CN112219850B - 一种蓟马引诱剂与杀虫剂联合诱杀方法 - Google Patents

一种蓟马引诱剂与杀虫剂联合诱杀方法 Download PDF

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CN112219850B
CN112219850B CN202011252023.3A CN202011252023A CN112219850B CN 112219850 B CN112219850 B CN 112219850B CN 202011252023 A CN202011252023 A CN 202011252023A CN 112219850 B CN112219850 B CN 112219850B
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田厚军
魏辉
陈勇
陈艺欣
林硕
林涛
曾兆华
游泳
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Abstract

本发明提供一种蓟马引诱剂与杀虫剂联合诱杀方法,引诱剂是以液体石蜡为溶剂,将100μg/ml的烟酸甲酯、反‑β‑法尼烯、辛醛、3‑蒈烯、异丁香酚、β‑苯乙醇进行相同质量混合而成。本发明诱杀剂含有特定的寄主植物源挥发性物质,种类多样,作用靶标位点广,互补性强,通过一定比例配制成对蓟马具有较强诱集作用的复合物,诱集蓟马到粘虫色板上,同时在粘虫板上喷施蓟马类害虫高效低残留杀虫剂,利用杀虫剂的毒性将蓟马杀死,彻底将蓟马消灭,防止二次逃脱为害。本发明诱杀剂具有成本低廉、环保高效、不易产生抗药性等特点;而且诱集物质来源于植物,安全可靠,可在蔬菜、果树等作物上广泛应用,具有广阔的应用前景。

