Evergreen alpine shrubs have high freezing resistance in spring,irrespective of snowmelt timing and exposure to frost: an investigation from the Snowy Mountains,Australia

Over winter, alpine plants are protected from low-temperature extremes by a blanket of snow. Climate change predictions indicate an overall reduction in snowpack and an earlier thaw; a situation which could expose the tips of shrubs which extend above the snowpack to freezing events in early spring, and cause foliar frost damage during the onset of physiological activity. We assessed the photosynthetic responses of freezing-damaged shrub leaves from an assay of freezing temperatures in the Snowy Mountains in south-eastern Australia, using chlorophyll fluorometery ex situ. We sampled leaves that were exposed early during the spring thaw and leaves that were buried in snow for up to two extra weeks, from four evergreen shrub species at monthly intervals following the period of snowmelt. Freezing resistance (estimated from LT₅₀) was poorest at the earliest spring sampling time, in both exposed above-snow and protected below-snow foliage in all species. Protected foliage in early spring had lower freezing resistance than exposed foliage, but not significantly so. By the third sampling time, freezing resistance was significantly better in the lower protected foliage (LT₅₀ of ? 14) compared with the upper exposed foliage (LT₅₀ of ? 10) in one species. Over the course of spring, freezing resistance improved significantly in all species, with LT₅₀ values of between ? 10 and ? 15 °C by the third sampling time, which is lower than the minimum air temperatures recorded at that time (> ? 5 °C). The results indicate that the dominant evergreen shrub species in this area may only be susceptible to freezing events very early in spring, before a period of frost-hardening occurs after snowmelt. Later in spring, these alpine shrubs appear frost hardy, thus further perpetuating the positive feedbacks surrounding shrub expansion in alpine areas.     

Dehydrin, alcohol dehydrogenase, and central metabolite levels are associated with cold tolerance in diploid strawberry (Fragaria spp.)

The use of artificial freezing tests, identification of biomarkers linked to or directly involved in the low-temperature tolerance processes, could prove useful in applied strawberry breeding. This study was conducted to identify genotypes of diploid strawberry that differ in their tolerance to low-temperature stress and to investigate whether a set of candidate proteins and metabolites correlate with the level of tolerance. 17 Fragaria vesca, 2 F. nilgerrensis, 2 F. nubicola, and 1 F. pentaphylla genotypes were evaluated for low-temperature tolerance. Estimates of temperatures where 50 % of the plants survived (LT₅₀) ranged from ?4.7 to ?12.0 °C between the genotypes. Among the F. vesca genotypes, the LT₅₀ varied from ?7.7 °C to ?12.0 °C. Among the most tolerant were three F. vesca ssp. bracteata genotypes (FDP821, NCGR424, and NCGR502), while a F. vesca ssp. californica genotype (FDP817) was the least tolerant (LT₅₀ ?7.7 °C). Alcohol dehydrogenase (ADH), total dehydrin expression, and content of central metabolism constituents were assayed in select plants acclimated at 2 °C. The LT₅₀ estimates and the expression of ADH and total dehydrins were highly correlated (r _adh = ?0.87, r _dehyd = ?0.82). Compounds related to the citric acid cycle were quantified in the leaves during acclimation. While several sugars and acids were significantly correlated to the LT₅₀ estimates early in the acclimation period, only galactinol proved to be a good LT₅₀ predictor after 28 days of acclimation (r _galact = 0.79). It is concluded that ADH, dehydrins, and galactinol show great potential to serve as biomarkers for cold tolerance in diploid strawberry.     

Freezing avoidance by supercooling in Olea europaea cultivars: the role of apoplastic water,solute content and cell wall rigidity

Plants can avoid freezing damage by preventing extracellular ice formation below the equilibrium freezing temperature (supercooling). We used Olea europaea cultivars to assess which traits contribute to avoid ice nucleation at sub‐zero temperatures. Seasonal leaf water relations, non‐structural carbohydrates, nitrogen and tissue damage and ice nucleation temperatures in different plant parts were determined in five cultivars growing in the Patagonian cold desert. Ice seeding in roots occurred at higher temperatures than in stems and leaves. Leaves of cold acclimated cultivars supercooled down to ?13 °C, substantially lower than the minimum air temperatures observed in the study site. During winter, leaf ice nucleation and leaf freezing damage (LT₅₀) occurred at similar temperatures, typical of plant tissues that supercool. Higher leaf density and cell wall rigidity were observed during winter, consistent with a substantial acclimation to sub‐zero temperatures. Larger supercooling capacity and lower LT₅₀ were observed in cold‐acclimated cultivars with higher osmotically active solute content, higher tissue elastic adjustments and lower apoplastic water. Irreversible leaf damage was only observed in laboratory experiments at very low temperatures, but not in the field. A comparative analysis of closely related plants avoids phylogenetic independence bias in a comparative study of adaptations to survive low temperatures.     

