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

EAGER SitS：使用工程孢子进行土壤监测的智能长寿命生物传感器

• 批准号: 1841419
• 负责人: Sylvia Daunert
• 金额: $ 30万
• 依托单位: University of Miami School of Medicine
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1841419&HistoricalAwards=false
• 关键词: EAGER; SitS; Smart; Long; Lived

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.

迈阿密大学与佛罗里达大学之间的合作项目，以制造新颖，廉价，长寿的传感器，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的生长。这些传感器基于工程细菌孢子，只有在发现有害化学物质和/或微生物在土壤中存在有害化学物质和/或微生物时才发出光。这些传感器可能会引起农业工人的警报，以保护农作物免受这些威胁。这些传感器可能通过新创建的技术的商业化在农业领域中广泛使用。研究人员将利用该项目的结果吸引和培训来自工程领域不同背景的学生。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动态化学和微生物的变化，这对于了解土壤生态系统和改善土壤健康至关重要。将在土壤中开发并部署用于内生孢子细菌的分析物的全细胞生物传感器（WCB），以监测化学和微生物特征。将孢子用作存储元素的使用提供了一种新的手段，可以在轻度和极端环境中保存，包装，运输，存储和使用WCB，而在领域和远程位置都无法轻松访问实验室。将实现三个目标以实现该项目的目标。第一个目标是为目标分析物制备WCB，然后将其转化为形成孢子的细菌。这些生物传感器将用于模拟农田来监测土壤化学特征。对于第二个目标，现有的用于法定人数传感分子的WCB将掺入内孢子形成细菌中。所得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学的变化。最终目标是在模拟的农田中验证在可变条件下使用孢子的生物传感器用于土壤监测，并具有不同类型的土壤。传感后，将诱导孢子形成，并保持感测孢子休眠，直到需要另一个检测周期为止。孢子传感器平台以及最先进的检测将产生可运输，具有成本效益，易于使用的，可远程寻址或原位现场测试。孢子传感器（1）可作为检测土壤化学/微生物特性的工具； （2）可以合并WCB用于分析物检测； （3）是便携式和坚固的，能够承受土壤变化； （4）不需要特殊的存储和使用条件； （5）可以埋在土壤中，并在需要时对分析物响应； （6）可以在低成本的情况下以高效的方式制备； （7）提供了一个平台，用于制备WCB的准备，存储，运输和使用，以监测农业土壤生态系统中生长的土壤和植物的健康。该研究使K-12，本科，研究生和博士后生的教育在跨学科研究中。研究进度将通过会议上的出版物和演讲来传播。来自不同背景的研究生，包括来自代表性不足的种族和社会经济群体的研究生，将通过以非技术性的方式介绍核心思想，以有效地与全球受众进行培训。孢子传感器将通过学校，博物馆和科学博览会的演讲来吸引学生进入STEM领域。在农业农场进行现场访问，将传感器的使用将在Bionanotechnology课程中纳入现场。该奖项反映了NSF的法定任务，并认为使用基金会的知识分子和更广泛的影响评估标准，认为值得通过评估值得支持。

项目成果

Isothermal Amplification and Lateral Flow Nucleic Acid Test for the Detection of Shiga Toxin-Producing Bacteria for Food Monitoring

• DOI: 10.3390/chemosensors10060210
• 发表时间: 2022-06-01
• 期刊: CHEMOSENSORS
• 影响因子: 4.2
• 作者: Petrucci，Sabrina;Dikici，Emre;Deo，Sapna K.
• 通讯作者: Deo，Sapna K.

On-site detection of food and waterborne bacteria - current technologies, challenges, and future directions.

• DOI: 10.1016/j.tifs.2021.06.054
• 发表时间: 2021-09
• 期刊: Trends in food science & technology
• 影响因子: 15.3
• 作者: Petrucci S;Costa C;Broyles D;Dikici E;Daunert S;Deo S
• 通讯作者: Deo S

Molecular Aptamer Beacons and Their Applications in Sensing, Imaging, and Diagnostics

• DOI: 10.1002/smll.201902248
• 发表时间: 2019-07-17
• 期刊: SMALL
• 影响因子: 13.3
• 作者: Moutsiopoulou, Angeliki;Broyles, David;Deo, Sapna K.
• 通讯作者: Deo, Sapna K.

Multiplexing cytokine analysis: towards reducing sample volume needs in clinical diagnostics

• DOI: 10.1039/c9an00297a
• 发表时间: 2019-05-21
• 期刊: ANALYST
• 影响因子: 4.2
• 作者: Yu，Xiaowen;Scott，Daniel;Daunert，Sylvia
• 通讯作者: Daunert，Sylvia

Orally Administrable Therapeutic Synthetic Nanoparticle for Zika Virus

• DOI: 10.1021/acsnano.9b02807
• 发表时间: 2019-10-01
• 期刊: ACS NANO
• 影响因子: 17.1
• 作者: Surnar, Bapurao;Kamran, Mohammad Z.;Dhar, Shanta
• 通讯作者: Dhar, Shanta