Abstract
A method is described to measure photochemical activity in intact cells of Euglena under in vivo conditions. The method employs a cell wall digesting enzyme (cellulysin) to induce enough permeability in the cell walls and membranes in order to allow dyes, commonly used to investigate light-dependent electron transport reactions to enter, but without inducing a concomittant efflux of metabolites. Between 1 and 2 h of incubation in 5% (w/v) cellulysin provided conditions which allowed measurement of photosystem I-, II- and I+II-dependent electron transport with rates up to 600% higher than in control cells; whereas other cell wall degrading enzymes (cellulase and pectinase) still did not increase the entry of the dyes. Cellulysin up to 2 h of incubation had little or no effect on whole cell respiration, photosynthetic O2 evolution, or the export of potassium and (14C) labeled compounds out of cells; therefore cellulysin obviously did not change the normal habit or physiology of Euglena. Cellulysin (4 h digestion), cellulase and pectinase (2–4 h of incubation) on the other hand led to a lowering of respiration and light-dependent O2 evolution, and increased the efflux of K+, but apparently decreased that of (14C)labeled fixation products.
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Abbreviations
- DBMIB:
-
dibromothymoquinone
- DCPIP:
-
2,6-dichlorophenol-indophenol
- DCMU:
-
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- DMMIB:
-
2,3-dimethyl-5,6-methylenedioxy-p-benzoquinone
- MV:
-
methylviologen
- PSI:
-
photosystem I
- PS II:
-
photosystem II
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De Filippis, L.F., Hampp, R. An improved method of measuring photosynthetic electron transport in Euglena under in vivo conditions. Arch. Microbiol. 126, 237–243 (1980). https://doi.org/10.1007/BF00409926
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DOI: https://doi.org/10.1007/BF00409926