Freezing tolerance, cold acclimation and oxidative stress in potato. Paraquat tolerance is related to acclimation but is a poor indicator of freezing tolerance

Potato is a species commonly cultivated in temperate areas where the growing season may be interrupted by frosts, resulting in loss of yield. Cultivated potato, Solanum tuberosum, is freezing sensitive, but it has several freezing-tolerant wild potato relatives, one of which is S. commersonii. Our study was aimed to resolve the relationship between enhanced freezing tolerance, acclimation capacity and capacity to tolerate active oxygen species. To be able to characterize freezing tolerant ideotypes, a potato population (S1), which segregates in freezing tolerance, acclimation capacity and capacity to tolerate superoxide radicals, was produced by selfing a somatic hybrid between a freezing-tolerant Solanum commersonii (LT₅₀=-4.6°C) and -sensitive S. tuberosum (LT₅₀=-3.0°C). The distribution of non-acclimated freezing tolerance (NA-freezing tolerance) of the S1 population varied between the parental lines and we were able to identify genotypes, having significantly high or low NA-freezing tolerance. When a population of 25 genotypes was tested both for NA-freezing and paraquat (PQ) tolerance, no correlation was found between these two traits (R = 0.02). However, the most NA-freezing tolerant genotypes were also among the most PQ tolerant plants. Simultaneously, one of the NA-freezing sensitive genotypes (2022) (LT₅₀=-3.0°C) was observed to be PQ tolerant. These conflicting results may reflect a significant, but not obligatory, role of superoxide scavenging mechanisms in the NA-freezing tolerance of S. commersonii. The freezing tolerance after cold acclimation (CA-freezing tolerance) and the acclimation capacity (AC) was measured after acclimation for 7 days at 4/2°C. Lack of correlation between NA-freezing tolerance and AC (R =-0.05) in the S1 population points to independent genetic control of NA-freezing tolerance and AC in Solanum sp. Increased freezing tolerance after cold acclimation was clearly related to PQ tolerance of all S1 genotypes, especially those having good acclimation capacity. The rapid loss of improved PQ tolerance under deacclimation conditions confirmed the close relationship between the process of cold acclimation and enhanced PQ tolerance. Here, we report an increased PQ tolerance in cold-acclimated plants compared to non-acclimated controls. However, we concluded that high PQ tolerance is not a good indicator of actual freezing tolerance and should not be used as a selectable marker for the identification of a freezing-tolerant genotype.     

Foliar freezing resistance of Australian alpine plants over the growing season

We assessed the freezing resistance of leaves ex situ of 25 Australian alpine plant species. We compared the freezing resistance of forb, graminoid and shrub species from three alpine summits of different altitudes; from a low altitude site just above treeline, to a fully alpine tundra site. Foliar freezing resistance (LT₅₀) in spring varied from ?5.9°C to ?18.7°C and standardized LT₅₀ values within species were significantly related to site altitude. Additionally, when comparing all the species in the study, freezing resistance was significantly related to site; the LT₅₀ of samples from a low‐altitude summit (1696 m) were significantly lower than those of samples from mid‐ (1805 m) and high‐altitude (1860 m) summits. The LT₅₀ of juvenile foliage did not differ significantly from that of adult foliage. Shrubs were highly resistant to freezing. At the highest summit, we examined the course of seasonal freezing resistance from early summer to early autumn across three alpine plant communities that differed in the time of natural snowmelt; from sheltered (snowpatch) to exposed (open heath). No differences in freezing resistance over the growing season were detected for exposed or sheltered communities and there were no consistent trends indicating frost hardening over the growing season. Overall, the common Australian alpine species we investigated appear well adapted to freezing conditions throughout the snow‐free growing season. We have no evidence to suggest that freezing temperatures soon after snowmelt in spring are especially damaging to the alpine plants at these summits.     

