Microanalysis of Plant Cell Wall Polysaccharides Obel Nicolai , Erben Veronika , Schwarz Tatjana , Kühnel Stefan , Fodor Andrea , Pauly Markus   Molecular Plant  Volume 2, Issue 5, Pages 922-932 (September 2009) DOI: 10.1093/mp/ssp046 Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

Figure 1 OLIMP Spectra of Wall Material Derived from 8-Week-Old Rosette Leaves. (A) Spectrum of solubilized XyG oligosaccharides (XyGOs) facilitating a xyloglucanase (XEG). The structures of the various ion signals are shown using the nomenclature described by Fry et al. (1993). Underlined structures represent O-acetylated sidechains. (B) Spectrum of solubilized oligogalacturonides using a combination of endopolygalacturonase (endoPG) and pectinmethylesterase (PME). Putative structures are shown. GalA, galacturonic acid; Me, methylester; Ac, O-acetyl substituent. Molecular Plant 2009 2, 922-932DOI: (10.1093/mp/ssp046) Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

Figure 2 Principal Component Analysis of OLIMP Derived from Different Arabidopsis Plant Organs. Various plant organs were analyzed by OLIMP with XEG (A) and the combination of endoPG and PME (B). Each point represents the average of at least three replicates from a single harvest, and each plant tissue was harvested three independent times. Black circles, trichomes; open triangles, petals; open circles, rosette leaf; black triangles, cauline leaves; open diamond, roots; open square, top stem segment; black square, middle stem segment; gray square, bottom stem segment; closed square, siliques; gray circle, petiole. Molecular Plant 2009 2, 922-932DOI: (10.1093/mp/ssp046) Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

Figure 3 Laser Dissection and Catapulting of Specific Leaf Cells. 20 μm thick cross-sections of paraffin-embedded leaves were used for laser dissecting. (A–E) show 10× magnification of a leaf cross-section. (A) Leaf cross-section. (B) Leaf cross-section after removal and collection of whole epidermis cells. (C) Same as (B) after removal of only the outer layer of the epidermis. (D) Removal of palisade cells. (E) Mesophyl cells sampling. (F) and (G) show a 40× magnification of the midrib before (F) and after (G) removal of the vascular tissue. Molecular Plant 2009 2, 922-932DOI: (10.1093/mp/ssp046) Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

Figure 4 PCA and Pair-Wise Comparison of Specific Plant Tissues as Shown in Figure 3. (A) Each point in the PCA represents the OLIMP profile of material collected using laser dissecting. The following material was collected: vascular tissue (star); outer wall of epidermis (circle), entire epidermis layer (diamond), palisade (triangle), and mesophyl (square). (B) Pair-wise comparison of vascular (gray bars) and mesophyl (white bars) samples.***(C) Pair-wise comparison of outer epidermis (gray bars) and whole epidermis (white bars) samples. Significant different abundance levels (p < 0.05) are indicated with an asterisk, n  =  3. Molecular Plant 2009 2, 922-932DOI: (10.1093/mp/ssp046) Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

Figure 5 In Situ OLIMP Analysis of Various Plant Tissue. (A) Roots of Arabidopsis WT and mur1 mutant placed on the target plate with added 0.5 μl of enzyme drops at various positions (halos, white triangles). (B) Etiolated 5-day-old Arabidopsis hypocotyl placed on the target plate with added enzyme drops at various positions (halos, white triangles). (C) OLIMP spectra obtained from the two encircled spots (white circles, (A)) of WT and mur1. (D) OLIMP spectra obtained from the two encircled spots (white circles, (B)) of cotyledons and root. The structures of the various ion signals are shown using the nomenclature described by Fry et al. (1993) including the J sidechain, where an L sidechain is extended with an α-L-Galp residue (Zablackis, 1996). Molecular Plant 2009 2, 922-932DOI: (10.1093/mp/ssp046) Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

Figure 6 XyG OLIMP Analysis of a Golgi-Enriched Fraction. (A) XyG OLIMP spectrum of an isolated Golgi-enriched fraction obtained from etiolated Arabidopsis hypocotyls. (B) Pair-wise comparison of XyGO profiles from cell walls (gray) and Golgi-enriched fraction (white). Significant different abundance levels (p < 0.05) are indicated with an asterisk, n  =  3. Molecular Plant 2009 2, 922-932DOI: (10.1093/mp/ssp046) Copyright © 2009 The Authors. All rights reserved. Terms and Conditions