Burge Environmental, Inc./Groundswell Technologies, Inc.Scott R. Burge, Ph.D. • Burge Environmental, Inc.

Mark Kram, Ph.D. • Groundswell Technologies, Inc.Kiril Hristovski Ph.D. • Arizona State University

• Burge Environmental will develop an automated real-time plant rhizosphere sensor probe capable of detecting plant and rhizome stimulus-response signals due to adverse changes in the ecosystem (i.e. biological, chemical, nuclear agents).

• The proprietary microbial sensor technology is already in early commercialization phases for water, wastewater, well monitoring, and aquaculture industries.Lab investigations indicate potential applications in dry soil soils and for direct plant health measurement.

• Project Goal: Develop a sensor monitoring system for collaboration with groups working on Phase I, II, or III program for plant selection and stimulus detection.

Dr. Scott Burge & Robert Harding, Burge Environmental, Inc.• Developed Sr-90, I-129, TC-99, Tritium, TCE, Cr (VI), Uranium, and VOC Sensors

since 1995 Dr. Mark Kram, Groundswell Technologies, Inc.• Developed real-time sensor telemetry and data visualization platformDr. Kiril Hristovski, Arizona State University• Experience with rhizosphere and wastewater technology commercialization

Objective:Develop and fabricate sensor probes for real-time direct rhizosphere and plant monitoring

Technical Challenges:• Partner with appropriate facilities required to test sensor monitoring system for

chemical, biological, and nuclear agents• Partners to provide genetically modified plants to determine microbial signal

response to biological, chemical, and nuclear agents

Accelerate collaborative research with real-time signal platform

Potential Applications:• Precision Agriculture• Agriculture R&D Acceleration• Energy & Resource optimization of biotechnology industries

Milestones & Metrics:• Identify rhizosphere and plant stimulus response signals• Compare sensor signal response to standard analytical methods• Deploy plant & rhizosphere sensors in the field• Demonstrate reduced analysis cost and turn around time

Technology Transition:• Team is experienced with commercialization and collaborative research of new

technologies

Project Overview

Teaming Overview and Objectives Impacts

Joel G. Burken, Missouri S&T, MO Plant-Explo

Teaming Overview and Objectives

Project Overview

• Missouri S&T– PI J.G. Burken, CoPI Z.Yin, Sr.Pers. C.E. Johnson, Grad.Res. P.V. Manley, Sr. Pers. M.E. Bookout-MinerFly group (S&T), St. Louis Univ (SLU)–Dr. A. ‘Wasit’ Ghulam, Danforth Plant Science Center (DPSC) – CoPI T. C. Mockler, Sr.Pers. N. Shakoor, University of York, UK (York) – Sr.Pers. E.L. Rylott

• Assets: Greenhouse and chambers, integrated with imaging and analytic facilities. Plant-contaminant interactions Explosives program and range facility, with all permitting & licensing UAV fleet with FAA certified pilot, manned aircraft - (S&T) (Advanced remote sensing laboratory and facilities (SLU); Plant Genome laboratory (DPSC); and LemnoTech high throughput plant phenotyping, sensing (DPSC);

• Missouri Plant-Explo team will advance plant-based, threat identification through UAV-based sensing of WT-plants and advance to applications with enhance bio-sentinels via precision gene editing.

• Highspeed data fusion of multiple plant-stimuli response hyperspectral, thermal, & geosphere data integrated real time for threat projection, assessment and visualization for rapid response.

Impact

J.G. Burken – Burken@mst.edu – (573)-201-9106 1

GIS-CV GIS-VIs

IR Data

Real Time data fusion, anomaly identification and

analysis, geo-integration and visualization

Trait ID

• Deployable plant-stimuli response sensing for native, WT species of target interest. Genetic modification -specific target promotors of visible stimuli-response, in conjunction with UAV and manned aircraft.

• Sensitive plant stimuli-responses indicate presence of explosives and developed Explosives-Specific Indices can be used in multi-spectral filters for a multitude of sensors. These could be deployed on cell phones, airborne sensors, or even a soldier’s HUD.

• The potential to save thousands of lives will be developed.