Induced expression of the class II chitinase gene during cold acclimation and dehydration of bermudagrass (Cynodon sp.)

Bermudagrass cultivars vary greatly in their ability to survive freezing temperatures as a result of a differential ability to cold acclimate (CA) at temperatures slightly above 0°C. Little information exists on the genetic and physiological mechanisms associated with the cold acclimation process in bermudagrass. Experiments were conducted to study the changes in chitinase gene expression during cold acclimation of freeze-tolerant bermudagrass cultivars. A chitinase gene (CynCHT1) was isolated from ’Midiron’ bermudagrass. Because the hydrophilic protein putatively encoded by the gene lacked an N-terminal cysteine-rich domain and a hydrophobic C-terminal extension, it was classified a class II chitinase. The expression patterns of this and related chitinase genes in response to CA, drought, and ABA were investigated in freeze-tolerant ’MSU’ (LT₅₀=?11°C), Midiron (LT₅₀=?10°C) and ’Uganda’ (LT₅₀=?8°C) bermudagrasses. Northern-blot analysis indicated expression in the crown tissues induced by CA at 8°C/2°C day/night temperature cycles. Induction of gene expression was evident in tissues sampled at 2 and 28 days after initiating CA. Expression after 2-days de-acclimation at 28°C/24°C was similar to control levels. Significantly higher levels of CA-induced chitinase gene expression were observed in MSU and Midiron, compared to Uganda. Similar expression patterns were observed among the cultivars in responses to drought and ABA. These results suggest that chitinases have important roles in bermudagrass response to low temperature and dehydration stresses.     

Freezing tolerance of ectomycorrhizal fungi in pure culture

The ability to survive freezing and thawing is a key factor for the existence of life forms in large parts of the world. However, little is known about the freezing tolerance of mycorrhizal fungi and their role in the freezing tolerance of mycorrhizas. Threshold temperatures for the survival of these fungi have not been assessed experimentally. We grew isolates of Suillus luteus, Suillus variegatus, Laccaria laccata, and Hebeloma sp. in liquid culture at room temperature. Subsequently, we exposed samples to a series of temperatures between +5°C and −48°C. Relative electrolyte leakage (REL) and re-growth measurements were used to assess the damage. The REL test indicated that the lethal temperature for 50% of samples (LT₅₀) was between −8.3°C and −13.5°C. However, in the re-growth experiment, all isolates resumed growth after exposure to −8°C and higher temperatures. As many as 64% of L. laccata samples but only 11% in S. variegatus survived −48°C. There was no growth of Hebeloma and S. luteus after exposure to −48°C, but part of their samples survived −30°C. The fungi tolerated lower temperatures than was expected on the basis of earlier studies on fine roots of ectomycorrhizal trees. The most likely freezing tolerance mechanism here is tolerance to apoplastic freezing and the concomitant intracellular dehydration with consequent concentrating of cryoprotectant substances in cells. Studying the properties of fungi in isolation promotes the understanding of the role of the different partners of the mycorrhizal symbiosis in the freezing tolerance.     

Frost tolerance and biochemical changes during hardening and dehardening in contrasting white clover populations

Successful winter survival of perennial plants, like white clover, is dependent on proper timing of both hardening and dehardening. The purpose of this study was to investigate the regulation of these processes in two cultivars (AberCrest and AberHerald) and two Norwegian ecotypes (Særheim collected at 58°46′N lat. and Bodø at 67°20′N lat.) of white clover (Trifolium repens L.). For hardening and dehardening, plants were exposed to controlled temperature conditions and frost hardiness of stolons was tested by programmed freezing at the rate of 3°C per hour. In addition, stolons were analysed for starch, soluble sugars and soluble amino acids. Cultivars AberCrest and AberHerald, selected for growth at low temperature and winter hardiness in the United Kingdom, were significantly less hardy than the Norwegian populations. After six weeks of hardening (2 weeks at 6°C and 4 weeks at 0.5°C), estimated LT₅₀ values were ?13.8, ?13.0, ?17.8 and ?20.3°C for AberCrest, AberHerald, Saerheim and Bodø, respectively. The rate of dehardening increased with increasing temperature. At low temperature (6°C), the northern ecotype from Bodø was more resistant to dehardening than AberHerald. However, at 18°C the absolute rate of dehardening (°C day^?1) was twice as high in Bodø as in AberHerald plants. Stolon elongation during dehardening was initiated at lower temperatures in AberHerald than in plants of the Bodø ecotype. The content of total soluble sugars, sucrose and the amino acids proline and arginine were significantly higher in hardy plants of Bodø than in those of AberHerald. Sucrose levels decreased during dehardening and correlations between sucrose content and LT₅₀ during this process were statistically highly significant for both Bodø and AberHerald. The least hardy populations of white clover were characterized by thick stolons, long internodes and large leaves.     

