George Hines1, Cyrus Modavi2, Keni Jiang3, Andrew Packard1, Kameshwar Poolla1, Lewis Feldman4. 1. Department of Mechanical Engineering, University of California, Berkeley, CA, 94720-3102, USA. 2. Department of Bioengineering, University of California, Berkeley, CA, 94720-3102, USA. 3. Department of Plant and Microbial Biology, University of California, Berkeley, CA, 94720-3102, USA. 4. Department of Plant and Microbial Biology, University of California, Berkeley, CA, 94720-3102, USA. ljfeldman@berkeley.edu.
Abstract
MAIN CONCLUSION: The activation and level of expression of an endogenous, stress-responsive biosensor (bioreporter) can be visualized in real-time and non-destructively using highly accessible equipment (fluorometer). Biosensor output can be linked to computer-controlled systems to enable feedback-based control of a greenhouse environment. Today's agriculture requires an ability to precisely and rapidly assess the physiological stress status of plants in order to optimize crop yield. Here we describe the implementation and utility of a detection system based on a simple fluorometer design for real-time, continuous, and non-destructive monitoring of a genetically engineered biosensor plant. We report the responses to heat stress of Arabidopsis thaliana plants expressing a Yellow Fluorescent Protein bioreporter under the control of the DREB2A temperature-sensing promoter. Use of this bioreporter provides the ability to identify transient and steady-state behavior of gene activation in response to stress, and serves as an interface for novel experimental protocols. Models identified through such experiments inform the development of computer-based feedback control systems for the greenhouse environment, based on in situ monitoring of mature plants. More broadly, the work here provides a basis for informing biologists and engineers about the kinetics of bioreporter constructs, and also about ways in which other fluorescent protein constructs could be integrated into automated control systems.
MAIN CONCLUSION: The activation and level of expression of an endogenous, stress-responsive biosensor (bioreporter) can be visualized in real-time and non-destructively using highly accessible equipment (fluorometer). Biosensor output can be linked to computer-controlled systems to enable feedback-based control of a greenhouse environment. Today's agriculture requires an ability to precisely and rapidly assess the physiological stress status of plants in order to optimize crop yield. Here we describe the implementation and utility of a detection system based on a simple fluorometer design for real-time, continuous, and non-destructive monitoring of a genetically engineered biosensor plant. We report the responses to heat stress of Arabidopsis thaliana plants expressing a Yellow Fluorescent Protein bioreporter under the control of the DREB2A temperature-sensing promoter. Use of this bioreporter provides the ability to identify transient and steady-state behavior of gene activation in response to stress, and serves as an interface for novel experimental protocols. Models identified through such experiments inform the development of computer-based feedback control systems for the greenhouse environment, based on in situ monitoring of mature plants. More broadly, the work here provides a basis for informing biologists and engineers about the kinetics of bioreporter constructs, and also about ways in which other fluorescent protein constructs could be integrated into automated control systems.
Authors: Rajeev K Varshney; Kailash C Bansal; Pramod K Aggarwal; Swapan K Datta; Peter Q Craufurd Journal: Trends Plant Sci Date: 2011-04-15 Impact factor: 18.313
Authors: Andreas Milias-Argeitis; Sean Summers; Jacob Stewart-Ornstein; Ignacio Zuleta; David Pincus; Hana El-Samad; Mustafa Khammash; John Lygeros Journal: Nat Biotechnol Date: 2011-11-06 Impact factor: 54.908