CN112889819B - 一种咖啡脊虎天牛性引诱剂 - Google Patents
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Abstract
本申请涉及一种咖啡脊虎天牛性引诱剂,包含该性引诱剂的诱芯,及该性引诱剂和诱芯用于防治咖啡脊虎天牛的用途。特别地,所述咖啡脊虎天牛性引诱剂包含长叶环烯和荜澄茄油萜烯以及任选的辅料,其中长叶环烯和荜澄茄油萜烯以特定的比例存在。所述咖啡脊虎天牛性引诱剂能有效诱杀大量咖啡脊虎天牛雄性成虫,可以用于咖啡脊虎天牛种群监测和防治,其对该虫的诱杀和预测预报具有重要的作用和意义。
Description
技术领域
本发明属于农业害虫防治领域,具体涉及一种咖啡脊虎天牛性引诱剂,包含其的诱芯,及其用于防治咖啡脊虎天牛的用途。
背景技术
咖啡脊虎天牛(Xylotrechus grayii)是金银花的主要蛀干害虫,属鞘翅目(Coleoptera)天牛科(Cerambycidae)天牛亚科(Cerambycinae)虎天牛族(Clytini)脊虎天牛属(Xylotrechus)。在中国广泛分布于华北、甘肃、西藏、四川、江苏、福建、广东、台湾。该虫是重要药用植物金银花的重要蛀干害虫。
咖啡脊虎天牛在山东等北方地区一年发生一代,以蛹或成虫越冬,越冬成虫于4月中旬开始出孔,4月下旬到5月上旬为出孔盛期,成虫出孔并在金银花老枝条基部产卵,卵孵化后,幼虫先在表皮下为害,逐渐向木质层蛀食,危害期长达3-5个月,虫粪不排出虫孔,危害非常隐蔽,使受害枝条枯萎,一经发现其对金银花危害已很严重。在金银花主产区山东平邑地区咖啡脊虎天牛的危害率高达70-80%,严重威胁金银花产业。
目前的防治方法以化学防治为主,辅以人工除虫和生物防治。化学防治中使用农药包括氧化乐果、敌敌畏、辛硫磷乳油等,但由于该虫幼虫在树皮下串食为害,具有隐蔽性,高浓度大剂量化学农药灌注或用药棉堵塞虫孔的方法不但效果差,也会伤及害虫天敌,且容易造成农药残留,严重影响金银花药品安全性,急需寻求无公害防治措施。
昆虫性信息素在害虫监测和防治中具有很大的应用潜力,越来越受到人们重视。申请人等发现咖啡脊虎天牛存在挥发性信息素,雌成虫的挥发物对雄成虫表现出显著的引诱作用。因此,鉴定咖啡脊虎天牛性信息素的活性成分,并在春季该天牛成虫出孔羽化的窗口期田间喷施含活性成虫的物质的性引诱剂来对该虫进行无公害防治及预测预报,具有显著的生态、经济和社会效益。
发明内容
申请人研究发现,长叶环烯(Longicyclene)和荜澄茄油萜烯(β-Cubebene)是两种咖啡脊虎天牛的挥发性信息素,可以用作咖啡脊虎天牛性引诱剂。
长叶环烯的化学名为(1S,2S)-1,2,6,6-四甲基四环[8.1.0.02.8.07.11]十一烷,其结构如下:
荜澄茄油萜烯的化学名为(3aS,3bR,4S,7R,7aR)-7-甲基-3-亚甲基-4-(丙-2-基)八氢-1H-环戊二烯并[1,3]环丙并[1,2]苯,其结构如下:
因此,本发明涉及一种咖啡脊虎天牛性引诱剂,其包含长叶环烯和荜澄茄油萜烯以及任选的辅料。
在本发明的一个优选的实施方案中,所述咖啡脊虎天牛性引诱剂中长叶环烯和荜澄茄油萜烯的质量比例为1:6-2:3。
在本发明的一个优选的实施方案中,所述辅料为可以溶解长叶环烯和荜澄茄油萜烯的有机溶剂,例如正己烷。
本发明还涉及一种诱芯,其包含上述咖啡脊虎天牛性引诱剂、有机溶剂和释放载体。
所述诱芯可以通过将咖啡脊虎天牛性引诱剂溶解于适量有机溶剂中,并将所得的性引诱剂溶液装载到释放载体上而得到。
在本发明的一个优选的实施方案中,所述释放载体可以为有机硅胶芯。
本发明还涉及上述咖啡脊虎天牛性引诱剂或诱芯用于防治咖啡脊虎天牛的用途。
本发明的咖啡脊虎天牛性引诱剂能有效诱杀大量咖啡脊虎天牛雄性成虫,可以用于咖啡脊虎天牛种群监测和防治,其对该虫的诱杀和预测预报具有重要的作用和意义。
附图说明
图1:提取物的总离子流图。
图2:长叶环烯(Longicyclene)谱库(mainlib)和提取物离子碎片图(text file)对比。
