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

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.

doi:10.1038/s41598-017-14988-4

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

Journal Article · 08 November 2017 · Scientific Reports

DOI: https://doi.org/10.1038/s41598-017-14988-4 · OSTI ID:1507735

^[1]; Zehfroosh, Nina ^[2]; Hancock, Brandon L. ^[2]; Hines, Kevin ^[2]; Fang, Wenjuan ^[2]; Kilfoil, Maria ^[2]; Learned-Miller, Erik ^[2]; Sanguinet, Karen A. ^[2]; Goldner, Lori S. ^[2]; Baskin, Tobias I. ^[2]

Univ. of Massachusetts, Amherst, MA (United States); University of Massachusetts
Univ. of Massachusetts, Amherst, MA (United States)

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.

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Research Organization:: Univ. of Massachusetts, Amherst, MA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22). Chemical Sciences, Geosciences & Biosciences Division

Grant/Contract Number:: FG02-03ER15421

OSTI ID:: 1507735

Journal Information:: Scientific Reports, Journal Name: Scientific Reports Journal Issue: 1 Vol. 7; ISSN 2045-2322

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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The Fragile Fiber1 Kinesin Contributes to Cortical Microtubule-Mediated Trafficking of Cell Wall Components Zhu, Chuanmei; Ganguly, Anindya; Baskin, Tobias I. Plant Physiology, Vol. 167, Issue 3 https://doi.org/10.1104/pp.114.251462	journal	February 2015
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Visualization of Cellulose Synthase Demonstrates Functional Association with Microtubules Paredez, A. R. Science, Vol. 312, Issue 5779 https://doi.org/10.1126/science.1126551	journal	June 2006
Visualization of cellulose synthases in Arabidopsis secondary cell walls Watanabe, Y.; Meents, M. J.; McDonnell, L. M. Science, Vol. 350, Issue 6257 https://doi.org/10.1126/science.aac7446	journal	October 2015
Brachypodium distachyon and Setaria viridis : Model Genetic Systems for the Grasses Brutnell, Thomas P.; Bennetzen, Jeffrey L.; Vogel, John P. Annual Review of Plant Biology, Vol. 66, Issue 1 https://doi.org/10.1146/annurev-arplant-042811-105528	journal	April 2015
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