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Detector Plants for Agriculture, Food andEnvironmental MonitoringJune MedfordColorado State UniversityFort Collins, Coloradojune.medford@colostate.edu(No manuscript.)[Editors’ note: Dr. Medford described how techniques in synthetic biology—with fundingfrom the Defense Advanced Research Projects Agency and the Department ofDefense—are being developed to modify plants to detect with high sensitivity the presenceof chemicals (e.g. pollutants, explosives) or pathogens and reveal their presence byrapid de-greening of chlorophyll, for use in agriculture and at high-population terroristtargets such as airports.]Some of the information presented by Dr. Medford at NABC 23 has been published:Antunes MS et al. (2006) A synthetic de-greening gene circuit provides a reportingsystem that is remotely detectable and has a re-set capacity. Plant Biotechnology Journal4 605–622. doi: 10.1111/j.1467-7652.2006.00205.xSummary: Plants have evolved elegant mechanisms to continuously sense and respondto their environment, suggesting that these properties can be adapted to make inexpensiveand widely used biological monitors, or sentinels, for human threats. For a plant tobe a sentinel, a reporting system is needed for large areas and widespread monitoring.The reporter or readout mechanism must be easily detectable, allow remote monitoringand provide a re-set capacity; all current gene reporting technologies fall short of theserequirements. Chlorophyll is one of the best-recognized plant pigments with an alreadywell-developed remote imaging technology. However, chlorophyll is very abundant, withlevels regulated by both genetic and environmental factors. We designed a synthetic degreeningcircuit that produced rapid chlorophyll loss on perception of a specific input.With induction of the de-greening circuit, changes were remotely detected within 2 h.83
Analyses of multiple de-greening circuits suggested that the de-greening circuit functioned,in part, via light-dependent damage to photosystem cores and the production of reactiveoxygen species. Within 24–48 h of induction, an easily recognized white phenotyperesulted. Microarray analysis showed that the synthetic de-greening initiated a processlargely distinct from normal chlorophyll loss in senescence. Remarkably, syntheticallyde-greened white plants re-greened after removal of the inducer, providing the first easilyre-settable reporter system for plants and the capacity to make re-settable biosensors.Our results showed that the de-greening circuit allowed chlorophyll to be employed as asimple but powerful reporter system useful for widespread areas.Antunes MS et al. (2009) Engineering key components in a synthetic eukaryotic signaltransduction pathway. Molecular Systems Biology 5; Article number 270; doi:10.1038/msb.2009.28.Abstract: Signal transduction underlies how living organisms detect and respond tostimuli. A goal of synthetic biology is to rewire natural signal transduction systems. Bacteria,yeast, and plants sense environmental aspects through conserved histidine kinase(HK) signal transduction systems. HK protein components are typically comprised ofmultiple, relatively modular, and conserved domains. Phosphate transfer between thesecomponents may exhibit considerable cross talk between the otherwise apparently linearpathways, thereby establishing networks that integrate multiple signals. We show thatsequence conservation and cross talk can extend across kingdoms and can be exploitedto produce a synthetic plant signal transduction system. In response to HK cross talk,heterologously expressed bacterial response regulators, PhoB and OmpR, translocate tothe nucleus on HK activation. Using this discovery, combined with modification of PhoB(PhoBVP64), we produced a key component of a eukaryotic synthetic signal transductionpathway. In response to exogenous cytokinin, PhoB-VP64 translocates to the nucleus,binds a synthetic PlantPho promoter, and activates gene expression. These results showthat conserved-signaling components can be used across kingdoms and adapted to producesynthetic eukaryotic signal transduction pathways.Antunes MS et al. (2011) Programmable ligand detection system in plants through asynthetic signal transduction pathway. PLoS ONE 6(1): e16292. doi:10.1371/journal.pone.0016292.Background: There is an unmet need to monitor human and natural environments forsubstances that are intentionally or unintentionally introduced. A long-sought goal is toadapt plants to sense and respond to specific substances for use as environmental monitors.Computationally re-designed periplasmic binding proteins (PBPs) provide a meansto design highly sensitive and specific ligand sensing capabilities in receptors. Input fromthese proteins can be linked to gene expression through histidine kinase (HK) mediatedsignaling. Components of HK signaling systems are evolutionarily conserved betweenbacteria and plants. We previously reported that in response to cytokinin-mediated HK84 Food Security: The Intersection of Sustainability, Safety and Defense
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2011National Agricultural Biotechno
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NABC Report 23Food Security: The In
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AcknowledgmentsNABC’s twenty-thir
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Participants in the Student Voice a
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123 Preparing for Emerging and Unkn
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Food Security: The Intersection of
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These interconnected threads demand
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In the developed world, it will req
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On the other hand, his definition o
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Plenary Session 4. Emerging Biotech
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12 Food Security: The Intersection
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The participants’ responses did n
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they are safe. Cultural barriers ca
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• Hydrological mapping, as in Min
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• Food security is a multidiscipl
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awareness beyond anything that advo
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• Some research results indicate
Page 43 and 44: change, reduce or remove GHG emissi
Page 45 and 46: perspectives and priorities differ
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Page 49 and 50: Water• Withdrawals from the Color
Page 51 and 52: Efficient Use of Resources• We ne