图3:荜澄茄油萜烯(β-Cubebene)谱库(mainlib)和提取物离子碎片图(textfile)对比。
图4:提取物的气相色谱与触角电位分析图。
具体实施方式
以下结合实施例对本发明作进一步描述。
实施例1、提取信息素
挥发性性信息素的提取:利用动态顶空抽提法对成熟的咖啡脊虎天牛雌雄成虫挥发物进行抽提,雌雄成虫各选择30头,分3次进行提取,每次各10头,抽提8小时,抽提时间每天9:00到17:00,大气采样仪(QC-1B,北京劳动保护科学研究所)流速300ml/min,抽提装置按以下顺序连接:压缩空气-流量计-粗活性炭-细活性炭-蒸馏水-流量计-样品瓶-吸附剂采样管-大气采样仪。吸附剂选择Porapak Q(50~80目,Waters Corporation,USA),每枝吸附管填充100mg。二氯甲烷(天津市天昊化工有限公司)600μl分3次进行洗脱,每次200μl。
实施例2、性信息素鉴定
挥发性性信息素的分析:成熟咖啡脊虎天牛雌雄成虫的挥发物用气相色谱-质谱联用仪(美国安捷伦HP7890A/5875C),色谱柱DB-Wax(30m×250μm×0.25μm)分析。气相色谱的工作条件,升温程序:初始温度40℃,保留1min,以10℃/min升至190℃,然后以5℃/min升至250℃,保留5min。加热器温度为220℃,以氦气为载气,流速为1ml/min,不分流进样,进样量1μl。质谱的工作条件,电离方式为EI,电子能量为70eV,离子源温度230℃,质谱扫描范围(m/z)50~500。所得质谱图与标准质谱图库(NIST08)进行比对分析,确定化合物种类,通过比较化合物的质谱离子峰面积确定各成分的相对含量。
图1示出咖啡脊虎天牛雌性成虫提取物的气相色谱联用分析,提取物的总离子流图。图2和图3示出长叶环烯和荜澄茄油萜烯谱库(mainlib)和提取物离子碎片图(textfile)对比。结果表明,咖啡脊虎天牛的性信息素组分为长叶环烯和荜澄茄油萜烯。
实施例3、提取物气相色谱和触角电位分析
利用气相色谱与触角电位联用仪(GC-EAD)进行提取物气相色谱-触角电位分析。触角电位仪(SYNTECH,Hilversum,the Netherlands)由微动操作台(MP-15),气流控制器(CS-55)和数据采集控制器(IDAC-4)组成。将咖啡脊虎天牛雄性成虫的触角自头部剪下,用手术刀快速去除触角顶端约0.1mm长的一段,然后用导电胶(Spectra 360Electrode Gel,Parker Laboratories Inc Orange,NJ,USA)将其固定在金属电极上并安装于触角电位仪上。色谱柱DB-Wax,气相色谱出口端的分流比1:7,分流的化合物一部分进入FID氢火焰离子检测器,另一部分经过加热套吹向触角。用气相色谱与触角电位联用仪软件同步记录气相色谱和触角电位图。
图4示出咖啡脊虎天牛雄性成虫触角对雌性成虫提取物气相色谱-触角电位分析。雌性提取物中的长叶环烯和荜澄茄油萜烯均引起雄性成虫触角电位反应,进一步表明咖啡脊虎天牛的性信息素组分为长叶环烯和荜澄茄油萜烯。
实施例4、室内电生理实验
使用不同比例构成的性引诱剂溶液在室内进行触角电位实验(EAG)。利用气相色谱与触角电位联用仪(GC-EAD),触角电位仪(SYNTECH,Hilversum,the Netherlands)由微动操作台(MP-15),气流控制器(CS-55)和数据采集控制器(IDAC-4)组成。
将咖啡脊虎天牛触角自头部剪下,用手术刀快速去除触角顶端约0.1mm长的一段,然后用导电胶(Spectra 360Electrode Gel,Parker Laboratories Inc Orange,NJ,USA)将其固定在金属电极上并安装在于触角电位仪。色谱柱DB-Wax,气相色谱的条件同实施例3,气相色谱出口端的分流比1:7,分流的化合物一部分进入FID氢火焰离子检测器,另一部分经过加热套吹向触角,用气相色谱与触角电位联用仪软件同步记录气相色谱和触角电位图。
实验组为长叶环烯(购自sigma公司)和荜澄茄油萜烯(购自sigma公司)按以下质量比例1:6、1:3、1:2、2:3配制的混合物。对照组为正己烷,不含长叶环烯和荜澄茄油萜烯。
