Effects of Algal Extracellular Polysaccharides on the Formation of Filamentous Manganese Oxide Particles in the Near-Bottom Layer of Lake Biwa.

Filamentous manganese (Mn) oxide particles, which occur in the suboxic zone of stratified waterbodies, are important drivers of diverse elemental cycles. These particles are considered to be bacteriogenic; despite the importance of biogeochemical implications, however, the environmental factor responsible for their formation has not been identified. The aim of this study was to demonstrate the involvement of algal extracellular polysaccharides in Mn oxide particle formation. Based on this study of laboratory cultures of a model Mn(II)-oxidizing bacterium, the supply of algal extracellular mucilage was shown to stimulate Mn(II) oxidation and thus the production of filamentous Mn oxide particles. This observation was consistent with the results obtained for naturally occurring particles collected from a near-bottom layer (depth of approximately 90 m) in the northern basin of Lake Biwa, Japan, that is, most Mn particles resembling δ-MnO2 were associated with an extracellular mucilage-like gelatinous matrix, which contained dead algal cells and was lectin-stainable. In the lake water column, polysaccharides produced by algal photosynthesis sank to the bottom layer. The analysis of the quality of water samples, which have been collected from the study site for 18 years, reveals that the annual average total phytoplankton biovolume in the surface layer correlates with the density of filamentous Mn particles in the near-bottom layer. Among different phytoplankton species, green algae appeared to be the key species. The results of this study suggest that algal extracellular polysaccharides serve as an important inducer for the formation of filamentous Mn oxide particles in the near-bottom layer of the northern basin of Lake Biwa.

Identification of 13 Spirogyra species (Zygnemataceae) by traits of sexual reproduction induced under laboratory culture conditions.

The genus Spirogyra is abundant in freshwater habitats worldwide, and comprises approximately 380 species. Species assignment is often difficult because identification is based on the characteristics of sexual reproduction in wild-collected samples and spores produced in the field or laboratory culture. We developed an identification procedure based on an improved methodology for inducing sexual conjugation in laboratory-cultivated filaments. We tested the modified procedure on 52 newly established and genetically different strains collected from diverse localities in Japan. We induced conjugation or aplanospore formation under controlled laboratory conditions in 15 of the 52 strains, which allowed us to identify 13 species. Two of the thirteen species were assignable to a related but taxonomically uncertain genus, Temnogyra, based on the unique characteristics of sexual reproduction. Our phylogenetic analysis demonstrated that the two Temnogyra species are included in a large clade comprising many species of Spirogyra. Thus, separation of Temnogyra from Spirogyra may be untenable, much as the separation of Sirogonium from Spirogyra is not supported by molecular analyses.

Studies on conjugation of Spirogyra using monoclonal culture.

We succeeded in inducing conjugation of Spirogyra castanacea by incubating algal filaments on agar plate. Conjugation could be induced using clone culture. The scalariform conjugation was generally observed, while lateral conjugation was rarely. When two filaments formed scalariform conjugation, all cells of one filament behaved as male and those of other filament did as female. Very rarely, however, zygospores were formed in both of pair filaments. The surface of conjugation tube was stained with fluorescently labeled-lectins, such as Bandeiraea (Griffonia) simplicifolia lectin (BSL-I) and jacalin. BSL-I strongly stained the conjugation tubes, while weakly did the cell surface of female gamete first and then that of male gamete. Jacalin stained mainly the conjugation tubes. Addition of jacalin inhibited the formation of papilla, suggesting some important role of jacalin-binding material at the initial step of formation of the conjugation tubes.

Rhizoid differentiation of Spirogyra is regulated by substratum.

Some species of Spirogyra can anchor to substratum with rod- or rosette-shaped rhizoid (hapteron). The rhizoid differentiation can be induced by cutting algal filaments in a laboratory. Requirement of contact stimulation for rhizoid differentiation has been reported (Nagata in Plant Cell Physiol 14:531-541, 1973a). However, the control mechanism of rhizoid morphology has not been elucidated. When cut filaments were incubated on the glass surface, start of tip growth, secretion of lectin-binding material and callose synthesis were observed. In the absence of contact to the glass surface, none of above phenomena was induced. Systematic analysis showed that rosette-shaped rhizoid was formed only on the hydrophobic substratum. On the hydrophobic substratum, both Bandeiraea (Griffonia) simplicifolia lectin and jacalin strongly stained the rhizoids. On the hydrophilic substratum, however, only Bandeiraea (Griffonia) simplicifolia lectin strongly stained the rhizoids.

Saponin-induced release of single cells from filaments and rhizoid differentiation in Spirogyra.

Some species of Spirogyra living in streams can anchor to the substratum by differentiating a rhizoid from a terminal cell of a filament. Rhizoid differentiation occurs in the light but not in the dark. When a filament of Spirogyra sp. competent for rhizoid differentiation was incubated in a medium containing 0.1% saponin, terminal cells were released one by one, forming single cells. Single cells effectively differentiated to be rhizoids when saponin in the incubation medium was removed. The single-cell system developed in the present study seems suitable for analysis of gene expression during rhizoid differentiation of Spirogyra.