The Freezing Response of an Arctic Cushion Plant, Saxifraga caespitosa L.: Acclimation, Freezing Tolerance and Ice Nucleation

Cold hardiness in actively growing plants of Saxifraga caespitosaL., an arctic and subarctic cushion plant, was examined. Plantscollected from subarctic and arctic sites were cultivated ina phytotron at temperatures of 3, 9, 12 and 21 °C undera 24-h photoperiod, and examined for freezing tolerance usingcontrolled freezing at a cooling rate of 3–4 °C eitherin air or in moist sand. Post-freezing injury was assessed byvisual inspection and with chlorophyll fluorescence, which appearedto be well suited for the evaluation of injury in Saxifragaleaves. Freezing of excised leaves in moist sand distinguishedwell among the various treatments, but the differences werepartly masked by significant supercooling when the tissue wasfrozen in air. Excised leaves, meristems, stem tissue and flowerssupercooled to –9 to –15 °C, but in rosettesand in intact plants ice nucleation was initiated at –4to –7 °C. The arctic plants tended to be more coldhardy than the subarctic plants, but in plants from both locationscold hardiness increased significantly with decreasing growthtemperature. Plants grown at 12 °C or less developed resistanceto freezing, and excised leaves of arctic Saxifraga grown at3 °C survived temperatures down to about –20 °C.Exposure to –3 °C temperature for up to 5 d did notsignificantly enhance the hardiness obtained at 3 °C. Whenwhole plants of arctic Saxifraga were frozen, with roots protectedfrom freezing, they survived –15 °C and –25°C when cultivated at 12 and 3 °C, respectively, althougha high percentage of the leaves were killed. The basal levelof freezing tolerance maintained in these plants throughoutperiods of active growth may have adaptive significance in subarcticand arctic environments. Saxifraga caespitosa L., arctic, chlorophyll fluorescence, cold acclimation, cushion plant, freezing stress, freezing tolerance, ice nucleation, supercooling     

Short-Term Changes in Heat Tolerance in the Alpine Cushion Plant Silene acaulis ssp. excapa [All.] J. Braun at Different Altitudes

Abstract: The habit of cushion growth positively affects plant temperature but at the same may increase the risk of occasional overheating. In order to determine the adaptive response to short-term heat stress, we exposed S. acaulis cushions at field sites to controlled heat treatments using infrared lamps. Natural diurnal changes in heat tolerance were monitored at alpine sites and at a site distinctly below the natural distribution boundary, where higher temperatures were expected. The range of heat tolerance limits in summer, 45.5 - 54.5 °C (9 K), exceeded that reported for other alpine species (0.1 - 5 K) and even that for total seasonal changes (5 - 8 K). Heat tolerance either increased or decreased on most days (80 %). The maximum diurnal increase was + 4.7 K. Under the experimental conditions heat hardening started at leaf temperatures around 30 °C and proceeded at mean rates of 1.0 ± 0.5 K/h. The onset of functional disturbances in photosystem II also occurred at 30 °C. Heating rates exceeding those naturally found above 30 °C (> 10 K/h) appeared to retard heat hardening. During summer average leaf temperature maxima were 12.4 K (600 m) and 13.0 K (1945 m) higher than air temperature which corroborates the heat trapping nature of cushion plants. At 600 m, as compared to 1945 m, cushions experienced significantly higher leaf temperature maxima (+ 8.8 K) and exceeded 30 °C on most days (80 %). This resulted in a significantly higher heat tolerance (LT₅₀) at 600 m (51.7 ± 0.2 °C) than at 1945 m (49.8 ± 0.2 °C). The fast short-term changes of heat tolerance in summer help S. acaulis to cope with the occasional diurnal short-term heat stress associated with cushion growth.     

Muskoxen Modify Plant Abundance,Phenology, and Nitrogen Dynamics in a High Arctic Fen

Ecosystems - Herbivores are key drivers of vegetation dynamics in most ecosystems. However, the effect of high arctic herbivores on vegetation dynamics throughout a growing season is not well...     

