杨树云斑天牛成虫寄主定位中的信息化学物质
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摘要
云斑天牛Batocera horsfieldi(Hope),属鞘翅目Coleoptera,天牛科Cerambycidae,是我国重要的一种林木蛀干害虫。这类害虫生活隐蔽,研究和防治比较艰难。成虫是其唯一裸露活动、有行为导向的虫态。因此,从云斑天牛大面积治理及工业用材品质的要求出发,成虫裸露活动期是研究和防治的关键。本课题在分析云斑天牛成虫寄主行为选择及分离鉴定其补充营养寄主和幼虫寄主挥发性化合物的基础上,利用触角电位仪(EAG)和Y型嗅觉仪鉴定出对成虫具有电生理及行为活性的化合物;并利用扫描电镜(SEM)观察了成虫头部附器的感器类型及分布,以期为开发利用天然活性物质来防治天牛提供一定的理论基础和技术支持。主要研究结果如下:
     1.云斑天牛成虫对寄主植物的嗅觉反应
     在室内利用Y型嗅觉仪测定云斑天牛不同生理状态的成虫对寄主植物的嗅觉反应,结果表明:以空气为对照时,补充营养寄主野蔷薇对未交配的雌雄成虫及已交配雌虫均有明显的吸引作用;幼虫(产卵)寄主杨树只对已交配雌虫具有明显的吸引作用。野蔷薇和杨树比较时,云斑天牛成虫对这两种寄主挥发性气味的趋性反应均没有显著差异。
     2.寄主植物挥发性化合物的分离鉴定
     利用顶空动态采集法和热脱附-气相色谱-质谱联用(TCT-GC-MS)分离鉴定了野蔷薇和杨树的挥发性化合物。结果表明:野蔷薇挥发性气味有10种组分,柠檬烯相对含量最高,3-蒈烯和β蒎烯次之;杨树挥发性气味有16种组分,反-2-己烯醛相对含量最高,己醛和顺-3-己烯醇次之。两者有5种共有组分,分别是己醛、反-2-己烯醇、顺-3-己烯醇、R型α蒎烯和β蒎烯,但相对含量上存在很大差异。
     3.云斑天牛成虫对挥发性化合物的EAG反应
     根据不同生理状态的云斑天牛成虫对浓度为100μg/μl的21种化合物溶液的EAG反应结果,筛选了8种电生理活性较高的化合物分别是顺-3-己烯醇、顺-2-戊烯醇、3-戊醇、3-蒈烯、己醛、反-2-己烯醛、反-2-己烯醇和1-戊烯-3-醇;云斑天牛成虫对上述8种化合物的EAG反应值绝大多数与浓度呈正相关,且顺-3-己烯醇、顺-2-戊烯醇、己醛、反-2-己烯醇和1-戊烯-3-醇在高浓度下引起的EAG反应在不同生理状态成虫之间存在一些显著差异。
     4.云斑天牛成虫对挥发性化合物的行为反应
     利用Y型嗅觉仪测试了云斑天牛成虫对8种电生理活性较强的化合物的行为反应,结果表明:有6种具有电生理活性的化合物具有行为活性,其中浓度为1和0.1μg/μl的顺-3-己烯醇以及1μg/μl的反-2-己烯醇对雄虫对有明显的吸引作用,浓度为1μg/μl的顺-3-己烯醇和0.1μg/μl的3-戊醇对雌虫有明显的吸引作用;而浓度为100μg/μl的3-蒈烯和1-戊烯-3-醇对两性成虫均有趋避作用,浓度为1和100μg/μl的反-2-己烯醛对雄虫有趋避作用。
     5.云斑天牛头部附器感器的扫描电镜观察
     利用扫描电镜观测云斑天牛成虫的触角、下颚须和下唇须,结果表明:云斑天牛成虫触角上有5类感器,即毛形感器、锥形感器(5种亚型)、刺形感器(2种亚型)、凹槽钉形感器、乳突形感器,且感器的类型、数量和分布存在明显的性二型现象。下颚须和下唇须均有毛形感器、长刺形感器、末梢锥形感器(2个亚型)、钟形感器和隙缝感器等5类感器,而感器的数量在雌雄之间、下唇须与下颚须之间存在着差异。
The longhorned beetle,Batocera horsfieldi(Hope)(Coleoptera:Cerambycidae),is one of most abundant and destructive cerambycids widely distributed in China.The larvae cryptic feeding habit makes the control of such wood-boring pests difficult.So to be effective,control strategies have to be based on controlling or killing longhorned beetle adults that are exposed to wild to feed and copulate.Our studies researched the behavioral and electrophysiological responses of B.horsfieldi adults to feeding- or larval-host plant odors present in headspace collections.In order to identify the range of feeding- or larval-host-associated volatiles that the longhomed beetles can detect,therefore,maybe develop attractants or reples for adults that could be used in a push-push control program. And we also researched the morphology,ultrastructure and distribution of sensilla on the antennae,mandible palp and labial palp of B.horsfieldi adults by scanning electron microscopy,to realize more about the olfaction recept mechanism in this species.The main results as follows:
     1.Olfactometer Bioassays to host plant volatiles
     When compared with the control air,both virgin sexes were significantly attracted to odors from Rosa multiflora Thunb.volatiles in olfactometer bioassays,mated females were attracted to volatiles from the two host plants.But all of the beetles tested showed no significant different responding between R.multiflora and Populus sp.
