基于远程寄主定位机理的假眼小绿叶蝉化学生态和物理调控
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摘要
假眼小绿叶蝉（Empoasca vitis G the）是我国茶树的重要害虫，防治难度很大，现阶段以化学农药防治为主。环境保护和食品安全问题的日益突出，使得假眼小绿叶蝉的无公害防治越来越被重视。本文基于假眼小绿叶蝉远程寄主定位的视、嗅觉机理，通过研究假眼小绿叶蝉对特定光、颜色和植物挥发物的趋性，筛选出对假眼小绿叶蝉具有显著诱集作用的光、颜色以及植物挥发物引诱剂，最后在田间采用诱虫灯、颜色诱捕器和植物挥发物引诱剂对假眼小绿叶蝉进行诱杀，为茶园中假眼小绿叶蝉的无公害防治提供参考。
首先通过扫描电镜对叶蝉的视、嗅觉器官进行观察，并结合生测实验明确假眼小绿叶蝉远程寄主定位中视觉和嗅觉的作用和关系，以及视觉刺激对叶蝉寄主定位的干扰作用，结果显示叶蝉在远程寄主定位中主要依靠视觉和嗅觉，其中视觉在叶蝉寄主定位中起主导作用，外界的颜色和光对叶蝉的寄主定位具有强烈的干扰作用。
利用色差计和颜色分析方法研究了寄主植物茶树的颜色特征，茶枝上不同叶龄的叶片颜色具有显著差异，叶龄越大叶片颜色中的黄色成分越低。叶蝉对特殊的光和颜色表现出显著的趋性，在RGB颜色模式下，采用正交试验设计对颜色的诱捕色进行筛选，金色(RGB:255,255,0)对叶蝉的吸引作用最强；叶蝉对不同波长的单色光趋性不同，其中长波紫外光和紫光对叶蝉表现出显著的吸引作用。
利用HS-SPME-GC-MS和顶空吸附法鉴定了不同生理状态的茶梢挥发物的释放特征，嫩叶丰富的茶梢以萜烯类挥发物为主，β-linalool的相对释放量最大，成熟叶丰富的茶梢以绿叶挥发物为主，(Z)-3-hexen-1-ol和(Z)-3-hexen-1-ol acetate的相对释放量最大，挥发物释放总量最高；老叶较多的茶梢β-linalool相对释放量最低，挥发物释放总量也最小；昼夜茶梢挥发物的成分与释放量具有显著差别，茶梢在白天释放的挥发物种类多且总量大，以酯类和大分子萜烯类物质为主，夜间挥发物释放量降低，以小分子萜烯类物质为主；茶梢被叶蝉持续危害后，挥发物的释放特征会发生明显的变化，一方面一些挥发物的释放量明显增大，另一方面诱导型挥发物大量释放，诱导型挥发物的释放具有明显的昼夜节律，夜间的释放量比白天显著降低，并且不同挥发物的释放高峰期也不一致。风洞生测实验显示叶蝉对不同生理状态的茶梢挥发物的趋性差异显著，其中对嫩叶丰富的未受虫害的茶梢趋性最大。基于茶梢挥发物的成分，通过Y型嗅觉仪对叶蝉的挥发物引诱剂进行筛选，结果显示挥发物的成分以及成分之间的比例是影响引诱剂效果的关键因素，(Z)-3-hexen-1-ol、(Z)-3-hexenyl acetate和β-linalool以合适的比例混合后对叶蝉具有显著的吸引效果。
缓释材料是挥发物引诱剂田间应用的基础，采用溶胶-凝胶法以四甲基正硅烷和甲基三甲氧基硅烷为单聚体，筛选出对植物挥发物具有良好缓释能力的合成材料，并比较了溶剂萃取法、动态吸附法和电子鼻法在植物挥发物缓释速率测定上的特点，其中动态吸附法的精确性和准确度最好。
最后，通过田间试验对诱虫灯、色板和挥发物引诱剂的效果进行了测定，紫光LED单波诱虫灯和金色数字化色板对叶蝉具有显著的诱捕效果，单波诱虫灯与常规杀虫灯相比显著降低了对天敌的诱杀数量，数字化色板与市售黄板相比显著提高了对叶蝉的诱杀数量；(Z)-3-hexen-1-ol、(Z)-3-hexen-1-ol acetate、β-linalool、ocimene和nonanal以合适成分及比例混合后配制的引诱剂对田间叶蝉有显著的生态调控功能，并对色板诱杀叶蝉起到显著的增效作用。
Empoasca vitis G the is a major pest and difficult to control in Chinese tea gardens. Application ofpesticides is the main way to control this pest. With the increasingly prominent situation inenvironmental pollution and food safety, people have attached more attention to the pollution-freecontrol of tea leafhoppers. In the paper, based on the mechanism of olfaction and vision in thelong-range hostlocation, the tropism of leafhoppers to monochromatic light, color and plant volatile wasstudied. The light trap, color trap and plant volatile attractant, which were significant attractive toleafhoppers, used to trap leafhoppers in the field would provide reference for the pollution-free controlof leafhoppers in tea gardens.
