Changes in vacuolation in the root apex cells of soybean seedlings in microgravity.

Changes in the vacuolation in root apex cells of soybean (Glycine max L. [Merr.]) seedlings grown in microgravity were investigated. Spaceflight and ground control seedlings were grown in the absence or presence of KMnO4 (to remove ethylene) for 6 days. After landing, in order to study of cell ultrastructure and subcellular free calcium ion distribution, seedling root apices were fixed in 2.5% (w/v) glutaraldehyde in 0.1 M cacodylate buffer and 2% (w/v) glutaraldehyde, 2.5% (w/v) formaldehyde, 2% (w/v) potassium antimonate K[Sb(OH)6] in 0.1 M K2HPO4 buffer with an osmolarity (calculated theoretically) of 0.45 and 1.26 osmol. The concentrations of ethylene in all spaceflight canisters were significantly higher than in the ground control canisters. Seedling growth was reduced in the spaceflight-exposed plants. Additionally, the spaceflight-exposed plants exhibited progressive vacuolation in the root apex cells, particularly in the columella cells, to a greater degree than the ground controls. Plasmolysis was observed in columella cells of spaceflight roots fixed in solutions with relatively high osmolarity (1.26 osmol). The appearance of plasmolysis permitted the evaluation of the water status of cells. The water potential of the spaceflight cells was higher than the surrounding fixative solution. A decrease in osmotic potential and/or an increase in turgor potential may have induced increases in cell water potential. However, the plasmolysed (i.e. non-turgid) cells implied that increases in water potential were accompanied with a decrease in osmotic potential. In such cells changes in vacuolation may have been involved to maintain turgor pressure or may have been a result of intensification of other vacuolar functions like digestion and storage.     

Microorganisms and enzyme activity in permafrost after removal of long-term cold stress

Associations of immobilized microbial cells and organic-mineral complexes containing active enzymes are resistant to long-term (from tens of thousands to millions of years) effects of extremely low temperatures. This association enables the cells to restore their metabolic activity during permafrost thawing, because interactions with the heterogenous medium is made possible by the availability of active immobilized enzymes. The long-term effect of the cold probably favors an adaptational change of microbial metabolism that activates enzymes and cells during thawing.     

Long-term preservation of microbial ecosystems in permafrost.

It has been established that significant numbers (up to 10 million cells per gram of sample) of living microorganisms of various ecological and morphological groups have been preserved under permafrost conditions, at temperatures ranging from -9 to -13 degrees C and depths of up to 100 m, for thousands and sometimes millions of years. Preserved since the formation of permafrost in sand-clay sediments of the Pliocene-Quaternary period and in paleosols and peats buried among them, these cells art the only living organisms that have survived for a geologically significant period of time. The complexity of the microbial community preserved varies with the age of the permafrost. Eukaryotes are found only in Holocene sediments; while prokaryotes are found to greater ages, i.e., Pliocene and Pleistocene. The diversity of microorganisms decreases with increasing age of sediments, and as a result cocci and corynebacteria are predominant. Enzyme activity (catalase and hydrolytic enzymes) and photosynthetic pigments (chlorophyll and pheophytin have also been detected in permafrost sediments. These results permit us to outline some approaches to the search for traces of life in the permafrost of Martian sediments by borehole core sampling. It is in the deep horizons (and not on the planet surface), isolated by permafrost from the external conditions, that results similar to those obtained on Earth can be expected.     

Cytochemical localization of calcium in soybean root cap cells in microgravity.