The Influence of Altered Rainfall Regimes on Early Season N Partitioning Among Early Phenology Annual Plants,a Late Phenology Shrub,and Microbes in a Semi-arid Ecosystem

In Mediterranean-type ecosystems, nitrogen (N) accumulates in soil during dry summer months and rapidly becomes available during early season rain events. The availability of early season N could depend on the size of rainfall events, soil microbial activity, and phenology of the plant community. However, it is poorly understood how precipitation patterns affect the fate of early season N. Microbes and plants with early phenology may compete strongly for early season N but theory suggests that microbial N storage can meet plant N demands later in the season. Using a ¹⁵N tracer and rainfall manipulation we investigated the fate of early season N. N allocation patterns differed substantially between microbes, early and late phenology plants. As expected early phenology annuals and microbes took up ¹⁵N, within 1 day, whereas a late-phenology shrub allocated ¹⁵N to leaves later in the season. We saw no evidence for microbial storage of early season N; the peak of ¹⁵N in shrub leaves did not coincide with detectable levels of ¹⁵N in the microbial biomass or labile soil pool. This suggests that shrubs were able to access early season N, store and allocate it for growth later in the season. Although we saw no evidence of microbial N storage, N retention in soil organic matter (SOM) was high and microbes may play an important role in sequestering N to SOM. Plant N uptake did not respond significantly to 1 year of rainfall manipulation, but microbes were sensitive to dry conditions. 1 year after ¹⁵N addition shrubs had resorbed up to half of the N from leaves whereas N in annuals remained as dead leaf litter. Differences in end-of-season N partitioning between dead and living biomass in the two vegetation types suggest that plant species composition could affect N availability in the following growing season, but it may take several years of altered precipitation patterns to produce rainfall-dependent changes.     

Cryoprotectants and Extreme Freeze Tolerance in a Subarctic Population of the Wood Frog

Wood frogs (Rana sylvatica) exhibit marked geographic variation in freeze tolerance, with subarctic populations tolerating experimental freezing to temperatures at least 10-13 degrees Celsius below the lethal limits for conspecifics from more temperate locales. We determined how seasonal responses enhance the cryoprotectant system in these northern frogs, and also investigated their physiological responses to somatic freezing at extreme temperatures. Alaskan frogs collected in late summer had plasma urea levels near 10 μmol ml^-1, but this level rose during preparation for winter to 85.5 ± 2.9 μmol ml^-1 (mean ± SEM) in frogs that remained fully hydrated, and to 186.9 ± 12.4 μmol ml^-1 in frogs held under a restricted moisture regime. An osmolality gap indicated that the plasma of winter-conditioned frogs contained an as yet unidentified osmolyte(s) that contributed about 75 mOsmol kg^-1 to total osmotic pressure. Experimental freezing to –8°C, either directly or following three cycles of freezing/thawing between –4 and 0°C, or –16°C increased the liver’s synthesis of glucose and, to a lesser extent, urea. Concomitantly, organs shed up to one-half (skeletal muscle) or two-thirds (liver) of their water, with cryoprotectant in the remaining fluid reaching concentrations as high as 0.2 and 2.1 M, respectively. Freeze/thaw cycling, which was readily survived by winter-conditioned frogs, greatly increased hepatic glycogenolysis and delivery of glucose (but not urea) to skeletal muscle. We conclude that cryoprotectant accrual in anticipation of and in response to freezing have been greatly enhanced and contribute to extreme freeze tolerance in northern R. sylvatica.     

