【作者】 袁秀洁；

【导师】 张真；

【作者基本信息】 中国林业科学研究院 ， 森林保护学， 2013， 博士

【摘要】 松阿扁叶蜂Acantholyda posticalis是一种重要的松树害虫。世界范围内主要分布于欧洲和亚洲东部，并且已经对大面积松林造成了严重危害。了解昆虫由于适应不同地理环境条件，在形态学、生物学等方面产生的地理变异，对于深入研究和管理害虫种群十分重要。本文对不同地理种群松阿扁叶蜂的形态学差异进行了研究，利用分子标记技术对我国不同地理种群的松阿扁叶蜂进行了遗传变异分析，并与欧洲地区报道的松阿扁叶蜂的遗传序列信息结合，研究揭示了各种群间的遗传分化及亲缘关系。另外，本文还针对遗传距离最远的两个地理种群进行了触角感器超微结构观察和寄主挥发物的主要成分及其对松阿扁叶蜂的生理活性研究。研究结果如下：（1）形态学观察发现不同地理种群松阿扁叶蜂在虫体大小，雄性生殖器和阳茎瓣大小，头部色斑，胸部盾片色斑，触角梗节色斑及雄性生殖器颜色方面均存在差异。其中中部种群的虫体最大，而西南部种群虫体最小，雌雄虫体长均随纬度增加而增加，符合Bergmann法则，同时雌虫体长随经度增加而增加，雌雄虫体长和翅展与海拔均无显著相关。雌雄虫展翅与经度均无显著相关。中部种群虫体长度和宽度的二型性指数最大，西南部种群最小，性体型二型性指数与纬度具有显著相关性。（2）8个采样点的138个松阿扁叶蜂样品中共检测到了56个单倍型。其中有4个单倍型是由多个地理种群共享的，有52个单倍型是单个地理种群所独享的。单倍型HT12和HT43是两个主要的单倍型，分别被3个采样点和2个采样点所共享。8个地理种群的单倍型多样度（h）为0.962±0.006；核苷酸多样度（π）为0.04365±0.00224。在区域内种群间（FSC=0.21046，p<0.001）和种群内个体间（FST=0.26826，p<0.001）两个层次均存在显著的遗传分化，其中种群内部的遗传变异率最高，为73.17%。8个采样点中西南部地区与中部地区种群的遗传距离最远（0.079-0.107），北部地区的抚顺种群与欧洲种群的亲缘关系最近，遗传距离为（0.016-0.021）。网络结构图显示56个单倍型明显分为三组，西南部地区的单倍型形成一组A，北部和东部地区的单倍型形成一组B，中部地区种群的单倍型形成一组C。其中A组以HT12为中心，其它单倍型均由HT12进化而来。（3）电镜观察发现松阿扁叶蜂的触角为丝状，包括32-35节，由柄节、梗节和鞭节组成。松阿扁叶蜂触角表面共分布有6种感受器，分别是刺形感器、毛形感器、锥形感器I、锥形感器II、腔锥形感器和钟形感器。刺形感器基部有臼状窝，外形刚直如刺，向顶部渐尖细，厚壁表面有明显的斜螺纹，内部没有神经树突；毛形感器呈毛状突起，表面有纵向平行的沟槽，顶部圆钝有一顶孔，内部有4个神经树突；锥形感器I呈拇指状突起，基部着生于垫状凸起，表面有纵向平行的沟槽，顶部有多个裂缝状开口，内部的树突鞘内有大量神经树突；锥形感器II呈短圆锥状突起，基部着生于触角表面较浅的圆形凹陷内，壁上布满小孔，顶端一侧有顶孔，内部有许多神经树突及分支；腔锥形感器表面光滑，着生于触角表面较深的圆形凹陷内，顶部中央有顶孔，四周分布纵向开裂，内部有多个神经树突；钟形感器着生于触角表面凹陷的圆形小穴内，顶端与触角表皮基本持平，顶部有一小孔，内部布满神经束。锥形感器I、锥形感器II、腔锥形感器和钟形感器集中分布于触角鞭节的腹侧面上，而刺形感器和毛形感器则广泛分布于整个触角的表面。雌虫触角显著长于雄虫触角。雄虫触角上锥形感器I的数量明显多于雌虫触角。两个地理种群之间进行比较，阳泉种群触角上的刺形感器和锥形感器II数量显著多于万荣种群，但锥形感器I的数量明显少于万荣种群。结合外部形态、内部结构特点以及相关文献对各种感器的功能进行了讨论。（4）两个地理种群所在的油松挥发物都包括11种主要的萜烯类物质：R-(+)-α-蒎烯、S-(-)-α-蒎烯、S-(-)-β-蒎烯、(+)-3-蒈烯、香叶烯、(+)-柠檬烯、(-)-柠檬烯、γ-萜品烯、α-萜品烯、崁烯和萜品油烯。其中R-(+)-α-蒎烯、S-(-)-α-蒎烯、香叶烯和(-)-柠檬烯为主要成分，S-(-)-β-蒎烯、(+)-3-蒈烯、(+)-柠檬烯的相对含量较少，γ-萜品烯、α-萜品烯、崁烯和萜品油烯为微量成分。两地理种群之间各个成分的含量没有显著性差异。EAG测试显示，两个地理种群的松阿扁叶蜂对(+)-3-蒈烯、香叶烯、γ-萜品烯、(+)-柠檬烯、(-)-柠檬烯、α-萜品烯和萜品油烯7种具有显著的生理活性，以(+)-柠檬烯活性最高。两个地理种群松阿扁叶蜂对11种寄主挥发物的反应均无显著性差异。剂量反应测试显示，两个地理种群的松阿扁叶蜂雌雄成虫对(+)-3-蒈烯、(-)-柠檬烯和香叶烯3种化合物的反应强度与化合物的剂量成正相关，对(+)-柠檬烯、γ-萜品烯和萜品油烯3种化合物的剂量反应在625ug达到最大饱和点，随后剂量增加时反应值下降，万荣种群雌雄成虫对α-萜品烯的剂量反应在625ug时达到最大饱和点，而阳泉种群雌雄成虫对α-萜品烯的剂量反应强度与化合物的剂量成正相关。更多还原

【Abstract】 Acantholyda posticalis（Hymenoptera: Pamohiliidae） is an important pine pest andwidely distributed throughout the Europe and Asia. It had caused serious damage to largeexpanses of forests worldwide. Understanding geographical variation of insect in themorphology, biology and other aspects, to adapt to different environmental conditions, is veryimportant for further study and management of pest populations. In this paper, we studied themorphologiacl difference of A. posticalis from different populations. At the same time thegenetic variation between different geographical populations of