松果梢斑螟信息化学物质的研究
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摘要
松果梢斑螟Dioryctria pryeri是一种重要的针叶树蛀食类害虫。它的分布与针叶树寄主植物分布相一致。分布于中国的黑龙江、吉林、辽宁、内蒙古、山东、河北、北京、河南、山西、陕西、甘肃、青海、新疆、安徽、江苏、浙江、四川和台湾等地,以及日本。近几年,在黑龙江小兴安岭及吉林省长白山林区的露水河、黄泥河及二道白河等地爆发成灾;危害红松的球果及枝梢,造成严重的经济损失,并引起了生态环境破坏。本文以松果梢斑螟生物生态学特性研究为起点;从信息化学物质寄主挥发性物质入手,分析其不同危害期红松球果挥发性物质、越冬幼虫转移危害期不同组织挥发性物质及成虫产卵期球果挥发性萜类物质时间变化规律;同时开展了幼虫对红松不同组织选择特点及挥发性物质中有效的引诱成分测试;在此基础上分析了红松球果中可溶性糖及总氮含量与幼虫危害关系;扫描电镜对其成虫触角、幼虫触角与口器化学感器的研究,为幼虫对寄主挥发性物质选择机理及幼虫取食危害时化学通讯联系提供了形态学依据;为松果梢斑螟的无公害治理工作奠定了基础。
     采用室内饲养与林间观测相结合方法观测了松果梢斑螟生物学特性,同时采用林间样地对角线和随机取样法及生物统计学中昆虫空间分布型聚集度指标和回归分析法分析了松果梢斑螟果幼虫空间分布型生态学特性,发现其于红松林间、树冠不同方位、树冠不同层次及球果上不同部位均具有聚集分布特性。
     利用动态顶空活体吸附剂采样法收集样品,采用热脱附、气相色谱与质谱联用技术(TCT-GC/MS)分析了红松挥发性物质组成及相对含量,结合嗅觉仪反应测试结果,探讨了越冬幼虫转移危害期幼虫对红松不同组织的选择,发现此时幼虫显著趋向于健康球果挥发性物质标样乙醇溶液,而对红松球果挥发性物质单组分a-蒎烯、柠檬烯及双组分的a-蒎烯+β-蒎烯各自百分含量标样乙醇溶液的选择显著高于红松球果挥发性物质标样乙醇溶液本身。幼虫对3种引诱源选择强弱顺序为a-蒎烯、柠檬烯、a-蒎烯+β-蒎烯乙醇溶液。越冬幼虫转移危害期,单组分物质在越冬幼虫对寄主选择中起主要作用。成虫产卵期则分析了红松健康球果与受害球果挥发性萜类释放时间变化规律。一天中挥发性萜类含量的测定结果表明,一天中大多数挥发性萜类的含量上午不同采样时间含量差异不显著,下午时差异显著;而上、下午之间含量差异显著。而采样时起不同日龄红松健康球果与受害球果a-蒎烯、β-蒎烯和柠檬烯含量较高,且变化规律一致。越冬幼虫转移危害期与成虫产卵期红松球果释放的挥发性萜类物质不同;前者多于后者,且均具有特异性组分。
     嗅觉盆与Y形嗅觉仪测试幼虫对红松不同类型球果选择行为发现,幼虫对受害球果选择的百分率大于健康球果,这一结果在红松人工林、混交林球果引诱源的研究中得到同样的结果。而对于红松混交林该虫危害球果不同部位形成的不同类型受害球果的选择结果表明,上部受害球果>下部受害球果>中部受害球果>整体受害球果;对红松人工林该虫危害球果不同部位形成的不同类型受害球果的选择的结果表明,整体受害球果>下部受害球果>上部受害球果>中部受害球果。在此基础上采用了蒽酮法及凯氏定氮法研究了红松球果整体、果鳞及果轴中可溶性糖及总氮的含量。发现幼虫对不同类型红松球果取食喜好程度与可溶性糖含量关系密切,随可溶性糖含量增加而加强,但取食球果蛀孔多少与其对幼虫引诱及取食喜好成分相关。
     蒽酮测试法与凯氏定氮法测试了松果梢斑螟不同危害期红松球果可溶性糖及总氮含量,幼虫转移危害期、成虫产卵期球果可溶性糖的含量低于总氮;幼虫转移越冬期可溶性糖含量高于总氮。红松果鳞与果轴中这两类物质含量变化与球果整体一致。此外,随着红松球果生长量的增加可溶性糖含量呈上升趋势,而氮含量呈下降趋势,与球果生理状态密切相关。而且幼虫转移越冬期红松健康球果可溶性糖含量低于受害球果,而总氮含量高于受害球果,是球果防御幼虫取食危害的一种策略。幼虫主要取食红松球果中幼嫩且质地柔软的果轴,可溶性糖含量红松球果整体>果鳞>果轴,氮素的含量为红松球果整体>果轴>果鳞。
     扫描电镜观察红松上松果梢斑螟成虫触角上感器及幼虫头部触角和口器上感器。成虫触角上分布有毛形、刺形、耳形和腔锥感器。毛形、刺形和腔锥感器为化学感器,依据资料推测分别执行感受昆虫性信息素的功能;感受机械刺激和味觉;感受水蒸气、二氧化氮、湿度变化、植物气味等作用。而五龄幼虫触角与口器上未发现任何感器,四龄幼虫触角末端2个栓锥形感器、2个锥形感器和1个毛形感器完成感受气味的功能;及触角第2节上刺形感器完成感受机械刺激与物质质地的功能。四龄幼虫口器的下颚及下唇须上分布化学感器。外颚内侧壁上具有2个栓锥形感器;下颚须端节端部有4个栓锥形感器;幼虫取食时,负责感受取食物质营养成分和气味感受作用;下唇须上有1栓锥形感器和1锥形感器,栓锥感器较锥形感器长且粗。
Dioryctria pryeri is one of the most destructive pests of conifer,distributed in Heilongjiang,Liaoning,Jilin,Neimenggu,Shandong,Hebei,Beijing,Henan,Shanxi,Gansu, Qinghai,Xinjang,Anhui,Jiangsu,Zhejiang,Sichuan,and Taiwan Provnce in China,and Japan, whose distribution matches to the the distributiong of host plants.Recent years,Outbreaks of Dioryctria pryeri have caused great damages to economy and ecological envioronment in Xing'an Mountins,Lushuihe,Huangnihe,Erdanbai of Changbai Mountain areas.The paper began with its biological and ecological characteristic.Host volatiles of semiochemicals were studied,in order to analyse of volatiles from cones on different damage periods,volatiles from different tissues on overwinter larvae transfer damage period,and time variational rule on adults oviposition period,larvae forage different tissues of Pinus koraiesis characteristic and tested active components of volatiles.Soluble surgar and total nutrigen contents of cones were analysd,so as to clarify the relationship between nutrition substance contents and damage of larvae.Sensillum of adult antenna,larvae antenna and mouthparts were observed by scanning electron microscope,which supply data to larvae forage host selection,and chemical communication.All of them will establish basis of control Dioryctria pryeri without social effects of pollution.
     Observe the biological characteristic with culture in laboratory and observation in fields. Adopted diagonal and random sample in fields,and aggregate index,regression analysis to analyse spatial distribution of ecological characteristic,found that the larvae distribution in fields,different positions and layers of tree crown,and different parts of cones is aggregation.