The function and relationship between olfaction and vision and the interference effect of visualstimulation in the long-range hostlocation process of leafhoppers were investigated through theobservation of visual and olfactory organs with scanning electron microscope and the relative bioassays.The results showed that the long-range hostloction of leafhoppers depended mostly on the vision andsmell. The vision played leading roles in this process, and the ambient color and light showed stronginterference. The color character of host plant was studied by the color-difference meter and coloranalysis method. The colors of leaves in different leafage had significantly differences. The leaves ofolder leafage had lower level of yellow intensity. Leafhoppers exhibited highly significant tropism tospecific light and color. With RGB color mode, the most attractive color was screened by the orthogonalexperimental design. The color gold (RGB:255,255,0) was the most attractive color to leafhoppers.Moreover, light of different wavelengths exhibited different attraction, and UVA and violet light weremore attractive to leafhoppers.
The release characteristics of volatile compounds in tea plant under varied physical condition wasdetermined by HS-SPME-GC-MS and headspace absorption method. Firstly, the tea shoots with moretender leaves released more terpenoids, in which the amount of β-linalool was the highest. Secondly, thetea shoots with more mature leaves released more green leaf volatiles, in which the amount of(Z)-3-hexen-1-oland (Z)-3-hexen-1-ol acetate were the highest. Finally, the amount of total volatilesreleased by tea shoots with more old leaves was minimum and the amount of β-linalool was the lowest.The composition and release rates of tea plant volatiles in day and night were significantly different inday and night. On one hand, the tea shoots released more types and larger amounts of volatiles duringthe day, mainly esters and macromolecular terpenoids. On the other hand, volatiles released in the nightwere mainly micromolecular terpenoids and the total amounts were lower. Visible changes in thereleasing characteristics of volatiles would happen when tea shoots were damaged by leafhoppers. Therelease amount of total volatiles and induced volatiles clearly increased. The induced volatiles releasedwith a clear day-night rhythm, the release amounts during night was lower than those in daytime.Various volatiles showed different releasing peak period. Tea leafhoppers showed significantly differenttropism to tea shoots with various physiological status. The most attractive tea shoots had more tenderleaves. Based on the composition of volatiles and Y shaped olfactometer bioassays, the key factorsaffected the attractant effects were the composition and the proportion of volatiles. The attractant mixed with (Z)-3-hexen-1-ol、(Z)-3-hexenyl acetate and β-linalool in appropriate proportions could attract tealeafhoppers.
Slow release material was the foundation for the application of volatile attractants in the field. Thesuitable formula for plant volatile attractants was screened with sol-gel method in the lab. Thecomparison of three methods: solvent extraction, dynamic adsorption and zNoseTM, was carried out toselected an suitable method for the releasing characteristic of attractants in the sol-gel dispensers.Results showed that the dynamic adsorption method exhibited better precision and accuracy.
