Sustainable Development Goals
Currently, 925 million people worldwide face food insecurity and consequently have limited access to affordable and nutritious produce [1]. With the rapidly rising population, the United Nations (UN) projects that we will need to increase current food production by 60% . [2]. As the amount of arable land decreases and urban sprawl increases, communities are required to identify means to grow crops in low-light, low-maintenance settings. Compact vertical hydroponics systems produce nutrient-dense crops and are ideal for heavily populated areas. AgroSENSE encourages those affected by food insecurity to take advantage of hydroponics systems to produce locally-sourced, nutritious food. However, a barrier to wider public adoption of small-scale hydroponics systems is nutrient fluctuations . Low concentrations of phosphate and nitrate can hinder the plant’s ability to grow. Conversely, high levels of phosphate reduce the plants ability to uptake micronutrients [3], primarily iron and zinc, both of which are critical for plant growth; similarly, high levels of nitrate affect the plant’s ability to photosynthesize [4], leading to poor growth. To consistently monitor nutrient levels, we developed cell-free nitrate biosensors and whole-cell phosphate biosensors which provide specific nutrient concentrations that can eliminate errors arising from commercial test strips, allowing hydroponics users to adjust nutrient levels earlier. Additionally, nutrient concentrations outside optimal ranges can decrease plant health, ultimately increasing its susceptibility to plant pathogens, including Fusarium oxysporum and Phytophthora cryptogea . Currently, both pathogens are detected via visual cues, such as wilting leaves, as late as eight to ten days after initial infections [5]. By the time these symptoms arise, the pathogen causes irreversible damage to the entire crop population. To address this, we engineered two riboregulated switches that detect F. oxysporum and P. cryptogea in their spore stage while still treatable, allowing farmers to increase crop yield. By 2050, the UN estimates that 68% of the world’s population will live in urban areas [6]. With increasing urban sprawl, limited arable land hinders urbanites from accessing fresh produce in their vicinity. AgroSENSE increases the sustainability of hydroponics by providing an effective system to monitor nutrient levels and pathogen presence, and increase crop yield, thereby contributing to the Zero Hunger Sustainable Development Goal . To educate future generations about synthetic biology and its progressing applications, we collaborated with ONASI Bilingual College in Ebolowa, Cameroon . Their principal, Mr. Onana Messi, informed us that while students were eager to learn about synthetic biology, they often avoided the science pathway at their school due to limited lab opportunities and resources. This prompted us to develop a standards-based supplementary synthetic biology series that expanded off their curriculum . We referred to the Biotechnology pathway curriculum provided by the Georgia Department of Education from the Georgia Standards of Excellence [11] as a basis for our lessons. Once we had a clear understanding of the curriculum, we arranged to develop a series of virtual workshops and send materials for a teacher training that would take place in early September. In the future, we anticipate Mr. Onana will be able to share the training framework with the ministry of education in Ebolowa and promote the use of a community lab kit that would provide materials to expand this opportunity to other schools in the Mvila school district. To introduce synthetic biology to the next generation of scientists, our team wrote and illustrated a storybook targeted towards elementary school students . Our goal was to produce an engaging story aligned with the Georgia Department of Education Standards of Excellence for 3rd through 5th grade to give students early exposure to the fundamentals of biotechnology and encourage academic interest [12]. We consulted with Dr. Brittney Cantrell, the Science Specialist for grades 6-12 in Forsyth County Schools, and she offered insight regarding which educational standards to refer to and how to tie in elements of synthetic biology into the existing curriculum. Our storybook, Grow and Glow , was inspired by an article published by the Massachusetts Institute of Technology that described how researchers plan to engineer bioluminescent trees [13]. Grow and Glow introduces Enrique, a student on a field trip, to Dr. Rosalind, a synthetic biologist who engineers bioluminescent trees. She invites Enrique into her lab and describes the process of bacterial cloning using explanatory graphics and analogies. Enrique also learns about the potential of bioluminescent trees to encourage sustainability by saving electricity [13]. To distribute Grow and Glow into our local community, we explored two main avenues: partnering with our local public library and collaborating with Forsyth County elementary school teachers . Post survey data from teachers revealed that our storybook made synthetic biology “very approachable” and that students were “engaged and enjoyed reading about the topic.” After our Q&A session, over 88% of students understood the fundamentals of synthetic biology as compared to the initial 30%. Lambert iGEM hosted a collaboration to teach newly developed iGEM teams about the restriction enzyme cloning workflow . We had the opportunity to meet with 7 teams: RUM- UPRM, IISER Tirupati, IISER Berhampur, Istanbul_Tech, UESTC-China, and two unconstructed teams . This webinar introduced each