HSI Data

Wolfgang Busch, Salk Institute, Team: TBA

Teaming Overview and Objectives

Project Overview

Haves:• High-throughput root phenotyping platforms to map plant

sensing and response mechanisms (Slovak et al., 2014; Satbai et al. 2017)

• Robot for phenotyping (Feb. 2018)• Machine vision for root growth response quantification • Quantitative biology and systems genetics expertise• Expertise in the genetics of local adaptation• Large collections of fully sequenced strains that can be used

for sensor identification and provide locally adapted backgrounds

How can we get highly specific and robust molecular Sensing Circuits for a variety of compounds?How do we make sure that Reporter Plants survive permanently in target areas?

How to tackle this:• High-throughput phenotyping of natural strains and mutants for identification of sensory modules• Perennial (Lotus japonicus) and annual (Arabidopsis thaliana) species that are environmentally highly

adaptable and can be easily genetically engineered

Impact• Identify molecular sensors for specific compounds• Deploy in multiple target species in genetic backgrounds that are locally adapted

Wolfgang Busch – wbusch@salk.edu – (858) 453-4100 x1765 1

Wants:• assessment for the most impactful molecules to

sense, their time-scales for detection and the appropriate read-out for it (satellite, drones)

• wiring sensing modules to suitable read-outs

Sean Cutler cutler@ucr.edu– 951-367-7096

PI: Sean Cutler, UC-Riverside; Team: Whitehead (MSU), Wheeldon (UCR), Brutnell, Nusinow, Gehan, Zhang (Danforth Plant Science Center)

Teaming Overview and Objectives

• Cutler/Whitehead/Wheeldon: protein engineering, computational protein design, plant and microbial synthetic biology, rapid prototyping.

•Redesigned a plant hormone receptors to respond to non-native ligands enabling orthogonal control of leaf temperature.

•New pipelines for protein engineering that coupling saturationmutagenesis & next gen sequecing.

• Brutnell/Nusinow/Gehan/Zhang: grass systems, synthetic circuits, plant growth, development, light perception, imaging and analysis software.

• Collaborators needed: remote sensing, ecology modeling.

Impact• Field-ready technologies to for designing

multisensor sentinel plants.

• New orthogonal genetic circuits for controling plant traits in the field.

• High throughput platforms for developing sensors of emergent threats.

• Rapid pathway to crops, limiting gene flow to native species.

1

Modules portable to crops & suited for field deployment

Outputs: Height • Leaf temperatureFlowering • Leaf color • Fluorescense

Modules insulated from endogenous signals

Rapid module design/characterization pipelineWT

Sentinel

Native signaling module

Reprogrammed module

Reprogrammed physiology

Orthogonal modules can be multiplexed

We will use computational design to reengineer plant hormone sensors to function as orthogonal sense-and-response modules that report environmental threats by regulating physiological outputs.

Doherty, North Carolina State University, Timing & Detection

Teaming Overview and Objectives

Project Overview

• Doherty: Signaling expertise:• Circadian control of signaling, resources, & metabolism; • Oscillatory pathways; • Light (and temperature) signaling; • Novel pigment pathways; • Experience w/ Setaria viridis.

• Kudenov: Imaging expertise. Has developed: • Snapshot Imaging Spectrometer:• Fraunhofer Line Discrimination (FLD): • Spectropolarimetry. deployment at NCSU’s agricultural research

stations.• Other NCSU Researchers: CRISPR Lab, Ralph Dean, PHHI- expertise in

plant pigment Genetics Environment and Society Center with strong modellers.

• NCSU facilities: • Plant transformation lab, transformation of grasses and ”unusual

plants” Institutional assets • CRISPR Lab- novel genome editing tools• GES Center- interdisciplinary team with modelling expertise

• Looking for collaborations: Interested in being Sub / Co-I

• Plants that detect abnormal lighting patterns, detection of chemical with nucleotide aptamers (potential targets- Ricin A-chain, aflatoxin), detection of pathogens

• Unique aspects: 1) Paired response-detector combinations 2) Temporally coordinate signal detection, resource usage, and signal emission to ensure consistency with existing rhythms (control of homeostasis) 3) Oscillatory signals

• Stimulus-response: Detecting Exogenous pigments; Response-optics pairing; Oscillatory signals; • Resources- Working within the existing resource allocation networks by coordinating timing of activities; • Technical Challenges- transformation of plants in Phase II time window.

Impact• Pairing of response - detection will reduce

resource burden. • Optimized detection can facilitate

screening for events with desired sensitivity.

