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Glutathione biosynthesis in Arabidopsis trichome cells

By Gloria Gutiérrez-Alcalá, Cecilia Gotor, Andreas J. Meyer, Mark Fricker, José M. Vega and Luis C. Romero

Abstract

In <em>Arabidopsis thaliana</em>, trichome cells are specialized unicellular structures with uncertain functions. Based on earlier observations that one of the genes involved in cysteine biosynthesis (Atcys-3A) is highly expressed in trichomes, we have extended our studies in trichome cells to determine their capacity for glutathione (GSH) biosynthesis. First, we have analyzed by <em>in situ</em> hybridization the tissue-specific expression of the genes Atcys-3A and sat5, which encode O-acetylserine(thio)lyase (OASTL) and serine acetyltransferase (SAT), respectively, as well as gsh1 and gsh2, which encode γ-glutamylcysteine synthetase and glutathione synthetase, respectively. The four genes are highly expressed in leaf trichomes of <em>Arabidopsis</em>, and their mRNA accumulate to high levels. Second, we have directly measured cytoplasmic GSH concentration in intact cells by laser-scanning microscopy after labeling with monochlorobimane as a GSH-specific probe. From these measurements, cytosolic GSH concentrations of 238 ± 25, 80 ± 2, and 144 ± 19 μM were estimated for trichome, basement, and epidermal cells, respectively. Taking into account the volume of the cells measured using stereological techniques, the trichomes have a total GSH content more than 300-fold higher than the basement and epidermal cells. Third, after NACl treatment, GSH biosynthesis is markedly decreased in trichomes. Atcys-3A, sat5, gsh1, and gsh2 mRNA levels show a decrease in transcript abundance, and [GSH] <sub>cyt</sub> is reduced to 47 ± 5 μM. These results suggest the important physiological significance of trichome cells related to GSH biosynthesis and their possible role as a sink during detoxification processes.The full-text of this article is not currently available in ORA, but you may be able to access the article via the publisher copy link on this record page. Citation: Gutiérrez-Alcalá, G. et al. (2000). 'Glutathione biosynthesis in <em>Arabidopsis</em> trichome cells', PNAS 97(20), 11108-11113. [Available at http://www.pnas.org/]

Topics: Plant Sciences, cysteine, glutathione, two-photon laser-scanning microscopy, O-acetylserine(thio)lyase, sulfate assimilation
Year: 2000
DOI identifier: 10.1073/pnas.0900206106
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