The investigation of spatial heterogeneity within the thylakoid membrane (TM) proteins has gained increasing attention in photosynthetic research. The recent advances in live-cell imaging have allowed the identification of heterogeneous organisation of photosystems in small cyanobacterial cells. These sub-micrometre TM regions, termed microdomains in cyanobacteria, exhibit functional similarities with granal (Photosystem II dominant) and stromal (Photosystem I dominant) regions observed in TM of higher plants. This study delves into microdomain heterogeneity using super-resolution Airyscan-based microscopy enhancing resolution to approximately ~125 nm in x-y dimension. The new data reveal membrane areas rich in Photosystem I within the inner TM rings. Moreover, we identified analogous dynamics in the mobility of Photosystem II and phycobilisomes; countering earlier models that postulated differing mobility of these complexes. These novel findings thus hold significance for our understanding of photosynthesis regulation, particularly during state transitions.

• Keywords
• Airyscan, FRAP, cyanobacteria, microdomain, photosystem, protein mobility, super-resolution microscopy, thylakoid membrane heterogeneity,
• Publication type
• Journal Article MeSH

The Trichodesmium genus comprises some of the most abundant N2-fixing organisms in oligotrophic marine ecosystems. Since nitrogenase, the key enzyme for N2 fixation, is irreversibly inhibited upon O2 exposure, these organisms have to coordinate their N2-fixing ability with simultaneous photosynthetic O2 production. Although being the principal object of many laboratory and field studies, the overall process of how Trichodesmium reconciles these two mutually exclusive processes remains unresolved. This is in part due to contradictory results that fuel the Trichodesmium enigma. In this review, we sift through methodological details that could potentially explain the discrepancy between findings related to Trichodesmium's physiology. In doing so, we exhaustively contrast studies concerning both spatial and temporal nitrogenase protective strategies, with particular attention to more recent insights. Finally, we suggest new experimental approaches for solving the complex orchestration of N2 fixation and photosynthesis in Trichodesmium.

• Keywords
• O2-scavenging mechanisms, cyanobacteria, diazocyte, immunolabelling, method comparison, microscopy,
• Publication type
• Journal Article MeSH
• Review MeSH

While photosynthetic processes have become increasingly understood in cyanobacterial model strains, differences in the spatial distribution of thylakoid membranes among various lineages have been largely unexplored. Cyanobacterial cells exhibit an intriguing diversity in thylakoid arrangements, ranging from simple parietal to radial, coiled, parallel, and special types. Although metabolic background of their variability remains unknown, it has been suggested that thylakoid patterns are stable in certain phylogenetic clades. For decades, thylakoid arrangements have been used in cyanobacterial classification as one of the crucial characters for definition of taxa. The last comprehensive study addressing their evolutionary history in cyanobacteria was published 15 years ago. Since then both DNA sequence and electron microscopy data have grown rapidly. In the current study, we map ultrastructural data of >200 strains onto the SSU rRNA gene tree, and the resulting phylogeny is compared to a phylogenomic tree. Changes in thylakoid architecture in general follow the phylogeny of housekeeping loci. Parietal arrangement is resolved as the original thylakoid organization, evolving into complex arrangement in the most derived group of heterocytous cyanobacteria. Cyanobacteria occupying intermediate phylogenetic positions (greater filamentous, coccoid, and baeocytous types) exhibit fascicular, radial, and parallel arrangements, partly tracing the reconstructed course of phylogenetic branching. Contrary to previous studies, taxonomic value of thylakoid morphology seems very limited. Only special cases such as thylakoid absence or the parallel arrangement could be used as taxonomically informative apomorphies. The phylogenetic trees provide evidence of both paraphyly and reversion from more derived architectures in the simple parietal thylakoid pattern. Repeated convergent evolution is suggested for the radial and fascicular architectures. Moreover, thylakoid arrangement is constrained by cell size, excluding the occurrence of complex architectures in cyanobacteria smaller than 2 μm in width. It may further be dependent on unknown (eco)physiological factors as suggested by recurrence of the radial type in unrelated but morphologically similar cyanobacteria, and occurrence of special features throughout the phylogeny. No straightforward phylogenetic congruences have been found between proteins involved in photosynthesis and thylakoid formation, and the thylakoid patterns. Remarkably, several postulated thylakoid biogenesis factors are partly or completely missing in cyanobacteria, challenging their proposed essential roles.

• Keywords
• SSU rRNA gene, cyanobacteria, evolution, photosynthesis, phylogenomics, taxonomy, thylakoid pattern,
• Publication type
• Journal Article MeSH