• Handheld spectrophotometer will facilitate greenhouse measurements.

• Coordination between time and sensing / response will improve consistency and reduce resource burden.

• Oscillatory signals offer opportunities for measuring duration of stimulus presence, integrating multiple stimuli, signal amplification.

• Novel pigments offer minimal feedback into existing pathways, simplified detection

Contact Information – colleen_doherty@ncsu.edu 919.515-5802 1

Plant Scientists and Phenotyping Researchers at Agricultural College of Purdue University

Teaming Overview and Objectives

Project Overview

Existing team members• Prof. Jian Jin, Plant Phenotyping and Sensor Data Modeling• Prof. Melba Crawford, Remose Sensing• Prof. Mitch Tuinstra, Agronomist, Biologist and Plant

Scientist• Prof. Cankui Zhang, Plant Transformation, Plant Scientist Major accomplishments:• Numerous Plant Assays Experiences in Greenhouse and field • Expertise and experiences in plant transformationInstitutional assets • Fully Automatic Greenhouse with High-throughput Plant Phenotyping

Systems• Field Phenotyping Platforms (UAVs, PhenoRover in DOE’s ARPA-E

TERRA Program)• Plant Transformation Laboratory in Agronomy• A new Handheld Device with Geo-Spacial Map Service

• The goal is to establish a high-throughput plant screening, evaluation, and sorting protocol in Purdue’s greenhouse, which can evaluate and select the best plant sensor candidates for the APT program.

• Purdue’s greenhouse phenotyping systems have shown the capability of measuring plants responses to various stimuli (chemical sprays, pathogens, insects, etc.), as well as abiotic stresses (drought, nutrition, heat, etc.) through numerous greenhouse plants assays. We can establish a protocol to generate plants candidates and precisely evaluate/sort the candidates for the best performance in sensing and reporting, tolerance to lack of resources and ecology challenges.

Impact• The fastest research pipeline to speed up and advance the

discovery of the next “plant sensors” for APT • Examples of potential applications:

• Predicting chemical types and # of Days-After-Spraying • Prediction of aerial polutants• If a transgene construct in which the promoter of MTs is fused

in front of an ABA synthesis gene is transformed into a plant, the stomata of the transgenic plants will be partially closed when confronting metals, e.g., Zn, Cu.

• Metrics:• Screening Throughput• Plant’s Performances in Sensing and Stress

Tolerance • Milestones:

• Establish of the screening protocol• Delivery of the first plant sensor accepted by APT

Jian Jin – jinjian@purdue.edu – 765-494-11821

Bogumil Karas, University of Western Ontario/Designer Microbes Inc., “Western-DMI”

Teaming Overview and Objectives

Project Overview

• Team: Dr. Bogumil Karas (PI), Dr. Igor Kolotilin (Co-applicant), Dr. Preetam Janakiram (Staff Scientist, DMI), Ryan Cochrane and Stephanie Brumwell (graduate students), and additional personnel (hired as needed).

• The proposed work is based on Dr. Karas’ prior work: Direct genome transfer from bacteria to yeast (Nature Methods, 2013), Development of transformation method for algae (Nature Communications, 2015), Contribution to creation of the minimal synthetic cell (Science, 2016).

• Assets: Biotron research facility, a state-of-the-art climate-controlled space at the University of Western Ontario.

• We are seeking collaborators for remote sensing but also for identification of appropriate stimulus and response strategy.

• We are developing efficient methods for the construction and installation of natural or synthetic genomes (nuclear, mitochondrial, and chloroplast) in plant and algal cells.

• Our approach will facilitate each of the three APT objectives (stimulus-response, resources, ecology). The anticipated technical challenges are: optimization of our yeast-plant cell fusion protocol and targeted installation of organellar genomes in plant or algal cells.

• We propose a project in three phases:• Phase 1 - cloning/assembly of whole organelle genomes/metabolic

pathways in yeast• Phase 2 - install these genomes/pathways in plant/algal cells• Phase 3 - verify functional installation in whole plants and algal cultures

Impact• Successful and robust genome transfer into plant and algal

cells would provide a powerful approach to engineering eukaryotic cells.

• Application: Plants controlled by designer synthetic nuclear or organellar genomes, which facilitate rapid installation of designer pathways.

