We have used 16S ribosomal genes to estimate the bacterial contents of Lake Vostok accretion ice samples at 3551 m and 3607 m, both containing sediment inclusions and formed 20000–15000 yr ago. Decontamination proved to be a critical issue, and we used stringent ice chemistry-based procedures and comprehensive biological controls in order to restrain contamination. As a result, up to now we have only recognized one 16S rDNA bacterial phylotype with confident relevance to the lake environment. It was found in one sample at 3607 m depth and represents the extant thermophilic facultative chemolithoautotroph Hydrogenophilus thermoluteolus of beta-Proteobacteria, and until now had only been found in hot springs. No confident findings were detected in the sample at 3551 m, and all other phylotypes revealed (a total of 16 phylotypes, 336 clones including controls) are presumed to be contaminants. It seems that the Lake Vostok accretion ice is actually microbe-free, indicating that the water body should also be hosting a highly sparse life. The message of thermophilic bacteria suggests that a geothermal system exists beneath the cold water body of Lake Vostok, what is supported by the geological setting, the long-term seismotectonic evidence from ⁴He degassing and the ‘¹⁸O shift’ of the Vostok accretion ice. The seismotectonic activity that seems to operate in deep faults beneath the lake could sustain thermophilic chemolithoautotrophic microbial communities. Such a primary production scenario for Lake Vostok may have relevance for icy planets and the approaches used for estimating microbial contents in accretion ice are clearly relevant for searching for extraterrestrial life.

We developed a PCR detection method that selectively recognizes a single biological control agent and demonstrated that universally primed PCR (UP-PCR) can identify strain-specific markers. Antagonistic strains of Clonostachys rosea (syn. Gliocladium roseum ) were screened by UP-PCR, and a strain-specific marker was identified for strain GR5. No significant sequence homology was found between this marker and any other sequences in the databases. Southern blot analysis of the PCR product revealed that the marker represented a single-copy sequence specific for strain GR5. The marker was converted into a sequence-characterized amplified region (SCAR), and a specific PCR primer pair was designed. Eighty-two strains, isolated primarily from Danish soils, and 31 soil samples, originating from different localities, were tested, and this specificity was confirmed. Two strains responded to the SCAR primers under suboptimal PCR conditions, and the amplified sequences from these strains were similar, but not identical, to the GR5 marker. Soil assays in which total DNA was extracted from GR5-infested and noninoculated field soils showed that the SCAR primers could detect GR5 in a pool of mixed DNA and that no other soil microorganisms present contained sequences amplified by the primers. The assay developed will be useful for monitoring biological control agents released into natural field soil.