Low-temperature recovery strategies for the isolation of bacteria from ancient permafrost sediments (original) (raw)

In the Arctic there are lenses of overcooled water brines (cryopegs) sandwiched within permafrost marine sediments 100–120 thousand years old. We have investigated the physiological properties of the pure cultures of anaerobic Clostridium sp. strain 14D1 and two strains of aerobic bacteria Psychrobacter sp. isolated from these cryopegs. The structural and physiological characteristics of new bacteria from water brines have shown their ability to survive and develop under harsh conditions, such as subzero temperatures and high salinity.

Despite the presence of well-documented changes in vegetation and faunal communities at the Pleistocene-Holocene transition, it is unclear whether similar shifts occurred in soil microbes. Recent studies do not show a clear connection between soil parameters and community structure, suggesting permafrost microbiome-climate studies may be unreliable. However, the majority of the permafrost microbial ecological studies have been performed only in either Holocene-or Pleistocene-aged sediments and not on permafrost that formed across the dramatic ecosystem reorganization at the Pleistocene-Holocene transition. In our study, we used permafrost recovered in proximity to the Pleistocene-Holocene transition subsampled under strict sterile conditions developed for ancient DNA studies. Our ordination analyses of microbial community composition based on 16S RNA genes and chemical composition of the soil samples resulted into two distinct clusters based on whether they were of late Pleistocene or Holocene age, while samples within an epoch were more similar than those across the boundary and did not result in age based separation. Between epochs, there was a statistically significant correlation between changes in OTU composition and soil chemical properties, but only Ca and Mn were correlated to OTU composition within Holocene aged samples; furthermore, no chemical parameters were correlated to OTU composition within Pleistocene aged samples. Thus, the results indicate that both soil chemical and microbial parameters are fairly stable until a threshold, driven by climate change in our study, is crossed, after which there is a shift to a new steady state. Modern anthropogenic climate change may lead to similar transitions in state for soil biogeochemical systems and microbial communities in Arctic regions.

Microbial metabolic activity occurs at subzero temperatures in permafrost, an environment representing B25% of the global soil organic matter. Although much of the observed subzero microbial activity may be due to basal metabolism or macromolecular repair, there is also ample evidence for cellular growth. Unfortunately, most metabolic measurements or culture-based laboratory experiments cannot elucidate the specific microorganisms responsible for metabolic activities in native permafrost, nor, can bulk approaches determine whether different members of the microbial community modulate their responses as a function of changing subzero temperatures. Here, we report on the use of stable isotope probing with 13 C-acetate to demonstrate bacterial genome replication in Alaskan permafrost at temperatures of 0 to À 20 1C. We found that the majority (80%) of operational taxonomic units detected in permafrost microcosms were active and could synthesize 13 C-labeled DNA when supplemented with 13 C-acetate at temperatures of 0 to À 20 1C during a 6-month incubation. The data indicated that some members of the bacterial community were active across all of the experimental temperatures, whereas many others only synthesized DNA within a narrow subzero temperature range. Phylogenetic analysis of 13 C-labeled 16S rRNA genes revealed that the subzero active bacteria were members of the Acidobacteria, Actinobacteria, Chloroflexi, Gemmatimonadetes and Proteobacteria phyla and were distantly related to currently cultivated psychrophiles. These results imply that small subzero temperature changes may lead to changes in the active microbial community, which could have consequences for biogeochemical cycling in permanently frozen systems Q1. Subject Category: Geomicrobiology and microbial contributions to geochemical cycles