Regulation of photosynthesis during heterocyst differentiation in Anabaena sp. strain PCC 7120 investigated in vivo at single-cell level by chlorophyll fluorescence kinetic microscopy.

Changes of photosynthetic activity in vivo of individual heterocysts and vegetative cells in the diazotrophic cyanobacterium Anabaena sp. strain PCC 7120 during the course of diazotrophic acclimation were determined using fluorescence kinetic microscopy (FKM). Distinct phases of stress and acclimation following nitrogen step-down were observed. The first was a period of perception, in which the cells used their internally stored nitrogen without detectable loss of PS II activity or pigments. In the second, the stress phase of nitrogen limitation, the cell differentiation occurred and an abrupt decline of fluorescence yield was observed. This decline in fluorescence was not paralleled by a corresponding decline in photosynthetic pigment content and PS II activity. Both maximal quantum yield and sustained electron flow were not altered in vegetative cells, only in the forming heterocysts. The third, acclimation phase started first in the differentiating heterocysts with a recovery of PS II photochemical yields [Formula: see text] Afterwards, the onset of nitrogenase activity was observed, followed by the restoration of antenna pigments in the vegetative cells, but not in the heterocysts. Surprisingly, mature heterocysts were found to have an intact PS II as judged by photochemical yields, but a strongly reduced PS II-associated antenna as judged by decreased F 0. The possible importance of the functional PS II in heterocysts is discussed. Also, the FKM approach allowed to follow in vivo and evaluate the heterogeneity in photosynthetic performance among individual vegetative cells as well as heterocysts in the course of diazotrophic acclimation. Some cells along the filament (so-called "superbright cells") were observed to display transiently increased fluorescence yield, which apparently proceeded by apoptosis.

Acclimation of Trichodesmium erythraeum ISM101 to high and low irradiance analysed on the physiological, biophysical and biochemical level.

As the nonheterocystous diazotrophic cyanobacterium Trichodesmium lives both at the ocean surface and deep in the water column, it has to acclimate to vastly different irradiances. Here, we investigate its strategy of light acclimation in several ways. In this study, we used spectrally resolved fluorescence kinetic microscopy to investigate the biophysics of photosynthesis in individual cells, analysed cell extracts for pigment and phycobiliprotein composition, measured nitrogenase activity and the abundance of key proteins, and assayed protein synthesis/degradation by radioactive labelling. After acclimation to high light, Trichodesmium grew faster at 1000 micromol m(-2) s(-1) than at 100 micromol m(-2) s(-1). This acclimation was associated with decreasing cell diameter, faster protein turnover, the down-regulation of light-harvesting pigments and the outer part of the phycobiliprotein antenna, the up-regulation of light-protective carotenoids, changes in the coupling of phycobilisomes to the reaction centres and in the coupling of individual phycobiliproteins to the phycobilisomes. The latter was particularly interesting, as it represents an as yet unreported light acclimation strategy. Only in the low light-acclimated culture and only after the onset of actinic light did phycourobilin and phycoerythrin contribute to photochemical fluorescence quenching, showing that these phycobiliproteins may become quickly (in seconds) very closely coupled to photosystem II. This fast reversible coupling also became visible in the nonphotochemical changes of the fluorescence quantum yield.

Iron limitation in the marine cyanobacterium Trichodesmium reveals new insights into regulation of photosynthesis and nitrogen fixation.

* As iron (Fe) deficiency is a main limiting factor of ocean productivity, its effects were investigated on interactions between photosynthesis and nitrogen fixation in the marine nonheterocystous diazotrophic cyanobacterium Trichodesmium IMS101. * Biophysical methods such as fluorescence kinetic microscopy, fast repetition rate (FRR) fluorimetry, and in vivo and in vitro spectroscopy of pigment composition were used, and nitrogenase activity and the abundance of key proteins were measured. * Fe limitation caused a fast down-regulation of nitrogenase activity and protein levels. By contrast, the abundance of Fe-requiring photosystem I (PSI) components remained constant. Total levels of phycobiliproteins remained unchanged according to single-cell in vivo spectra. However, the regular 16-kDa phycoerythrin band decreased and finally disappeared 16-20 d after initiation of Fe limitation, concomitant with the accumulation of a 20-kDa protein cross-reacting with the phycoerythrin antibody. Concurrently, nitrogenase expression and activity increased. Fe limitation dampened the daily cycle of photosystem II (PSII) activity characteristic of diazotrophic Trichodesmium cells. Further, it increased the number and prolonged the time period of occurrence of cells with elevated basic fluorescence (F(0)). Additionally, it increased the effective cross-section of PSII, probably as a result of enhanced coupling of phycobilisomes to PSII, and led to up-regulation of the Fe stress protein IsiA. * Trichodesmium survives short-term Fe limitation by selectively down-regulating nitrogen fixation while maintaining but re-arranging the photosynthetic apparatus.

Copper-induced inhibition of photosynthesis: limiting steps of in vivo copper chlorophyll formation in Scenedesmus quadricauda.

The in vivo substitution of Mg2+ in chlorophyll by heavy metals is an important damage mechanism in heavy metal-stressed plants that leads to an inhibition of photosynthesis. In photosynthetic organisms with LHC II antennae, the in vivo substitution of Mg2+ by Cu2+ occurs particularly readily under low irradiance with a dark phase - a phenomenon referred to as 'shade reaction'. In the present study the limiting steps of the shade reaction were investigated with synchronised cultures of the chlorococcal green alga Scenedesmus quadricauda (Turp.) Bréb. The rate of copper chlorophyll formation during shade reaction was shown to be controlled by several factors; firstly, in some phases of the cell cycle, especially at the end of the light period, Mg2+ in chlorophyll was not accessible to substitution. This pattern is likely to be caused by cell cycle-dependent changes in photosynthesis and thylakoid ultrastructure, which were published earlier and are reconsidered in the discussion of the present work. Secondly, prolonged culture in a medium containing 3 µM Cu2+ reversibly increased the resistance of the strain to Cu2+. Culturing without added Cu2+ lowered the threshold concentrations of various deleterious effects more than 10-fold within 8 months of de-adaptation. Adaptation to high Cu2+ levels is discussed in the context of studies of the regulation of metal transporter proteins. In addition, it was also observed that toxic Cu2+ levels impaired photosynthesis sooner than cell division.