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Title: Imaging cellulose synthase motility during primary cell wall synthesis in the grass Brachypodium distachyon

Abstract

The mechanism of cellulose synthesis has been studied by characterizing the motility of cellulose synthase complexes tagged with a fluorescent protein; however, this approach has been used exclusively on the hypocotyl of Arabidopsis thaliana. Here we characterize cellulose synthase motility in the model grass, Brachypodium distachyon. We generated lines in which mEGFP is fused N-terminal to BdCESA3 or BdCESA6 and which grew indistinguishably from the wild type (Bd21-3) and had dense fluorescent puncta at or near the plasma membrane. Measured with a particle tracking algorithm, the average speed of GFP-BdCESA3 particles in the mesocotyl was 164 ± 78 nm min-1 (error gives standard deviation [SD], n = 1451 particles). Mean speed in the root appeared similar. For comparison, average speed in the A. thaliana hypocotyl expressing GFP-AtCESA6 was 184 ± 86 nm min-1 (n = 2755). For B. distachyon, we quantified root diameter and elongation rate in response to inhibitors of cellulose (dichlorobenylnitrile; DCB), microtubules (oryzalin), or actin (latrunculin B). Neither oryzalin nor latrunculin affected the speed of CESA complexes; whereas, DCB reduced average speed by about 50% in B. distachyon and by about 35% in A. thaliana. Evidently, between these species, CESA motility is well conserved.

Authors:
ORCiD logo [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1]
  1. Univ. of Massachusetts, Amherst, MA (United States)
Publication Date:
Research Org.:
Univ. of Massachusetts, Amherst, MA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22). Chemical Sciences, Geosciences & Biosciences Division
OSTI Identifier:
1507735
Grant/Contract Number:  
FG02-03ER15421
Resource Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 7; Journal Issue: 1; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Liu, Derui, Zehfroosh, Nina, Hancock, Brandon L., Hines, Kevin, Fang, Wenjuan, Kilfoil, Maria, Learned-Miller, Erik, Sanguinet, Karen A., Goldner, Lori S., and Baskin, Tobias I. Imaging cellulose synthase motility during primary cell wall synthesis in the grass Brachypodium distachyon. United States: N. p., 2017. Web. doi:10.1038/s41598-017-14988-4.
Liu, Derui, Zehfroosh, Nina, Hancock, Brandon L., Hines, Kevin, Fang, Wenjuan, Kilfoil, Maria, Learned-Miller, Erik, Sanguinet, Karen A., Goldner, Lori S., & Baskin, Tobias I. Imaging cellulose synthase motility during primary cell wall synthesis in the grass Brachypodium distachyon. United States. doi:10.1038/s41598-017-14988-4.
Liu, Derui, Zehfroosh, Nina, Hancock, Brandon L., Hines, Kevin, Fang, Wenjuan, Kilfoil, Maria, Learned-Miller, Erik, Sanguinet, Karen A., Goldner, Lori S., and Baskin, Tobias I. Wed . "Imaging cellulose synthase motility during primary cell wall synthesis in the grass Brachypodium distachyon". United States. doi:10.1038/s41598-017-14988-4. https://www.osti.gov/servlets/purl/1507735.
@article{osti_1507735,
title = {Imaging cellulose synthase motility during primary cell wall synthesis in the grass Brachypodium distachyon},
author = {Liu, Derui and Zehfroosh, Nina and Hancock, Brandon L. and Hines, Kevin and Fang, Wenjuan and Kilfoil, Maria and Learned-Miller, Erik and Sanguinet, Karen A. and Goldner, Lori S. and Baskin, Tobias I.},
abstractNote = {The mechanism of cellulose synthesis has been studied by characterizing the motility of cellulose synthase complexes tagged with a fluorescent protein; however, this approach has been used exclusively on the hypocotyl of Arabidopsis thaliana. Here we characterize cellulose synthase motility in the model grass, Brachypodium distachyon. We generated lines in which mEGFP is fused N-terminal to BdCESA3 or BdCESA6 and which grew indistinguishably from the wild type (Bd21-3) and had dense fluorescent puncta at or near the plasma membrane. Measured with a particle tracking algorithm, the average speed of GFP-BdCESA3 particles in the mesocotyl was 164 ± 78 nm min-1 (error gives standard deviation [SD], n = 1451 particles). Mean speed in the root appeared similar. For comparison, average speed in the A. thaliana hypocotyl expressing GFP-AtCESA6 was 184 ± 86 nm min-1 (n = 2755). For B. distachyon, we quantified root diameter and elongation rate in response to inhibitors of cellulose (dichlorobenylnitrile; DCB), microtubules (oryzalin), or actin (latrunculin B). Neither oryzalin nor latrunculin affected the speed of CESA complexes; whereas, DCB reduced average speed by about 50% in B. distachyon and by about 35% in A. thaliana. Evidently, between these species, CESA motility is well conserved.},
doi = {10.1038/s41598-017-14988-4},
journal = {Scientific Reports},
number = 1,
volume = 7,
place = {United States},
year = {2017},
month = {11}
}