The Effect of Abscisic Acid on the Freezing Tolerance of Callus Cultures of Lotus corniculatus L

The effects of growth temperature (2°C and 24°C), abscisic acid (ABA) concentration, duration of exposure to ABA, and light were assessed for their ability to induce acclimation to freezing temperatures in callus cultures of Lotus corniculatus L. cv Leo, a perennial forage legume. The maximal expression of freezing tolerance was achieved on B₅ media containing 10⁻⁵ molar ABA, at 24°C for 7 or 14 days. Under these culture conditions, the freezing tolerance of the callus approximated that observed in field grown plants. In contrast, low temperatures (2°C) induced only a limited degree of freezing tolerance in these cultures. Viability was assessed by tetrazolium reduction and by regrowth of the callus. The two assays often differed in their estimates of absolute freezing tolerance. Regression analysis of the temperature profile suggested that there may be two or more distinct populations of cells differing in freezing tolerance, which may have contributed to the variability between viability assays.     

水母雪莲愈伤组织和悬浮培养细胞的抗冻性研究

水母雪莲（Saussurea meduasa Maxim）是典型的高山雪线植物。本文研究了其愈伤组织及悬浮细胞的培养过程,并对其抗寒性做了初步研究,研究结果表明,水母雪莲愈伤组织和悬浮培养细胞分别可抵抗-6.5℃、-7.5℃的冰冻低温胁迫,水母雪莲愈伤组织细胞内丰富的蛋白质和淀粉粒多糖构成其较强抗冻能力的物质基础,低温锻炼后,悬浮细胞分泌蛋白中有新的多肽（76,48,27.5,19.5kD）合成,而33,51kD两条多肽合成增强,悬浮细胞抗冻能力的提高同蛋白质合成的增强是一致的。     

植物抗冻机理研究新进展：抗冻基因表达及其功能

综述了迄今业已分离和鉴定的低温诱导表达的抗冻基因,以及通过染色体基因制图方法褐示和证实的抗冻性基因,并着重地介绍了这些抗冻基因在提高植物抗冻能力上的功能。这些抗冻基因表达合成的新多肽具3有高度的亲水性,起着保护和稳定细胞膜结构的作用,从而防止冰冻伤害,提高植物的抗冻能力,同时还介绍了抗冻基因表达中的某些调节因子,其中尤其值得注意的是,CBF基因似乎有可能作为抗冻基因表达的“主开关”,以及Ca^2 作为植物细胞的第二信使在传递低温信号,启动和调节抗冻基因表达中可能的重要作用,这些研究结果,不仅为阐明植物的抗冻机理提供了新的证据,而且为改良农作物的抗寒性提供了新的启示。     

Pollen viability and limitation of seed production in a population of the circumpolar cushion plant, Silene acaulis (Caryophyllaceae)

Pollen viability among genders and limitation of female seed production in a natural trioecious population of the circumpolar cushion plant Silene acaulis was examined. Pollen viability was estimated by an in vitro pollen germination experiment. Both male and hermaphrodite flowers displayed large variation in pollen viability (0–53% in hermaphrodite and 0–54% in male flowers). There was a significant difference between genders in pollen viability: male plants had on average higher pollen viability than hermaphrodite plants. Resource and pollen limitation of seed production was studied by an experiment consisting of three treatments; (I) hand-pollination and removal of all other flowers on the cushion, (II) hand-pollination without removal of other flowers, and (III) open pollination without removal of flowers. Hand-pollination increased seed production, whereas removal of flowers had no effect on seed production. Abortion of pollinated ovules during seed development and seed mass did not differ among treatments. To control for effect of fruit number on seed production, data from naturally pollinated individuals was used. There was a positive correlation between both total number of seeds and fruit number, mean seed number per fruit and fruit number, respectively. These results indicate that seed production of 5. acaulis is mainly limited by pollen availability whereas resource competition between fruits is not important as a limiting factor. The possible role of male quality differences between genders and pollen limitation of seed production for maintenance of trioecious reproductive systems is discussed.     

