CN115885997B - 一种大粒横沟象信息素引诱剂及其应用 - Google Patents
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Abstract
本发明公开了一种大粒横沟象信息素引诱剂及其应用,属于信息素生物防治技术领域。大粒横沟象信息素引诱剂包括香叶醇乙酯、顺‑9‑十八碳烯醇、顺‑2‑(3,3‑二甲基亚环乙基)乙酸乙酯。采用本发明的大粒横沟象信息素引诱剂加入到释放载体中即制得大粒横沟象信息素引诱剂诱芯,可大量引诱到大粒横沟象成虫,为防治大粒横沟象提供了一种绿色安全的生物防控措施。
Description
技术领域
本申请涉及害虫防治技术领域,具体而言,涉及一种大粒横沟象信息素引诱剂及其应用。
背景技术
油橄榄Olea europaea L.是著名亚热带果树和重要经济林木,在中国主要分布在甘肃、广东、广西、云南、四川等省区。橄榄油含有丰富的营养和功能活性成分,具有预防心脑血管疾病和抗肿瘤等功能,素有“植物油皇后”的美誉。大粒横沟象Dysceruscribripennis为害油橄榄及多种林木、果树,主要以成虫和幼虫蛀茎食害主干、枝丫等韧皮及木质部,破坏输导组织,造成枝丫坏死,导致株干枯死。大粒横沟象多以成虫在土里、幼虫在树皮里越冬,有世代重叠现象。目前防治大粒横沟象的方法主要包括人工物理和化学农药防治,人工物理防治费时费工,大面积防治很不现实。化学农药防治虽然快速、高效,但长期大量使用,会增加农药残留,油橄榄最终的产品为食用的橄榄油,农药残留会影响油的品质。
随着城镇园林绿化面积的增长,大粒横沟象也从单一寄主油橄榄,向多寄主进军危害。2006年山东济南白蜡树、2010年四川香樟及2016年宁波象山桂花树均发生大粒横沟象危害。
利用昆虫自身的信息化学物质控制害虫的危害或者干扰和阻止害虫为害,可达到直接或间接防治害虫的目的。这类产品具有环境友好、无残留,对人畜安全无毒的优点,具有广泛的应用前景。
象甲科昆虫是许多农林业的重要害虫,已鉴定出的信息素成分的象甲约27种(靳秀芳等,林业与环境科学,2019,35(3):107-115;杨毅等,浙江农林大学学报,2012,29(1):5)。多数象甲的信息素由雄虫分泌,棉铃象甲的信息素由(+)-顺-2-异丙烯基-1-甲基环丁基乙醇、顺-2-(3,3-二甲基-环己亚基)乙醇、顺-2-(3,3-二甲基-环己亚基)乙醛和反-2-(3,3-二甲基-环己亚基)乙醛组成(HEDIN P A et al,Journal of Chemical Ecology,1979,5(4):617-627;MCKIBBN G H et al,Journal of Chemical Ecology,1977(3):331-335)。核桃象甲与棉铃虫象甲信息素成分相同(HEDIN P A et al,Journal of ChemicalEcology,1997,23(4):965-978)。白松木蠹象、北方松木蠹象和喜马拉雅杉木蠹象信息素相同,均为象甲醇和象甲醛两种物质,但是比例不同(BOOTH D C et al,Journal ofChemical Ecology,1983,9(1):1-12)。蔓越橘象甲信息素成分顺-2-(3,3-二甲基-环己亚基)乙醇、顺-(3,3-二甲基环己亚基)乙醛、反-(3,3-二甲基环己亚基)乙醛由雄虫释放,而香叶醇雌虫和雄虫均可释放(BIANCA G et al,Journal of Chemical Ecology,2012,38(3):272-277)。但利用信息素对象甲科昆虫进行综合防治的生产应用并不多,只有棉铃象甲、甘薯象甲、亚洲鼻隐喙象和红棕象甲的信息素用于害虫监测防治,取得了一定的防治效果。根据对大粒横沟象防治技术的了解和查证,大粒横沟象信息素尚未得到鉴定,国内外未有使用信息素防治大粒横沟象的案例和记载。因此,研究和应用大粒横沟象信息素产品进行防治具有重要意义。
