GFP-FABD2 fusion construct allows in vivo visualization of the dynamic actin cytoskeleton in all cells of Arabidopsis seedlings.
In vivo visualization of filamentous actin in all cells of Arabidopsis thaliana seedlings is essential for understanding the numerous roles of the actin cytoskeleton in diverse processes of cell differentiation. A previously introduced reporter construct based on the actin-binding domain of mouse talin proved to be useful for unravelling some of these aspects in cell layers close to the organ surface. However, cells more deeply embedded, especially stelar cells active in polar transport of auxin, show either diffuse or no fluorescence at all due to the lack of expression of the fusion protein. The same problem is encountered in the root meristem. Recently introduced actin reporters based on fusions between A. thaliana fimbrin 1 and GFP gave brilliant results in organs from the root differentiation zone upwards to the leaves, however failed to depict the filamentous actin cytoskeleton in the transition zone of the root, in the apical meristem and the root cap. To overcome these problems, we have prepared new transgenic lines for the visualization of F-actin in vivo. We report here that a construct consisting of GFP fused to the C-terminal half of A. thaliana fimbrin 1 reveals dynamic arrays of F-actin in all cells of stably transformed A. thaliana seedlings.
Voigt
Boris
B
Department of Plant Cell Biology, Institute of Cellular and Molecular Botany, Rheinische Friedrich-Wilhelms-University of Bonn, Kirschallee 1, D-53115 Bonn, Germany.
Timmers
Antonius C J
AC
Samaj
Jozef
J
Müller
Jens
J
Baluska
Frantisek
F
Menzel
Diedrik
D
D000199:Actins / Q000378:metabolism*; D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics; D018522:Gravitropism; D049452:Green Fluorescent Proteins / Q000235:genetics; D018613:Microscopy, Confocal; D018524:Phototropism; D019441:Plant Epidermis / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D018515:Plant Leaves / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D018517:Plant Roots / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D036226:Seedlings / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*
C110898:ATFIM1 protein, Arabidopsis; D000199:Actins; D029681:Arabidopsis Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins