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ABSTRACT: In this paper we report the results of measurements performed by FLIM on the photoreceptor of Euglenagracilis. This organelle consists of optically bistable proteins, characterized by two thermally stable isomeric forms: A(498,) non fluorescent and B(462), fluorescent. Our data indicate that the primary photoevent of Euglena photoreception upon photon absorption consists of two contemporaneous different phenomena: an intramolecular photo-switch (i.e., A(498) becomes B(462)), and a intermolecular and unidirectional Forster-type energy transfer. During the FRET process, the fluorescent B(462) form acts as donor for the non-fluorescent A(498) form of the protein nearby, which acts as acceptor. We hypothesize that in nature these phenomena follow each other with a domino progression along the orderly organized and closely packed proteins in the photoreceptor layer(s), modulating the isomeric composition of the photoreceptive protein pool. This mechanism guarantees that few photons are sufficient to produce a signal detectable by the cell.
Biochemical and Biophysical Research Communications 06/2009; 385(2):176-80. · 2.48 Impact Factor
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ABSTRACT: In microorganisms and plants, chromium (Cr) is not essential for any metabolic process, and can ultimately prove highly deleterious. Due to its widespread industrial use, chromium has become a serious pollutant in diverse environmental settings. The presence of Cr leads to the selection of specific algal populations able to tolerate high levels of Cr compounds. The varying Cr-resistance mechanisms displayed by microorganisms include biosorption, diminished accumulation, precipitation, reduction of Cr(6+) to Cr(3+), and chromate efflux. In this paper we describe the effects of Cr(6+) (the most toxic species) on the photosynthetic and photoreceptive apparatus of two fresh water microalgae, Eudorina unicocca and Chlorella kessleri. We measured the effect of this heavy metal by means of in vivo absorption microspectroscopy of both the thylakoid compartments and the eyespot. The decomposition of the overall absorption spectra in pigment constituents indicates that Cr(6+) effects are very different in the two algae. In E. unicocca the metal induced a complete pheophinitization of the chlorophylls and a modification of the carotenoids present in the eyespot after only 120 h of exposition at a concentration equal or greater than 40 microM, which is the limit for total Cr discharge established by US EPA regulations. In C. kessleri, chromium concentrations a hundred times higher than this limit had no effect on the photosynthetic machinery. The different tolerance level of the two algae is suggested to be due to the different properties of the mucilaginous envelope and cell wall covering, respectively, the colonies of Eudorina and the single cells of Chlorella, which binds chromium cations to a different extent.
Journal of Environmental Monitoring 12/2008; 10(11):1313-8. · 1.99 Impact Factor
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ABSTRACT: Chromium is a highly toxic non-essential metal for microorganisms and plants. Due to its widespread industrial use, chromium (Cr) has become a serious pollutant in diverse environmental settings. The presence of Cr leads to the selection of algal populations able to tolerate high levels of Cr compounds. The diverse Cr-resistance mechanisms displayed by microorganisms include biosorption, diminished accumulation, precipitation, reduction of Cr(6+) to Cr(3+), and chromate efflux. In this paper we describe the effects of Cr(6+) (the more toxic species) on the photosynthetic and photoreceptive apparatus of the fresh water unicellular alga Chlamydomonas reinhardtii. We measured the effect of the heavy metal by means of in vivo absorption microspectroscopy of both the thylakoid compartments and the eyespot. The decomposition of the overall absorption spectra in pigment constituents indicates that Cr(6+) induced a complete pheophinitization of the chrorophylls and a modification of the carotenoids present in the eyespot only when its concentration is equal or greater than 10 microM. Due to this low tolerance level, C. reinhardtii could be used as indicator of Cr pollution, but it is not feasible for bioremediation purposes.
Environmental Research 11/2007; 105(2):234-9. · 3.40 Impact Factor
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ABSTRACT: A microspectrophotometer is a digital microscope used to measure absorption and fluorescence spectra. In this paper we describe a polychromator-based microspectrophotometer that performs in vivo absorption or emission measurements at the same time on different subcellular compartments such as photoreceptive and photosynthetic structures of algal cells. In this system, a flat field imaging concave grating polychromator is connected to the slit-shaped exit pupil of a light-guide probe mounted onto a microscope equipped with an epifluorescence module. The subcellular components, on which the spectra will be measured, are placed in the microscope field and finely adjusted. The outer bundle of the probe is used for centering the objects, while the central bundle of the probe, containing 19 light guides, is used for acquiring either transmitted or emitted light (i.e. fluorescence). The light transmitted or emitted by the subcellular components is collected by the probe mounted in the back focal plane of the ocular. The exit pupil of this probe, connected to a flat field imaging concave grating polychromator, produces a dispersion image that in turn is focused onto a digital slow scan cooled CCD camera. Absorption and emission spectra of algal subcellular compartments are presented.
International journal of biological sciences 02/2007; 3(4):251-6. · 2.70 Impact Factor