短命植物角茴香的开花物候与生殖特征

为了研究角茴香的开花物候特征及其对生殖成功的影响,于2015年对其自然种群各水平的开花物候指数、开花过程中的花部特征变化和结实特性进行了观察和统计。结果表明：（1）角茴香种群花期为4月下旬至5月下旬,种群水平的花期持续时间为36 d。（2）单花具有独特的花瓣结构,其内层花瓣由白色的侧裂片和黄色的兜状中裂片构成,中裂片是角茴香植物花粉二次呈现的关键功能器官;开花进程依其外层花瓣开裂大小可分为4个阶段,即外层花瓣顶端未张口、外层花瓣顶端张口5~7 mm、外层花瓣顶端张口12~14 mm和外层花瓣顶端张口3~5 mm。（3）花序水平的开花振幅曲线呈渐进式单峰曲线,开花同步指数为0.748,个体水平表现出较高的相对开花强度,其分布频度的偏斜率为0.45,主要频度范围分别集中在20%~30%和50%~70%之间。（4）花序水平的始花日期与花期持续时间存在显著负相关关系,开花数与座果数存在显著正相关关系。（5）自然状态下,角茴香结实率和结籽率分别是82.7%±0.1%和93.6%±0.8%。角茴香开花强度分布频度上出现的高、低强度的分异趋势是吸引传粉者、促进结实的有效生殖对策。     

Glycemic Response to Exercise in the Subarctic Depends on Time of Day,Season and Sex

Between December 1992 and September 1996, a total of 62 healthy athletes (32 males and 30 females) exercised for 30 minutes, with an intensity of 60% VO 2, in one or more morning and afternoon sessions in the months December, February, April, June and September. Blood samples obtained for determinations of glucose were drawn at 0, 1, 5, 10 and 30 minutes of each exercise span. The total area under the glucose-time curves was calculated both in original units (mmol/l) and after normalization to percent of starting value for 263 exercise spans. These calculated areas, representing the glucose “response” during exercise, were tested for the effect of sex, time of day and season by ANOVA and by the least-squares fit of a 1-year cosine. During exercise a significant effect was found by ANOVA for sex (p < 0.001), time of exercise (p =0.006) and month (p =0.007). No significant interactions were found. Exercise in the morning beginning at 11:30h produced a smaller glucose response for both sexes, when compared with exercise in the afternoon beginning at 16:30h. With regard to sex, a smaller integrated glucose response to exercise occurred in females. The response was lowest for both sexes in Dec (winter) compared to other months, while the response was greatest for females in September, and for males in April and June. A circannual rhythm was found both for baseline glucose (p =0.005, acrophase= Feb 20) and glucose–time response areas in mmol/l (p <0.001, acrophase=Feb 4), and normalized values (p =0.05, acrophase=Dec 19). This finding is in accordance with previous reports that, at least in the subarctic area, the glycemic response to a standardized exercise is influenced by season and timing of exercise. This observation may be of importance for athletes involved in vigorous training and patients with Diabetes Mellitus, among others.     

The development of regular seasonal habitat shifts in a landlocked Arctic charr, Salvelinus alpinus L., population

Parts of the Arctic charr population of the subarctic Lake Visjön in north-west Sweden migrate upstream during the spring to two small, recently eutrophied and very productive lakes. Large repeat migrants arrive first, followed by young first-time migrants. Charr in the small lakes grow more rapidly than those resident in L. Visjön. In early September mature fish leave the lakes, followed by immature fish later in September and in October. Overwintering and spawning takes place in L. Visjön. Migratory females attain maturity at age 4 years and resident females at age 6 years. The migrant fish return annually until they are 5–6 years old. This limit may be due to reduced relative growth benefits of the habitat shift for larger individuals. The rapid development of these regular habitat shifts could be explained by an internally fixed exploratory behaviour in these Arctic charr that makes the detection and utilization of distant feeding resources possible. Migrants will possess a considerably higher fitness, if survival rates for migratory and resident fish are equal.     

