SBIR Phase I: Control of Lesion Nematodes by Transgenic RNA Interference

这项小型企业创新研究（SBIR）第一阶段研究将产生对植物寄生线虫具有抗性的转基因作物，重点是病变线虫（短体线虫）。植物寄生线虫每年在美国造成80亿美元的损失，是最难控制的植物病原体之一。转基因作物有很大的机会通过提高产量和降低投入成本为种植者提供经济效益，并取代有毒的杀线虫农药。 本项目将利用RNA干扰技术，通过沉默线虫基因来控制病斑线虫。我们将测试特异性双链RNA（dsRNA）抑制必需寄生虫基因的功效。Divergence建立了一个高效的转基因植物毛状根表达系统，用于筛选候选dsRNA。 已取得了防治其他植物寄生线虫的有希望的结果，并且相同的系统可用于病斑线虫。目的筛选、克隆和测序斯氏短体线虫中可能是所有病害线虫所必需的基因靶标。在目标II中，将在毛状根系统中测定靶基因的RNAi效力。在第I阶段结束时，预期将选择几个表现出病斑线虫繁殖减少的靶基因用于第II阶段整株植物转化。 第一次引入转基因病变线虫抗性的可能作物是玉米，这是美国最大面积和最有价值的作物之一。该项目的更广泛影响将是减少美国农业中广泛使用农药的需求，并对抗影响玉米生产的侵略性寄生虫。

This Small Business Innovation Reserach (SBIR) Phase I research will generate transgenic crops with resistance to plant parasitic nematodes with a focus on the lesion nematode (Pratylenchus). Plant parasitic nematodes annually cause damage of $8 billion in the U.S. and are among the most difficult plant pathogens to control. Transgenic crops have a significant opportunity to provide economic benefit to growers through improved yields and decreased input costs and to replace toxic nematicidal pesticides. This project will use RNA interference (RNAi) to control the lesion nematode by silencing nematode genes. We will test the efficacy of specific double-stranded RNAs (dsRNAs) to inhibit essential parasite genes. Divergence has established an efficient transgenic hairy root in planta expression system to screen candidate dsRNAs. Promising results controlling other plant parasitic nematodes have been achieved and the same system can be utilized for lesion nematode. Objective I will include the selection, cloning, and sequencing of gene targets from Pratylenchus scribneri that are likely to be essential in all lesion nematodes. In objective II, the RNAi potency of target genes will be assayed in the hairy root system. At the conclusion of Phase I, it is anticipated that several target genes demonstrating reduction in lesion nematode reproduction will be selected for Phase II whole plant transformation. A likely crop for the first introduction of transgenic lesion nematode resistance is corn, one of the largest acreage and most valuable crops in the U.S.The broader impact of this project will be to reduce the need for extensive use of pesticides in US agriculture and to counter an aggressive parasite that affects the production of corn.