The γ-subunits of chloroplast ATP synthases are about 30 amino acids longer than the bacterial or mitochondrial homologous proteins. This additional sequence is located in the mean part of the polypeptide chain and includes in green algae and higher plants two cysteines (Cys198 and Cys204 in Chlamydomonas reinhardtii) responsible for thiol regulation. In order to investigate its functional significance, a segment ranging from Asp-D210 to Arg-226 in the γ-subunit of chloroplast ATP synthase from C. reinhardtii was deleted. This deletion mutant called T2 grows photoautotrophically, but slowly than the parental strain. The chloroplast ATP synthase complex with the mutated γ is assembled, membrane bound, and as CF0CF1 displays normal ATPase activity, but photophosphorylation is inhibited by about 20 %. This inhibition is referred to lower light-induced transmembrane proton gradient. Reduction of the proton gradient is apparently caused by a disturbed functional connection between CF1 and CF0 effecting a partially leaky ATP synthase complex.
We recently developed a chlorophyll a fluorescence method (activated F0 rise) for estimating if a light wavelength preferably excites PSI or PSII in plants. Here, the method was tested in green microalgae: Scenedesmus quadricauda, Scenedesmus ecornis, Scenedesmus fuscus, Chlamydomonas reinhardtii, Chlorella sorokiniana, and Ettlia oleoabundans. The Scenedesmus species displayed a plant-like action spectra of F0 rise, suggesting that PSII/PSI absorption ratio is conserved from higher plants to green algae. F0 rise was weak in a strain of C. reinhardtii, C. sorokiniana, and E. oleoabundans. Interestingly, another C. reinhardtii strain exhibited a strong F0 rise. The result indicates that the same illumination can lead to different redox states of the plastoquinone pool in different algae. Flavodiiron activity enhanced the F0 rise, presumably by oxidizing the plastoquinone pool during pre-illumination. The activity of plastid terminal oxidase, in turn, diminished the F0 rise, but to a small degree.
We describe an instrument that allows the rapid measurement of fluorescence lifetime-resolved images of leaves as well as sub-cellular structures of intact plants or single cells of algae. Lifetime and intensity fluorescence images can be acquired and displayed in real time (up to 55 lifetime-resolved images per s). Our imaging technique therefore allows rapid measurements that are necessary to determine the fluorescence lifetimes at the maximum (P level) fluorescence following initial illumination during the chlorophyll (Chl) a fluorescence transient (induction) in photosynthetic organisms. We demonstrate the application of this new instrument and methodology to measurements of: (1) Arabidopsis thaliana leaves showing the effect of dehydration on the fluorescence lifetime images; (2) Zea mays leaves showing differences in the fluorescence lifetimes due to differences in the bundle sheath cells (having a higher amount of low yield photosystem 1) and the mesophyll cells (having a higher amount of high yield photosystem 2); and (3) single cells of wild type Chlamydomonas reinhardtii and its non-photochemical quenching mutant NPQ2 (where the conversion of zeaxanthin to violaxanthin is blocked), with NPQ2 showing lowered lifetime of Chl a fluorescence. In addition to the lifetime differences referred to in (1) and (2), structural dependent heterogeneities in the fluorescence lifetimes were generally observed when imaging mesophyll cells in leaves. and O. Holub ... [et al.].
Fluorescence parameters obtained during steady-state electron transport are frequently used to evaluate photosynthetic efficiency of plants. We studied the behaviour of those parameters as a function of irradiance-adapted fluorescence yields FS and F'M. Applied simulations showed that photochemical quenching evaluated by qP is greatly influenced by the steady-state fluorescence level (FS), and that its evolution is not complementary to non-photochemical quenching (qN). On the other hand, the relative photochemical and non-photochemical quenching coefficients (qP(rel) and qN(rel)) proposed by Buschmann (1995) represent better the balance between the energy dissipation pathways. However, these relative parameters are also non-linearly related when the FS level is varied. We investigated the application of a new parameter, the relative unquenched fluorescence (UQF(rel)) which takes into account the fraction of non-quenched fluorescence yield (FS), which is related to closed photosystem 2 reaction centres not participating in electron transport. By using computer simulations and real in vivo measurements, we found that this new parameter is complementary to qP(rel) and qN(rel), which may facilitate the use of PAM fluorescence as diagnostic tool in environmental studies. and P. Juneau, B. R. Green, P. J. Harrison.