Page 53 and 54: • Foley is working with Google to
Page 55 and 56: drinking water, stormwater, agricul
Page 57 and 58: Table 1A. Issues, strategies and de
Page 59 and 60: The Framework in Summary—A Ten- a
Page 61 and 62: If funded, Research Implementation
Page 63 and 64: If funded, Research Implementation
Page 65 and 66: to be initiated in the first five y
Page 67 and 68: Deborah Swackhamer is professor and
Page 69 and 70: These under-nutrition woes are not
Page 71 and 72: gies. Agricultural sustainability r
Page 73 and 74: Investment in TechnologyImproved pe
Page 75 and 76: Last November, the Forum announced
Page 77 and 78: Syngenta Foundation for Sustainable
Page 79 and 80: Food and Agriculture Organization (
Page 83 and 84: Richard Isaacson (University of Min
Page 85 and 86: possibility of cross-pollination th
Page 87 and 88: in part to companies like Cargill w
Page 89 and 90: Ralph Hardy (National Agricultural
Page 91 and 92: and the impacts those things have o
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Page 97 and 98: June Medford, a professor of biolog
Page 99 and 100: Genetically Modified CropsIt is cle
Page 101 and 102: the toxin levels are higher than wo
Page 103 and 104: • Separate common species• Sepa
Page 105 and 106: We have a similar situation in the
Page 107 and 108: ReferencesLövei GL et al. (2009) T
Page 109 and 110: Figure 1. Canada’s food-safety sy
Page 111 and 112: However, significant challenges rem
Page 113 and 114: confirmed cases in seven provinces,
Page 117 and 118: and from the IIASA Strategic Plan 2
Page 119 and 120: provided software support. We also
Page 121 and 122: Figure 4. Biological agents (classi
Page 125 and 126: Detlof von Winterfeldt is the direc
Page 127 and 128: Medford: For things like explosives
Page 129 and 130: Robert Buchanan (University of Mary
Page 131 and 132: Ishimaru: In particular with wheat,
Page 133 and 134: Duplessis: In terms of microfluidic
Page 137 and 138: Emergence/Re-EmergenceRegarding eme
Page 139 and 140: • Anticipate and prevent the emer
Page 141 and 142: AcuteWith a new emergence, the need
Page 143 and 144: Assessing Immune StatusDuring an ou
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Table 1. Recent zoonotic agents ide
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Figure 2. The impact of culling she
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ecognized as a new disease among ca
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The West Nile virus is another dram
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deer in an attempt to limit the tra
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disease centers for training, resea
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Lanciotti RS et al. (1999) Origin o
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Plant diseases still affect human h
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Climate ChangeThe predicted transit
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In addition to the NPDRS and the NP
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arely have a realistic understandin
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National Research Council (NRC) (20
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our universities we can increase aw
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evidence and no microbiologic evide
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food-preservation technologies and
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concern, particular products of con
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PreventionMuch can and should be do
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Figure 3. Selected recent multi-sta
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Figure 6. PulseNet electronic commu
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Figure 10. All Salmonella Typhimuri
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Figure 13. Listeriosis in the Unite
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Figure 15. Case definitions for clu
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Figure 18. The surveillance process
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Figure 20. Etiology of acute gastro
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I will discuss emerging intentional
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low-level contamination is on-going
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Sometimes these supply chains fall
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Clearly, the food system is globall
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Figure 13. 2010 Cocoa and cocoa-pre
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damage on those who profit from the
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target population. In their lives,
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it before it reaches the consumer.
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Shaun Kennedy is director of the Na
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ing replacements for cream, for exa
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safety system in reality by guarant
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Figure 2 shows the types of identif
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on better standards for Chinese sta
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Figure 6. Skim-milk-powder project:
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Jeffrey C. Moore is a scientific li
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John Besser: I was a little mystifi
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forensic purposes, as used with OJ
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my farmer friends thought I was cra
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Minnesota’s HeritageAs to why som
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in reality, legislation alone won
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The other aspect of this challenge
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private industry, enhancing the pro
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But it does seem that the United St
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Cathie Woteki is under secretary fo
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We offer the following comments and
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Ehlke, NancyUniversity of Minnesota
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Lumor, StephenUniversity of Minneso
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National Agricultural Biotechnology
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