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

迈阿密大学和佛罗里达大学的这一合作项目致力于制造新型坚固、廉价、寿命长的传感器，以安全、快速和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。这些传感器是基于经过改造的细菌孢子，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作者发出警报，保护农作物免受这些威胁。通过将这项新发明的技术商业化，这种传感器可能会在农业领域得到广泛应用。研究人员将利用这一项目的成果来吸引和培训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，进而帮助确保国家的粮食安全。这项研究旨在开发传感器来检测土壤中的动态化学和微生物变化，这些变化对于了解土壤生态系统和改善土壤健康至关重要。将开发用于内孢子形成细菌分析物的全细胞生物传感器(WCBS)，并在土壤中部署，以监测化学和微生物特征。使用孢子作为储存元件提供了一种新的保存、包装、运输、储存和使用WCBS的方法，无论是在温和和极端环境中，还是在野外和偏远地区，都不容易进入实验室。为实现该项目的目标，将采取三项目标。第一个目标是为目标分析物准备WCBS，然后转化为芽胞形成细菌。这些生物传感器将用于模拟农业领域，以监测土壤化学特性。对于第二个目标，现有的用于群体感应分子的WCBS将被整合到内孢子形成细菌中。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。最终目标是验证基于孢子的生物传感器在不同土壤类型的模拟农田中在不同条件下进行土壤监测的使用。在感应后，将诱导产孢子，感应孢子将保持休眠，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、经济高效、易于使用、可远程寻址或现场测试。孢子传感器(1)可用作检测土壤化学/微生物特征的工具；(2)可以结合WCBS进行分析物检测；(3)便携式和坚固耐用，能够承受土壤变化；(4)不需要特殊的储存和使用条件；(5)可以埋在土壤中，在需要时对分析物做出反应；(6)可以以高效、简便的方式低成本制备WCBS；以及(7)为监测农业土壤生态系统中生长的土壤和植物的健康状况提供WCBS的平台。这项研究有助于在跨学科研究中培养K-12、本科生、研究生和博士后。研究进展将通过出版物和在会议上的陈述来传播。将培训来自不同背景的研究生，包括那些来自代表性不足的族裔和社会经济群体的研究生，以便通过用非技术性语言介绍核心思想，有效地与全球受众沟通。孢子传感器将被用来通过在学校、博物馆和科学博览会上的演讲来吸引学生进入STEM领域。对农业农场的实地考察和现场传感器的使用将被纳入一门生物纳米技术课程。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.