Volume 4, Issue 5, Pages 794-804 (September 2011) Glow in the Dark: Fluorescent Proteins as Cell and Tissue-Specific Markers in Plants  Ckurshumova Wenzislava , Caragea Adriana E. , Goldstein Rochelle S. , Berleth Thomas   Molecular Plant  Volume 4, Issue 5, Pages 794-804 (September 2011) DOI: 10.1093/mp/ssr059 Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Box 1 Commonly Used Fluorescent Proteins in Plants. List of the most common variants, grouped by wavelength. * More then 30 mutations are involved in the optimization of the dsRed sequence (Campbell et al., 2002). ** The efficiency of the fluorophore in absorbing light, where a low extinction coefficient implies low levels of exciting light, reducing photo-oxidation due to free radical formation, and thus reducing damage of the fluorescent protein and neighboring molecules. 1 Data taken from Patterson et al. (2001). 2 Data taken from Shaner et al. (2005). 3 Data taken from Shaner et al. (2005). 4 Data taken from (Davidson and Campbel 2009). * More then 30 mutations are involved in the optimization of the dsRed sequence (Campbell et al., 2002). Molecular Plant 2011 4, 794-804DOI: (10.1093/mp/ssr059) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Box 2 Distinguishing Features of Commonly Used Imaging Systems. Molecular Plant 2011 4, 794-804DOI: (10.1093/mp/ssr059) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Figure 1 Specifying Procambial Cell Fate. (A) Morphologically distinct procambial cell (outlined in red), elongated along its longitudinal axis in the context of surrounding isodiametrical ground meristem cells in the Arabidopsis leaf. (B) Anatomically inconspicuous pre-procambial cells (note isodiametrical shape outlined in red) expressing the procambium specification marker ATHB8:HTA6:EYFP localized to the nucleus. (C) ATHB8::HTA6:EYFP. (D) SHR::mCherry-Nuc. (E) Co-localization of ATHB8 and SHR to the same anatomically inconspicuous pre-procambial cell. Images in (C-E) are color-coded with a dual-channel LUT, whereby fluorescence in each detection channel is shown in either magenta or cyan. Images have been modified with permission from Trends in Plant Science (A) and Developmental Dynamics (C–E) from references Berleth et al. (2000) and Sawchuk et al. (2007), respectively. Molecular Plant 2011 4, 794-804DOI: (10.1093/mp/ssr059) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Figure 2 Tracing Epidermal PIN1 Expression. Lateral convergence point (red arrowhead) in young leaf primordia visualized through confocal laser scanning microscopy using PIN1:GFP. (A) Overlay of fluorescene and transmitted light images. (B) Enlarged fluorescent image showing dual polarity. (C) LUT image of (B). Images were reproduced with permission from Genes and Development from reference Scarpella et al. (2006). m, mid vein. Molecular Plant 2011 4, 794-804DOI: (10.1093/mp/ssr059) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Figure 3 Presumptive Sites of Auxin Internalization–Epidermal Convergence Points. The direction of PIN1-dependent auxin flow (red arrows) is inferred from the polar subcellular localization of PIN1. (A) Epidermal auxin flow in late globular embryo, PIN1, localizes to establish two symmetrically positioned convergence points that coincide with the sites of future cotyledon emergence. (B) In the shoot apical meristem, auxin foci are associated with formation of leaf primordia. It is assumed that auxin is depleted from areas around emerging leaf primordia sub-epidermally through transport in the vascular system of the growing primordium, as well as epidermally through reversal of PIN1 polarity towards a forming leaf. (C) Deduced routes of auxin transport in early leaf primordia. (D) CUC2 is required for PIN1 relocalization leading to establishment of epidermal auxin maxima in the leaf margin. Elevated auxin concentrations in turn will down-regulate CUC2 to stabilize the location of the maximum. h, hypophyseal cell; s, suspensor. Molecular Plant 2011 4, 794-804DOI: (10.1093/mp/ssr059) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions

Figure 4 Use of GFP as a Tag. (A) The GAL4/GFP two-component system. Module 1, comprising a T-DNA harboring the GAL4 transcription factor and a GAL4-dependent GFP reporter. Upon random insertion in the genome, the GFP will reflect the expression pattern of a neighboring plant enhancer element. Module 2, used for targeting expression, comprises a T-DNA harboring a gene of interest under UAS control. (B) Schematic representation of a FACS experiment. Emission spectra of protoplasts individually passing through a photomultiplier tube is translated into computer-controlled charging and corresponding sorting of droplets according to charge. Empty droplets carry no charge. The method can produce sufficient amounts of RNA and derived cDNA for cell-type-specific full-transcriptome analyses. Molecular Plant 2011 4, 794-804DOI: (10.1093/mp/ssr059) Copyright © 2011 The Authors. All rights reserved. Terms and Conditions