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.

迈阿密大学和佛罗里达大学之间的这一合作项目的重点是制造新型坚固耐用、价格低廉、寿命长的传感器，以安全、快速、可靠地监测土壤的化学和微生物特征，从而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。这些传感器可以作为农业工人的警报，以保护作物免受这些威胁。通过新技术的商业化，这些传感器有可能广泛应用于农业领域。研究人员将利用该项目的结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一个宝贵的工具，从而有助于确保国家的粮食安全。该研究旨在开发用于检测土壤中动态化学和微生物变化的传感器，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中分析物的全细胞生物传感器（WCB）并部署在土壤中，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WCB在温和和极端的环境，现场和偏远地区，不易进入实验室。为实现本项目的目标，将实现三个目标。第一个目标是制备用于目标分析物的WCB，然后转化为孢子形成细菌。这些生物传感器将用于模拟农田监测土壤化学特性。对于第二个目标，现有的群体感应分子的WCB将被纳入内孢子形成细菌。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。最后的目标是验证使用孢子为基础的生物传感器的土壤监测在不同的条件下，在一个模拟的农业领域与不同类型的土壤。在感测之后，将诱导孢子形成，并且感测孢子将保持休眠，直到需要另一个检测循环。孢子传感器平台，沿着最先进的检测，将产生可运输的，具有成本效益的，易于使用的，可远程寻址的或原位的现场测试。该孢子传感器（1）可用作检测土壤化学/微生物特性的工具;（2）可结合WCB用于分析物检测;（3）便携且坚固，能够承受土壤变化;（4）不需要特殊的储存和使用条件;（5）可埋在土壤中并在需要时响应分析物;（6）可以以低成本以有效的简易方式制备;以及（7）为WCB的制备、储存、运输和使用提供平台，以监测农业土壤生态系统中土壤和植物的健康状况。这项研究有助于K-12，本科生，研究生和博士后学生在跨学科研究的教育。研究进展将通过出版物和在会议上的介绍加以传播。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，将接受培训，通过以非技术语言展示核心思想，与全球受众进行有效沟通。孢子传感器将用于通过在学校，博物馆和科学博览会上的演讲吸引学生到STEM领域。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。

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.

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.

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

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