• Metrics: We will demonstrate transfer of large DNA fragments from yeast to plant cells via cell fusion, and replacement of complete organellar genomes in plants and algae.

• This research will yield patentable technology and provide opportunities for starting up new biotech companies or licensing IP to existing ones.

Bogumil Karas – bkaras@uwo.ca – 519-719-0340 1

Carlos F MartinoFlorida Institute of TechnologyQuantum Life Science Group

Teaming Overview and Objectives

Project Overview

• PI: Carlos F Martino – theoretical and technical expertise inradiofrequency electromagnetic fields, link fundamentalbiochemical processes in the plants to growth responses

• Co-PI: Margaret Ahmad – radiofrequency magnetic field effectsin plants, static magnetic field effects in plants, cryptochromemechanisms and engineering

• Co-PI: Busov - Directional effects in plants exposed to magneticfields, transformation of novel plant varieties.

• Co-PI: Ritz Thorsten - theoretical considerations to optimize theresponse of identified plant components to man-made electro-magnetic fields.

ImpactImpact• Project will develop novel biosensors of electromagnetic radiation • Discoveries can lead to new technologies in magneto-sensing Applications• Electromagnetic noise as a consequence of human activity can be

detected in remote places by plant growth changes which can be visualized by satellite. Large metal stockpiles can be detected.

• Radiofrequency or high frequency transmission can be visualized by plant growth changes.

• Enables sensing of a directional magnetic fieldMetrics• Optimized cryptochrome receptors• Optimized transgenic plants• Varieties for field testing

Carlos F Martino – cmartino@fit.edu - 321.674.8497 1

Engineering plant cryptochromes as biosensors of electromagnetic fields• Goal: To alter plant growth responses to man-made electromagnetic fields, detectable by satellite imaging• Stimulus response: plant varieties optimized for different electromagnetic field types (radio frequency, weak static, extremely-

low frequency)• Resources and ecology: different plant species will be engineered for maximum resilience• Methodology

Phase I: Characterize magneto-sensing genes in existing plant modelsPhase II: Modify components for optimal signal recognitionPhase III: Introduce into plant species of interest for field tests

30 cm

N

10 MHz Static

High Frequency Magnetic Field Direction

June Medford David Baker Josh LeonardColorado State University University of Washington Northwestern University

Teaming Overview and ObjectivesOverview• Medford, Baker, Leonard Labs; others• Previously Developed plant sentinels to small molecules

with computational input, controllers, and readouts• Models and parts for in planta quantitative controls• Published & Patented Abilities to sense and move signal• Complementary platforms for engineering novel biosensors• Decades of experience with plant systemsObjectives• Establish and prove robust functions in planta

Impact• Ability of plants to serve as silent sensors

• Substantial expansion of sense & response ability,• Substantial expansion to quantitatively tune abilities

• Ability to sense in one portion of plant & respond where desired• Numerous synthetic biology applications– biofuels, orthogonal

control systems, more• Tech Transfer Plans

• Partner with Institute for Protein Design (UW)• Small Women-owned Business (Phytodetectors, Inc.)• For Government Applications, Expand Interactions with

Defense Contractors

1

De novo Computational Protein Designsfor Sensing Biologicals, Chemicals, More

Orthogonally Sourced Materials

MemoryAmplificationReset-ability

Quantitatively TunedControllers

Project Overview

As needed:Resource & Ecological Systems

INPUTs

Contact Information Email: June.Medford@colostate.edu; Phone: 970-491-7865

Develop Highly Modular Robust Plant Sentinels for All Potential Stimuli & Plant SpeciesApproach: Encode traits in genetic circuit & genome editing

In planta response from variable & transient stimuli

Remotely Detectable Responses

Brutnell(PI), Cutler, Gehan, Nusinow, Wheeldon, Whitehead, Zhang Donald Danforth Plant Science Center/UC Riverside/Michigan State University

Teaming Overview and Objectives

Project Overview

• Team: Brutnell/Cutler/Gehan/Nusinow/Wheeldon/ Whitehead/Zhang labs and associated personnel.

• Expertise in: grass systems, synthetic circuits, plant growth, abiotic stress, development, light perception, imaging, and image analytics.

• Access to 84 growth chambers with spectrally controlled lighting (0-1100 PAR), temperature (-10 to 50˚C), carbon (100-3000 ppm) and humidity to test technology stability.

