植物次生代谢产物与化学合成杀虫剂相比，具有对哺乳动物低毒，环境友好，昆虫不易产生抗性等优点。吴茱萸（Evodia rutaecarpa）为传统中药材，可与动物类药材共贮防虫，以此为切入点，本文利用化学生态学和地理学相关研究技术，对吴茱萸属（Evodia）植物进行了防治仓储害虫的抗虫活性物质基础研究，驱避活性作用机理研究，以及利用最大熵模型（MaxEnt）和ArcGIS技术基于挥发性抗虫活性物质对吴茱萸进行植物资源分布预测，以期从中药材资源合理开发利用和可持续发展的角度为吴茱萸属植物对仓储害虫的绿色防控提供新的研究思路。

本文主要研究内容和结果如下：

1. 吴茱萸属植物抗虫活性研究

（1）基于抗虫活性的化学成分分析。用水蒸气蒸馏法提取吴茱萸果实等6个样本的挥发油，并通过GC-MS分析化学成分及相对百分含量；对吴茱萸果实等8个植物样本进行甲醇提取物制备，利用高效液相（HPLC）和核磁氢谱（¹H-NMR）技术获取8个样本的主要化合物类型及其丰富程度；根据甲醇提取物谱学信息、活性测试结果和文献调研，从吴茱萸果实的甲醇提取物中选择性分离得到5个特征活性化合物（2个生物碱和3个柠檬苦素类化合物）。分析结果显示，吴茱萸属植物中含有丰富的抗虫活性物质，具有很好的开发利用价值。

（2）三种仓储害虫的抗虫活性研究。对6种挥发油及其8个主要化学成分（β-石竹烯、氧化石竹烯、月桂烯、罗勒烯、柠檬烯、β-蒎烯、3-蒈烯和芳樟醇）进行赤拟谷盗、烟草甲和嗜卷书虱的熏蒸、触杀和驱避活性测试；对8个甲醇提取物进行三种靶标昆虫的驱避活性测试；对5个分离化合物进行赤拟谷盗成虫的拒食活性测试。结果表明，不同类型的活性物质在不同的作用方式下对特定的靶标昆虫均具有一定的抗虫活性。例如吴茱萸叶挥发油在78.63 nL/cm²，2h时对赤拟谷盗的驱避率达80%以上；蜜楝吴萸果实挥发油对赤拟谷盗和烟草甲均具有较好的熏蒸和触杀毒性；甲醇提取物在213.89 nL/cm²，2h时对赤拟谷盗的驱避率基本均在60%以上；5个分离的化合物在最高浓度1500 ppm时对赤拟谷盗的拒食率在18.57%~39.29%之间。测试结果验证了前期化学成分分析阶段的推测，为吴茱萸属植物对三种靶标昆虫的防治奠定了研究基础，丰富了该属植物的生物活性信息。

（3）主要挥发性物质驱避机理研究。根据前一阶段的活性测试结果，筛选吴茱萸果实和蜜楝吴萸果实挥发油，及8个主要挥发性化合物分别进行赤拟谷盗雌雄成虫的触角电位（EAG）和驱避活性测试，结合上述试验结果对赤拟谷盗的驱避活性作用机理进行研究和探讨。结果表明，低浓度0.005 μg/μL和0.05 μg/μL对应的不同时间点，雌雄虫驱避率存在显著差异的情况较多，对应EAG反应也存在显著差异。高浓度驱避率普遍较高，且雌雄虫之间驱避活性一般没有显著差异，但EAG反应差异显著。测试样品均具有一定的熏蒸毒性，试虫驱避属于躲避行为，该行为与诸多因素有关，目前的研究结果还不能阐释清楚，但为仓储害虫的行为学研究提供了一定的研究基础。

2. 基于抗虫活性物质的吴茱萸区划研究。由抗虫活性测试结果可知，吴茱萸的药用和非药用部位的挥发油均具有很好的抗虫活性，且在我国资源分布广泛，具有很好的潜在开发利用价值。通过文献调研收集与吴茱萸挥发油研究相关的分布点21个，结合37个环境因子，利用最大熵模型MaxEnt与ArcGIS技术，对吴茱萸在中国的潜在生态适宜性分布区域进行预测；并筛选出主导环境因子为最干季度降水量（bio 17）、最冷月最低温度（bio 6）和降水量变异系数（bio 15）；利用ArcGIS将预测结果划分出高、中、低和不适生区4个等级，并计算得出高、中、低适生区覆盖的13个省份各生态适宜区占陆地总面积的比例和面积。结果显示，高适生区主要分布在湖南、湖北和江西省。分布主要受与温度和降水量相关的气候因子影响，预测结果可指导基于抗虫活性的吴茱萸植物资源的栽培种植和合理开发利用。

本工作丰富了吴茱萸属植物的化学和生物活性信息，为该属植物的充分合理利用提供了科学依据；测定了赤拟谷盗雌雄虫对中药材挥发油及主要活性成分的EAG反应值，初步研究探讨了其驱避活性作用机理，充实了传统“对抗同贮”法的科学依据；将最大熵模型与ArcGIS技术引入本工作，基于仓储害虫的绿色防控，对药用植物资源的潜在分布区域进行预测，为药用植物资源的充分和合理利用提供了的研究基础。

Comparing with synthetic insecticides, plant secondary metabolites are low mammalian toxicity, environment friendly and less insect resistance. Evodia rutaecarpa is a traditional Chinese medicine from Evodia genus plants. It is usually stored with the medicine of animals for anti-insect purpose. According to this, we proceeded our work with the related technology of chemical ecology and geography, including research of anti-insect active substances of Evodia genus plants against stored product insects, preliminary study on the mechanism of repellent activity, and predition of potential geographic distribution area of Evodia genus plants in China by using Maximum Entropy model (MaxEnt) and ArcGIS. This work would provide a more valuable scientific basis and research ideas for prevention and control of stored product insects with Evodia genus plants from the perspective of rational development and utilization of Chinese medical herbs.

