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 (RubA) 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, 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 chlorophyll was detected in D1mod but not D2mod, formation of RCII appears to be important for stable binding of most of the chlorophylls and both pheophytins. We suggest that chlorophyll can be delivered to RCII from either monomeric PSI or Ycf39/Hlip complexes

Pre-symptomatic detection of Plasmopara viticola infection in grapevine leaves using chlorophyll fluorescence imaging

Plasmopara viticola is an economically important pathogen of grapevine. Early detection of P. viticola infection can lead to improved fungicide treatment. Our study aimed to determine whether chlorophyll fluorescence (Chl-F) imaging can be used to reveal early stages of P. viticola infection under conditions similar to those occurring in commercial vineyards. Maximum (F(V)/F(M)) and effective quantum yield of photosystem II (I broken vertical bar(PSII)) were identified as the most sensitive reporters of the infection. Heterogeneous distribution of F(V)/F(M) and I broken vertical bar(PSII) in artificially inoculated leaves was associated with the presence of the developing mycelium 3 days before the occurrence of visible symptoms and 5 days before the release of spores. Significant changes of F(V)/F(M) and I broken vertical bar(PSII) were spatially coincident with localised spots of inoculation across the leaf lamina. Reduction of F(V)/F(M) was restricted to the leaf area that later yielded sporulation, while the area with significantly lower I broken vertical bar(PSII) was larger and probably reflected the leaf parts in which photosynthesis was impaired. Our results indicate that Chl-F can be used for the early detection of P. viticola infection. Because P. viticola does not expand systemically in the host tissues and the effects of infection are localised, Chl-F imaging at high resolution is necessary to reveal the disease in the field