Freezing cytorrhysis and critical temperature thresholds for photosystem II in the peat moss Sphagnum capillifolium

Leaflets of Sphagnum capillifolium were exposed to temperatures from ?5°C to +60°C under controlled conditions while mounted on a microscope stage. The resultant cytological response to these temperature treatments was successfully monitored using a light and fluorescence microscope. In addition to the observable cytological changes during freezing cytorrhysis and heat exposure on the leaflets, the concomitant critical temperature thresholds for inactivation of photosystem II (PS II) were studied using a micro fibre optic and a chlorophyll fluorometer mounted to the microscope stage. Chlorophyllous cells of S. capillifolium showed extended freezing cytorrhysis immediately after ice nucleation at ?1.1°C in the water in which the leaflets were submersed during the measurement. The occurrence of freezing cytorrhysis, which was visually manifested by cell shrinkage, was highly dynamic and was completed within 2 s. A total reduction of the mean projected diameter of the chloroplast containing area during freezing cytorrhysis from 8.9 to 3.8 μm indicates a cell volume reduction of approximately ?82%. Simultaneous measurement of chlorophyll fluorescence of PS II was possible even through the frozen water in which the leaf samples were submersed. Freezing cytorrhysis was accompanied by a sudden rise of basic chlorophyll fluorescence. The critical freezing temperature threshold of PS II was identical to the ice nucleation temperature (?1.1°C). This is significantly above the temperature threshold at which frost damage to S. capillifolium leaflets occurs (?16.1°C; LT₅₀) which is higher than observed in most higher plants from the European Alps during summer. High temperature thresholds of PS II were 44.5°C which is significantly below the heat tolerance of chlorophyllous cells (49.9°C; LT₅₀). It is demonstrated that light and fluorescence microscopic techniques combined with simultaneous chlorophyll fluorescence measurements may act as a useful tool to study heat, low temperature, and ice-encasement effects on the cellular structure and primary photosynthetic processes of intact leaf tissues.     

Photosynthesis in cold acclimated leaves of plants with various degrees of freezing tolerance

The objective of this study was to compare the photosynthetic changes during cold acclimation in various plant types able to acquire different degrees of freezing tolerance. Four herbaceous and six woody plants were hardened under natural or artificial conditions and – after determination of their frost resistance (LT₅₀) – the net photosynthetic rate at an ambient CO₂ of 33 Pa (P_n33), the dependencies of P_n to light and to CO₂ and the room temperature chlorophyll a fluorescence were recorded under optimal conditions. Herbaceous plants acquired freezing tolerances to temperatures between ?10 and ?15°C when hardened at temperatures around 0°C. Most leaves fully developed prior to frost hardening exhibited typical symptoms of senescence after frost hardening. In non-senescing leaves P_n33 was reduced by 15 to 50% mainly due to a reduced stomatal conductance. After hardening at temperatures around ?10°C Brassica survived down to ?24°C, but P_n33 was almost abolished as a result of disturbances in the chloroplasts. After transferring the plants to 20/15°C P_n33 recovered completely within a few days. Woody plants hardened at temperatures around 0°C tolerated – 15 to ?36°C: P_n33 was reduced by 25 to 60% and hardly recovered at 20/15°C. Hardening at ?10°C induced a tolerance of ?32 to n33 was almost totally blocked, but at 20/15°C it returned to the values of the plants hardened at 0°C within a few days. In woody plants disturbances were invariably localized in the chloroplasts. Thus, conifers, and especially Pinus cembra, can survive much lower temperatures than herbaceous plants and, at the same level of freezing tolerance, exhibit appreciably less restriction in relative P_n33.     