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Cited by: 6 works
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Figures / Tables:

Figure 1 Figure 1: B. distachyon transgenic lines expressing GFP-BdCESA proteins. (Top) Representative plants at the heading stage grown in the greenhouse. T1 and T7 indicate independent transgenic lines. For GFP-CESA6, line T6 is shown. Scale bars = 3 cm. (Middle) Growth parameters for plants at the stage shown above. Data aremore » mean ± SD for 8 to 10 plants from a single experiment. (Bottom) RT-PCR detection of mRNA from the transgenes. The first two lanes (T1, T7) are the two GFP-CESA3 lines. UBC18 stands for ubiquitin-conjugating enzyme 18.« less

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Works referenced in this record:

In Vitro Versus in Vivo Cellulose Microfibrils from Plant Primary Wall Synthases: Structural Differences
journal, July 2002

  • Lai-Kee-Him, Joséphine; Chanzy, Henri; Müller, Martin
  • Journal of Biological Chemistry, Vol. 277, Issue 40
  • DOI: 10.1074/jbc.M203530200

Arabidopsis cortical microtubules position cellulose synthase delivery to the plasma membrane and interact with cellulose synthase trafficking compartments
journal, June 2009

  • Gutierrez, Ryan; Lindeboom, Jelmer J.; Paredez, Alex R.
  • Nature Cell Biology, Vol. 11, Issue 7
  • DOI: 10.1038/ncb1886

The roles of the cytoskeleton during cellulose deposition at the secondary cell wall
journal, June 2008


The Cell Biology of Cellulose Synthesis
journal, April 2014


Perturbation of Brachypodium distachyon CELLULOSE SYNTHASE A4 or 7 results in abnormal cell walls
journal, January 2013

  • Handakumbura, Pubudu P.; Matos, Dominick A.; Osmont, Karen S.
  • BMC Plant Biology, Vol. 13, Issue 1
  • DOI: 10.1186/1471-2229-13-131

A Set of Modular Binary Vectors for Transformation of Cereals
journal, November 2007

  • Himmelbach, Axel; Zierold, Uwe; Hensel, Götz
  • Plant Physiology, Vol. 145, Issue 4
  • DOI: 10.1104/pp.107.111575

Brachypodium distachyon as a Genetic Model System
journal, November 2015


Rapid formin-mediated actin-filament elongation is essential for polarized plant cell growth
journal, July 2009

  • Vidali, L.; van Gisbergen, P. A. C.; Guerin, C.
  • Proceedings of the National Academy of Sciences, Vol. 106, Issue 32
  • DOI: 10.1073/pnas.0901170106

Cellulose microfibril crystallinity is reduced by mutating C-terminal transmembrane region residues CESA1A903V and CESA3T942I of cellulose synthase
journal, February 2012

  • Harris, D. M.; Corbin, K.; Wang, T.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 11
  • DOI: 10.1073/pnas.1200352109

Genetic Evidence That Cellulose Synthase Activity Influences Microtubule Cortical Array Organization
journal, June 2008

  • Paredez, Alexander R.; Persson, Staffan; Ehrhardt, David W.
  • Plant Physiology, Vol. 147, Issue 4
  • DOI: 10.1104/pp.108.120196

Phytochrome Regulation of Cellulose Synthesis in Arabidopsis
journal, November 2011


Microrheology of Microtubule Solutions and Actin-Microtubule Composite Networks
journal, May 2009


Auxin, actin and growth of the Arabidopsis thaliana primary root: Auxin and actin interaction
journal, April 2007


A Mechanism for Sustained Cellulose Synthesis during Salt Stress
journal, September 2015


Visualization of cellulose synthases in Arabidopsis secondary cell walls
journal, October 2015


A Molecular Description of Cellulose Biosynthesis
journal, June 2015


Brachypodium: a promising hub between model species and cereals
journal, September 2014

  • Girin, Thomas; David, Laure C.; Chardin, Camille
  • Journal of Experimental Botany, Vol. 65, Issue 19
  • DOI: 10.1093/jxb/eru376

Quinclorac does not inhibit cellulose (cell wall) biosynthesis in sensitive barnyard grass and maize roots
journal, March 2003


Mutations of cellulose synthase (CESA1) phosphorylation sites modulate anisotropic cell expansion and bidirectional mobility of cellulose synthase
journal, September 2010

  • Chen, S.; Ehrhardt, D. W.; Somerville, C. R.
  • Proceedings of the National Academy of Sciences, Vol. 107, Issue 40
  • DOI: 10.1073/pnas.1012348107

Organization of cellulose synthase complexes involved in primary cell wall synthesis in Arabidopsis thaliana
journal, September 2007

