Mosquito Vector Control Using Bt Coupled to Seed Oil Bodies

描述(由申请人提供)：这个小型企业创新研究第一阶段项目开发了一种更有效和更环保的方法来生产和运送苏云金芽孢杆菌(Bti)毒素和BtBooster(BTB)用于控制蚊子。BTI毒素具有很强的蚊子物种特异性活性，比化学杀虫剂更安全。BTB蛋白通过增加毒素摄取来增强Bti的毒性。需要一种新的和廉价的BTI和BTB生产和交付系统来接触各种水环境中的蚊子。该项目的生物技术假设是：BTI和BTB偶联到富油作物种子油体表面的油球蛋白，可以作为磨制的Bt面粉有效地输送，这些面粉将漂浮在水柱中的不同高度，并杀灭取食蚊子的幼虫。这个第一阶段的应用将通过将bti和btb与富含油脂的拟南芥种子中的油球蛋白偶联来检验这一假说。种子将被磨成Bt面粉，其颗粒大小接近于滤食性幼虫消耗的食物的大小。Bt面粉将被分散在水中，并喂给作为疾病媒介和昆虫死亡率量化的三种蚊子的幼虫。这将证明BTB和BTI之间的协同作用可以导致增加对传播疾病的蚊子物种的控制。蚊子是许多通过血液传播的人类疾病的昆虫媒介，包括疟疾、西尼罗热、黄热病和登革热，这些疾病每年在全球范围内造成数百万人死亡。蚊子幼虫是滤食性动物，生活在安静的开阔水域环境中，根据物种的不同，它们的幼虫生活在水柱的不同深度。在所有人畜共患传播传染病的媒介中，蚊子是最难控制的媒介之一。这是可能的，但只有在水环境中添加大量化学或生物杀虫剂。该项目将证明，BTB和BTI之间的协同作用可以显著改善生物杀虫剂对传播疾病的蚊子物种的控制，降低进入水环境的化学杀虫剂的数量。美国的蚊子幼虫防治剂市场每年约为4500万美元，全球市场要大得多。市场机会既来自传统的蚊子控制领域，也来自新兴的监管领域。计划进行第二阶段的应用，其中油球蛋白偶联的bti和btb将在油菜中表达(油菜)。表达Bti/BTB的油菜籽可以生产并以较低的成本碾磨成Bt-粉。公共卫生相关性：蚊子是多种血液传播疾病的媒介，包括疟疾、黄热病、西尼罗河病毒、丝虫病、日本脑炎和登革热，每年在全世界造成数百万人死亡。该项目旨在开发一种新的、更高效、廉价和环境安全的解决方案，用于生产和交付与Bti相关的毒素和BtBooster，用于在幼虫阶段控制蚊子。Bti和BtBooster作为漂浮在水柱中不同高度的研磨面粉之间的协同作用，将导致生物杀虫剂大幅改善对传播疾病的蚊子物种的控制，降低进入水环境的化学杀虫剂的数量，从而通过更好地控制媒介传播的疾病，产生与公共卫生有关的重大好处。

DESCRIPTION (provided by applicant): This Small Business Innovation Research Phase I project develops a more efficient and environmentally safe method to produce and deliver Bacillus thuringiensis israelensis (Bti) toxins and BtBoosters (BtB) for mosquito control. Bti toxins have strong mosquito species- specific activities, and are safer than chemical insecticides. BtB proteins enhance Bti toxicity by increasing toxin uptake. A novel and inexpensive system for Bti and BtB production and delivery is needed to reach mosquitoes in various aquatic environments. This project's biotechnical hypothesis is: Bti and BtB coupled to oleosins on the surface of seed oil bodies of oil-rich crops can be delivered effectively as milled Bt-flour that will float at various levels in the water column and kill feeding mosquito larvae. This Phase I application will test this hypothesis by coupling Bti and BtB to oleosins in the oil-rich seeds of Arabidopsis thaliana. Seeds will be milled into Bt-flour, with a particle size approximating that of food consumed by filter-feeding larvae. The Bt-flour will be dispersed in water and fed to larvae of three mosquito species that act as disease vectors and insect mortality quantified. This will demonstrate the synergism between BtB and Bti can lead to increased control of disease vectoring mosquito species. Mosquitoes are the insect vectors for numerous blood-borne human diseases including malaria, West Nile fever, yellow fever, and dengue that cause millions of deaths each year worldwide. Mosquito larvae are filter-feeders that inhabit quiet open-water environments, and depending upon species, their larvae live at various depths in the water column. Of all vectors for zoonotic transmission of infectious agents, mosquitoes are one of the more difficult to control. This is possible, but only by adding large amounts of chemical or biological insecticides to the aquatic environment. This project will demonstrate that the synergy between BtB and Bti can lead to substantially improved control of disease vectoring mosquito species by biological pesticides, lowering the amount of chemical pesticides entering aquatic environments. The U.S. market for mosquito larvicide control agents is about $45 million per year, with a much larger worldwide market. The market opportunity will be from both the traditional segment of mosquito control and also the emerging regulatory segment. A phase II application is planned in which oleosin coupled Bti and BtB will be expressed in canola (Brassica napus). The Bti/BtB expressing canola seeds can be produced and milled into Bt-flour at relatively low cost. PUBLIC HEALTH RELEVANCE: Mosquitoes are vectors for numerous blood-borne diseases, including malaria, yellow fever, West Nile virus, filariasis, Japanese encephalitis, and dengue fever and cause millions of deaths worldwide annually. This project is designed to develop a novel, more efficient, inexpensive, and environmentally safe solution to the production and delivery of Bti-related toxins and BtBoosters for mosquito control at the larval stage. The synergy between Bti and BtBooster as milled flours that float at various levels in the water column will lead to substantially improved control of disease-vectoring mosquito species by biological pesticides, lowering the amount of chemical pesticides entering aquatic environments and thereby producing significant public health-related benefits through better control of vector-borne diseases.