发明内容
本发明的目的在于提供了一种大粒横沟象信息素引诱剂及其制备方法和应用。
本发明所描述的大粒横沟象信息素成分是通过气相色谱及气相-触角电位的方法进行鉴定。
本发明提供一种大粒横沟象信息素引诱剂,所述信息素引诱剂包括香叶醇乙酯、顺-9-十八碳烯醇和顺-2-(3,3-二甲基亚环乙基)乙酸乙酯。
优选的,香叶醇乙酯、顺-9-十八碳烯醇、顺-2-(3,3-二甲基亚环乙基)乙酸乙酯的质量比为1:0.1~3:0.1~3。
优选的,香叶醇乙酯、顺-9-十八碳烯醇、顺-2-(3,3-二甲基亚环乙基)乙酸乙酯的质量比为1:2:2。
本发明还提供一种前述大粒横沟象信息素引诱剂在大粒横沟象的生物防治中的应用。具体的,所述应用为将香叶醇乙酯、顺-9-十八碳烯醇和顺-2-(3,3-二甲基亚环乙基)乙酸乙酯加入到载体中制得大粒横沟象信息素引诱剂诱芯使用。所述载体为聚乙烯管、聚氯乙烯管、聚乙烯瓶、聚乙烯缓释袋、聚氯乙烯缓释袋、橡胶塞中的任意一种。优选的,所述载体为聚乙烯瓶。
使用时,将大粒横沟象信息素引诱剂制成诱芯,将装有大粒横沟象信息素引诱剂诱芯的诱捕器悬挂于大粒横沟象危害区域,用于大粒横沟象的虫情监测、大量诱捕。所述的诱捕器为网状诱捕器。优选的,所述引诱剂的悬挂高度高于1.5m的高度。每个所述诱芯含有5000ug所述信息素引诱剂,所述诱芯之间的平均间距为50m。
本发明的有益效果包括:
本发明提供的大粒横沟象信息素引诱剂可大量引诱到大粒横沟象成虫,为防治大粒横沟象提供了一种无公害的信息素防控技术,可进行大粒横沟象虫情监测和大量诱捕,具有环境友好,无污染的优点,是解决油橄榄等经济林生产有机或绿色食品技术关键之一。
附图说明
图1为大粒横沟象成虫对挥发性有机物的GC-EAD电生理反应图;
图2为大粒横沟象信息素引诱剂对雌、雄成虫诱捕能力分析图。
具体实施方式
下面结合实施例对本发明进行进一步说明和描述,但所描述的实施例仅仅是本发明的一部分实施例,而不是全部的实施例。基于本发明和实施例中,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他发明和实施例,都属于本发明保护的范围。
下述实施例中所使用的实验方法如无特殊说明,均为常规方法。
下述实施例中所用的材料、试剂等,如无特殊说明,均可从商业途径得到。信息素成分顺-2-(3,3-二甲基亚环乙基)乙酸乙酯的合成方法是按照公开文献“KazuhikoTanaka,et al.,“New Methods for Synthesis ofα,β-Unsaturated Carboxylic Estersfrom Carbonyl Compounds Using Monoanions of Dithiocarbonates,and Dianions ofEthyl Mercaptoacetate and Ethyl 2-Mercaptopropionate”,Bulletin of theChemical Society of Japan,1979,52(12),3619-3625”中所述的合成方法制备获得。
实施例1大粒横沟象信息素成分鉴定
(1)挥发性有机成分萃取
溶剂萃取法采样:将大粒横沟象雌雄成虫后肠用尖头镊子扯出,每10条成虫后肠放置1ml玻璃瓶中,加入100ul正己烷萃取40min后,将正己烷萃取液浓缩成1ul后封存于毛细玻璃管中,-20℃保存备用。雌雄处理重复5次。
(2)GC-MS鉴定化合物成分
后肠提取物经气质联用(安捷伦7890B-5977)分析,色谱柱是DB-5MS