     Volatiles from host were collected by Headspace Sample,components and relative contents were analysed by TCT-GC/MS.Olfactory response were used to test larvae esponse to different tissues on larvae transfer damage period,to find that larvae reponse standard sample compounds of volatiles from cones to alcohol,while reponse sigle component a-pinene、Limonene,and double components a-pinene+β-pinene to standard sample compounds of volatiles from cones,which can draw the conclusion that single component substance play a key role in larvae host selection on overwinter larvae transfer damage period.Terpenes from healthy and damaged cones varied on adult oviposition period;there is significant difference between contents of most volatiles in pm,significant difference between am and pm,no significant difference in am.a-pinene,β-pinene and limnene has the most contents and same change of volatiles from healthy and damaged cones on different days.Voltile components released from cones on overwinter larvae transfer damage period are more than that on adult oviposition period.
     Olfactory basin and Y tube olfactory were used to test larvae response to Pinus koraiesis different kinds of cones,find that those response damage cones to healthy cones in mixture and plant forests.That response to different kinds of damaged cones which form from the different parts of cones damaged by Dioryctria pryeri,is Pinus koraiesis cones above damaged>cones below damaged>cones middle damged>cones total damaged in mixture forests, while is Pinus koraiesis cones total damage>cones below damaged>cones above damaged>cones middle damaged in plant forests.Anthranone method and Kjeldahl Determination were adopted to test soluble surgar and notrigen contents of Pinus koraiesis cones,cone axis, cone scale,find that there is tight correlation between larvae forage apt and soluble surgar,and related with the number of eat hole and quality of Pinus koraiesis cones.
     Adopting Anthranone method and Kjeldahl Determination test soluble surgar and total nutrigen contents of cones on different damage periods.The content of soluble surgar is lower than nutrigen on larvae transfer damage period and adult oviposition period,which is revised on transfer overwinter period,which is the same in cone axis and cone scale.In addition,the content of soluble surgar is increased and total nitrogen is decreased with cones growth.And, soluble content of healthy cones is lower than that of damaged cones,which is revised of the total notrigen content,which was the recovery strategy of cones to larvae forage.Larvae forage cone axis,soluble conent is that cones>cone scale>cone axis,and total nutrigen is cones>cone axis>cone scale.
     Scanning electron microscope was used to observe sensillum of adult antenna and larvae mouthparts and antenna.Sensillum trichoidea,sensillum chaetica,sensillum auricillica and sensillum coeloconica distributed on adult antenna,and sensillum trichoidea,sensillum chaetica,sensillum coeloconicais are chemical sensillum which recept sex pheromone, mechanical stimulate,and water vapor,nutrigen dioxide,humidity and odor of plants, respectively.There is no sensillum on antenna and mouthparts of the fifth age larvae,while there are 2 sensillum styloconica,2 sensillum basiconica and 1 sensillum trichoidea on the fourth age larvae attenna terminal that recept ordor,sensillum chaetica on the second antenna that recept ordor,2 sensillum styloconicaon on inner side of outer galea and 4 sensillum styloconicaon on terminal of palpus maxillaris that recept odor nutrition component when forage,1 sensillum styloconica on palpus labialis is longer than sensillum basiconica on palpus labialis.
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