  • Desprez, T.; Juraniec, M.; Crowell, E. F.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 39
  • DOI: 10.1073/pnas.0706569104

Cellulose synthase interactive protein 1 (CSI1) links microtubules and cellulose synthase complexes
journal, December 2011

  • Li, S.; Lei, L.; Somerville, C. R.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 1
  • DOI: 10.1073/pnas.1118560109

Pausing of Golgi Bodies on Microtubules Regulates Secretion of Cellulose Synthase Complexes in Arabidopsis
journal, April 2009

  • Crowell, E. F.; Bischoff, V.; Desprez, T.
  • The Plant Cell, Vol. 21, Issue 4, p. 1141-1154
  • DOI: 10.1105/tpc.108.065334

The trafficking of the cellulose synthase complex in higher plants
journal, March 2014

  • Bashline, Logan; Li, Shundai; Gu, Ying
  • Annals of Botany, Vol. 114, Issue 6
  • DOI: 10.1093/aob/mcu040

3,7-Dichloroquinolinecarboxylic Acid Inhibits Cell-Wall Biosynthesis in Maize Roots
journal, November 1996

  • Koo, S. J.; Neal, J. C.; DiTomaso, J. M.
  • Plant Physiology, Vol. 112, Issue 3
  • DOI: 10.1104/pp.112.3.1383

Thaxtomin A affects CESA-complex density, expression of cell wall genes, cell wall composition, and causes ectopic lignification in Arabidopsis thaliana seedlings
journal, March 2009

  • Bischoff, Volker; Cookson, Sarah Jane; Wu, Shuang
  • Journal of Experimental Botany, Vol. 60, Issue 3
  • DOI: 10.1093/jxb/ern344

Precise Nanometer Localization Analysis for Individual Fluorescent Probes
journal, May 2002


Xyloglucan Deficiency Disrupts Microtubule Stability and Cellulose Biosynthesis in Arabidopsis, Altering Cell Growth and Morphogenesis
journal, November 2015

  • Xiao, Chaowen; Zhang, Tian; Zheng, Yunzhen
  • Plant Physiology, Vol. 170, Issue 1
  • DOI: 10.1104/pp.15.01395

Biosynthesis of (1→3)-β- d -glucan (callose) by detergent extracts of a microsomal fraction from Arabidopsis thaliana : Biosynthesis of (1→3)-β-
journal, September 2001


The Fragile Fiber1 Kinesin Contributes to Cortical Microtubule-Mediated Trafficking of Cell Wall Components
journal, February 2015

  • Zhu, Chuanmei; Ganguly, Anindya; Baskin, Tobias I.
  • Plant Physiology, Vol. 167, Issue 3
  • DOI: 10.1104/pp.114.251462

Plant cellulose synthesis: CESA proteins crossing kingdoms
journal, April 2015


Patterning and Lifetime of Plasma Membrane-Localized Cellulose Synthase Is Dependent on Actin Organization in Arabidopsis Interphase Cells
journal, April 2013

  • Sampathkumar, Arun; Gutierrez, Ryan; McFarlane, Heather E.
  • Plant Physiology, Vol. 162, Issue 2
  • DOI: 10.1104/pp.113.215277

On the alignment of cellulose microfibrils by cortical microtubules: A review and a model
journal, March 2001


Variable-angle epifluorescence microscopy: a new way to look at protein dynamics in the plant cell cortex
journal, January 2008


Biosynthesis of Callose and Cellulose by Detergent Extracts of Tobacco Cell Membranes and Quantification of the Polymers Synthesized in vitro
journal, February 2010


Brachypodium distachyon and Setaria viridis : Model Genetic Systems for the Grasses
journal, April 2015


Making parallel lines meet: Transferring information from microtubules to extracellular matrix
journal, September 2012

  • Baskin, Tobias I.; Gu, Ying
  • Cell Adhesion & Migration, Vol. 6, Issue 5
  • DOI: 10.4161/cam.21121

NIH Image to ImageJ: 25 years of image analysis
journal, June 2012

  • Schneider, Caroline A.; Rasband, Wayne S.; Eliceiri, Kevin W.
  • Nature Methods, Vol. 9, Issue 7
  • DOI: 10.1038/nmeth.2089

High-efficiency Agrobacterium-mediated transformation of Brachypodium distachyon inbred line Bd21-3
journal, November 2007


Morlin, an inhibitor of cortical microtubule dynamics and cellulose synthase movement
journal, March 2007

  • DeBolt, S.; Gutierrez, R.; Ehrhardt, D. W.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 14
  • DOI: 10.1073/pnas.0700789104

Distribution fields for tracking
conference, June 2012

  • Sevilla-Lara, L.; Learned-Miller, E.
  • 2012 IEEE Conference on Computer Vision and Pattern Recognition (CVPR)
  • DOI: 10.1109/CVPR.2012.6247891

Herbicides that inhibit cellulose biosynthesis
journal, December 1999


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