Description

一种蓟马引诱剂与杀虫剂联合诱杀方法
技术领域
本发明涉及农林昆虫生物技术领域,具体地说,涉及一种蓟马引诱剂与杀虫剂联合诱杀方法。
背景技术
蓟马是缨翅目(Thysanoptera)昆虫的统称,虫体微小,常隐匿于花与嫩叶中为害。目前,全世界已记录的蓟马达7700 余种,中国占570 余种,是果蔬、花卉等多种农作物的重要害虫,也是一种具有危险性的世界性海关检疫害虫,可危害600多种植物,严重为害多种重要的农业经济作物。
目前国内外对蓟马的防治仍然主要采用化学防治方法。但是,化学药剂的大量使用给人类健康和环境造成极大危害。另外,西花蓟马和棕榈蓟马这两种蓟马对多种化学药剂产生抗性,因而导致单一的化学防治方法难以取得理想的防控效果。
蓟马类害虫种类繁多,至今还未找到对多种蓟马类害虫有效的诱杀组分。植物-昆虫信号联系是当前科学研究的热点问题。植食性昆虫通过叶子、花朵和果实散发出的化学线索来定位寄主植物。识别寄主植物对于植食性昆虫满足其营养需求、寻找合适的产卵场所、以及成功交配至关重要。在自然界中,植物释放的挥发物有数百种之多,但是真正起作用的可能是一种或几种,以某种比例混合(Buttery and Ling,1984)。对害虫起作用的主导化合物中每一种成分都与昆虫触角上相应的嗅觉气味结合蛋白受体在某种程度上具有较强的结合特性,而且还存在结合阈值,一旦超过这个阈值,效果可能会适得其反,因此随意增加某种化合物的配比含量都会影响最终的诱集效果。由于实际环境下很难模仿自然条件下植物释放的挥发物之间的比例,因此,本发明人在实验室内将尽量筛选主要化合物的单体诱集活性,通过大量的组合配比,最终确定对西花蓟马和棕榈蓟马具有高效诱集作用的有效配方,结合杀虫剂的毒杀作用,将蓟马杀死,防止其二次逃脱。
发明内容
本发明提供一种蓟马引诱剂与杀虫剂联合诱杀方法,引诱剂是以液体石蜡为溶剂,将100μg/ml的烟酸甲酯、反-β-法尼烯、辛醛、3-蒈烯、异丁香酚、β-苯乙醇进行相同质量混合而成。本发明诱杀剂含有特定的寄主植物源挥发性物质,通过一定比例配制成对蓟马具有较强诱集作用的复合物,诱集蓟马到粘虫色板上,同时在粘虫板上喷施蓟马类害虫常用杀虫剂,利用杀虫剂的毒性将蓟马杀死,彻底将蓟马消灭,防止二次逃脱为害。本发明诱杀剂具有成本低廉、环保高效、不易产生抗药性等特点;而且诱集物质来源于植物,安全可靠,可在蔬菜、果树等作物上广泛应用,具有广阔的应用前景。
为实现上述目的,本发明采用如下技术方案:
一种蓟马引诱剂与杀虫剂联合诱杀方法,将引诱剂悬挂于喷施有杀虫剂的粘虫色板中上部,通过引诱剂诱集蓟马到粘虫色板上,同时在粘虫板上喷施杀虫剂,利用杀虫剂的毒性将蓟马杀死,彻底将蓟马消灭,防止二次逃脱为害;所述的引诱剂是以液体石蜡为溶剂,将100μg/ml的烟酸甲酯、反-β-法尼烯、辛醛、3-蒈烯、异丁香酚和β-苯乙醇的单一组分溶液按相同质量混合而制得。
所述的蓟马引诱剂和杀虫剂联合诱杀方法,具体包括如下步骤:
(1)100μg/ml的单一组分溶液的制备
100μg/ml的烟酸甲酯溶液的制备:将事先配制好的10μg/μl的烟酸甲酯标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的反-β-法尼烯溶液的制备:将事先配制好的10μg/μl的反-β-法尼烯标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的辛醛溶液的制备:将事先配制好的10μg/μl的辛醛标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的3-蒈烯溶液的制备:将事先配制好的10μg/μl的3-蒈烯标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的异丁香酚溶液的制备:将事先配制好的10μg/μl的异丁香酚标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的β-苯乙醇溶液的制备:将事先配制好的10μg/μl的β-苯乙醇标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
(2)将步骤(1)得到的100μg/ml的烟酸甲酯溶液、反-β-法尼烯溶液、辛醛溶液、3-蒈烯溶液、异丁香酚溶液和β-苯乙醇溶液进行相同质量混合,制得引诱剂;
(3)将步骤(2)获得的蓟马引诱剂装于5ml离心管中,离心管悬挂于喷施有杀虫剂的粘虫色板中上部。
所述的离心管上盖为数个直径为0.1mm-1mm的圆孔。
本发明的显著优点在于:(1)筛选的物质中包含酯类、醇类、萜烯类、醛类和酚类物质,作用靶标位点广,而且每种物质都是从众多同类成分中筛选的,可以起到增效互补的作用;(2)对环境安全、无污染,引诱剂组分均为植物源天然化合物,不容易引起昆虫产生抗性;且在诱杀昆虫的过程中不存在化学杀虫剂给生态环境带来污染的问题;(3)本发明研制的引诱剂不仅能高效诱集多种蓟马类害虫,而且与杀虫剂联合诱杀,能彻底将蓟马消灭,防止二次逃脱为害。
附图说明
图1为西花蓟马成虫对各处理的定向行为反应示意图;
图2为棕榈蓟马成虫对各处理的定向行为反应示意图;不同小写字母表示蓟马的定向选择行为反应各处理间差异显著(P<0.05);
图3为采用不同处理的诱杀剂处理3天时对棕榈蓟马的诱集数量;
图4为采用不同处理的诱杀剂处理7天时对棕榈蓟马的诱集数量;
图5为采用不同处理的诱杀剂处理14天时对棕榈蓟马的诱集数量,不同小写字母表示处理和对照间诱集量差异显著(P<0.05)。
具体实施方式
本发明用下列实施例来进一步说明本发明,但本发明的保护范围并不限于下列实施例。
实施例1
一、蓟马诱杀剂的制备方法
1.1 100μg/ml的单一组分溶液的制备