• Access to high-throughput phenotyping technologies including fluorescence, NIR, and hyperspectral imaging.

• We are interested in collaborators to help with ecological modeling and robustness.

• Develop orthogonal light-activated circuits to induce sensors/outputs at-a-distance in a widely deployable grass species.

• Technical challenges: Developing robust, sensitive, and selective orthogonal induction and sensor systems in planta

Impact• Delivery of sensor systems to detect biological, chemical,

and environmental threats with minimal drag on plant growth and development.

• Several agriculture solutions for pests, pathogens and environmental challenges envisioned with this technology

• Delivery of light activated circuits to deconvolute threats when multiple sensors used.

• We have identified corporate partners in image analytics and in-field testing and will draw on our connections in plant biotech for transitioning technology

Dmitri Nusinow– dnusinow@danforthcenter.org– 314-587-1489 1

Ecology: environmentally robust species, capable of wide deployments, non-native species to limit gene flow

Response: anatomical, colorimetric and fluorescence detection of elicited response.

1

• Follow the general guidance to include the information requested.• DARPA strongly encourages establishing teams to address all technical

objectives/phases to ensure the expertise and capabilities necessary to meet program goals.

• Provide a concise and informative summary of your proposal interest.• Unclassified information only.• Only one slide will be presented.• Please submit in MS PowerPoint (preferred) or equivalent file format.

Guidance for Proposers Day Slide

Shawn Kaeppler, UW-Madison, Wisconsin Crop Innovation Center

Teaming Overview and Objectives

Project Overview

• Dr. Shawn Kaeppler (plant genomics, genetics), Dr. Heidi Kaeppler (plant transformation), Dr. Edgar Spalding (ion transport/plant imaging), Michael Petersen/Brian Martinell/Edward Williams/Amy Miyamoto (>90 combined years of tissue culture and plant transformation experience in private industry), Dr. Ray Collier (gene expression, cloning), UW Radiation Center (EM source), Middleton Spectral Vision (plant imaging)

• WCIC is a 100K ft2 R&D facility with 34K ft2 of high quality greenhouse and light room space and over 17K ft2 of laboratory facilities. USDA/APHIS stewardship, biocontainment, biosafety, and permitting experience. Access to UW-Madison core facilities for potential stimulus treatments. The Center has securekey card access and has 24/7 environmental monitoring

• Seeking collaborators with receptor binding or

Our team is looking to partner with others that have a key stimulus/response path but need expertise in efficiently deploying it to a particular plant species, challenge the transgenic plant with the stimuli, and

monitor results in a controlled and ecologically relevant environment. We are considering a project

using EM responsive promoters and aerially detectable reporters but are open to alternatives

Impact• Our team will be able to deploy transgene/CRISPER

cassettes directly to a plant species of interest and has high throughput cloning and transformation capacity.

• Our team has the ability to transform relevant genotypes at a substantially shortened timeframe (6-9 months) than that of conventional transformation technologies

Michael Petersen – mwpetersen@wisc.edu – 608-338-3168 2

Phase I (6 months) Procure plant species/validate tissue culture and growth

parameters Begin transformation control experiments to validate

transformation methodology

Phase II (18 months) Plant transformation of

stimulus/response vectors into key species

Treat plants with specific stimuli and monitor response in controlled

environmental settings (isolated internal growth rooms or greenhouses) Monitoring of response could include RGB, hyperspectral, and

fluorescence imaging Gene copy number, expression

analysis, and insertional site sequencing

Validate positive events, collect seed, and plant progeny for T1

greenhouse testing

Phase III (24 months) Pressure testing of homozygous T2+ events

under complex greenhouse environments for response,

vigor, and survivability Multiple generational testing

to include expression analysis, phenotype

monitoring and imaging, testing stimuli dose/response

curves

Daniel Sabo, Georgia Tech Research Institute

Teaming Overview and Objectives

Project Overview

GTRI:• Research scientists and graduate and co-op students• Experience in engineering, robotics, and sensor

development• Access to resources devoted to sensor development• Publications and presentations for sensor

development, robotic systems, and presymptomatic disease detection using VOCs

UGA:• Research scientists, post-docs, and grad students• Experience in plant biology, bioinformatics, and

molecular genetics• Access to resources to genetically modify plants• Publications and presentations contributing to the