The effects of light trap, color trap and attractants in capturing tea leafhoppers were investigatedunder field experiments. Compared with conventional light trap, violet monochromatic LED light trapcaptured less natural enemies. Compared with yellow sticky trap obtained from the market, gold stickytrap captured more leafhoppers. The attractants, mixed with (Z)-3-hexen-1-ol、(Z)-3-hexenyl acetate,β-linalool, ocimene and nonanal in appropriate formulas and proportions, showed an effectiveecological regulating function for tea leafhoppers in the field.
首先通过扫描电镜对叶蝉的视、嗅觉器官进行观察，并结合生测实验明确假眼小绿叶蝉远程寄主定位中视觉和嗅觉的作用和关系，以及视觉刺激对叶蝉寄主定位的干扰作用，结果显示叶蝉在远程寄主定位中主要依靠视觉和嗅觉，其中视觉在叶蝉寄主定位中起主导作用，外界的颜色和光对叶蝉的寄主定位具有强烈的干扰作用。
利用色差计和颜色分析方法研究了寄主植物茶树的颜色特征，茶枝上不同叶龄的叶片颜色具有显著差异，叶龄越大叶片颜色中的黄色成分越低。叶蝉对特殊的光和颜色表现出显著的趋性，在RGB颜色模式下，采用正交试验设计对颜色的诱捕色进行筛选，金色(RGB:255,255,0)对叶蝉的吸引作用最强；叶蝉对不同波长的单色光趋性不同，其中长波紫外光和紫光对叶蝉表现出显著的吸引作用。
利用HS-SPME-GC-MS和顶空吸附法鉴定了不同生理状态的茶梢挥发物的释放特征，嫩叶丰富的茶梢以萜烯类挥发物为主，β-linalool的相对释放量最大，成熟叶丰富的茶梢以绿叶挥发物为主，(Z)-3-hexen-1-ol和(Z)-3-hexen-1-ol acetate的相对释放量最大，挥发物释放总量最高；老叶较多的茶梢β-linalool相对释放量最低，挥发物释放总量也最小；昼夜茶梢挥发物的成分与释放量具有显著差别，茶梢在白天释放的挥发物种类多且总量大，以酯类和大分子萜烯类物质为主，夜间挥发物释放量降低，以小分子萜烯类物质为主；茶梢被叶蝉持续危害后，挥发物的释放特征会发生明显的变化，一方面一些挥发物的释放量明显增大，另一方面诱导型挥发物大量释放，诱导型挥发物的释放具有明显的昼夜节律，夜间的释放量比白天显著降低，并且不同挥发物的释放高峰期也不一致。风洞生测实验显示叶蝉对不同生理状态的茶梢挥发物的趋性差异显著，其中对嫩叶丰富的未受虫害的茶梢趋性最大。基于茶梢挥发物的成分，通过Y型嗅觉仪对叶蝉的挥发物引诱剂进行筛选，结果显示挥发物的成分以及成分之间的比例是影响引诱剂效果的关键因素，(Z)-3-hexen-1-ol、(Z)-3-hexenyl acetate和β-linalool以合适的比例混合后对叶蝉具有显著的吸引效果。
缓释材料是挥发物引诱剂田间应用的基础，采用溶胶-凝胶法以四甲基正硅烷和甲基三甲氧基硅烷为单聚体，筛选出对植物挥发物具有良好缓释能力的合成材料，并比较了溶剂萃取法、动态吸附法和电子鼻法在植物挥发物缓释速率测定上的特点，其中动态吸附法的精确性和准确度最好。
最后，通过田间试验对诱虫灯、色板和挥发物引诱剂的效果进行了测定，紫光LED单波诱虫灯和金色数字化色板对叶蝉具有显著的诱捕效果，单波诱虫灯与常规杀虫灯相比显著降低了对天敌的诱杀数量，数字化色板与市售黄板相比显著提高了对叶蝉的诱杀数量；(Z)-3-hexen-1-ol、(Z)-3-hexen-1-ol acetate、β-linalool、ocimene和nonanal以合适成分及比例混合后配制的引诱剂对田间叶蝉有显著的生态调控功能，并对色板诱杀叶蝉起到显著的增效作用。
Empoasca vitis G the is a major pest and difficult to control in Chinese tea gardens. Application ofpesticides is the main way to control this pest. With the increasingly prominent situation inenvironmental pollution and food safety, people have attached more attention to the pollution-freecontrol of tea leafhoppers. In the paper, based on the mechanism of olfaction and vision in thelong-range hostlocation, the tropism of leafhoppers to monochromatic light, color and plant volatile wasstudied. The light trap, color trap and plant volatile attractant, which were significant attractive toleafhoppers, used to trap leafhoppers in the field would provide reference for the pollution-free controlof leafhoppers in tea gardens.