     2.The identified compounds of host plant volatiles
     10 and 16 compounds were identified from R.multiflora and P.sp.by TCT-GC/MS respectively,and 5 compounds common to both two host plant species included:hexanal, E-2-hexenal,Z-3-hexen-1-ol,1R-α-Pinene andβ-Pinene,but the relative proportion were different for each species.The mostly compounds of R.multiflora Thunb.are Limonene, 3-carene andβ-Pinene.E-2-Hexenal is the most abundant component of the floral scent of P.sp.,hexanal,Z-3-Hexen-1-ol and E-2-Hexen-1-ol acetate in the next place.
     3.EAG activity of synthetic compounds
     All of the identified compounds were tested for EAG responses of the B.horsfieldi antennae.According to the electrophysiological assessment of 100μg/μl compounds,we selected eight higher EAG-active components for EAG dose-response:Z-3-Hexen-1-ol, 3-pentanol,Z-2-Penten-1-ol,3-carene,Hexanal,E-2-Hexen-1-al,E-2-Hexen-1-ol and 1-penten-3-ol.Most of EAG responses of B.horsfieldi increased with stimulus strength. The EAG responses of Z-3-Hexen-1-ol,Z-2-Penten-1-ol,Hexanal,E-2-Hexen-1-ol and 1-penten-3-ol at the higher dose were significantly differences among mated female, virgin female and male.
     4.Bioassay for behavioral activity of EAG-active compounds
     When compared with the liquid paraffin(blank control),males showed a significantly preference for the Z-3-Hexen-1-ol at a dose of 1 and 0.1μg/μl and E-2-Hexen-1-ol at a dose of 1μg/μl,females showed a significantly preference to Z-3-Hexen-1-ol at a dose of 1μg/μl and to 3-Pentanol at a dose of 0.1μg/μl.Both of males and females showed a significantly preference for blank control over the 3-Carene and 1-penten-3-ol at a dose of 100μg/μl,and males showed a preference for blank control over the E-2-Hexen-1-al at a dose of 1μg/μl.
     5.Observation of sensilla on the cephalic appendage of B.horsfieldi with SEM
     The antennae,mandible palp and labial palp of B.horsfieldi adults were observed by scanning electron microscopy.The antennae contained five types sensilla,including sensilla trichodea,s.basiconica(including five subtypes:BⅠ~BⅤ),s.cheatica (including two subtypes:CHⅠand CHⅡ),s.grooved peg,s.cavitata-peg,respectively. Sexual dimorphism was found in B.horsfieldi,the length of the antennae of male were significantly longer than that of females;s.grooved peg was only found on the female antenna,and female antenna contained more s.BⅠ,s.BⅡand s.ChⅡthan males,while the number of s.BⅢwas fewer than that of males;the average length of s.cheatica of female was significantly longer than that of males.Both mandible palp and labial palp contained five types of sensilla,including s.trichodea,s.ChⅡ,s.twig basiconca,s. campullacea and s.aperture.Mandible palp contained more s.twig basiconca and fewer s. campullacea than labial palp.The number of s.twig basiconca was higher in female antennae,and s.campullacea was higher in male.
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