触角电位试验表明:咖啡脊虎天牛雄性成虫触角对不同比例性信息素的反映存在差异,实验组显著高于对照组。
实施例5、室内行为学实验
利用Y型嗅觉仪(两臂长23cm,主臂长16cm,两臂夹角60度,直径3cm)测定成虫的求偶定向行为,试验在室内观察笼(长100cm,宽60cm,高60cm)中进行,温度28℃,测试时间9:00到17:00。测定时,将Y型嗅觉仪放置在绘王A3智能烤台上,该烤台从下部提供光源,保证两臂光线均匀。测试装置按如下顺序连接:压缩空气-流量计-粗活性炭-细活性炭-蒸馏水-气味源/空气对照-流量计-Y型管。气流接通后,两臂管内的气流速度调节为500mL/min。将2ml长叶环烯和荜澄茄油萜烯按1:6、1:3、1:2、2:3比例配制的混合物滴加到不同缓释袋里的载体过滤纤维上作为气味源,对照为不含长叶环烯和荜澄茄油萜烯的正己烷。
每次测定释放一头雄性成虫,释放后,在主臂正对的分叉处放置小型手电并打开,待释放的成虫爬行至分叉处时关闭,测试时间15min。测试成虫爬行超过某臂长1/2并停留超过30s或到达某臂末端记为选择,否则记录为无选择。每测试1头成虫,用无水乙醇清洗Y型嗅觉仪的各部分,再用蒸馏水冲洗,电吹风吹干后用无水乙醇擦拭一遍再进行下一头的试验。每测试3头成虫反转一次Y型管。
室内行为学实验表明,含长叶环烯和荜澄茄油萜烯的气味源具有显著的引诱活性。
表1咖啡脊虎天牛雄性成虫对引诱剂成分的行为反应
*长叶环烯:荜澄茄油萜烯比例
ab:同一行标同一字母表示在0.05水平上的差异不显著,标不同字母表示差异显著。
结论
截止目前,国内外无咖啡脊虎天牛性信息素的报道。本发明人首次发现并报道了咖啡脊虎天牛性信息素组分,即长叶环烯和荜澄茄油萜烯。包含这两种成分的咖啡脊虎天牛性引诱剂能有效诱杀大量雄性成虫,可以用于咖啡脊虎天牛种群监测和防治。
Claims (6)
1.一种咖啡脊虎天牛性引诱剂,其包含长叶环烯和荜澄茄油萜烯以及任选的辅料,其中长叶环烯和荜澄茄油萜烯的质量比例为1:6-2:3。
2.根据权利要求1所述的咖啡脊虎天牛性引诱剂,其中辅料为可以溶解长叶环烯和荜澄茄油萜烯的有机溶剂。
3.根据权利要求2所述的咖啡脊虎天牛性引诱剂,其中辅料为正己烷。
4.一种诱芯,其包含根据权利要求1至3中任一项所述的咖啡脊虎天牛性引诱剂、有机溶剂和释放载体。
5.根据权利要求4所述的诱芯,其中释放载体为有机硅胶芯。
6.根据权利要求1至3中任一项所述的咖啡脊虎天牛性引诱剂或者根据权利要求4或5所述的诱芯用于防治咖啡脊虎天牛的用途。
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Non-Patent Citations (3)
Title |
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Acaricidal activity and repellency of essential oil from Piper aduncum and its components against Tetranychus urticae;Araujo, Mario J. C.;Experimental and Applied Acarology;第57卷(第2期);139-155 * |
Toxicity of some terpenoids of essential oils of Xylopia aethiopica from Cameroon against Sitophilus zeamais Motschulsky;Kouninki, H.;Journal of Applied Entomology;第131卷(第4期);269-274 * |
天牛嗅觉感受相关蛋白研究进展;詹文会;张苏芳;耿红卫;王野影;郭昆;陈君;;河南农业科学(第03期);全文 * |
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