The function and relationship between olfaction and vision and the interference effect of visualstimulation in the long-range hostlocation process of leafhoppers were investigated through theobservation of visual and olfactory organs with scanning electron microscope and the relative bioassays.The results showed that the long-range hostloction of leafhoppers depended mostly on the vision andsmell. The vision played leading roles in this process, and the ambient color and light showed stronginterference. The color character of host plant was studied by the color-difference meter and coloranalysis method. The colors of leaves in different leafage had significantly differences. The leaves ofolder leafage had lower level of yellow intensity. Leafhoppers exhibited highly significant tropism tospecific light and color. With RGB color mode, the most attractive color was screened by the orthogonalexperimental design. The color gold (RGB:255,255,0) was the most attractive color to leafhoppers.Moreover, light of different wavelengths exhibited different attraction, and UVA and violet light weremore attractive to leafhoppers.
The release characteristics of volatile compounds in tea plant under varied physical condition wasdetermined by HS-SPME-GC-MS and headspace absorption method. Firstly, the tea shoots with moretender leaves released more terpenoids, in which the amount of β-linalool was the highest. Secondly, thetea shoots with more mature leaves released more green leaf volatiles, in which the amount of(Z)-3-hexen-1-oland (Z)-3-hexen-1-ol acetate were the highest. Finally, the amount of total volatilesreleased by tea shoots with more old leaves was minimum and the amount of β-linalool was the lowest.The composition and release rates of tea plant volatiles in day and night were significantly different inday and night. On one hand, the tea shoots released more types and larger amounts of volatiles duringthe day, mainly esters and macromolecular terpenoids. On the other hand, volatiles released in the nightwere mainly micromolecular terpenoids and the total amounts were lower. Visible changes in thereleasing characteristics of volatiles would happen when tea shoots were damaged by leafhoppers. Therelease amount of total volatiles and induced volatiles clearly increased. The induced volatiles releasedwith a clear day-night rhythm, the release amounts during night was lower than those in daytime.Various volatiles showed different releasing peak period. Tea leafhoppers showed significantly differenttropism to tea shoots with various physiological status. The most attractive tea shoots had more tenderleaves. Based on the composition of volatiles and Y shaped olfactometer bioassays, the key factorsaffected the attractant effects were the composition and the proportion of volatiles. The attractant mixed with (Z)-3-hexen-1-ol、(Z)-3-hexenyl acetate and β-linalool in appropriate proportions could attract tealeafhoppers.
Slow release material was the foundation for the application of volatile attractants in the field. Thesuitable formula for plant volatile attractants was screened with sol-gel method in the lab. Thecomparison of three methods: solvent extraction, dynamic adsorption and zNoseTM, was carried out toselected an suitable method for the releasing characteristic of attractants in the sol-gel dispensers.Results showed that the dynamic adsorption method exhibited better precision and accuracy.
The effects of light trap, color trap and attractants in capturing tea leafhoppers were investigatedunder field experiments. Compared with conventional light trap, violet monochromatic LED light trapcaptured less natural enemies. Compared with yellow sticky trap obtained from the market, gold stickytrap captured more leafhoppers. The attractants, mixed with (Z)-3-hexen-1-ol、(Z)-3-hexenyl acetate,β-linalool, ocimene and nonanal in appropriate formulas and proportions, showed an effectiveecological regulating function for tea leafhoppers in the field.
引文
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