step of the workflow and briefed participants on how to build an effective plasmid and insert. Due to the complexity of bacterial cloning, Lambert iGEM believed it would be beneficial for newer iGEM teams to grasp a theoretical understanding of the workflow before beginning their project. After participants signed a waiver allowing use of data collection we asked comprehension questions throughout the webinar and conducted a post-survey after the workshop. Using our poll questions asked every few minutes to identify if we needed to supplement our explanations. The post-survey at the end contained a series of comprehension questions regarding the overall workflow. With the majority of the participants answering all of the questions correctly, indicating that they were comprehending and grasping the concepts taught . With our synthetic biology workshops and storybook, AgroSENSE aims to address the Quality Education Sustainable Development Goal . Across the United States, 23.5 million people lack easy access to fresh produce [7] and are limited to highly processed food from local convenience stores. Consequently, these Americans are vulnerable to diet-related health issues including cardiovascular disease, diabetes, and even premature death [8]. With AgroSENSE, people living in food deserts are able to implement hydroponics systems and produce nutrient-dense crops as a result of close mechanical monitoring, eventually increasing their good health and wellbeing [9]. Due to the COVID-19 pandemic, fresh produce prices have skyrocketed, further hindering food insecure communities from accessing affordable and nutritious food . As of August 2021, the Food Price Index has been on the rise and still remains alarmingly high as a result of current transportation, labor, and fresh produce costs (see Fig. 1). Food insecurity is an increasing issue in the United States. AgroSENSE aims to help people facing food insecurity by encouraging the use of hydroponics, an affordable agricultural solution to sustainably create nutrient-dense crops. Our biosensors allow users to maintain hydroponics systems easier , thereby addressing the Good Health and Wellbeing Sustainable Development Goal by making the use of hydroponics more accessible. As vertical hydroponics is a compact, modular form of agriculture, AgroSENSE is city-friendly as it does not require large amounts of arable land . Due to urban sprawl, traditional farms tend to be farther away from population centers and transportation of produce typically utilizes high amounts of economic resources while emitting high amounts of greenhouse gases [15] [16] [17] [18]. By consuming homegrown produce, people reduce toxic emissions resulting from mass transportation, support the “eat local” movement , and allow money to flow back into the local economy . Moreover, hydroponic systems create fewer greenhouse gases, a smaller carbon footprint, more renewable resources, and an overall sustainable future. In a 2014 study “Up, Up and Away! The Economics of Vertical Farming,” published in the Journal of Agricultural Studies, estimated yields of a CEA vertical farm were compared to that of a field farm. The study stated that traditional methods of agriculture can grow up to 16 crops in one square foot, while hydroponics and vertical farms produce an average of 516 times as many tons of produce per 10,000 meters [19], showing that hydroponics systems are more compact and produce higher yields . Urbanization is foreseen to continue at an accelerating pace, and cities are expected to account for 68% of the world population in 2050 [6]. On the other hand, rural areas, after peaking sometime in the next decade, are projected to decline in population density. By encouraging the use of locally-grown hydroponics, we promote sustainable and environmentally-friendly communities for current and future generations of urbanites, addressing the Sustainable Cities and Communities Goal . Goal 2 - Zero Hunger Goal 4 - Quality Education Goal 3 - Good Health and Wellbeing Goal 11 - Sustainable Cities and Communities Goal 17 - Partnership for the Goals Frugal Lyophilizer and Plate Reader Phosphate Biosensor Plant Pathogen Biosensors Model Nitrate Biosensor and Cell-free Legislation Human Practices Dr. Bhamla held biweekly meetings to guide our Lyophilizer and Plate Reader teams to improve the designs and provide constructive feedback on our work in 2020 and 2021. Dr. Styczynski provided us with wet lab guidance throughout our project,which helped us lower our phosphate concentration ranges, resulting in refined characterization data. Dr. Paul elaborated upon the concept of microneedle patches and how to apply them to extract DNA from pathogens on plant roots. He also introduced alternatives to microneedle patches and gave us the idea of grinding roots, which ultimately led to our sink strainer extraction protocol. Ms. McSweeney taught the team how to use NUPACK software for our pathogen toehold designs Ms. McSweeney and Ms. Han assisted us with parameter estimations on MATLAB. Ms. McSweeny gave us advice on how to improve our lysate preparation by visiting our lab and helped us troubleshoot our experimental testing. She lent us several reagents to use for our cell-free experiments, as well as providing ongoing support and troubleshooting. Dr. Silverman introduced us to cell-free protein synthesis by providing his publications and protocols. He guided us through preparing enriched lysates, as well providing continuous feedback to improve our extraction performance. In addition, he shared test plasmids for us to test our initial lysates. Ms. Zhang assisted us with obtaining the proper