(30m×0.25mm×0.25μm),载气为高纯氦气,流速为1ml/min,不分流进样,分流阀1min后打开。进样量1ul,进样口温度250℃,升温程序:初温80℃,保持1min,以10℃/min升至280℃,保持10min。MS分析条件:EI离子源,电子能量70ev,传输线温度250℃,离子源温度220℃,扫描范围41-650amu。化合物的鉴定是通过与标准化合物比较保留时间和质谱、与Nist02谱库中的标准谱图进行比较,对化合物进行结构鉴定,结果如下表1所示。
表1挥发性有机物的鉴定
实施例2大粒横沟象成虫对挥发性有机物的GC-EAD分析
将上述鉴定成分分别称取1000ug,溶于1ml正己烷中配置成浓度为1000ug/ml的混合溶液,然后取25ul混合溶液加入到475ul的正己烷中,稀释20倍,每种化合物浓度为50ug/ml。取大粒横沟象触角,两端连接在装有银丝的玻璃电极和参比电极上,玻璃电极内充满昆虫生理盐水。触角悬在L型的玻璃管口0.5cm处,处于连续、湿润的空气流中(空气流速:900ml/min-1)。进样量是1ul。使用B-5MS柱子,GC的升温程序是初始温度80℃,保持1分钟,然后每分钟升高10℃直到280℃,保持10分钟。高纯度氦气作为载气,流速20mL/min,后检测器温度300℃。FID温度为300℃,将氦气以39mL/min的速度添加到分流器中,并通过压装Y分离器(Agilent7890)在GC的火焰电离检测器(FID)和EAD之间平均分配。用于EAD的GC流出物通过加热器,并与湿空气(300mL/min,20℃)混合。通过GC-EAD软件显示并分析从接口获取的EAD和FID信号。对大粒横沟象四头雄虫和四头雌虫分别测试混合物,每个混合物重复3次,结果如图1所示。从图1中可以看出,大粒横沟象雌、雄触角均对香叶醇乙酯:顺-9-十八碳烯醇:顺-2-(3,3-二甲基亚环乙基)乙酸乙酯有明显的触角电位反应,说明这三种成分可能是大粒横沟象信息素活性成分。
实施例3筛选大粒横沟象信息素引诱剂最佳配比
试验地设在四川利州区油橄榄产业园区,为保证诱捕效果,诱捕器之间的平均间距为50m,诱芯高度设置为离地面高度1.5m,诱捕器类型为网状诱捕器。根据雌雄成虫触角对挥发性成分的敏感性设计诱芯进行实验,共设置如下表2中所示的共11个处理组和1个空白组,每个处理3个重复。按照质量比将信息素成分装在聚乙烯瓶中(PE bottle,15ml,Contech Enterprises,Delta,BC,Canada),每个诱芯含有5000ug的信息素。每3天调查一次,收集并记录诱捕的虫数量和种类,连续调查1个月。选天气晴朗的上午9-11点悬挂引诱剂。悬挂引诱剂时将封口膜去掉,用细铁丝将引诱剂悬挂于诱捕器中央,记录诱捕实验数据。
表2不同物质质量比的大粒横沟象引诱剂处理组诱捕实验数据
表2中数据表明:(1)单元组分信息素(处理1-3)诱捕效果一般;(2)二元组分信息素(处理4-6)诱捕效果强于单元组分信息素;(3)三元信息素组分((处理7-11))的诱捕效果最好,优于单元组分和二元组分的配方;其中,三种成分配置的复合引诱剂中以三种成分比例为1:2:2时的引诱效果最好,在大粒横沟象的监测和防控方面有重要的应用价值。
实施例4信息素引诱剂最佳释放载体筛选
采用实施例3中处理组10制备的大粒横沟象信息素引诱剂,分别采用不同的缓释载体,包括橡胶塞、聚乙烯瓶和聚乙烯缓释袋三种释放载体,按照与实施例3相同的方法分别制成诱芯,放入与实施例3相同结构的诱捕装置中进行试验,诱捕器之间的平均间距为50m,诱芯高度设置为离地面高度1.5m,诱捕器类型为网状诱捕器,3次重复。诱捕器放置后,每隔3天调查诱捕到的大粒横沟象数量,并随机更换诱捕器的位置,持续调查一个月,结果如表3。
