EAGER: Enhancing plant immunity and growth with cell-penetrating peptides for organic agriculture

EAGER：利用有机农业的细胞穿透肽增强植物免疫力和生长

• 批准号: 2154863
• 负责人: Guo-Liang Wang
• 金额: $ 30万
• 依托单位: Ohio State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2154863&HistoricalAwards=false
• 关键词: EAGER; Enhancing; plant; immunity; growth

To feed the growing world population, the agricultural sector needs to increase food production even while being attentive to the impact of new strategies on ecosystems and human health. Although fertilizers and pesticides play a crucial role in modern agriculture, their adverse impact on the environment and animal/human health have recently inspired a rapid rise in sustainable crop production. A promising environment-friendly alternative is the use of biologics. However, many of the plant defense activators, biostimulants, and biopesticides currently used in sustainable agriculture are proteins and peptides that have low entry efficiency into plant cells, thus necessitating the use of high concentrations and consequently high costs. To address this limitation, this project will leverage a powerful membrane translocation domain (MTD) technology to improve the delivery of biological agents. HrpZ, a proven defense activator, will be fused to MTD4, one of the most potent MTDs in hand, and tested in tomato for its efficacy in eliciting immune responses to bacterial and fungal pathogens. This approach should decrease the effective concentration of defense activators and thereby increase the economic viability of plant biologics for sustainable agriculture and farmers’ income. In addition, the team will explore outreach activities to demonstrate how biodefense molecules can help plants ward off pathogens.Although cell-penetrating peptides (CPPs) have been widely explored for delivering therapeutic agents to treat human diseases/conditions, their applications in plants have been more limited. The goal of this project is to examine the utility of a newly discovered class of CPPs, MTDs, to efficiently deliver HrpZ, a known biodefense activator/biostimulant, into crop plants to protect them from pathogen infection and insect infestation. The three specific aims of this project are: [1] improve the efficiency of cellular entry and metabolic stability of MTD4; [2] determine the effects of MTD4-HrpZ and MTD4-N21 on tomato and rice defense, growth, yield, and drought tolerance; and [3] optimize and develop low-cost recombinant protein expression and chromatography-free purification strategies for large-scale field applications. After completion of the three objectives, this project will provide insights into how CPPs enter plant cells and how their different cellular structures (e.g., lipid composition in membranes) affect the cellular entry mechanism and efficiency of CPPs. Importantly, the use of CPPs to promote plant immunity is likely to be transformative for plant disease control in sustainable agriculture. Our new strategies for overexpression and chromatography-free purification of recombinant proteins are also likely to be of broad interest. In addition, identification of powerful CPPs that can efficiently deliver peptides/proteins into plant cells will provide new tools for biological studies that seek to uncover in planta interactions between different proteins, including pathogen effectors.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.

为了养活不断增长的世界人口，即使注意到新策略对生态系统和人类健康的影响，农业部门也需要增加粮食产量。尽管肥料和农药在现代农业中起着至关重要的作用，但它们对环境和动物/人类健康的不利影响最近激发了可持续作物生产的迅速增长。承诺的环境友好的替代方法是使用生物制剂。但是，当前在可持续农业中使用的植物防御激活剂，生物刺激剂和生物农药是蛋白质和胡椒粉，它们对植物细胞的进入效率较低，因此对于使用高浓度并因此高成本所必需。为了解决这一限制，该项目将利用强大的膜易位域（MTD）技术来改善生物学剂的输送。 HRPZ是一种经过验证的防御激活剂，将与MTD4融合，MTD4是最有效的MTD，并在番茄中测试了其在引起对细菌和真菌病原体的免疫反应方面的有效性。这种方法应减少国防激活者的有效集中，从而增加植物生物制剂对可持续农业和农民收入的经济可行性。此外，该团队将探索外展活动，以证明生物性分子如何帮助植物避免病原体。尽管细胞穿透肽（CPP）已被广泛探索用于输送治疗剂以治疗人类疾病/状况，但其在植物中的应用受到了更大的限制。该项目的目的是检查新发现的CPPS类MTD的实用性，以有效地将HRPZ（一种已知的生物化激活剂/生物刺激剂）传递到农作物植物中，以保护它们免受病原体感染和昆虫侵袭的影响。该项目的三个特定目的是：[1]提高MTD4的细胞进入和代谢稳定性的效率； [2]确定MTD4-HRPZ和MTD4-N21对番茄和水稻防御，生长，产量和干旱耐受性的影响； [3]为大规模现场应用优化和开发低成本重组蛋白表达和无色谱纯化策略。完成三个目标后，该项目将提供有关CPP如何进入植物细胞以及它们不同的细胞结构（例如机制中的脂质组成）如何影响CPP的细胞进入机制和效率的见解。重要的是，使用CPP来促进植物免疫学可能是可持续农业中植物性疾病控制的变革。我们对重组蛋白的过表达和无色纯化纯化的新策略也可能引起广泛关注。此外，鉴定可以有效地将肽/蛋白质传递到植物细胞中的强大CPP将为生物学研究提供新的工具，以寻求在包括病原体效应在内的植物学相互作用（包括病原体效应）之间进行植物学相互作用。该奖项反映了NSF的法定任务，并通过使用该基金会的知识分子优点和广泛的影响来评估NSF的法定任务，并被认为是珍贵的支持。