EAGER: Non-integrative transient delivery of reagents into plant cells via the type IV secretion system of A. tumefaciens

EAGER：通过根癌农杆菌的 IV 型分泌系统将试剂非整合瞬时输送到植物细胞中

• 批准号: 1759445
• 负责人: Thomas Clemente
• 金额: $ 29.9万
• 依托单位: University of Nebraska-Lincoln
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1759445&HistoricalAwards=false
• 关键词: EAGER; Non; integrative; transient; delivery

Innovations developed through public sector investments over the past few decades have translated to technologies that have benefited U.S. agriculture productivity significantly, for example, reducing agriculture inputs through precision application of water, nutrients, and pesticides. Genetic technologies have led to better predictive models for plant breeding and introduction of novel traits into crops that could not be added by conventional means. However, the ability to introduce novel traits using biotechnology is often hampered by low efficiencies. This project seeks to improve the efficiency with which Agrobacterium tumefaciens can serve as a tool to introduce novel traits into crop plants by exploiting the bacterium's ability to secrete proteins into plant cells. The transient delivery of these proteins will impact both the ability of the plant cell to detect and fight off the bacterium, and the ability of the plant cell to regenerate a new plant once it has been altered. To meet this goal, a multidisciplinary approach will be taken that brings together expertise in genetics, microbiology and molecular biology. Importantly, the underlying biology of the project will be communicated through outreach efforts to the non-science community on all aspects of food production and on how innovations in agriculture help maintain the U. S. strategic advantage through its capacity to produce a plentiful and safe food supply. The research to be conducted in this project lays out an innovative set of studies to design and test novel Agrobacterium tumefaciens strains that can help mitigate two major inefficiencies of plant transformation: the competency of the cell to receive a genetic reagent and the ability of that same cell to lead to germline transmission of the induced genetic change. Pseudomonas effector proteins that suppress the plant innate immune response will be transiently expressed to improve the efficiency of gene transfer from bacterium to plant cell. In addition, plant morphogenic proteins will be transiently expressed to improve the efficiency of regenerating plants from transformed tissue culture cells. The results of these studies will also provide insight into the size limit of a protein that can be shuttled through the Agrobacterium type IV secretion system. Overall, this project should lead to A. tumefaciens strains with enhanced transformation capacity across a broader array of genotypes within a plant species and that are easily transferable to the plant science community.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.

在过去几十年中，通过公共部门投资开发的创新已经转化为使美国农业生产力显著受益的技术，例如，通过精确应用水，养分和农药来减少农业投入。遗传技术为植物育种和将传统方法无法添加的新性状引入作物带来了更好的预测模型。然而，利用生物技术引入新性状的能力往往受到低效率的阻碍。该项目旨在通过利用细菌向植物细胞分泌蛋白质的能力来提高农杆菌作为将新性状引入作物的工具的效率。这些蛋白质的瞬时递送将影响植物细胞检测和抵抗细菌的能力，以及植物细胞一旦被改变就再生新植物的能力。为实现这一目标，将采取多学科方法，汇集遗传学、微生物学和分子生物学方面的专门知识。重要的是，该项目的基础生物学将通过外展工作传达给非科学界，涉及粮食生产的各个方面，以及农业创新如何帮助维持美国的粮食安全。S.通过其生产充足和安全粮食供应的能力而具有战略优势。在该项目中进行的研究列出了一系列创新的研究，以设计和测试新的根癌农杆菌菌株，这些菌株可以帮助减轻植物转化的两个主要效率低下：细胞接受遗传试剂的能力和同一细胞导致诱导的遗传变化的种系传播的能力。 假单胞菌效应蛋白抑制植物的先天免疫反应，将瞬时表达，以提高从细菌到植物细胞的基因转移的效率。 此外，植物形态发生蛋白将被瞬时表达以提高从转化的组织培养细胞再生植物的效率。 这些研究的结果还将提供对可以穿梭通过农杆菌IV型分泌系统的蛋白质的大小限制的了解。总的来说，这个项目应该导致A。该奖项反映了NSF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。