areas of plant genetic modification

The GTRI/UGA team seeks to utilize presymptomatic disease detection techniques (plant volatile organic compounds (VOCs), hyperspectral imaging, and advanced perception techniques) to aid in the detection and location of chemical, biological, radiological, and nuclear (CBRN) threats using both genetically modified and unmodified plants. A multidisciplinary approach that brings together experts in engineering, sensor development, genetic modification, and plant biology will be key to accomplishing this goal and to overcoming several challenges, including the identification and modification of plant genes; the development of sensors that can observe target responses (VOCs); the development of robotic systems to enable large area monitoring; and the creation of advanced perception techniques.

Impact• Modification of plants that produce a range of measurable

responses, specifically VOCs, when in the presence of CBRN threats

• The development of a suite of sensors and sensing technology to detect these responses not only in modified plants but also unmodified local vegetation

• A list of potential applications:• Genetic modification of plants for rapid response

(VOCs) to targeted stimuli• Sensors for detection of plant responses• Development of advanced perception techniques

Daniel Sabo – daniel.sabo@gtri.gatech.edu – 404-407-6730

Ludovico Cademartiri, Iowa State University

Teaming Overview and Objectives

Project Overview

Impact

Ludovico Cademartir lcademar@iastate.edu 515-294-4549i – – 1

≈ →Typical Method

Model ecosystemsReal ecosystems

• Benchtop approaches for creation of model ecosystems as physical networks (quantitative control of signaling dynamics, individual environments, and selective stimulation of ecosystem members)

• Transparent soil allows for the phenotyping of plants in vivo in the presence of external stimuli

We are interested in collaboration for:- Phenotyping of plants/microbes in model ecosystems - Using transparent soil and model ecosystem approaches to

study microbiome and collective effects - Development of custom tools for phenotyping of intricate

plant phenomena (e.g. hydrotropism, developmental shifts)

PI: Ludovico Cademartiri Materials ChemistryNanomaterialsBiological Environments by Design

Benchtopmodel

ecosystemsfor detection

Biotic Stimuli

Chemical Stimuli

Radioactivity

Mechanical Stress

Phenotypes

Collective Behaviors

Nanoparticles Transparent Soil

Neal Stewart, University of Tennessee, Phytosensors 2.0

Teaming Overview and Objectives

Phytosensors 2.0 Project Description

• A multidisciplinary Stewart-Lenaghan team with approximately 15 postdocs/grad students/science staff is currently working in the domain of APT. We have a history of broad collaborations with universities and national labs; currently Stanford, Penn, Berkeley, ORNL, and PNNL in the area of synthetic biology for high risk-reward research.

• Over two dozen phytosensor research papers have been published by Stewart et al. since 2003, including stand-off detection, specific inducibility, and field deployment.

• We have over 20 years of experience in fluorescence, field testing regulated engineered plants, and are at the forefront of plant synthetic biology.

• Collaborators in UAV- and satellite-based platforms, as well as machine learning collaborations could be useful for quantifiable remote multiplex sensing.

• The team seeks to build and deploy next-gen “phytosensors” for targeted microbiome detection. Stewart coined the term around the time of his first DARPA seedling grant in 2001 to describe genetically engineered plant-based environmental sensors.

• Using synthetic DNA regulatory elements, we will genetically engineer field-deployable plants to sense and report key microbiome and radiological signatures. Fluorescent proteins, pigments, or other spectral signatures will be detected by UAVs and/or satellites.

• Synthetic beta systems will then be multiplexed in arrays and scored using appropriate binary platforms, prior to scalar sensing of multiple targets simultaneously.

Impact• APT represents the perfect opportunity to design/engineer

plants for multiplex sensing capabilities in both field- and the built environment for key targets. We will design synthetic promoters, enhancers, and transcription factors for broad and specific molecular signal detection by the plant, then report the presence and quantity of each target utilizing multiple spectral outputs.

• In the field, plants will be engineered to detect plant pathogens, food-borne pathogens, and ionizing radiation. In the built environment, the detection of mold and other microbiome signatures will be targeted.

• Required is multiplexed and quantitative detection of targets in 48 hours of application in two plant species.

