Multidimensional mapping of root responses to soil environmental cues using a luminescence-based imaging system

Multidimensional mapping of root responses to soil environmental cues using a luminescence-based imaging system

Root systems develop different root types that individually sense cues from their local environment and integrate them with systemic signals. This complex multi-dimensional amalgam of inputs leads to continuous adjustment of root growth rates, direction and metabolic activity to define a dynamic physical network. Current methods for analyzing root biology balance physiological relevance with imaging capability. To bridge this divide, we developed an integrated imaging system called Growth and Luminescence Observatory for Roots (GLO-Roots) that uses luminescence-based reporters to enable studies of root architecture and gene expression patterns in soil-grown, light-shielded roots. We have developed image analysis algorithms that allow the spatial integration of soil properties such as soil moisture with root traits. We propose GLO-Roots as a system that has great utility in both presenting environmental stimuli to roots in ways that evoke natural adaptive responses, and in providing tools for developing a multi-dimensional understanding of such processes.

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Rubén Rellán Álvarez

April 05, 2015
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    Multidimensional mapping of root responses to soil environmental cues ubén

    Rellán Álvarez bit.ly/glo-roots-preprint bit.ly/glo-roots-slides www.rrlab.org/glo_roots @rubenrellan rrellan@langebio.cinvestav.mx www.rrlab.org
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    Roots integrate a variety of signals and adjust function and

    shape accordingly Drew et al. 1978 JXB 109:435 Lima et al. 2011 PMID: 21119058 Tabata et al. 2014PMID: 25324386 AMT1:GFP
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    Yun et al. 2014 PMID: 24927545 From metabolite in vivo

    imaging to 3D in soil root architecture Grossman et al. 2011 PMID: 22186371 Soil Artificial Substrate
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    Yun et al. 2014 PMID: 24927545 From metabolite in vivo

    imaging to 3D in soil root architecture Grossman et al. 2011 PMID: 22186371 Soil Artificial Substrate
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    Soil Soil Properties Time Root Structure Gene Expression A multidimensional

    map of root responses to soil environmental cues
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    Soil Microbe Colonization Soil Properties Time Root Structure Gene Expression

    A multidimensional map of root responses to soil environmental cues
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    Soil Microbe Colonization Soil Properties Time Root Structure Gene Expression

    A multidimensional map of root responses to soil environmental cues
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    30 cm 15 cm 2 mm 100 cm3 of soil

    Rhizotrons: a 2D sample of soil
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    30 cm 15 cm 2 mm 100 cm3 of soil

    Rhizotrons: a 2D sample of soil
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    The expression profile of roots grown in pots and rhizotrons

    is very similar Fresh Weight FW (mg) 0 60 120 180 240 MS Pot Rhizotron
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    Different environmental cues can be recreated and quantified in rhizotrons

    water Phosphate 80 60 40 20 0 moisture content (%)
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    Different environmental cues can be recreated and quantified in rhizotrons

    water Phosphate 80 60 40 20 0 moisture content (%)
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    Different environmental cues can be recreated and quantified in rhizotrons

    water Phosphate 80 60 40 20 0 moisture content (%)
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    We use luciferase to visualize roots in soil www.firefly.org/firefly-pictures.html Luciferase

    is great! • Does not require any light excitation • No background luminescence • LUC can be used in temporal studies • Dynamic response to gene expression • Multiple color reporters are possible luciferin + ATP + O2 → oxyluciferin +
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    GLO-RIA enables semi-automatic quantification of RSA traits 11 12 13

    14 15 16 17 18 19 20 21 Angle Count 0 45 90 0 45 90 0 45 90 0 45 90 0 45 90 0 45 90 0 45 90 0 45 90 0 45 90 0 45 90 0 45 90 Root system area
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    If water deprivation starts later there is an induction of

    lateral root growth and a higher proliferation of tertiary roots
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    500 540 580 620 660 700 UBQ10:vLUC2 ACT2:PpyREo We have

    generated a golden gate ready collection of different luciferases Intensity 1005 1064 1123 1182 1241 1300 UBQ10:CBRo Intensity 1005 1064 1123 1182 1241 1300 500 540 580 620 660 700 UBQ10:LUC2o 500 540 580 620 660 700 UBQ10:CBGo LUC2o Maximum Intensity vLUC CBG99o PpyRE8o CBRo λ (nm)
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    Soil Microbe Colonization Soil Properties Time Root Structure Gene Expression

    A multidimensional map of root responses to soil environmental cues
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    Soil Microbe Colonization Soil Properties Time Root Structure Gene Expression

    A multidimensional map of root responses to soil environmental cues
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    Soil Microbe Colonization Soil Properties Time Root Structure Gene Expression

    A multidimensional map of root responses to soil environmental cues
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    Dinneny Lab Muh-Ching Yee Geng Yu Emilie Winfield Pierre-Luc Pradier

    Charlotte Trontin Thérèse LaRue Heike Lindner Guillaume Lobet Image Analysis Jose Sebastian John Vogel Julin Maloof Other Species Cara Haney
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