ABSTRACT
In plants, cellulose is synthesized at the cell surface by plasma membrane (PM)-localized cellulose synthase (CESA) complexes (CSCs). The molecular and cellular mechanisms that underpin delivery of CSCs to the PM, however, are poorly understood. Cortical microtubules have been shown to interact with CESA-containing compartments and mark the site for CSC delivery, but are not required for the delivery itself. Here, we demonstrate that myosin XI and the actin cytoskeleton mediate CSC delivery to the PM by coordinating the exocytosis of CESA-containing compartments. Measurement of cellulose content indicated that cellulose biosynthesis was significantly reduced in a myosin xik xi1 xi2 triple knockout (xi3KO) mutant. By combining genetic and pharmacological disruption of myosin activity with quantitative live-cell imaging, we observed decreased abundance of PM-localized CSCs and reduced delivery rate of CSCs in myosin-deficient cells. These phenotypes correlated with a significant increase in failed vesicle secretion events at the PM as well as an abnormal accumulation of CESA-containing compartments at the cell cortex. Through high spatiotemporal assays of cortical vesicle behavior, we identified defects in CSC vesicle tethering and fusion at the PM. Furthermore, disruption of myosin activity reduced the delivery of several other secretory markers to the PM and reduced constitutive and receptor-mediated endocytosis. These findings reveal a previously undescribed role for myosin in vesicle secretion and cellulose production at the cytoskeleton–PM–cell wall nexus.
Footnotes
↵2 Author for contact: Christopher J. Staiger; staiger{at}purdue.edu.
One sentence summary: Arabidopsis myosins XI are responsible for trafficking and insertion of cellulose synthase complexes at the plasma membrane.
W.Z., C.C. and C.J.S. designed the research. W.Z. and C.C. performed the experiments and data analysis. W.Z. and C.J.S. wrote the article.
This work was supported by an award from the Office of Science at the US Department of Energy, Physical Biosciences Program, under contract number DE-FGO2-09ER15526 to C.J.S.
