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Inhibitors of myosin, but not actin, alter transport through Tradescantia plasmodesmata

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Abstract

Actin and myosin are components of plasmodesmata, the cytoplasmic channels between plant cells, but their role in regulating these channels is unclear. Here, we investigated the role of myosin in regulating plasmodesmata in a well-studied, simple system comprising single filaments of cells which form stamen hairs in Tradescantia virginiana flowers. Effects of myosin inhibitors were assessed by analysing cell-to-cell movement of fluorescent tracers microinjected into treated cells. Incubation in the myosin inhibitor, 2,3-butanedione monoxime (BDM) or injection of anti-myosin antibodies increased cell–cell transport of fluorescent dextrans, while treatment with the myosin inhibitor N-ethylmaleimide (NEM) decreased cell–cell transport. Pretreatment with the callose synthesis inhibitor, deoxy-d-glucose (DDG), enhanced transport induced by BDM treatment or injection of myosin antibodies but did not relieve NEM-induced reduction in transport. In contrast to the myosin inhibitors, cell-to-cell transport was unaffected by treatment with the actin polymerisation inhibitor, latrunculin B, after controlling for callose synthesis with DDG. Transport was increased following azide treatment, and reduced after injection of ATP, as in previous studies. We propose that myosin detachment from actin, induced by BDM, opens T. virginiana plasmodesmata whereas the firm attachment of myosin to actin, promoted by NEM, closes them.

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Acknowledgements

This research was supported by the Australian Research Council (RW) and by an Australian Postgraduate Student Award (JR).

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  1. Department of Biological Sciences, Monash University, Melbourne, VIC, 3800, Australia

    Janine E. Radford & Rosemary G. White

  2. Computer Associates, 380 St Kilda Road, Melbourne, VIC, 3004, Australia

    Janine E. Radford

  3. Commonwealth Scientific and Industrial Research Organisation, Division of Plant Industry, GPO Box 1600, Canberra, ACT, 2601, Australia

    Rosemary G. White

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Supplementary Fig. 1

Tradescantia virginiana stamen hairs stained for 2 h with 2 μm ER-tracker white (Invitrogen) in buffer (0.5 mM HEPES, 0.1 mM KCl, 0.1 mM CaCl2, 0.1 mM MgCl2, 0.5 mM NaCl, pH 7) and observed under UV fluorescence. In a and c, fluorescent compartments were observed around the nucleus (n) and at the cell periphery b, d Corresponding brightfield images. a, b and c, d are at different planes of focus. (GIF 31 kb)

High resolution image (EPS 1999 kb)

Supplementary Fig. 2a

Movies showing a cytoplasmic streaming in freshly isolated Tradescantia virginiana stamen hair cells in buffer and b slow streaming after incubation in 30 mM BDM in buffer for 30 min total time elapsed = 1 min. (MP4 215 kb)

Supplementary Fig. 2b

(MP4 194 kb)

Supplementary Fig. 3a

Movies showing a cytoplasmic streaming in freshly isolated Tradescantia virginiana stamen hair cells in buffer and b absence of streaming after incubation in 1 mM NEM in buffer for 8 min total time elapsed = 1 min. (MP4 219 kb)

Supplementary Fig. 3b

(MP4 192 kb)

Supplementary Fig. 4a

Movies showing a cytoplasmic streaming in freshly isolated Tradescantia virginiana stamen hair cells in buffer and b absence of streaming and breakdown of cytoplasmic strands after incubation in 1 mM latrunculin B in buffer for 45 min total time elapsed = 1 min. (MP4 212 kb)

Supplementary Fig. 4b

(MP4 185 kb)

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Radford, J.E., White, R.G. Inhibitors of myosin, but not actin, alter transport through Tradescantia plasmodesmata. Protoplasma 248, 205–216 (2011). https://doi.org/10.1007/s00709-010-0244-3

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