The antimonate precipitation technique was used to evaluate the effects of microgravity and ethylene on the cellular and subcellular distribution of free calcium ions in soybean root apices. Soybean (Glycine max L. [Merr.]) dry seeds were launched, activated by hydration, and germinated in the presence of KMnO4 (to remove ethylene) and in its absence onboard the space shuttle Columbia during the STS-87 mission. Primary root apices of 6-day old seedlings were fixed for electron microscopy after landing. Ultrastructural studies indicated that antimonate precipitation appeared as individual electron-dense particles which were more or less round in shape and varied in diameter from 10 nm (minimum size beginning from which the particles were well identified) to 90 nm. It was revealed that analyzed root cap cells varied in both the precipitate particle sizes and the amount particles per unit of the cellular area. In both flight and ground control treatments, antimonate precipitation level increases from apical meristem cells to peripheral (secretory) cells of root apices. In root cap statocytes, subcellular localization of precipitate particles was revealed in the cytoplasm, nucleus and small vacuoles. The quantitative analysis showed a reduction of precipitate density in the cytoplasm and the nucleus, and an increase in precipitate density in the vacuoles from statocytes of both spaceflight treatments in comparison with ground controls.     

Strategies for detection of putative life on Europa

Planned future exploration missions to the Jovian satellite Europa have a strong astrobiological motivation. Characterization of the potential habitability of the liquid water environments, and searching for life signals are the main astrobiological objectives of these missions. To meet these objectives specific strategies and instrumentation are required. Here we discuss some scenarios for the development of Europa potential biospheres. These scenarios are based on assumptions of the life similarity concept and knowledge about terrestrial life in extreme environments. Since the potential habitable environments on Europa are in the interior of the satellite it is not possibly to directly detect life. However, there are processes that link aqueous sub-surface environments with the near-surface environment, such as tectonism or magmatism. Therefore, by analysing endogenous materials that arise from the interior it is possible to make predictions about what is in the sub-surface. We propose some measurements and instrumentation for future missions to detect biosignatures on the upper layers of Europa, including the simple physico-chemical traces of metabolism to complex biomolecules or biostructures. Raman spectroscopy or biosensor technologies are the future for in situ exploration of the Solar System.     

The structure of resting bacterial populations in soil and subsoil permafrost

The structure of individual cells in microbial populations in situ of the Arctic and Antarctic permafrost was studied by scanning and transmission electron microscopy methods and compared with that of cyst-like resting forms generated under special conditions by the non-spore-forming bacteria Arthrobacter and Micrococcus isolated from the permafrost. Electron microscopy examination of microorganisms in situ revealed several types of bacterial cells having no signs of damage, including "dwarf" curved forms similar to nanoforms. Intact bacterial cells in situ and frozen cultures of the permafrost isolates differed from vegetative cells by thickened cell walls, the altered structure of cytoplasm, and the compact nucleoid, and were similar in these features to cyst-like resting forms of non-spore-forming "permafrost" bacterial strains of Arthrobacter and Micrococcus spp. Cyst-like cells, being resistant to adverse external factors, are regarded as being responsible for survival of the non-spore-formers under prolonged exposure to subzero temperatures and can be a target to search for living microorganisms in natural environments both on the Earth and on extraterrestrial bodies.     

Role of cell differentiation in high tolerance by prokaryotes of long-term preservation in permafrost

The effect of low temperature on the cell structure of bacteria isolated from permafrost results in structural changes leading to cell differentiation into types of resting cells rangign from spores showing a high endogenous dormancy to typical “dormant” cells of non-spore-forming bacteria showing exogenous dormancy, which is considered to be less highly resistant to extreme conditions in laboratory experiments. In permafrost, dormant cells of non-spore-forming bacteria may demonstrate considerable resistance to long-term freezing and as a result a higher survival level than spore-forming bacteria.     

Microbial populations in Antarctic permafrost: biodiversity, state, age, and implication for astrobiology

Antarctic permafrost soils have not received as much geocryological and biological study as has been devoted to the ice sheet, though the permafrost is more stable and older and inhabited by more microbes. This makes these soils potentially more informative and a more significant microbial repository than ice sheets. Due to the stability of the subsurface physicochemical regime, Antarctic permafrost is not an extreme environment but a balanced natural one. Up to 10(4) viable cells/g, whose age presumably corresponds to the longevity of the permanently frozen state of the sediments, have been isolated from Antarctic permafrost. Along with the microbes, metabolic by-products are preserved. This presumed natural cryopreservation makes it possible to observe what may be the oldest microbial communities on Earth. Here, we describe the Antarctic permafrost habitat and biodiversity and provide a model for martian ecosystems.