In vitro screening for cold hardiness of raspberry cultivars

Raspberry (Rubus idaeus L.) cultivars ‘Festival’, ‘Titan’ and ‘Willamette’ were cultured in vitro on three different media: (A) MS medium supplemented with 1.0 mg l^-1 BAP and 0.1 mg l^-1 IBA, (B) MS medium without growth regulators, and (C) MS medium with reduced sucrose (10 g l^-1), and exposed to different low temperature acclimation treatments: (1) control, no acclimation, (2) 1 week at +15 °C, 1 week at +2 °C, 24 h at -2 °C and 3 days at +2 °C, and (3) 2 weeks at +15 °C, 2 weeks at +2 °C, 24 h at −2 °C and 3 days at +2 °C. After acclimation, shoot moisture content was measured, and cold hardiness (LT₅₀) was determined by controlled freezing. Shoot moisture content was generally lower on culture medium B compared to the other media, but not affected by acclimation treatment. In non-acclimated plants, medium composition had no effect on cold hardiness and no cultivar differences in hardiness were observed. After acclimation, plants on culture medium B were on average more cold hardy than on the other media. Acclimation treatment 3 on media A and B allowed the best discrimination between the hardy cultivar ‘Festival’ and less cold hardy ‘Titan’ and ‘Willamette’. When acclimation treatments were tested further using 11 raspberry cultivars with different levels of cold hardiness, discrimination between cultivars was satisfactory only after acclimation treatment 3 on culture medium B. This revised version was published online in June 2006 with corrections to the Cover Date.     

Olive (Olea europaea L.) freezing tolerance related to antioxidant enzymes activity during cold acclimation and non acclimation

The changes in the antioxidant enzymes activity, total protein and proline content and their correlations with freezing tolerance (FT) (expressed as LT₅₀) were investigated at 11 different olive cultivars at cold-acclimation (CA, in February) and non-acclimation (NA, in August) stages. Leaf samples were collected from each cultivar and were divided into two groups. The first group was immediately frozen in liquid nitrogen for further biochemical analysis. The second ones was subjected to different freezing temperatures (?5, ?10, ?15 and ?20 °C) for 10 h, in order to determine their FT. The unfrozen control samples were kept at 4 °C. The results showed that Fishomi, Mission and Shengeh were the most freezing tolerant among other cultivars. In contrast, Zard, Manzanilla and Amigdalolia were the most sensitive ones. The cold acclimation enhanced the activities of superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), catalase (CAT), polyphenol oxidase (PPO) and total protein content. However, proline content and phenylalanine ammonia-lyase (PAL) activity did not change or even decreased slightly at CA stage, compare to those samples at NA stage. It was found that LT₅₀ to be closely correlated to POD, CAT, and PPO activity at CA and NA stages. Overall, higher leaf POD, CAT, and PPO activity could be used as important selection criteria in screening tolerant olive cultivars for cold zone climatic.     

Early Developments in RNA, Protein, and Sugar Levels during Cold Stress in Winter Rye (Secale cereale) Leaves

Changes in freezing tolerance of winter rye (Secale cerealeL. cv. Voima) were determined for leaf tissues during a 1-weekcold stress, which was performed by transferring the 7-d-oldseedlings from a greenhouse (25°C, long day) to 3°Cand short day conditions. The development of cold hardeningwas shown by using an ion leakage test and by determining theamounts of carbohydrates, soluble proteins and RNA. The firstevidence of the development of freezing resistance was foundafter 1 d at low temperature, i.e. an LT₅₀ value increased from-5 to -7°C. Plants cold treated for 7 d reached an LT₅₀value of -9°C. This increase in freezing tolerance was foundto be associated with the increased levels of soluble carbohydrates,total RNA and soluble proteins. These metabolic changes indicatethe association with adjustment of growth and cell metabolismto low temperatures at the beginning of cold acclimation ofwinter rye.Copyright 1994, 1999 Academic Press Secale cereale L., winter rye, cold stress, proteins, RNA, sugars     

Seasonal changes in cold tolerance,water relations and accumulation of cations and compatible solutes in Atriplex halimus L.

