RESUMEN
Cryopreservation is the long-term, indefinite storage of living biological resources at ultralow temperatures. It is almost universally assumed that cryogenic storage supports genetic and phenotypic stability of organisms. However, certain components of the cryopreservation process, particularly some cryoprotective additives (CPAs) and free radical mediated cryoinjury, may potentially cause genetic alterations. Genetic integrity in cryopreserved microalgae was assessed using a very sensitive molecular fingerprinting technique, AFLP, on 28 terrestrial microalgal strains. In about half of all investigated strains the AFLP fingerprints revealed, with high levels of reproducibility, clearly detectable genomic differences after cryopreservation employing a widely used standard two-step cooling protocol. Differences ranged from a single fragment position to multiple fragment changes and were compared to differences found between wild-type and UV-light- or radioisotope-induced mutants of Parachlorella kessleri. The basis of the changes are discussed in terms of their reversibility, as may be the case if they are attributed to DNA methylation and/or whether they are true mutations that may potentially manifest in the phenotype. The possibility that cryopreservation selects for genotypically different subpopulations of microalgae is also considered.
RESUMEN
Abstract The methanogenic archaeal communities in the sediment of two basins of an artificially partitioned acidic bog lake were studied. In the northeast basin, which was separated from a peat bog, a high methane production rate was measured only in the upper layers of the sediment. In contrast, methane production was detected at various depths of the sediment in the southwest basin, which continuously receiving humic acids from the bog. Ten bands were observed in the denaturing gradient gel electrophoresis (DGGE) fingerprint of the 16S rDNA archaeal amplicons from the NE basin top (0-5 cm) sediment layer, which reflected the presence of at least 10 ribotypes. Seventy clones of the 16S rDNA amplicons were obtained from the NE basin top sediment layer, and were grouped into 10 operational taxonomy units (OTUs) according to their positions on the DGGE gel. Seven of these OTUs could be matched with the bands of the community fingerprint. Phylogenetic analyses indicated that the sequences clustered into three groups: five of the OTUs were related to Methanosaeta, four OTUs to Methanomicrobiales and one OTU to Methanobacterium. Among the OTUs, sequences with high similarities (>96%) were retrieved. The sequence data suggested that the diversity of the methanogenic archaeal community was limited. Bands corresponding to those three phylogenetic groups were found in the DGGE fingerprints of both NE and SW basins, which reflected the presence of the same dominant methanogenic archaeal groups in both basins. However, differences in the distribution of the ribotypes that had high 16S rRNA sequence identities were observed in the fingerprints between the two basins at various depths. The microdiversity decreased along the sediment depth in the NE basin, and vice versa in the SW basin. The greater variety of the archaeal ribotypes, apparently, correlated with the higher methanogenesis rate.
RESUMEN
The complete 18S rRNA gene sequences of three Oocystis A. Braun species (Oocystaceae) and three other chlorococcal algae, Tetrachlorella alternans (G. M. Smith) Kors. (Scenedesmaceae), Makinoella tosaensis Okada (Scenedesmaceae), and Amphikrikos cf. nanus (Fott & Heynig) Hind. (Chlorellaceae) were determined and subjected to four different phylogenetic analysis algorithms. Independent of the reconstruction method, these taxa clustered together as a monophyletic group (Oocystaceae) within the Trebouxiophyceae. This result was supported by high bootstrap values. A comparison of morphological data with the phylogenetic reconstructions indicated that the evolution of Oocystaceae was accompanied by a reduction in the number of plastids. This study fully supports the taxonomic assignment of the Oocystaceae as a distinct family. The diacritic criterion that the cell walls are composed of several cellulose layers with perpendicular fibril orientations is in accordance with the molecular data.
