EAGER SitS:Smart Long-Lived Biosensors for Soil Monitoring Using Engineered Spores

迈阿密大学和佛罗里达大学之间的这个合作项目的重点是制造新型坚固、廉价、长寿命的传感器，以安全、快速、可靠地监测土壤的化学和微生物特征，以改善土壤健康和作物生长。这些传感器基于工程细菌孢子，只有当它们检测到土壤中存在有害化学物质和/或微生物时才会发光。这些传感器可以向农业工人发出警报，保护农作物免受这些威胁。通过这项新技术的商业化，这种传感器有可能被广泛应用于农业领域。研究人员将利用这一项目的成果来吸引和培养来自不同工程背景的学生。如果成功，该项目将为保护土壤健康提供宝贵的工具，进而有助于确保国家的粮食安全。本研究旨在开发用于检测土壤化学和微生物动态变化的传感器，这些变化对了解土壤生态系统和改善土壤健康至关重要。全细胞生物传感器（WCBs）的分析物在孢子形成细菌将开发和部署在土壤监测化学和微生物特征。孢子作为储存元素的使用为在温和和极端环境下（野外和难以进入实验室的偏远地区）保存、包装、运输、储存和使用wcb提供了一种新的手段。为实现该项目的目标，将采取三个目标。第一个目标是为目标分析物制备wcb，然后转化为孢子形成细菌。这些生物传感器将应用于模拟农田，监测土壤化学特性。对于第二个目标，现有的用于群体感应分子的wcb将被纳入孢子内形成细菌中。由此产生的生物传感器将用于确定微生物指纹，并优化以监测土壤微生物生态的变化。最后的目标是验证基于孢子的生物传感器在具有不同类型土壤的模拟农田中可变条件下土壤监测的使用。感应后，将诱导产孢，感应孢子将保持休眠状态，直到需要另一个检测周期。孢子传感器平台，连同最先进的检测，将产生可运输，具有成本效益，易于使用，远程寻址或现场测试。孢子传感器(1)是检测土壤化学/微生物特征的有用工具；(2)可合并wcb用于分析物检测；(3)便于携带，坚固耐用，能承受土壤变化；（四）不需要特殊贮存、使用条件的；(5)可埋于土壤中，并在需要时响应分析物；(6)可以低成本、高效、简便地制备；(7)为wcb的制备、储存、运输和使用提供平台，用于监测农业土壤生态系统中土壤和植物的健康状况。这项研究适用于K-12、本科生、研究生和博士后的跨学科研究教育。研究进展将通过出版物和在会议上的发言来传播。来自不同背景的研究生，包括那些来自代表性不足的种族和社会经济群体的研究生，将接受培训，通过非技术语言的核心思想的展示，有效地与全球受众沟通。孢子传感器将通过在学校、博物馆和科学博览会上的演示来吸引学生进入STEM领域。生物纳米技术课程将包括对农场的实地考察和现场传感器的使用。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。

This collaborative project between the University of Miami and the University of Florida centers on the manufacture of novel rugged, inexpensive, long-lived sensors to safely, quickly, and reliably monitor chemical and microbial characteristics of soil for improved soil health and crop growth. These sensors are based on engineered bacterial spores that emit light only when they detect the presence of harmful chemicals and/or microorganisms in soil. These sensors can serve as an alarm to agricultural workers to protect crops from these threats. The sensors could potentially be employed widely in agricultural fields via commercialization of the newly created technology. The researchers will use the results of this project to attract and train students from diverse backgrounds in engineering. If successful, this project will provide a valuable tool to protect soil health, and in turn, help ensure the food security of the Nation. This research aims to develop sensors for detecting dynamic chemical and microbial changes in soil, changes which are critical for understanding soil ecosystems and improving soil health. Whole cell biosensors (WCBs) for analytes in endospore-forming bacteria will be developed and deployed in soil to monitor chemical and microbial characteristics. The use of spores as storage elements provides a new means of preserving, packaging, transporting, storing, and using WCBs in both mild and extreme environments the field and remote locations with no easy access to laboratories. Three objectives will be undertaken to achieve the goal of the project. The first objective is to prepare WCBs for target analytes, followed by transformation into spore-forming bacteria. These biosensors will be employed in simulated agricultural fields to monitor soil chemical characteristics. For the second objective, existing WCBs for quorum sensing molecules will be incorporated into endospore-forming bacteria. The resulting biosensors will be used for determining a microbial fingerprint and optimized to monitor changes in the soil microbial ecology. The final objective is to validate the use of spore-based biosensors for soil monitoring under variable conditions in a simulated agricultural field with different types of soils. After sensing, sporulation will be induced, and the sensing spores will be kept dormant until there is need for another detection cycle. The spore sensor platform, along with state-of-the-art detection, will yield transportable, cost-effective, easy-to-use, remotely addressable or in-situ field tests. The spore sensors (1) are useful as tools for detection of soil chemical/microbial characteristics; (2) can incorporate WCBs for analyte detection; (3) are portable and rugged, capable of withstanding soil changes; (4) require no special conditions for storage and use; (5) can be buried in soil and respond to analytes when desired; (6) can be prepared at low-cost in an efficient facile manner; and (7) provide a platform for preparation, storage, transport and use of WCBs for monitoring the health of soils and plants growing in agricultural soil ecosystems. The research lends itself to education of K-12, undergraduate, graduate, and postdoctoral students in interdisciplinary research. Research progress will be disseminated through publications and presentations at conferences. Graduate students from diverse backgrounds, including those from underrepresented ethnic and socioeconomic groups, will be trained to effectively communicate to a global audience through presentation of core ideas in non-technical language. The spore sensors will be used to attract students to the STEM field through presentations in schools, museums, and science fairs. A site visit at an agricultural farm and use of sensors on-site will be incorporated in a Bionanotechnology course.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.