The Y-segment of novel cold dehydrin genes is conserved and codons in the PR-10 genes are under positive selection in Oxytropis (Fabaceae) from contrasting climates

While the arctic flora is particularly threatened by climate changes, the molecular aspects allowing colonization of this harsh environment remain largely enigmatic. Genes with a likely functional or evolutive role for arctic Oxytropis (Fabaceae) were previously discovered given a sharp differential expression between arctic and temperate species, but the evolutionary forces in action were unknown within the respective species. Here, we analyze gene duplication patterns and positive and negative selection between genes from species of contrasting environments, which can reveal potential gene functions. Genes were amplified and sequenced from two arctic (Oxytropis arctobia and O. maydelliana) and two temperate (O. campestris subsp. johannensis and O. splendens) species. Detection of codons under positive or negative selection and phylogenetic analyses were used to further elucidate pathogenesis-related class 10 (PR-10), ripening-related proteins, cold dehydrins gene families and light-harvesting complex (lhcaIII and lhcbI) genes from Oxytropis. Overall, results showed that the three gene families duplicated in tandem prior to the Oxytropis genus diversification; that genes overexpressed in arctic species evolve under higher constraints at the sequence level in these species; that evolving novel protein variants in PR-10 genes were required for initial adaptation to the Arctic, and that Oxytropis cold dehydrins are of a novel (K-like–Y₄–K–S) structure, where the Y-segment is under stringent evolutive constraints in the arctic species. This suggests a scenario not previously described for arctic plants, where tandem duplications precede gene recruitment that later become both highly expressed and under stringent constraints in the arctic species.     

Effects of low temperatures on nitrogenase activity in sainfoin (Onobrychis viciifolia) nodulated by arctic rhizobia

The temperate forage legume sainfoin (Onobrychis viciifolia) is readly nodulated by rhizobia isolated from arctic legumes (Astragalus and Oxytropis species). We have investigated the effects of low temperatures on nitrogenase activity in sainfoin nodulated by arctic and temperate (homologous) rhizobia. At low temperatures, nitrogenase activity of arctic rhizobia measured either with detached nodules or with whole plants, was higher than that of temperate rhizobia. At 5°C and 10°C, nitrogenase activity values of arctic rhizobia represented 12% and 33% of those measured at 20°C, while lower values of 3.7% and 22.4% were observed with temperate rhizobia. This cold adaptation was also reflected on bacterial growth where, at 5°C and 10°C, arctic rhizobia showed a shorter doubling time and synthesized more protein than temperate rhizobia.     

Symbiotic effectiveness of indigenous arctic rhizobia on a temperate forage legume: Sainfoin (Onobrychis viciifolia)

Sainfoin (Onobrychis viciifolia), a temperate perennial forage legume, can be nodulated by rhizobia isolated from 3 arctic legume species:Astragalus alpinus, oxytropis maydelliana andOxytropis arctobia. Arctic rhizobia, which are adapted to growth at low temperatures, may be useful in improving symbiotic nitrogen fixation during cold phases of the growing season, if they are effective on a temperate legume. In this study, we report on the symbiotic effectiveness of arctic rhizobia on sainfoin, as appraised by the total shoot dry matter yield obtained from 2 harvests. Under N-free conditions, 5 arctic strains at the first harvest and 8 at the second harvest were as effective as temperate standard strains. In the presence of 30 mgl⁻¹ NO₃-N, 7 arctic strains gave significantly higher yields than temperate strains at the second harvest. These results indicate that effective arctic rhizobia have a potential for use as inoculants on sainfoin. Contribution no 325 of Agriculture Canada Research Station a Sainte-Foy.     