The specific research contents and results are as follows:

1. Research on anti-insect activity of Evodia genus plants.

(1) Analysis of chemical components base on anti-insect activity. The essential oils (EOs) of six samples (E. rutaecarpa fruits, etc.) were produced by steam distillation and analyzed by GC-MS. Eight samples (E. rutaecarpa fruits, etc.) were extracted by methanol and analyzed by HPLC and ¹H-NMR for identification of the dominant compounds and their contents. According to the spectrum information,, anti-insect test and literature research, five compounds (2 alkaloids and 3 limonins) are selected separation from E. rutaecarpa fruits. The results showed that there were abundant anti-insect active substances in Evodia genus plants, which had a good value for development and utilization.

(2) Research on anti-insects activity of active substances. The fumigant, contact and repellent activity were tested on Tribolium castaneum, Lasioderma serricorne and Liposcelis bostrychophila with six EOs from fruits and leaves of E. Lenticellata and E. rutaecarpa, leaves of Evodia calcicola and Evodia trichotoma, and eight dominant compounds, including β-caryophyllene, caryophyllene oxide, myrcene, ocimene, limonene, linalool, β-pinene and 3-carene. Eight methanol extracts were tested on three target insects for their repellent activity. Five separated compounds were tested on T. castaneum for their antifeedant activity. The results demonstrated that different active substances exhibit specific anti-insect activity on target insects under different mode of action. For example, EO of E. lenticellata fruits displayed fumigant and contact toxicity on T. castaneum and L. serricorne. EOs from leaves of E. rutaecarpa exhibited strong repellent against T. castaneum (>80%) at the concentration of 78.63 nL/cm² when exposure time is 2 hours. All the methanol extracts have good repellent activity (>60%) on T. castaneum at the concentration of 213.89 nL/cm² when exposure time is 2 hours. Five pure separated nonvolatile compound showed a range of 18.57%~39.29% antifeedant activity against T. castaneum. The results validated aforementioned prediction about the anti-insect constituents, and also are laying research foundation on the control of three target insects. Enriched the bioactivity information of this genus.

(3) Research on mechanism of repellent of substances against T. castaneum. According to the results of active tests, EOs from fruits of E. rutaecarpa and E. Lenticellata and eight dominant volatile compounds were selected for electroantennogram (EAG) and repellent test on female and male T. castaneum adults. The results demonstrated that ten samples exhibited different levels of EAG response. At the lower concentrations (0.005μg/μL and 0.05μg/μL) of specific different exposure time, the differences among repellency of female and male T. castaneum adults are remarkable. Samples generally showed high repellecy at higher concentration. The differences among repellency of female and male T. castaneum adults at higher concentration are not remarkable, but EAG responses are not remarkable. The repellent activity might be due to the escape behavior of T. castaneum because of the fumigant toxicity of the samples on target insect. The mechanism of repellent avtivity can not be explained only with current research results. It might be related with many factors. Nevertheless, the results of this work is still laying a research foundation on the study of behavior of stored product insects.

2. Study of potential distribution of E. rutaecarpa in China. According to the results of anti-insect activity test, we got that EOs of E. rutaecarpa displayed good anti-insect activity on target insects. Distribution points of E. rutaecarpa based on research of EOs of E. rutaecarpa and 37 environmental factors were imported into the MaxEnt model and ArcGIS were used to study potential distribution of E. rutaecarpa. Dominant environmental factors are bio 17, bio 6 and bio 15. Four levels of potential distribution area, unsuitable region, lowly suitable region, moderately suitable region and highly suitable region, were divided by MaxEnt and ArcGIS. The four levels of potential distribution of 13 provinces were extracted by mask. Area and propotion of the total land area of were calculated by ArcGIS. The results showed that highly suitable region are mainly distributed in Hunan, Hubei and Jiangxi provinces. The distribution was mainly affected by climatic factors related to temperature and precipitation. The predicted results can guide the cultivation and rational utilization of E. rutaecarpa plant resources based on insect resistance activity.

  This work enriches chemical and biological activity information of genus Evodia. It provides scientific basis for the full and rational utilization of this genus. The scientific basis of traditional “anti-storage” is enriched by preliminary study on mechanism of repellent activity through testing EAG of EO from traditional Chinese medicinal herbs to female and male T. castaneum adults. MaxEnt and ArcGIS are introduced for this work. Predicting potential suitable distribution of medicinal plants base on ecological prevention and control of stored product insects in Chinese medicinal materials. Providing scientific basis for the purpose of sustainable development and rational utilization of Chinese medicinal herbs.