Journal Article

Immunolocalization of actin in root statocytes of <i>Lens culinaris</i> L.

D. Driss‐Ecole, J. Vassy, J. Rembur, A. Guivarc'h, M. Prouteau, W. Dewitte and G. Perbal

in Journal of Experimental Botany

Published on behalf of Society for Experimental Biology

Volume 51, issue 344, pages 521-528
Published in print March 2000 | ISSN: 0022-0957
Published online March 2000 | e-ISSN: 1460-2431 | DOI: http://dx.doi.org/10.1093/jexbot/51.344.521
Immunolocalization of actin in root statocytes of Lens culinaris L.

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Lentil root statocytes show a strict structural polarity of their organelles with respect to the g vector. These cells are involved in the perception of gravity and are responsible for the orientation of the root. Actin filaments take part in the positioning of their organelles and could also be involved in the transduction of the gravitropic signal. A pre‐embedding immunogold silver technique was carried out with a monoclonal antibody in order to study the distribution of actin cytoskeleton in the statocytes at the electron microscopic level. Some areas were never labelled (cell wall, vacuole, nucleoplasm, mitochondria, starch grains of the amyloplasts) or very slightly labelled (stroma of the amyloplasts). The labelling was scattered in the cytoplasm always close to, or on the nuclear and amyloplast envelopes and the tonoplast. Associations of 2 to 6 dots in file were observed, but these short files were not oriented in one preferential direction. They corresponded to a maximum distance of 0.9 μm. This work demonstrated that each statocyte organelle was enmeshed in an actin web of short filaments arranged in different ways. The images obtained by rhodamine‐phalloidin staining were in accordance with those of immunogold labelling. The diffuse fluorescence of the cytoplasm could be explained by the fact that the meshes of the web should be narrow. The vicinity of actin and of the amyloplasts envelope could account for the movement of these organelles that was observed in spatial microgravity.

Keywords: Actin; cytoskeleton; immunolocalization; lentil root; pre‐embedding technique; statocyte.; BB, Blocking buffer; BCIP/NBT, 5‐bromo‐4‐chloro‐3‐indolyl phosphatase/nitroblue tetrazolium; BSA, bovine serum albumin; CRA, cress root actin antibody; DMSO, dimethylsulphoxide; EDTA, ethylenediaminetetraacetic acid; EGTA, ethyleneglycol‐bis‐(β‐aminoethylether)‐N,N,N′,N′‐tetraacetic acid; ICN, anti‐chicken gizzard actin IgG, Biomedicals, Meckenheim/Germany; MBS, m‐maleimidobenzoyl N‐hydroxysuccinimide ester; NaN3, sodium azide; NBT, (p‐nitroblue tetrazolium chloride); NGS, normal goat serum; PIPES, piperazine‐N,N′‐bis(2‐ethane‐sulphonic acid); PBS, phosphate‐buffered saline; PMSF, phenylmethylsulfonyl fluoride; TBS, Tris‐buffered saline.

Journal Article.  4313 words.  Illustrated.

Subjects: Plant Sciences and Forestry

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