Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 110078
Title Determining Apoplastic pH Differences in Pea Roots by Use of the Fluorescent Dye Fluorescein
Author(s) Dorhout, R.; Kollöffel, C.
Source Journal of Experimental Botany 43 (1992)4. - ISSN 0022-0957 - p. 479 - 486.
Department(s) Laboratory of Nematology
Publication type Refereed Article in a scientific journal
Publication year 1992
Abstract The uptake of the fluorescent dye fluorescein (uranin) was used to determine apoplastic pH differences in pea roots. Fluorescein, an acidic dye (pKa≊4·9), is able to pass cell membranes in its undissociated form. The uptake was studied in roots of young and older pea seedlings growing in solution or in sand, respectively. Following 16 h of incubation in an unbuffered 0·01% (w/v) fluorescein solution with a pH of 7·9, the dye only accumulated in cells of the root apex, as was visible under ultraviolet irradiation. Dye accumulation was prevented when the proton buffering strength of the incubation medium was increased. Under more acidic conditions (pH 5·0) it also accumulated in the root base. Fluorescein uptake was strongly suppressed by CCCP, reduced by DNP and stimulated by KCl. The results indicate that net proton extrusion activity is the main mechanism for generating a low apoplastic pH. Cross-sections of roots of seedlings, incubated in the unbuffered fluorescein-containing medium, showed discriminative dye uptake by the different root tissues and cell types, suggesting that net proton extrusion activity may vary with the cell type and its location in the root. The method can be used to localize root regions, tissues, and cells inside the root, with a relatively low apoplastic pH.
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