We determined the cold (freezing) tolerance for field-grown plants of Atriplex halimus L. (Chenopodiaceae) in relation to plant ploidy level, leaf water relations and accumulation of osmolytes. Plants were grown at two sites in Murcia (Spain), having average minimum temperatures in the coldest month of 0.6 and 12.1 °C, respectively. LT₅₀ values derived from laboratory freezing tests, using leaves taken from the plants in early winter and in spring, showed greater tolerance for winter-harvested leaves; the acclimation was more pronounced at the cold-winter site. Cold tolerance was related positively with leaf K and/or Na accumulation. Analysis of compatible organic solutes (soluble sugars, total amino acids and quaternary ammonium compounds) showed that cold tolerance (measured both as LT₅₀ and as winter freezing damage in situ) was related most closely with leaf concentrations of soluble sugars. The leaf percentage dry matter content was related to both in vitro and in vivo tolerance, while tolerance in vitro was correlated also with the osmotic (potential ψ_s) and the relative water content. The two diploid (2n = 2x = 18) populations, from Spain, showed greater cold tolerance than the three tetraploid (2n = 4x = 36) populations, from North Africa and Syria, which may be related to the latter's greater cell size and consequent dilution of osmolytes. In this halophytic species, cold tolerance, like salinity and drought tolerance, seems to depend on osmotic adjustment, driven by vacuolar accumulation of K and Na and cytoplasmic accumulation of compatible solutes.     

Seasonal changes in cold tolerance,water relations and accumulation of cations and compatible solutes in Atriplex halimus L.

We determined the cold (freezing) tolerance for field-grown plants of Atriplex halimus L. (Chenopodiaceae) in relation to plant ploidy level, leaf water relations and accumulation of osmolytes. Plants were grown at two sites in Murcia (Spain), having average minimum temperatures in the coldest month of 0.6 and 12.1 °C, respectively. LT₅₀ values derived from laboratory freezing tests, using leaves taken from the plants in early winter and in spring, showed greater tolerance for winter-harvested leaves; the acclimation was more pronounced at the cold-winter site. Cold tolerance was related positively with leaf K and/or Na accumulation. Analysis of compatible organic solutes (soluble sugars, total amino acids and quaternary ammonium compounds) showed that cold tolerance (measured both as LT₅₀ and as winter freezing damage in situ) was related most closely with leaf concentrations of soluble sugars. The leaf percentage dry matter content was related to both in vitro and in vivo tolerance, while tolerance in vitro was correlated also with the osmotic (potential ψ_s) and the relative water content. The two diploid (2n = 2x = 18) populations, from Spain, showed greater cold tolerance than the three tetraploid (2n = 4x = 36) populations, from North Africa and Syria, which may be related to the latter's greater cell size and consequent dilution of osmolytes. In this halophytic species, cold tolerance, like salinity and drought tolerance, seems to depend on osmotic adjustment, driven by vacuolar accumulation of K and Na and cytoplasmic accumulation of compatible solutes.     

Heat tolerance of early developmental stages of glacier foreland species in the growth chamber and in the field

In glacier forelands, seeds readily germinate, however, a high proportion of seedlings die shortly after their appearance. We hypothesized that besides drought, frost and missing safe sites, heat on the ground surface could be one of the major threats for seedlings. The heat strain in different ground strata was assessed from 2007 to 2010. The heat tolerance (LT₅₀) of eleven alpine species from different successional stages was tested considering imbibed (G1) and germinated seeds (G2) as well as seedlings (G3). Additionally, the heat hardening capacity of seedlings was determined in the field. Across all species, LT₅₀ decreased significantly by 9 K from G1 (55 °C) to G3 (46 °C), similarly in all species of the successional stages. Field-grown seedlings had mostly an increased LT₅₀ (2K). Intraspecifically, LT₅₀ of seedlings varied between 40.6 and 52.5 °C. Along the chronosequence, LT₅₀ in G1 was similar, but was higher in G2 and G3 of early successional species. The highest temperatures occurred at 0–0.5 cm in air (mean/absolute maximum: 42.6/54.1 °C) posing a significant heat injury risk for seedlings when under water shortage transpirational cooling is prevented. Below small stones (0–0.5 cm), maxima were 4 K lower, indicating heat safer microsites. Maxima >30 °C occurred at 32.3, >40 °C at 6.2 %. Interannually, 2010 was the hottest year with heat exceeding LT₅₀ at all microsites (0–0.5 cm). Temperature maxima on sandy surfaces were lower than on microsites with gravel (diameter <5–10 mm). The hot summer of 2010 may be a small foretaste of in future more severe and frequent heat waves. Ground surface temperature maxima at the pioneer stage are already now critical for heat survival and may partly explain the high seedling mortality recognized on recently deglaciated terrain.     