A. posticalis was analized bymolecular marker technology. The genetic relationships were revealed between Chinapopulations and Europen population by combining the genetic sequence information of A.posticalis from Europen population. Meanwhile, ultrastructure of the antennal sensilla and themain components of host volatiles were investigated for two populations with farthest geneticdistance. Finally, respones of the two sawfly population to the volatiles of their host pines weretested. The main results are as follows:(1) Morphology study shows there were signification difference in the body size, hadpatch, antenna pedicel patch, scute patch and the size of the male genitalia. The largst body sizeoccured in the central population and the smallest body size in the southwest population. Bodylenth of both male and female incresed with the increse of latitude which follow theBengermann’s rule. The body lenth of female incresed with the increse of longgtitude as well.Body lenth, wing expanse of both male and female had no significant correlation with altitude.The wing expanses of both male and female had no significant correlation with longtitude.Sexual size dimorphism index (SSDI) of central popilation is the largest and which ofsouthwest population is smallest. There was significant correlation between SSDI and latitude.(2) There were56mitochondrial DNA haplotypes in138samples from8populations.There were4haplotypes shared by multi-populations and52haplotypes belongs a single population. The HT12and HT43were main haplotypes and occurred in3populations and2populations respectively. The haplotype and nucleotide diversity of the eight geographicpopulations were0.962±0.006and0.04365±0.00224respectively. Analyses of molecularvariance (AMOVA) revealed that eight populations exhibited significant genetic differentiationwithin population (FST=0.26826， p<0.001) and among populations from single region(FSC=0.21046，p<0.001). The variability within population was73.17%. The genetic distancebetween southwest region and the middle region was the farthest (0.079-0.107) while thegenetic distance betwenn European population and population in northern region was closest(0.016-0.021). In haplotype networks, all haplotyes divided into three groups obviously. GroupA was consisted of haplotyes from southwest region and with the haploty HT12as its centre.Group B was consisted of north and eastern region. Group C was consisted of middle region.All of other haploty were evolved from HT12.