100μg/ml的烟酸甲酯溶液的制备:将事先配制好的10μg/μl的烟酸甲酯标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的反-β-法尼烯溶液的制备:将事先配制好的10μg/μl的反-β-法尼烯标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的辛醛溶液的制备:将事先配制好的10μg/μl的辛醛标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的3-蒈烯溶液的制备:将事先配制好的10μg/μl的3-蒈烯标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的异丁香酚溶液的制备:将事先配制好的10μg/μl的异丁香酚标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的β-苯乙醇溶液的制备:将事先配制好的10μg/μl的β-苯乙醇标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
1.2蓟马引诱剂的制备
将步骤(1)得到的100μg/ml的烟酸甲酯溶液、反-β-法尼烯溶液、辛醛溶液、3-蒈烯溶液、异丁香酚溶液和β-苯乙醇溶液进行相同质量混合,制得本发明引诱剂;
1.3蓟马引诱剂配方1的制备
将步骤(1)得到的100μg/ml的烟酸甲酯溶液、反-β-法尼烯溶液、辛醛溶液、3-蒈烯溶液、异丁香酚溶液和β-苯乙醇溶液按照质量比2:2:1:1:2:1混合,制得引诱剂配方1;
1.4蓟马引诱剂配方2的制备
将步骤(1)得到的100μg/ml的烟酸甲酯溶液、反-β-法尼烯溶液、辛醛溶液、3-蒈烯溶液、异丁香酚溶液和β-苯乙醇溶液按照质量比1:1:2:2:1:2混合,制得引诱剂配方2;
将获得的本发明蓟马引诱剂、配方1、配方2分别装于5ml离心管中,离心管悬挂于喷施有杀虫剂的粘虫色板中上部。
二、蓟马对各组分及本发明引诱剂、配方1和配方2的触角电位反应
20µl样品溶液均匀滴在长5cm、宽0.5cm的“V”字形滤纸条上,放入巴斯德滴管中,滴管末端连接刺激气流控制装置,送气管管口与触角纵向垂直,并与触角相距1cm。调节连续气体流量为124ml/min,刺激气体流量为20ml/min,每次刺激时间为0.2s。两次刺激之间间隔40s,以保证触角感觉器官功能的完全恢复。各组分及复合物在一根触角上平行刺激3次取平均值,至少重复试验5根触角。各组分及复合物的EAG相对反应值以液体石蜡作为对照。对于每一待测样品,刺激顺序为液体石蜡、待测样品、液体石蜡。前后两次对照取平均值。
EAG相对反应值=待测样品的EAG相对反应值-液体石蜡平均EAG相对反应值。
触角电位反应结果表明,如表1所示,西花蓟马和棕榈蓟马对所测的6种单个组分均具有较强的触角电位反应(相对反应最高值为烟酸甲酯-110.15μV,最低值为3-蒈烯-53.31μV),西花蓟马对烟酸甲酯、反-β-法尼烯、辛醛、异丁香酚的EAG反应值略高于棕榈蓟马。西花蓟马对本发明引诱剂的触角电位反应相对值结果达到了-215.90μV,显著高于对配方1(-164.56μV)和配方2(-155.31μV)的EAG反应值;棕榈蓟马对本发明引诱剂的触角电位反应相对值结果达到了-194.90μV,高于对配方1(-158.24μV),显著高于配方2(-149.51μV)的EAG反应值。
表1 西花蓟马和棕榈蓟马对单一组分及本发明引诱剂、配方1、配方2的触角电位反应,不同小写字母表示不同处理间差异显著(P<0.05)
Figure DEST_PATH_IMAGE002
实施例2
采用Y型嗅觉仪测定棕榈蓟马对复合物的定向行为反应(由本实验室根据嗅觉仪基本原理进行改进,授权专利号为:ZL201020239767.7)。Y 型管主臂 20cm,两臂长度各为10cm,两臂夹角 90℃。两臂管末用硅胶管按序接上蒸馏水瓶和活性炭瓶,最后活性炭瓶开口于大气中,昆虫释放瓶处用纱布连于抽气装置。将滤纸条剪成5 cm × 1 cm,折成“V”形,置于陷阱入口玻璃管中。
将Y形嗅觉仪倾斜30°,温度控制在25±2℃。分别取100μl本发明引诱剂、配方1、配方2引诱剂均匀滴于滤纸条上放入一边的味源瓶中,另一边以滴有100μl的液体石蜡的滤纸作为参照,抽气气流流速为1L/min,分别进入两管的气流经过活性炭过滤和蒸馏水加湿。取3日龄西花蓟马或者棕榈蓟马成虫置于昆虫释放瓶中,西花蓟马和棕榈蓟马每次各30头,分开测定,重复3次,每次测完即用棉球蘸取无水乙醇Y形管内外壁并烘干,然后调换Y形管两臂与味源瓶、对照,以消除两臂的几何误差对蓟马行为可能造成的影响。将试虫放入Y形管的昆虫释放瓶中开始试验,每头试虫爬过选择臂1/2处,停留10 s以上,即算做出选择,停留在Y形管主臂上的蓟马均视为无反应,统计30 min内的实验结果。结果表明,本发明引诱剂、配方1、配方2引诱剂对西花蓟马的引诱量为19.0±1头、14.33±2.08头、13±1头,本发明引诱剂的诱集量显著高于配方1、配方2和对照(如图1所示);本发明引诱剂、配方1、配方2对棕榈蓟马的引诱量分别为17.0±1头、12±1头、10.67±2.08头,本发明引诱剂诱集量显著高于配方1、配方2和对照(如图2所示)。
由图1和图2结果可以看出,本发明引诱剂对蓟马具有显著的引诱作用。通过对花蓟马、大蓟马等蓟马的室内行为选择试验也有类似的引诱效果。
实施例3
试验地点选择福建省闽侯果园大棚,农作物为草莓。采用蓝色粘虫板作为诱捕装置,将本发明引诱剂、配方1、配方2,分别置于离心管中,悬挂于色板上,离心管上盖为5个直径0.2mm的圆孔。以蓝色粘虫板悬挂含有纯液体石蜡的离心管为对照,以悬挂诱集剂和喷施乙基多杀菌素的蓝色粘虫板为处理,设3个重复,不同处理之间随机排列,将色板悬挂于植株上方20 cm,跨度为6-8 m,每棚悬挂3列,列间距1.5 m,色板间距3 m。分别调查悬挂诱虫板3d、7 d、14d的结果,记录数据后换上新的色板,每小区选取3点,分别调查色板上棕榈蓟马的数量,计算每块色板的诱虫量。
如图3-图5所示,结果表明,悬挂本发明引诱剂的蓝板分别在3d、7d、14d对棕榈蓟马的诱集量显著高于配方2和蓝板对照,特别是7d、14d时诱集的蓟马多数在引诱剂复合物周围,位于蓝板的中上部及边缘较为密集,对照蓝板上尚有20%蓟马未完全死亡,随时有可能借助外力而挣扎脱离蓝板束缚,而喷施有乙基多杀菌素的蓝板上蓟马全部死亡。本发明引诱剂在第3天对棕榈蓟马的诱集量高于配方1的诱集量,但未达到显著水平,但是本发明引诱剂在第7d和14d对棕榈蓟马的诱集量显著高于配方1、配方2的诱集量(图4、图5)。
以上所述仅作为本发明的实施方式和例子,在本专业技术领域范围内并不脱离本发明原理的前提下,对其进行等效变换以及改进,均视为本发明的保护范围。