Long-Term Effects of Crude Oil on Uptake and Respiration of Glucose and Glutamate in Arctic and Subarctic Marine Sediments

The effects of crude oil on uptake and respiration (mineralization) of glucose and glutamate in marine sediments were investigated. After the sediments were treated with crude oil, they were replaced at or near the collection site by scuba divers. These sediments remained in situ until they were retrieved for analysis. Glucose and glutamate uptake rates were found to decrease, and the percent respired was found to increase in Arctic and subarctic marine sediments that had been exposed to fresh crude oil. These same changes were also observed when “weathered” crude oil was used and when untreated sediments were overlaid with oiled sediments. When the kinetics of glutamate uptake were determined, both the maximum potential uptake rate and the turnover time were significantly affected. A comparison between the proportion of glucose taken into the cells and that respired as CO₂ indicated that crude oil affected biosynthetic mechanisms. A study of sediments that had been exposed to crude oil for at least 5 months showed that glutamate transport into the cells was affected more extensively than biosynthetic mechanisms. In the initial months of exposure, bacterial concentrations and total adenylate concentrations were found to decrease in the presence of crude oil. Our data suggest that secondary productivity in the marine environment could be adversely affected by the presence of crude oil in marine sediments.     

Heat Tolerance of Cold Acclimated Puma Winter Rye Seedlings and the Effect of a Heat Shock on Freezing Tolerance

An increase in tolerance to one form of abiotic stress oftenresults in an increase in tolerance to another stress. The heattolerance of Puma rye (Secale cereale) was determined for seedlingseither not cold hardened or hardened under either controlledenvironmental or natural conditions. The heat tolerance wasdetermined either as a function of time at 42°C or the abilityto tolerate a maximum temperature. The seedlings were eithernot heat preconditioned or heat preconditioned before the heatstress. In all cases cold hardened seedlings were more heattolerant than non or partially cold hardened seedlings. Heatpreconditioning had no effect on the heat tolerance of naturallycold hardened seedlings. In contrast, seedlings cold hardenedin a controlled environment chamber, then heat preconditioned,were more heat tolerant than non preconditioned seedlings. Aheat shock of 36°C for 2 h increased the freezing toleranceof non hardened seedlings from –2.5°C to –4.5°C.Analysis of heat shock protein 70 (HSP70) gene expression indicatedthat the HSP70 gene was not induced by cold acclimation andtherefore not directly involved in the increased thermo toleranceobserved. A number of heat stable proteins, simple sugars andlong chain carbohydrate polymers accumulated during the coldacclimation process and may have a role in increasing heat toleranceas well as freezing tolerance. These data suggest cold hardeningincreases heat tolerance, however, a heat shock to non acclimatedseedlings only marginally increased the freezing tolerance ofPuma rye seedlings. ³Present address: Agriculture and Agri-Food Canada, 107 SciencePlace, Saskatoon SK S7N 0X2, Canada.     

Freezing Tolerance and Solute Changes in Contrasting Genotypes of Lolium perenne L. Acclimated to Cold and Drought

Clonal ramets of 12 contrasting genotypes of Lolium perenneL. were grown in sand or soil-based compost and maintained underwell-watered conditions at 20/15°C or acclimated to lowtemperature (2°C) or to a restricted water supply. Freezingtolerance was measured as LT₅₀ following exposure to sub-zerotemperatures in a freezing tank. Measurements were also madeof osmotic potential, water-soluble carbohydrates, free proline,free amino acids, and minerals in entire tillers. Acclimationto both drought and cold lowered LT₅₀, induced osmotic adjustment,and increased concentrations of proline and amino acids, Rootingmedium had little effect on LT₅₀, but caused large differencesin osmotic potential and in proline and amino-acid concentrations.There was considerable genetic diversity for all charactersmeasured, except for mineral contents. There was, however, norelationship between LT₅₀ and osmotic potential or solute contentthat was consistent across the three sources of variation (growingmedium, acclimation, genotype). Furthermore, the diverse genotypicvalues of cold-induced freezing tolerance were not correlatedwith those of drought-induced tolerance. It is concluded thatmore precise measurements are needed of the partitioning ofsolutes during acclimation and of the sensitivity of differentorgans and tissues to freezing.Copyright 1993, 1999 AcademicPress Perennial ryegrass, hardening, acclimation, osmotic potential, solute potential, carbohydrates, proline     