• Technology could be transitioned to “big ag” for field sensing and ‘houseplants 2.0’ to DIY/tech markets.

nealstewart@utk.edu 865 974 6487 1

Christopher Voigt, MIT

Teaming Overview and Objectives

Interests and Capabilities

• Relevant Experience:• Build sense-and-respond in the chloroplast (with

Ralph Bock)• Theophylline to T7 RNAP to nif expression in

chloroplast (19 genes)• Sensor design (12 in one cell), porting to eukaroytic

systems• Light sensors, including far-IR and UV

• Synthetic Biology Center at MIT leader in design in sensors and circuits

• MIT-Broad Foundry for rapid prototyping (including plant chloroplast TX-TL systems)

• Seeking collaborators in plant biotechnology and detection (e.g., by satellite)

Impact• Rapid design, testing, and optimization of plant sensors for

chemicals and other signals• Genetic circuits in plants for signal processing sensory

signals (e.g., integrating multiple sensors)• Control over multi-gene pathways for remote detection• Quantitative approaches to optimizing resource utilization

and measuring long-term performance / evolutionary stability

Contact Information – cavoigt@gmail.com - 617-324-4851 1

“Smart Plants”

Plastid rapid prototyping• Chemical sensing and control of large pathways (plastid)

• Sensor and circuit design

Nigel Wallbridge, Vivent sarl, Team under formation

Teaming Overview and Objectives

Project Overview

• Vivent is a Swiss-based startup with extensive experience of plant electrical signaling in-situ

• Lead institution of current RadioBio DARPA grant • Strong reputation with key plant research institutes• Goals are to:

• Enable standard and modified plants to be used as environmental sensors with immediate and measurable electrical responses.

• Reliably determine electrical signal characteristics that define plants’ responses to radiological, electromagnetic, chemical and physical stimuli

• We seek to partner with organization's that have:• Sophisticated plant testing and plant development

capabilities• Electronics / data processing capabilities

• Use electrical signals from plants to gauge their response to chemical, radiological, electromagnetic and physical (damage and unusual circumstances) stimuli.

• All plants are learning organisms (Venus Flytrap, Mimosa Pudica are known examples). Plants’ responses to stimuli are complex and often hidden and slow. For example

• visual response to Glyphosates can take days to appear.

• response to sudden light changes takes 45 minutes and remains invisible.

• But internal electrical signals reveal complex response behaviors quickly and reliably.

Impact

• Use of both standard and modified plants as sensors in a variety of situations.

• Standard plants can be used without disruption of agriculture and without ecological impact.

• Broader understanding of electrical signaling in plants willenable more reactive sensors and positively impact agricultural production, blight detection and efficient use of inputs such as energy and water

• For Ag-tech applications Vivent will work with partners to commercialise innovations.

• For defence and other applications Vivent would seek to license technology to suitable organisations.

Nigel Wallbridge, Vivent sarl– nigel.wallbridge@Vivent.ch – +41 79 511 3743 1

PI: Pamela Silver/Jeffrey Way, Harvard, Wyss Institute

Teaming Overview and Objectives

Project Overview

• Existing team members: PIs, ~2-3 PhD students, post-docs

• Relevant experience:• Construction of genetic memory circuits and

oscillators in bacteria and yeast• Kill switches for biocontainment• Single-step introduction of complex genetic systems

into plants• Manipulation of metazoan gene expression

• Institutional assets • Access to BSL2 plant facilities, harsh environments• Cell, systems and synthetic biology

• Seeking collaborators: With additional plant expertise

• We want to achieve: Plants that can report, remember and measure time from a stimulus• Approaches to sense and report: Genetic memory and oscillator/counter systems adapted to plants • Technical challenges:

• Adaptation of genetic circuit design that thus far has only been (partially) achieved in microbes• Genetic systems that work in variable temperature, nutritional environments• Containment and plant species-independence• (Detection-resistance)

• Program phase structure:• (Modular genetic parts for plants)• Proof of concept for separate circuits• Integration• Quasi-field testing

Impact• Anticipated impact:

• Arbitrary stimulus Detection Time Recording• Potential applications:

• Forensic plant-based recording systems• Reproducible plant engineering

• Metrics and milestones: • (1) Detection; (2) Memory; (3) Timing

• Technology transition: • Employ peacetime applications of broader

technology in a spinoff company.

Contact Information: jeff.way@wyss.harvard.edu 617-432-7782 1