Claims (3)

1.一种蓟马引诱剂与杀虫剂联合诱杀方法,其特征在于:将引诱剂悬挂于喷施有杀虫剂的粘虫色板中上部,通过引诱剂诱集蓟马到粘虫色板上,利用杀虫剂的毒性将蓟马杀死,彻底将蓟马消灭,防止二次逃脱为害;
所述引诱剂是以液体石蜡为溶剂,将100μg/ml的烟酸甲酯、反-β-法尼烯、辛醛、3-蒈烯、异丁香酚和β-苯乙醇的单一组分溶液按相同质量混合而制得。
2.根据权利要求1所述的蓟马引诱剂与杀虫剂联合诱杀方法,其特征在于:具体包括如下步骤:
(1)100μg/ml的单一组分溶液的制备
100μg/ml的烟酸甲酯溶液的制备:将事先配制好的10μg/μl的烟酸甲酯标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的反-β-法尼烯溶液的制备:将事先配制好的10μg/μl的反-β-法尼烯标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的辛醛溶液的制备:将事先配制好的10μg/μl的辛醛标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的3-蒈烯溶液的制备:将事先配制好的10μg/μl的3-蒈烯标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的异丁香酚溶液的制备:将事先配制好的10μg/μl的异丁香酚标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
100μg/ml的β-苯乙醇溶液的制备:将事先配制好的10μg/μl的β-苯乙醇标准化合物取10μl加入到990μl的液体石蜡中,并均匀混合;
(2)将步骤(1)得到的100μg/ml的烟酸甲酯溶液、反-β-法尼烯溶液、辛醛溶液、3-蒈烯溶液、异丁香酚溶液和β-苯乙醇溶液进行相同质量混合,制得引诱剂;
(3)将步骤(2)获得的蓟马引诱剂装于5ml离心管中,离心管悬挂于喷施有杀虫剂的粘虫色板中上部。
3.根据权利要求2所述的蓟马引诱剂与杀虫剂联合诱杀方法,其特征在于:所述的离心管上盖为数个直径0.1mm-1mm的圆孔。
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