Notes: Abstract Addition of the arginine analogue, canavanine, to cultures of nitrogen-fixing Anabaena cylindrica at the onset of akinete formation, resulted in the development of akinetes randomly distributed within the filament, in addition to those adjacent to heterocysts. The total frequency of akinetes increased up to five-fold. A feature of akinetes is their increased content of cyanophycin granules (an arginine-aspartic acid polymer) and addition of canavanine to cultures at an earlier stage resulted in entire filaments becoming agranular and containing agranular akinetes. The effects on akinete pattern appeared to be specific for canavanine since other amino acid analogues, although increasing the frequency of akinetes (approximately two-fold), had no effect on their position relative to heterocysts. In ammonia-grown, stationary phase cultures of A. cylindrica, akinetes were observed adjacent to proheterocysts and in positions more than 20 cells from any heterocyst. These observations indicate that nitrogen fixation and heterocysts are not essential for akinete formation in A. cylindrica, although the availability of a source of fixed nitrogen does appear to be a requirement. These results suggest that during exponential growth some aspect of the physiology of vegetative cells suppresses their development into akinetes and that the role of the heterocyst may not be one of direct stimulation of adjacent vegetative cells to form akinetes, but the removal or negation of the inhibition within them. A model for akinete formation and the involvement of canavanine is given.

Notes: Silicified deposits, such as sinters, occur in several modern geothermal environments, but the mechanisms of silicification (and crucially the role of microorganisms in their construction) are still largely unresolved. Detailed examination of siliceous sinter, in particular sections of microstromatolites growing at the Krisuvik hot spring, Iceland, reveals that biomineralization contributes a major component to the overall structure, with approximately half the sinter thickness attributed to silicified microorganisms. Almost all microorganisms observed under the scanning electron microscope (SEM) are mineralized, with epicellular silica ranging in thickness from 〈 5 μm coatings on individual cells, to regions where entire colonies are cemented together in an amorphous silica matrix tens of micrometres thick. Within the overall profile, there appears to be two very distinct types of laminae that alternate repeatedly throughout the microstromatolite: ‘microbial’ layers are predominantly consisting of filamentous, intact, vertically aligned, biomineralized cyanobacteria, identified as Calothrix and Fischerella sp.; and weakly laminated silica layers which appear to be devoid of any microbial component. The microbial layers commonly have a sharply defined base, overlying the weakly laminated silica, and a gradational upper surface merging into the weakly laminated silica. These cyclic laminations are probably explained by variations in microbial activity. Active growth during spring/summer allows the microorganisms to keep pace with silicification, with the cell surfaces facilitating silicification, while during their natural slow growth phase in the dark autumn/winter months silicification exceeds the bacteria’s ability to compensate (i.e. grow upwards). At this stage, the microbial colony is probably not essential to microstromatolite formation, with silicification presumably occurring abiogenically. When conditions once again become favourable for growth, recolonization of the solid silica surface by free-living bacteria occurs: cell motility is not responsible for the laminations. We have also observed that microbial populations within the microstromatolite, some several mm in depth, appear viable, i.e. they still have their pigmentation, the trichomes are not collapsed, cell walls are unbroken, cytoplasm is still present and they proved culturable. This suggests that the bulk of silicification occurred rapidly, probably while the cells were still alive. Surprisingly, however, measurements of light transmittance through sections of the microstromatolite revealed that photosynthetically active light (PAL) only transmitted through the uppermost 2 mm. Therefore the ‘deeper’ microbial populations must have either: (i) altered their metabolic pathways; (ii) become metabolically inactive; or (iii) the deeper populations may be dominated by different microbial assemblages from that of the surface. From these collective observations, it now seems unequivocal that microstromatolite formation is intimately linked to microbial activity and that the sinter fabric results from a combination of biomineralization, cell growth and recolonization. Furthermore, the similarities in morphology and microbial component to some Precambrian stromatolites, preserved in primary chert, suggests that we may be witnessing contemporaneous biomineralization processes and growth patterns analogous to those of the early Earth.

Notes: Abstract The cyanophage AN-15 was found to have a requirement for either 1 mM calcium or 1 mM magnesium ions to maintain viral stability, whereas 1 mM calcium ions alone were essential for the infection process to proceed in Anabaena sp. strain PCC 7120. Following prolonged incubation, phage-resistant cells were detected at a high frequency (approximately 10-5) in lysates, as either renewed growth in liquid cultures, or as colonies in confluently lysed lawns. Southern hybridisation failed to detect AN-15 DNA in any of the resistant strains, implying that resistance is unlikely to be due to the presence of temperate phages. A high rate of spontaneous mutation is therefore likely to be the cause of resistance. Two classes of resistant cells were identified; those in which AN-15 failed to attach to host cells, and those in which attachment occurred, but subsequent replication was defective. However, it was possible to overcome phage resistance by the isolation of spontaneous mutants of AN-15, capable of infecting phage-resistant cells. These observations imply that if cyanophages are to be assessed as a means of controlling cyanobacterial blooms in freshwater bodies, the ionic (notably calcium) concentration of the water must be considered, together with the possible need to employ alternative cyanophage strains if resistance to the original one arises.