussuriensis Kom.)、小青杨(P.pseudo-simonii Kitag.)、小叶杨(P.simonii Carr.)、小青×黑杨(P.pseudo-simoniiKitag.×P.nigra L.)和抗虫水平较高的树种,中东杨(P.berolinensis Dipp.)、小黑杨(P.simonii Carr.×P.nigra L.)、俄罗斯杨(P.russkii Jabl.)、银中杨(P.alba L.×P.berolinensis Dipp.)、白城2号(Populus×xiaozhuanica W.Y Hsu et Liang cv.’Baicheng-2’)和斯大林杨(P.stalintz Jabl.)的树皮挥发物;采用苏玛罐收集了小叶杨、中东杨、银中杨、小青杨和小黑杨5种杨树树干的挥发性气味,利用预浓缩系统与气相色谱-质谱技术(Gas chromatography-mass spectrometry GC-MS)联用分析其挥发物的化学组成。分析结果表明,在天牛成虫非产卵期内,十种杨树树皮挥发物中都含有己醛、邻羟基苯甲醛、十四醛、十六醛、2,6-二叔丁基-4-甲基苯酚、亚油酸、十六酸和11,14,17-二十碳三烯酸甲基酯化合物,但这些化合物存在于不同方法得到的提取液中;超声波提取液中化合物种类比水蒸气蒸馏提取液多。在抗性强的银中杨、中东杨中存在特异性组分,以姜烯、γ-蛇麻烯、别香木兰烯、a-蛇麻烯、d-杜松烯和樟脑为主。在成虫产卵期,采用苏玛罐收集的抗虫水平高的小黑杨、中东杨和银中杨中均含有1-[4-羟基苯甲基]-6-甲氧基,3,4-二氢异喹啉-7-醇,含量在20%以上,其中小黑杨含量最高,达到83.90%。(1R)-a蒎烯在抗虫水平低的小叶杨中含量最高,β-蒎烯和丙烯酸丁酯为抗虫水平低的小青杨中的主要组分。在水蒸气蒸馏液中,(1R)-a蒎烯、桉油精和乙酸乙酯含量在抗虫水平低的杨树挥发物中较高,β-蒎烯、2,4-癸二烯醛、1,2-环己二酮、乙氧基乙烯和吡唑在抗虫水平高的杨树中含量高。杨树树干挥发物的化学组成及其含量在不同抗虫水平树种之间的差异可能是昆虫行为以及树木抗虫性不同的重要因素。应用十种杨树树皮的水蒸气蒸馏液对青杨脊虎天牛雌雄成虫进行电生理和行为学测试,初步确定了小黑杨和小青×黑杨提取物均能引起雌雄两性显著的电生理反应,行为测试结果进一步指出,青杨脊虎天牛雌、雄成虫主要对抗虫性低的小青杨和小叶杨树皮提取液表现出趋向性,而抗虫水平高的白城二号杨和银中杨树皮提取液对其产生负趋向作用。该实验结论与青杨脊虎天牛的实际危害状况相一致。同时,我们对不同浓度的2,6-二叔丁基-4-甲基苯酚、苯酚、樟脑、苯甲酸、间二甲苯和糠醛6个单体进行了EAG测试,结果显示,2,6-二叔丁基-4-甲基苯酚对雌雄成虫都没有作用,糠醛在20μg的最高测试剂量下能对雌雄两性产生显著的电生理反应。同时,在相同测试浓度下,雌雄成虫对不同物质的敏感度也不相同。通过对青杨脊虎天牛交配行为机制的研究,初步判断青杨脊虎天牛在配偶寻找过程中,非接触性的嗅觉判断不起主导作用。青杨脊虎天牛可能存在来源于雌虫的接触型信息素。利用GC-EAD测试青杨脊虎天牛幼虫粪便挥发物对天牛的生理活性,初步确定引起雌、雄成虫电生理反应的成分为庚烷。但庚烷在实践中是否能够干扰青杨脊虎天牛成虫的行为则有待于日后大量的研究工作加以证明。不同抗虫水平的十种杨树树干韧皮部和木质部中均含有苯甲酸、咖啡酸、绿原酸、肉桂酸、阿魏酸、香豆酸、水杨酸、丁香酸和香草酸9种酚酸。存在于抗性较弱的树种中的香草酸含量最高,而抗性水平较高的树种中阿魏酸、咖啡酸、肉桂酸和香豆酸含量显著高于其他树种。抗虫水平高的小黑杨和中东杨的酚酸总量最高。同时测定了取食9种酚酸的青杨脊虎天牛幼虫体内过氧化物酶(POD)、超氧化物歧化酶(SOD)、GSH-ST(谷胱甘肽-S-转移酶)和乙酰胆碱酯酶(TChE)四种酶活力的变化。结果显示,除苯甲酸,其它8种酚酸都对乙酰胆碱酯酶活力有极显著的抑制作用,说明幼虫体内的乙酰胆碱酯酶对酚类物质极为敏感,酚类物质能够很好的抑制昆虫体内的这种解毒酶。绿原酸、咖啡酸和丁香酸都能显著抑制虫体内过氧化物酶和谷胱甘肽-S-转移酶的活力。更多还原

【Abstract】 Xylotrechus rusticus is a kind of newly important bark pests which break out to be the disaster in the Northeast in China in recent years.It has brought great difficulties to the control of this pest because they bore into the trunk of trees.Poplar is the main afforestation species in the Northeast area,nowadays some areas are suffering from X.rusticus,and they bring a serious threat to the development of local forestry.For searching the way of ecological control on X.rusticus,this paper focused on the semiochemicals involved in the communication between the longicorn beetles,the beetles and plants,to explore the function of these semiochemicals and strive to find a single compounds or mixtures of different compounds which have biological activity for X.rusticus,and to provide a theoretical basis for the ecological control of the pest.In this experiment,ten kinds of poplar bark volatiles were extracted from 4 sensitive poplar species which are Populus.ussuriensis,P.pseudo-simonii,P.simonii and P.pseudo-simonii×P.nigra,and 6 kinds of resistance species which are P.berolinensis,P.simonii×P. nigra,P.russkii,P.alba×P.berolinensis,Populus x xiaozhuanica W.Y.Hsu et Liang cv. ’Baicheng-2’ and P.stalintz,using steam distillation and supersonic extraction,respectively; Summa canister was adopt to collect trunk volatiles from 5 kinds of poplar,including P. simonii,P.berolinensis,P.alba L.×P.berolinensis,P.pseudo-simonii and P.simonii×P. nigra,applying the combination of pre-enrichment systems and gas chromatography-mass spectrometry(GC-MS) to analyse chemical composition of those volatiles.The analysis results showed that,in the non-oviposition period of longicorn beetles,all of those ten kinds of poplar barks extractions contains hexanal,salicylaldehyde,tetradecanal,hexadecanal,2,6-ditertbutyl-4-methyl phenol,linoleic acid,hexadecanoic acid and 11,14,17-eicosatrienoic acid,methyl ester,and these compounds were contained in the extracts obtained from different methods. The compounds contained in the volatiles obtained from ultrasonic extraction were more than that contained in those volatiles obtained by steam distillation extraction.Special compounds were found in those volatiles extracted from the insect resistance species such as P.alba L.