SBIR Phase I: Creating a Master Transformation Recipient Line for Chloroplast Engineering of Maize

这个小企业创新研究（SBIR）第一阶段项目的更广泛影响是开发环境可持续、高产、抗虫害的生物技术作物。未来的作物需要几十种性状来获得更高的产量，并保护自己免受病虫害和不断变化的环境条件的侵害。所提出的技术将使具有多性状基因的作物高效生产、所需性状的更高功效、更快的育种和种子生产以加速分配。与目前的解决方案不同，拟议的技术确保性状不会通过花粉转移到杂草或本地植物物种。最初的应用是玉米，这是美国最有价值的商业作物，其见解有望提高其他作物的产量。本课题旨在建立工程玉米叶绿体（质体）转化平台。虽然叶绿体工程（转质体）技术已在许多双子叶作物（如烟草、马铃薯）中成功开发，但尚未开发出一种可复制的技术用于任何单子叶作物（如玉米、高粱）。现有的证据表明，速率限制步骤是需要一个强大的选择系统，以便在单子房典型的胚性组织培养系统中鉴定和扩增罕见的转化质体。该项目将创建一个用于叶绿体工程平台的玉米主转化受体系（MTRL）。MTRL将在叶绿体编码的光合基因中包含一个靶向基因组缺失，从而允许开发一个新的选择系统，该系统可以导致单株作物叶绿体工程的成功。最终，相应的野生型叶绿体光合基因将与其他所需性状一起交付给MTRL，从而促进了对被删除的光合基因修复的转化质体的优雅选择，并携带了感兴趣的新性状。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact of this Small Business Innovation Research (SBIR) Phase I project is to develop environmentally sustainable, higher yielding, pest-resistant biotech crops. Crops of the future need dozens of traits to achieve higher yields and protect themselves against insects, pests, diseases and changing environmental conditions. The proposed technology will enable efficient production of crops engineered with multiple trait genes, higher efficacy of the desired traits, and faster breeding and seed production to accelerate distribution. Unlike current solutions, the proposed technology ensures that traits will not be transferred to weeds or native plant species via pollen. The initial application is corn, the most valuable commercial crop in the US, with insights expected to enhance other crops. The proposed project aims to create a chloroplast (plastid) transformation platform for engineering corn. Although chloroplast engineering (transplastomic) technology has been developed successfully in a number of dicot crop species (i.e., tobacco, potato), a reproducible technology has not been developed for any monocot crop (i.e., corn, sorghum). The available evidence indicates that the rate-limiting step is the need for a strong selection system to enable the identification and amplification of rare transformed plastids in the embryogenic tissue culture systems typical for monocots. This project will create a Master Transformation Recipient Line (MTRL) of corn for use in a chloroplast engineering platform. The MTRL will contain a targeted genome deletion in a chloroplast-encoded photosynthetic gene, allowing for development of a new selection system that can lead to successful chloroplast engineering of monocot crops. Ultimately, the corresponding wild-type chloroplast photosynthetic gene will be delivered to the MTRL with other desired traits, facilitating an elegant selection of transformed plastids with the deleted photosynthetic gene restored and as well as carrying the new traits of interest.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.