Identification of quantitative trait loci and associated candidate genes for low-temperature tolerance in cold-hardy winter wheat

Low-temperature (LT) tolerance is an important economic trait in winter wheat (Triticum aestivum L.) that determines the plants’ ability to cope with below freezing temperatures. Essential elements of the LT tolerance mechanism are associated with the winter growth habit controlled by the vernalization loci (Vrn-1) on the group 5 chromosomes. To identify genomic regions, which in addition to vrn-1 determine the level of LT tolerance in hexaploid wheat, two doubled haploid (DH) mapping populations were produced using parents with winter growth habit (vrn-A1, vrn-B1, and vrn-D1) but showing different LT tolerance levels. A total of 107 DH lines were analyzed by genetic mapping to produce a consensus map of 2,873 cM. The LT tolerance levels for the Norstar (LT₅₀=−20.7°C) × Winter Manitou (LT₅₀=−14.3°C) mapping population ranged from −12.0 to −22.0°C. Single marker analysis and interval mapping of phenotyped lines revealed a major quantitative trait locus (QTL) on chromosome 5A and a weaker QTL on chromosome 1D. The 5A QTL located 46 cM proximal to the vrn-A1 locus explained 40% of the LT tolerance variance. Two C-repeat Binding Factor (CBF) genes expressed during cold acclimation in Norstar were located at the peak of the 5A QTL.     

Differences in heat tolerance plasticity between supratidal and intertidal snails indicate complex responses to microhabitat temperature variation

Tropical intertidal gastropods that experience extreme and highly variable daily temperatures have evolved significant and complex heat tolerance plasticity, comprising components that respond to different timescales of temperature variation. An earlier study showed different plasticity attributes in snails from differently-heated coastlines, suggesting lifelong irreversible responses that matched habitat thermal regimes. To determine whether heat tolerance plasticity varied at a finer, within-shore spatial scale, we compared the responses of supratidal (predominantly shade-dwelling) and intertidal (frequently solar-exposed) populations of the tropical thermophilic gastropod, Echinolittorina malaccana. Snails modified lethal temperature (LT₅₀) under warm or cool laboratory acclimation, with the overall variation in LT₅₀ being greater in the supratidal (56.0–58.0 °C) than in the intertidal population (57.1–58.1 °C). Similar maximum LT₅₀s expressed by the populations after warm acclimation suggest a capacity limitation under these temperature conditons. The different minimum LT₅₀s after cool acclimation corresponded with microhabitat temperature and field acclimatization of the snails. Different responses to the same laboratory acclimation treatment imply long-term (and possibly lifelong) thermal acclimatization, which could benefit sedentary organisms that are randomly recruited as larvae from a common thermally-stable aquatic environment to thermally-unpredictable intertidal microhabitats. These findings provide another example of thermal tolerance plasticity operating at microhabitat scales, suggesting the importance of considering microhabitat thermal responses when assessing broad-scale environmental change.     

Induction of freezing tolerance in alfalfa cell suspension cultures

The effects of ABA, 2,4-D, kinetin and cold exposure on the cold hardiness of Medicago sativa L. cell suspensions were investigated. Cultures treated with 5×10^–5 M ABA at 2°C for 4 weeks in the absence of kinetin showed a 50% survival after freezing to –12.5°C, whereas cultures grown at 25°C under normal conditions tolerated freezing to only –3°C. The optimum ABA treatment of 5×10^–5 M for 4 weeks was effective only in combination with cold exposure. Of six cell lines tested, all showed different degrees of induced cold hardiness. The results suggest that ABA alone cannot induce freezing tolerance on alfalfa cell suspension cultures and that the deletion of kinetin and combination of low temperature and ABA is critical for the induction of cold hardiness in alfalfa cell suspension cultures.Abbreviations ABA abscisic acid - 2,4-D 2,4-dichlorophenoxyacetic acid - LT₅₀ 50% killing temperature