表3不同释放载体对大粒横沟象诱捕情况
平均诱捕量(头/诱捕器) | |
橡胶塞 | 33.67±0.57b |
聚乙烯瓶 | 56.33±0.32a |
聚乙烯缓释袋 | 40.00±0.26b |
从表3中可以看出,大粒横沟象信息素引诱剂以聚乙烯瓶载体最佳。
实施例5诱捕器最佳悬挂高度筛选
采用实施例4中的大粒横沟象聚乙烯瓶信息素引诱剂,研究不同悬挂高度对诱捕效果的影响。
试验方法:按照与实施例3处理组10相同的方法分别制成诱芯,放入与实施例3相同结构的诱捕装置中进行试验,诱捕器之间的平均间距为50m,诱捕器类型为网状诱捕器,3次重复。诱捕器放置后,每隔3天调查诱捕到的大粒横沟象数量,并随机更换诱捕器的位置,持续调查一个月,结果如表4。
表4诱捕器不同悬挂高度下大粒横沟象的诱捕情况
平均诱捕量(头/诱捕器) | |
1m | 42±0.27b |
1.5m | 57.33±0.23ab |
2.0m | 63±0.43a |
从表4中可以看出,诱捕器悬高度高于1.5m诱捕效果较好。
实施例6信息素引诱剂对大粒横沟象雌、雄成虫诱捕能力对比
采用实施例3中的大粒横沟象聚乙烯瓶信息素引诱剂,分析引诱剂对大粒横沟象雌、雄成虫的诱捕能力。
试验方法:按照与实施例3处理组10相同的方法分别制成诱芯,放入与实施例3相同结构的诱捕装置中进行试验,诱捕器之间的平均间距为50m,诱芯高度设置为离地面高度1.5m,诱捕器类型为网状诱捕器,共设置8套诱捕器。诱捕器放置后,每隔3天调查诱捕到的大粒横沟象数量,并随机更换诱捕器的位置,持续调查一个月,结果如表5、图2所示。
表5信息素引诱剂对大粒横沟象雌、雄成虫诱捕统计表
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 总数 | |
雌成虫 | 43 | 28 | 15 | 24 | 40 | 33 | 41 | 35 | 259 |
雄成虫 | 39 | 34 | 21 | 30 | 37 | 30 | 39 | 40 | 270 |
从表5和图2中数据分析得出,本发明大粒横沟象信息素引诱剂对大粒横沟象雌雄成虫之间的诱捕无显著性差异(P>0.05),说明大粒横沟象信息素引诱剂可同时实现对雌、雄成虫的诱杀,可满足大粒横沟象信息素大范围的防治。
Claims (8)
1.一种大粒横沟象信息素引诱剂,其特征在于,所述引诱剂包括香叶醇乙酯、顺-9-十八碳烯醇和顺-2-(3,3-二甲基亚环乙基)乙酸乙酯;
所述香叶醇乙酯、顺-9-十八碳烯醇、顺-2-(3,3-二甲基亚环乙基)乙酸乙酯的质量比为1:1~3:1~3。
2.根据权利要求1所述的引诱剂,其特征在于,所述香叶醇乙酯、顺-9-十八碳烯醇、顺-2-(3,3-二甲基亚环乙基)乙酸乙酯的质量比为1:2:2。
3.权利要求1或2所述的大粒横沟象信息素引诱剂在大粒横沟象的生物防治中的应用。
4.根据权利要求3所述的应用,其特征在于,所述应用为将香叶醇乙酯、顺-9-十八碳烯醇和顺-2-(3,3-二甲基亚环乙基)乙酸乙酯加入到载体中制得大粒横沟象信息素引诱剂诱芯使用。
5.根据权利要求4所述的应用,其特征在于,所述载体为聚乙烯管、聚氯乙烯管、聚乙烯瓶、聚乙烯缓释袋、聚氯乙烯缓释袋、橡胶塞中的任意一种。
6.根据权利要求5所述的应用,其特征在于,所述载体为聚乙烯瓶。
7.根据权利要求4所述的应用,其特征在于,将所述引诱剂悬挂于大粒横沟象危害区域高于1.5m的高度。
8.根据权利要求4所述的应用,其特征在于,每个所述诱芯含有5000ug所述信息素引诱剂,所述诱芯之间的平均间距为50m。
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