暗黑鳃金龟体内章鱼胺的时空分析及功能研究

发布时间：2018-07-17 02:35

【摘要】：暗黑鳃金龟Holotrichia parallela是一种世界性分布的重要地下害虫。其食性广杂,幼虫取食为害寄主植株幼苗的根系、咬食嫩茎和其他地下组织,成虫取食寄主的叶片、花等地上器官。近几年,随着免耕等种植模式的变更,暗黑鳃金龟在我国的发生日益严重,对农作物造成极大的危害。目前,随着人们对农药残留、食品及环境安全的关注,生物防治在控制地下害虫中的地位越来越重要。性信息素作为一种重要的生物防治手段已被研发并应用于田间防治暗黑鳃金龟。有研究发现,植物挥发物对于性信息素存在协同增效作用。作者针对章鱼胺在暗黑鳃金龟生长、繁殖中的作用进行了研究,为揭示章鱼胺的调控作用机理,以及寄主植物挥发物对于性信息素的增效机制提供依据。首先,筛选建立了一种简便、高效、快捷的成虫体内章鱼胺含量测定方法。然后对暗黑鳃金龟体内章鱼胺的含量和分布进行了分析。最后测定了章鱼胺对暗黑鳃金龟存活、交配、产卵以及卵孵化的影响等。主要试验结果如下:(1)建立了利用高效液相色谱检测暗黑鳃金龟神经组织内章鱼胺含量的方法。经方法学考察,其回归方程具有显著的正相关线性关系、试虫样品内章鱼胺在24 h内稳定性高、重现性良好,加样回收试验测定平均回收率为102.23%,表明该方法适用于暗黑鳃金龟神经组织内章鱼胺含量的准确分析与测定。(2)暗黑鳃金龟在不同生育期、不同性别、不同神经组织内的章鱼胺含量存在显著差异。雌、雄成虫之间,产卵初期的雌虫在腹部神经节和脑部神经组织中的章鱼胺含量均高于同期的雄虫,但在胸部神经节中显著低于雄虫;产卵后期的雌虫在脑部神经组织中章鱼胺含量显著高于雄虫,而在胸部和腹部神经组织中均显著低于雄虫。(3)利用Y型嗅觉仪测定章鱼胺对暗黑鳃金龟雄虫对性信息素的定位能力及选择性。通过Y型嗅觉仪的趋性行为测试表明,2μg/mL章鱼胺处理能够极显著的提高暗黑鳃金龟雄虫对性信息素的定位能力,对性信息素的选择系数显著高于对照组,而其拮抗剂米安色林则减弱了这种作用。(4)章鱼胺能够显著提高暗黑鳃金龟雌成虫单日单雌产卵量,而且可延长产卵期。其中喂食2μg/mL章鱼胺处理的单日单雌产卵量是对照组雌虫产卵量的2倍以上,当对照组停止产卵后,章鱼胺处理组依然保持产卵。章鱼胺对于暗黑鳃金龟雌雄成虫寿命无显著影响,而米安色林则显著延长了雌雄成虫寿命。而不同浓度章鱼胺及其拮抗剂米安色林处理对卵的孵化率无显著影响。
[Abstract]:Holotrichia parallela is a worldwide important underground pest. The larva eats the root system of the host plant seedling, bites the tender stem and other underground tissue, and the adult eats the leaf of the host plant, flower and other aboveground organs. In recent years, with the change of no-tillage and other planting patterns, the dark Gill tortoise is becoming more and more serious in our country, which causes great harm to crops. At present, with the concern of pesticide residues, food and environmental safety, biological control plays an increasingly important role in the control of underground pests. Sex pheromone as an important biological control method has been developed and applied to the field control of black-branched golden tortoise. Some studies have found that plant volatiles have synergistic effects on sex pheromones. In this paper, the role of octopamine in the growth and reproduction of dark branchial golden tortoise was studied in order to reveal the regulatory mechanism of octopine and the synergistic mechanism of host plant volatiles on sex pheromone. Firstly, a simple, efficient and fast method for the determination of octopamine in adults was established. Then the content and distribution of octopamine in the dark-branched golden tortoise were analyzed. Finally, the effects of octopamine on survival, mating, oviposition and egg hatching were determined. The main results are as follows: (1) A high performance liquid chromatography (HPLC) method was established for the determination of octopamine in the nerve tissues of the black Gill tortoise. The regression equation showed a significant positive correlation linear relationship. The octopamine in the sample was stable and reproducible within 24 hours. The average recovery rate was 102.23, which indicated that this method was suitable for the accurate analysis and determination of octopamine content in the nerve tissue of Black-Gill golden tortoise. (2) the content of octopamine was different in different growth stages. There were significant differences in octopamine content in different nerve tissues. The octopamine content in abdominal ganglion and brain nerve tissue of female, male adult and early oviposition was higher than that of male, but lower than that of male in thoracic ganglion. The contents of octopamine in brain nerve tissue of female in late oviposition were significantly higher than that in male. (3) the localization ability and selectivity of octopamine to sex pheromone in males were determined by Y type olfactometer. The sex behavior test of Y type olfactory instrument showed that the treatment of 2 渭 g / mL octopamine could significantly improve the ability of sex pheromone localization, and the selection coefficient of sex pheromone was significantly higher than that of the control group. The antagonist Mianserin weakened the effect. (4) Octopamine could significantly increase the number of female oviposition per day and prolong the oviposition period. The oviposition of single female treated with 2 渭 g / mL octopamine on a single day was more than 2 times as much as that of the control group. When the control group stopped oviposition, the oviposition was maintained in the octopamine treatment group. Octopamine had no significant effect on male and female adult life, but mianserin significantly prolonged male and female adult life. However, different concentrations of octopamine and its antagonist Mianserin had no significant effect on the hatching rate of eggs.
【学位授予单位】：中国农业科学院
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S433

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