×P. berolinensis and P.berolinensis.Those specials compounds are zingiberene,γ-muurolene, alloaromadendrene,a-muurolene,d-cadinene and camphor.During the oviposition period, using summa canister sampling,1-[4-hydroxy benzyl]-6-methoxy-3,4-Dihydroisoquinolin-7-ol were common component from P.simonii×P.nigra,P.berolinensis and P.alba×P. berolinensis with high insect-resistant,and the content were more than 20%,in which the content of P.simonii×P.nigra was the highest,reaching 83.90%.In volatiles from species with low insect-resistant,the content of(1R)-a-pinene from P.simonii was the highest,β- pinene and bornyl acetate were important components in P.pseudo-simonii.In the steam distillation extractions,the content of(1R)-a-pinene,eucalyptol and ethyl acetate was higher in volatiles from poplar species with low level of insect-resistant,β-pinene,2,4-Decadienal,1,2-cyclohexanedione,ethyloxy-ethylene and pyrazole usually have higher concentration in those volatiles from poplar species with higher insect-resistance.The differences of chemical composition and content in volatiles from poplar trunks with different insect-resistance level might be the main factors lead to the difference of insect behaviors and as well as the reason for insect-resistance of trees.The steam distillation extractions from bark of ten poplar species were used to test electrophysiological and behavioral response of both sexes of X.rusticus adults,the results primarily ascertained that volatiles from P.simonii×P.nigra and P.pseudo-simonii×P. nigra all aroused significant electrophysiological response of both sexes of X.rusticus.The behavioral test results further indicated that,the males and females exhibited attractive tendency to bark extraction of P.pseudo-simonii and P.simonii which have lower insect-resistant level,but showed repellent tendency to that of Populus×xiaozhuanica W.Y.Hsu et Liang cv.’Baicheng-2’ and P.alba×P.berolinensis which have higher insect-resistance ability.Those experiment conclusions were identical with the actual X.rusticus damage status. At the same time,EAG tests were performed to six single compounds,2,6-ditertbutyl-4-methyl phenol,phenol,camphor,benzoic acid,1,3-dimethylbenzene and furfural at different concentration.Results showed that 2,6-ditertbutyl-4-methyl phenol had no function on adults of both sexes,furfural at 200μg dosage,the highest dosageused in the experiment,aroused distinct electrophysiological response.Simultaneously,it was found that the female and male adults have different sensitivity to different compounds at the same concentration.Through the investigation of copulation mechanism of X.rusticus,the non-contacted olfactory distinguishing did not play the leading role in the search for spouse of X.rusticus adults,the contacted sex pheromone was likely secreted from female adults.Using GC-EAD to test physiological activity of volatiles from larval frass,the component rouse electrophysiological response of male and female adults was primarily identified as heptane. But whether the heptane could disturb the behavior of X..rusticus adults in practice still needs to do a lot of study works subsequently to confirm.In phloem and xylem of trunk of ten poplar species with different insect-resistant level,at least nine phenolic acids was contined,which are benzoic acid,caffeic acid,chlorogenic acid, cinnamic acid,feruic acid,p-coumaric acid,sylicylic acid,syringic acid and vanillic acid.The content of vanillic acid was the highest in popar species with low resistance ability.However, the level of feruic acid,caffeic acid,cinnamic acid and p-coumaric acid was much higher in those resistance species than in the other species.The total amount of phenolic acids of P. simonii×P.nigra and P.berolinensis with high insect resistance was the highest.The changes of the enzyme activity of POD,SOD,GSH-ST and TChE in X.rusticus larvae fed on food containing those nine phenolic acids was determined.Results indicated that,eight phenolic acids,except benzoic acid,had significant inhibitory effect on TChE activity.This showed that TChE in larvae body was sensitive to polyphenols and those polyphenols can effectively inhibit this detoxifying enzyme in tested insect.The evidently inhibitory function to the enzyme activity of POD and GSH-ST was observed in chlorogenic acid,caffeic acid and syringic acid in the larval body.更多还原