Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set
Article first published online: 27 MAR 2009
© 2009 The Authors. Journal compilation © 2009 Blackwell Publishing Ltd
The Plant Journal
Volume 59, Issue 1, pages 169–178, July 2009
How to Cite
Geldner, N., Dénervaud-Tendon, V., Hyman, D. L., Mayer, U., Stierhof, Y.-D. and Chory, J. (2009), Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set. The Plant Journal, 59: 169–178. doi: 10.1111/j.1365-313X.2009.03851.x
- Issue published online: 25 JUN 2009
- Article first published online: 27 MAR 2009
- Received 15 January 2009; revised 9 February 2009; accepted 19 February 2009; published online 27 March 2009.
- membrane compartments;
- vesicle trafficking;
- subcellular marker lines;
- live imaging
Plant membrane compartments and trafficking pathways are highly complex, and are often distinct from those of animals and fungi. Progress has been made in defining trafficking in plants using transient expression systems. However, many processes require a precise understanding of plant membrane trafficking in a developmental context, and in diverse, specialized cell types. These include defense responses to pathogens, regulation of transporter accumulation in plant nutrition or polar auxin transport in development. In all of these cases a central role is played by the endosomal membrane system, which, however, is the most divergent and ill-defined aspect of plant cell compartmentation. We have designed a new vector series, and have generated a large number of stably transformed plants expressing membrane protein fusions to spectrally distinct, fluorescent tags. We selected lines with distinct subcellular localization patterns, and stable, non-toxic expression. We demonstrate the power of this multicolor ‘Wave’ marker set for rapid, combinatorial analysis of plant cell membrane compartments, both in live-imaging and immunoelectron microscopy. Among other findings, our systematic co-localization analysis revealed that a class of plant Rab1-homologs has a much more extended localization than was previously assumed, and also localizes to trans-Golgi/endosomal compartments. Constructs that can be transformed into any genetic background or species, as well as seeds from transgenic Arabidopsis plants, will be freely available, and will promote rapid progress in diverse areas of plant cell biology.