Repeated colonization of alpine habitats by Arabidopsis arenosa viewed through freezing resistance and ice management strategies.

Success or failure of plants to cope with freezing temperatures can critically influence plant distribution and adaptation to new habitats. Especially in alpine environments, frost is a likely major selective force driving adaptation. In Arabidopsis arenosa (L.) Lawalrée, alpine populations have evolved independently in different mountain ranges, enabling studying mechanisms of acclimation and adaptation to alpine environments. We tested for heritable, parallel differentiation in freezing resistance, cold acclimation potential and ice management strategies using eight alpine and eight foothill populations. Plants from three European mountain ranges (Niedere Tauern, Fagaraș and Tatra Mountains) were grown from seeds of tetraploid populations in four common gardens, together with diploid populations from the Tatra Mountains. Freezing resistance was assessed using controlled freezing treatments and measuring effective quantum yield of photosystem II, and ice management strategies by infrared video thermography and cryomicroscopy. The alpine ecotype had a higher cold acclimation potential than the foothill ecotype, whereby this differentiation was more pronounced in tetraploid than diploid populations. However, no ecotypic differentiation was found in one region (Fagaraș), where the ancient lineage had a different evolutionary history. Upon freezing, an ice lens within a lacuna between the palisade and spongy parenchyma tissues was formed by separation of leaf tissues, a mechanism not previously reported for herbaceous species. The dynamic adjustment of freezing resistance to temperature conditions may be particularly important in alpine environments characterized by large temperature fluctuations. Furthermore, the formation of an extracellular ice lens may be a useful strategy to avoid tissue damage during freezing.

Critically reduced frost resistance of Picea abies during sprouting could be linked to cytological changes.

Frost resistance of sprouting Picea abies shoots is insufficient for survival of naturally occurring late frosts. The cellular changes during sprouting appeared to be responsible for frost damage as frost events that damaged sprouting shoots did not damage older needles and stems. Whilst resting buds showed initial frost damage at -15.0 degrees C, 20 days later, current year's growth was damaged at -5.6 degrees C. The decrease in frost resistance in sprouting shoots of P. abies was accompanied by a significant reduction of the cellular solute concentration, indicated by much less negative Psi(oSAT) values (increase from -2.8 to -1.2 MPa). psi(oSAT) decreased again after the final cell volume was reached and cell wall thickening began. After bud break, ice nucleation temperature increased from -4.7 degrees C to -1.5 degrees C. This increase was probably caused by the loss of bud scales, the onset of expansion growth of the central cylinder and the development of vascular tissue permitting the spread of ice from the stem into the growing needles. The onset of mesophyll cell wall thickening coincided with the lowest frost resistances. Cell wall thickening caused an increase in the modulus of elasticity, epsilon, indicating a decrease in tissue elasticity and after that frost resistance increased again. Metabolic and cytological changes that evidently leave little leeway for frost hardening are responsible for the low frost resistance in current year's growth of P. abies. This low frost resistance will be significant in the future as the risk of frost damage due to earlier bud break is anticipated to even further increase.

Cold-loving microbes, plants, and animals--fundamental and applied aspects.

Microorganisms, plants, and animals have successfully colonized cold environments, which represent the majority of the biosphere on Earth. They have evolved special mechanisms to overcome the life-endangering influence of low temperature and to survive freezing. Cold adaptation includes a complex range of structural and functional adaptations at the level of all cellular constituents, such as membranes, proteins, metabolic activity, and mechanisms to avoid the destructive effect of intracellular ice formation. These strategies offer multiple biotechnological applications of cold-adapted organisms and/or their products in various fields. In this review, we describe the mechanisms of microorganisms, plants, and animals to cope with the cold and the resulting biotechnological perspectives.

Frost resistance and ice nucleation in leaves of five woody timberline species measured in situ during shoot expansion.

Frost resistance and ice nucleation temperatures of leaves, from bud swelling until after full expansion, were measured in situ for five major woody timberline species with recently developed field freezing equipment. Frost resistance determined in situ on leaves of attached twigs was significantly higher than values determined on detached leaves in laboratory tests (e.g., the temperature at which incipient frost damage was observed (LTi) was 1.2 degrees C higher for detached leaves than for attached leaves of Picea abies (L.) Karst.). Frost resistance of leaves of all species changed significantly during shoot expansion (e.g., changes of 7.2 and 11 degrees C for Rhododendron ferrugineum L. and Larix decidua Mill., respectively). Expanding leaves (between 0 and 60% of full expansion) were the most sensitive to frost, with LTi values ranging from -3.4 degrees C in R. ferrugineum to -6.3 degrees C in L. decidua. Among the studied species, P. abies and R. ferrugineum were the most frost sensitive throughout the shoot elongation period. In situ freezing patterns of leaves of attached twigs also differed from those of leaves of excised twigs. During leaf expansion, two distinct freezing exotherms were always registered in situ. The first freezing event (E1, high-temperature exotherm) was recorded at -1.5 +/- 0.2 degrees C and reflected extracellular ice formation. Exposure of leaves to temperatures at which E1 occurred was, in all cases, noninjurious. The low-temperature exotherm (E2) mostly coincided with frost damage, except for some stages of leaf expansion in R. ferrugineum and P. abies, indicating that in situ freezing exotherms were not accurate estimators of frost damage in these species.

Frost resistance and susceptibility to ice formation during natural hardening in relation to leaf anatomy in three evergreen tree species from New Zealand.

Foliar frost resistance of three endemic New Zealand land trees, Nothofagus menziesii (Hook. f.) Oerst. (Fagaceae), Pittosporum eugenioides A. Cunn. (Pittosporaceae) and Griselinia littoralis Forst. f. (Cornaceae), was examined as the trees hardened from late summer to midwinter in a lowland forest site. The lowest temperatures causing 50% damage (LT(50)) occurred in late winter and were similar to those recorded for other forest trees native to New Zealand (-11.7 degrees C in N. menziesii, -10.7 degrees C in P. eugenioides, and -10.6 degrees C in G. littoralis). All three species hardened by 4-7 degrees C, with G. littoralis showing the least frost resistance in summer and hence the greatest degree of hardening. Thermal analysis during freezing indicated that all three species became more tolerant of extracellular ice formation in winter. Measurements of chlorophyll a fluorescence correlated well with visible injury. The differing patterns of frost damage development in the three species were related to leaf anatomy: visible injury was localized within the small compartments formed by the highly septate leaves of the most resistant species, N. menziesii, and was somewhat localized in the partially septate leaves of P. eugenioides, whereas damage could be initiated anywhere in the aseptate leaves of G. littoralis,which was the least frost resistant species, particularly in summer.

Cold-induced sudden reversible lowering of in vivo chlorophyll fluorescence after saturating light pulses : a sensitive marker for chilling susceptibility.

In chilling-sensitive plants (Glycine max, Saintpaulia ionantha, Saccharum officinarum) a sudden reversible drop in chlorophyll fluorescence occurs during photosynthetic induction immediately following saturating light pulses at low temperatures in the range 4 to 8 degrees C. A comparison of two soybean cultivars of different chilling sensitivities revealed that this phenomenon, termed lowwave, indicates specific thresholds of low temperature stress. Its occurrence under controlled chilling can be regarded as a quantitative marker for screening chilling susceptibility in angiosperms.