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Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set
Author(s) -
Geldner Niko,
DénervaudTendon Valérie,
Hyman Derek L.,
Mayer Ulrike,
Stierhof YorkDieter,
Chory Joanne
Publication year - 2009
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2009.03851.x
Subject(s) - arabidopsis , biology , golgi apparatus , microbiology and biotechnology , endosome , protein subcellular localization prediction , endomembrane system , context (archaeology) , subcellular localization , arabidopsis thaliana , plant cell , live cell imaging , transport protein , membrane protein , immunoelectron microscopy , protein targeting , cell , membrane , genetics , mutant , gene , endoplasmic reticulum , paleontology , cytoplasm , antibody , intracellular
Summary 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.