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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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Works referenced in this record:

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