(3) The antennae of A. posticalis are filiform, comprised32–35flagellomeres, and consistof scape, pedicel and a distal multi-segmented flagellum. Six sensillum types were found.Sensilla chaetica were straight setae with sharply-pointed tips and without dendrite in thelumen. Sensilla trichodea were characterized by a parallel-grooved wall and one terminal pore,innerved by4dendrites at the base. Sensilla basiconica I possessed longitudinally-groovedsurfaces and multiple terminal pores,5dendrites in the lumen. Sensilla basiconica II not onlyhad a distinct terminal pore but also had numerous tiny wall pores, numerous dendriticbranches within the sensillum lymph. Sensilla coeloconica had deep longitudinal grooves andone terminal pore, innerved by6dendrites, while s. campaniformia were thick-walled with aterminal opening and sensory nerve bundles in the lumen. Sensilla chaetica and s. trichodeawere most abundant and distributed over the entire antennae, while s. basiconica I and II, s.coeloconica, and s. campaniformia were restricted to the ventral flagellar surfaces. Althoughthe shape and structure of antennae were similar in males and females, females hadsignificantly longer antennae than males, and males had significantly more s. basiconica I thanfemales. The Yangquan population had more s. chaetica and s. basiconica II than the Wanrong population but substantially fewer s. basiconica I. In the Wanrong population, the s.coeloconica of males were longer and those of females shorter than in the Yangquan population.We compared the morphology and structure of these sensilla to other Hymenoptera anddiscussed their possible functions.(4) There were11main monoterpenes in host from two areas of two populations. Theywere R-(+)-α-pinene, S-(-)-α-pinene, S-(-)-β-pinene,(+)-3-carene, myrcene,(+)-limonene,(-)-limonene, γ-terpinene, α-terpinene, camphene, terpinolene. R-(+)-α-pinene, S-(-)-α-pinene,myrcene,(-)-limonene were main components. There were relative less amount ofS-(-)-β-pinene,(+)-3-carene,(+)-limonene and the γ-terpinene, α-terpinene, camphene,terpinolene were trace components. There was no significant difference between the twopopulations in term of monoterpene components and their contents. A. posticalis showedintensive response to (+)-3-carene, myrcene, γ-terpinene,(+)-limonene,(-)-limonene,α-terpinene and terpinolene, with the most significant response to (+)-limonene. There were nosignificant difference in response of two sawfly populations to11monoterpenes. There werepositive correlation between the response intensity of A. posticalis and the dose of (+)-3-carene,(-)-limonene and myrcene. When the dose of (+)-limonene, γ-terpinene and terpinolene was625ug, the response of A. posticalis reached saturation point and soon descend when the dosefurther increased. The response of A. posticalis from Wanrong population reach saturationpoint when the dose of α-terpinene was625ug while there was positive correlation between theresponse intensity of A. posticalis to the dose of α-terpinene from Yangquan population.更多还原