reagents for our phosphate biosensor characterization. She also helped us work on our protocol and adjust our phosphate solution ranges for the experimentation of our phosphate biosensor. Ms. Chilton provided us with proper reagents for characterization, ongoing feedback and support. Dr. Matsumara helped us alter our phosphate biosensor by changing the way we tested the data. He instructed us to discontinue working with the Pho A promoter sequence, and helped us improve our concentration ranges. Commissioner Black and his office made themselves available for multiple conversations and ongoing help. In one of our initial discussions with Commissioner Black he mentioned that there were no current legislative or regulatory frameworks concerning the use of biosensors in agricultural settings. Recognizing this gap, he helped network us with other Georgia State officials to research and develop appropriate action items. In addition, he provided pertinent information on the state of agriculture, economic impact of hydroponics, and potential future growth of the industry. Ms. Adan informed us that biosensors are an emerging innovation in agriculture, and therefore, have limited governmental barriers regarding their safe distribution. She helped us consider different methods to target this gap and helped us develop our initial legislative proposal. Ms. Haltom connected us with Chairman Todd Jones to further our legislative proposal. She also guided us through the process of developing a regulatory outline that mirrors established Georgian agricultural regulations. Chairman Jones encouraged our team to create a regulation rather than a legislation for biosensors to allow for smooth updates to the piece following technological developments. Dr. Dutta conveyed the consequences of Fusarium oxysporum in the agricultural field. He also informed us that our plant pathogen biosensors have the potential to positively impact many commercial farms that are impacted by plant pathogens. His feedback led us to narrow our focus to design a riboregulated toehold switch to detect the presence of Fusarium DNA in hydroponic systems. Mr. Crowe was instrumental in the progress of our project this year. He allowed us to visit the Sweetwater facility multiple times where we were able to obtain water samples and observe the prevalence and impact of the root rot Phytophthora cryptogea in tower farms. Visiting a commercial facility gave us valuable insight into the economics, sustainability, and viability of urban hydroponic systems. Mr. Crowe also provided feedback to our implementation team on the practicality of performing molecular based assays in the field as well as numerical data for our disease predictive model. Kristen Boscan, Director, and Megan Heaphy, Project Coordinator, worked with our team over the course of the year on topics related to survey development, science communication, standards alignment, and networking with life science professionals and educational initiatives. Dr. Brewer serves as the advisor for the GSU iGEM team who conducted tests with our frugal lyophilizer. They provided feedback on our prototype which led us to improve our protocols for other users. Dr. Devarapu’s continued feedback led us to modify our plate reader design based on light distribution and camera position, making the device more effective in fluorescence quantification. He helped our team coalesce ideas from multiple previous designs into a novel approach. Mr. Poorna led troubleshooting sessions that helped us solve issues such as air leakage on our lyophilizer, Optical Density, as well as fluorescence calculations on the plate reader, guiding us towards redesigning the system. Dr. Saad Bhamla Bhamla Lab, Georgia Institute of Technology Dr. Mark Styczynski Professor, Georgia Institute of Technology Dr. Rajesh Paul Researcher, North Carolina State University Megan McSweeney Graduate Student, Styczinski Lab at Georgia Institute of Technology Megan McSweeney and Yue Han Graduate Students, Styczinski Lab at Georgia Institute of Technology Megan McSweeney Graduate Student, Styczinski Lab at Georgia Institute of Technology Dr. Adam Silverman Researcher, Sherlock Biosciences Yan Zhang Styczynski Research Group, Georgia Institute of Technology Elizabeth Chilton Styczynski Research Group, Georgia Institute of Technology Dr. Ichiro Matsumara Professor, Emory University Gary Black Commissioner of Agriculture, Georgia Department of Agriculture Natalie Adan Food Safety Division, Georgia Department of Agriculture Ashley Haltom Vice President of Governmental Affairs, Georgia BioEd Institute Chairman Todd Jones Representative, Georgia House of Representatives Dr. Bhabesh Dutta Vegetable Disease Specialist, University of Georgia Mr. Clint Crowe Sweetwater Urban Farms Megan Heaphy and Kristen Boscan Rural Teacher Training Initiative, Georgia BioEd Institute Dr. Matthew Brewer Professor, Georgia State University Dr. Chinna Devarapu Professor, Postdoctoral Researcher, Tyndall International Institute Rajas Poorna PhD Student, Georgia Institute of Technology
Figure 1. Graph showing the overall Food Price Index for 2018-2021. (Food and Agriculture Organization of the United Nations)
FAO Food Price Index 2014-2016 = 100 130 2021 2018 2019 2020 121 112 103 94 85 J J J F M M A A S O N D
Dr. Styczynski provided online databases for rate constants and suggested that protein abundances were likely not necessary for constructing the model. Dr. Mark Styczynski Professor, Georgia Institute of Technology
REFERENCES
[1] Graziano Da Silva, J. (2021). Feeding the world sustainability. United Nations. www.un.org/en/chronicle/article/feeding-world-sustainably