Plant Phenology in a Cloud Forest on the Island of Maui,Hawaii1

We recorded the times of flowering, fruiting, and leafing for ten native canopy and subcanopy trees and shrubs (monthly from December 1994 through December 1997) in a montane cloud forest with relatively aseasonal rainfall on the island of Maui, Hawaii. These species represented the great majority of individual woody plants at the site. Flowers and fruits were available in the community year–round; however, all species exhibited annual patterns of flowering, and four species showed annual patterns of fruiting while the rest fruited in supra–annual patterns. Many species had protracted flowering or fruiting peaks, and some bore small numbers of flowers or fruit year‐round. Most species flowered in a monthly peak mainly between May and August, corresponding to the period of greatest solar irradiance and marginally higher temperatures. Fruit ripening followed at varying intervals. In contrast, the heaviest flowering occurred between November and March, resulting from bloom of the dominant tree, Metrosideros polymorpha. At the highest elevations, Metrosideros flowering was heaviest during September, but peak flowering of lower elevation trees occurred in late fall and winter. Two forms of this species differed in their temporal and spatial patterns of flowering. For M. polymorpha var. polymorpha and var. incana, bloom peaked annually between November and January; however, for M. polymorpha var. glaberrima, flowering peaked from April through July, with an earlier secondary peak in January.     

Freezing Tolerance and Alteration of Translatable mRNAs in Alfalfa (Medicago sativa L.) Hardened at Subzero Temperatures

We analyzed changes in populations of translatable mRNAs occurringin crowns of the cold-tolerant alfalfa (Medicago sativa L.)cv. Apica (CT) and the cold-sensitive cv. CUF-101 (CS) aftertheir acclimation at low nonfreezing temperatures and at subzerotemperatures. Both cultivars showed very similar translationprofiles under all treatments. Low temperatures induced significantchanges in the populations of translatable mRNAs. We observeda relationship between the accumulation of cold-regulated (COR)translation products and freezing tolerance within cultivars.Moreover, at least three COR translation products were specificto the CT and might be related to hardiness potential in alfalfa.Whereas extension of the cold acclimation period at 2C reducedcold tolerance, incubation at subzero temperatures increasedor maintained freezing tolerance. This increased hardiness wasassociated with enhanced translation of COR polypeptides andalso with the appearance of new translatable mRNAs. This is,to our knowledge, the first report of altered gene expressionin plants incubated at subzero temperatures. Marked changesin populations of translatable mRNAs at temperatures below freezingmight be related to previous reports that alfalfa achieves maximumhardiness under snow cover when the soil has frozen. Translationin the presence of [³H]glycine showed that a large proportionof the COR genes encode for glycine-rich proteins (GRPs) andthat some of the GRPs are specific to the CT. (Received May 29, 1992; Accepted October 13, 1992)     

Effects of experimental warming on arctic willows (Salix spp.): a comparison of responses from the Canadian High Arctic, Alaskan Arctic, and Swedish Subarctic

Three species of dwarf, prostrate willow ( Salix arctica , S. rotundifolia and S. herbacea ) were subjected to experimental summer warming in high arctic Canada, arctic Alaska, and subarctic Sweden, respectively, as part of the International Tundra Experiment. Phenological and growth responses of these species were compared for the second season of the experiment. Stigmas became receptive and pollen dispersal occurred significantly earlier for S. rotundifolia and S. herbacea in the ITEX open-top chambers, but not for S. arctica . Warming had no effect on the timing of seed dispersal, leaf yellowing, or leaf senescence. The length and dry weight of the largest leaves were greater for warmed plants, and was significant for S. rotundifolia . The number of catkins/plot did not differ among species or treatments, but the fruit : flower ratio was reduced in the experimental plots.