Assembly of D1/D2 complexes of photosystem II: Binding of pigments and a network of auxiliary proteins.

Photosystem II (PSII) is the multi-subunit light-driven oxidoreductase that drives photosynthetic electron transport using electrons extracted from water. To investigate the initial steps of PSII assembly, we used strains of the cyanobacterium Synechocystis sp. PCC 6803 arrested at early stages of PSII biogenesis and expressing affinity-tagged PSII subunits to isolate PSII reaction center assembly (RCII) complexes and their precursor D1 and D2 modules (D1mod and D2mod). RCII preparations isolated using either a His-tagged D2 or a FLAG-tagged PsbI subunit contained the previously described RCIIa and RCII* complexes that differ with respect to the presence of the Ycf39 assembly factor and high light-inducible proteins (Hlips) and a larger complex consisting of RCIIa bound to monomeric PSI. All RCII complexes contained the PSII subunits D1, D2, PsbI, PsbE, and PsbF and the assembly factors rubredoxin A and Ycf48, but we also detected PsbN, Slr1470, and the Slr0575 proteins, which all have plant homologs. The RCII preparations also contained prohibitins/stomatins (Phbs) of unknown function and FtsH protease subunits. RCII complexes were active in light-induced primary charge separation and bound chlorophylls (Chls), pheophytins, beta-carotenes, and heme. The isolated D1mod consisted of D1/PsbI/Ycf48 with some Ycf39 and Phb3, while D2mod contained D2/cytochrome b559 with co-purifying PsbY, Phb1, Phb3, FtsH2/FtsH3, CyanoP, and Slr1470. As stably bound, Chl was detected in D1mod but not D2mod, formation of RCII appears to be important for stable binding of most of the Chls and both pheophytins. We suggest that Chl can be delivered to RCII from either monomeric Photosystem I or Ycf39/Hlips complexes.

Functional coupling of duplex translocation to DNA cleavage in a type I restriction enzyme.

Type I restriction-modification enzymes are multifunctional heteromeric complexes with DNA cleavage and ATP-dependent DNA translocation activities located on motor subunit HsdR. Functional coupling of DNA cleavage and translocation is a hallmark of the Type I restriction systems that is consistent with their proposed role in horizontal gene transfer. DNA cleavage occurs at nonspecific sites distant from the cognate recognition sequence, apparently triggered by stalled translocation. The X-ray crystal structure of the complete HsdR subunit from E. coli plasmid R124 suggested that the triggering mechanism involves interdomain contacts mediated by ATP. In the present work, in vivo and in vitro activity assays and crystal structures of three mutants of EcoR124I HsdR designed to probe this mechanism are reported. The results indicate that interdomain engagement via ATP is indeed responsible for signal transmission between the endonuclease and helicase domains of the motor subunit. A previously identified sequence motif that is shared by the RecB nucleases and some Type I endonucleases is implicated in signaling.

Tracking viral movement in plants by means of chlorophyll fluorescence imaging.

Many techniques have been applied to understand viral cell-to-cell movement in host plants, but little progress has been made in understanding viral vascular transport mechanisms. We propose the use of chlorophyll fluorescence imaging techniques, not only to diagnose the viral infection, but also to follow the movement of the virus through the vascular system and its subsequent spread into the leaves. In Nicotiana benthamiana plants, imaging of chlorophyll fluorescence parameters such as Ф(PSII) and NPQ proved useful to follow infections with Pepper mild mottle virus. The results demonstrate a correlation between changes in the chlorophyll fluorescence parameters and the viral distribution analyzed by tissue printing.

Interactions between iron and titanium metabolism in spinach: a chlorophyll fluorescence study in hydropony.

One of the elements showing strong beneficial effect on plants at low concentrations and toxic effects at higher concentrations is titanium (Ti). We investigated the interconnection between the Fe uptake and the Ti intoxication in model experiment on Fe-deficient spinach (Spinacia oleracea) plants to help to elucidate the mechanism of the biological activity of titanium in plants. The two different Ti (0 and 20 mg L⁻¹) and two different Fe (0 and 1.35 mg L⁻¹) concentrations in hydroponic medium were used in all four possible combinations. We compared chemical analysis of Ti and Fe in roots and shoots with the changes of the in vivo chlorophyll fluorescence. Although Fe and Ti concentration found in shoots of Ti-non-treated Fe-deficient plants was comparable with that in Ti-treated Fe-deficient plants, the soluble form of Ti present in the growth media had a negative effect on photosynthetic activity monitored by chlorophyll fluorescence measurements. The presence of Fe in growth medium significantly decreased the Ti concentration in shoots and increased the photosynthetic activity. Here, we propose that Ti affect components of electron transport chain containing Fe in their structure (particularly photosystem I) and decrease the photosystem II efficiency.

Annual variation of the steady-state chlorophyll fluorescence emission of evergreen plants in temperate zone.

Remotely sensed passive chlorophyll fluorescence emission has a potential to become one of the major global-scale reporter signals on vegetation performance and stress. In contrast to the actively probed parameters such as maximal (FM') or minimal (F0') emission, the steady-state chlorophyll fluorescence, Chl-FS, (FM' > Chl-FS > F0') has not been adequately studied. Using fluorescence imaging of leaves, we explored the modulation of Chl-FS by actinic irradiance and by temperature in laboratory, as well as the changes that occurred in three coniferous and broadleaf plant species grown in field. The experiments revealed that Chl-FS is largely insensitive to the incident irradiance once this is above early morning or late evening levels. The characteristic, pre-noon measured Chl-FS correlated positively with the CO2 assimilation rate when measured in field during the year. It was low and stable in the cold winter months and steeply increased with the spring onset. The high values of the characteristic Chl-FS persisted throughout the vegetation season and rapidly decreased in the fall. The seasonal Chl-FS transitions coincided with the last spring frosts or the first fall frosts that persisted for several consecutive nights. The transitions were marked by an elevated variability of the Chl-FS signal. We propose that the signal variability occurring during the transition periods can be used to detect from satellites the beginning and the end of the photosynthetic activity in evergreen canopies of the temperate zone.