An Analysis of Plant Growth and its Control in Arctic Environments

The relative growth-rate of plants grown on a vermiculite culturemedium in an arctic climate during the growing season was abouta quarter of that of comparable plants on the same medium ina temperate climate. In both climates the relative growth-ratewas lower on natural soils than on vermiculite. Net assimilationrates and, to a lesser extent, leaf-area ratios were depressedby arctic climates and soils. Net assimilation rates of seven species in various habitatsin two arctic regions were about 0.1–0.3g dm^–2wk^–1.Previous suggestions that net assimilation rates in arctic regionsequal or exceed those in temperate regions are attributed tomisinterpretation of data or to inadequate methods. There is evidence that the depression of net assimilation ratesin arctic regions is due to the low temperatures, which, especiallywhen associated with soil nitrogen deficiency, reduce the rateat which assimilates are used in respiration and new growth;this causes sugars to accumulate to levels at which they depressassimilation.     

Cold stress and acclimation – what is important for metabolic adjustment?

As sessile organisms, plants are unable to escape from the many abiotic and biotic factors that cause a departure from optimal conditions of growth and development. Low temperature represents one of the most harmful abiotic stresses affecting temperate plants. These species have adapted to seasonal variations in temperature by adjusting their metabolism during autumn, increasing their content of a range of cryo‐protective compounds to maximise their cold tolerance. Some of these molecules are synthesised de novo. The down‐regulation of some gene products represents an additional important regulatory mechanism. Ways in which plants cope with cold stress are described, and the current state of the art with respect to both the model plant Arabidopsis thaliana and crop plants in the area of gene expression and metabolic pathways during low‐temperature stress are discussed.     

Identification and characterization of stress resistance related genes of Brassica rapa

Two biotic stress resistance related genes from the full-length cDNA library of Brassica rapa cv. Osome were identified from EST analysis and determined to be pathogenesis-related (PR) 12 Brassica defensin-like family protein (BrDLFP) and PR-10 Brassica Betv1 allergen family protein (BrBetv1AFP) after sequence analysis and homology study with other stress resistance related same family genes. In the expression analysis, both genes expressed in different organs and during all developmental growth stages in healthy plants. Expression of BrDLFP significantly increased and BrBetv1AFP gradually decreased after infection with Pectobacterium carotovorum subsp. carotovorum in Chinese cabbage. Expression of these two genes significantly changed after cold, salt, drought and ABA stress treatments. These two PR genes may therefore be involved in the plant resistance against biotic and abiotic stresses.     

Respiration rate of arctic plants as related to the production process

This work deals with two intertwined questions: (1) what are the factors underlying equally high respiration rates of arctic plants at low temperature and of temperate zone plants at 20–25°C and (2) whether this respiration feature would explain small size of the northern plants. In an attempt to answer these questions, we collected various hypotheses scattered in the current literature and experimentally examined the respiration- growth relationships by analyzing plant productivity characteristics in three representative species inhabiting Wrangel Island (lat. 71°N). The results show that the components of the production process stay in accord in the arctic plants so that their productivity characteristics at low temperatures are nearly the same as in the temperate zone plants at higher temperatures. Hence, respiration cannot account for small size of the northern plants. Upon the experimental results and general concepts for regulation of respiration, we conclude that the intense respiration of plants inhabiting cold climate regions is caused by higher metabolic demands for energy and intermediates under the northern conditions. The enhanced metabolic demands of plants at low temperature represent the main factor of intense respiration.     

Quantitative proteomic analysis of short photoperiod and low-temperature responses in bark tissues of peach (<Emphasis Type="Italic">Prunus persica</Emphasis> L. Batsch)

In the temperate climate of the northern hemisphere, winter survival of woody plants is determined by the ability to acclimate to freezing temperatures and to undergo a period of dormancy. Cold acclimation in many woody plants is initially induced by short photoperiod and low, non-freezing temperatures. These two factors (5°C and short photoperiod) were used to study changes in the proteome of bark tissues of 1-year-old peach trees. Difference in-gel electrophoresis technology, a gel-based approach involving the labeling of proteins with different fluorescent dyes, was used to conduct a quantitative assessment of changes in the peach bark proteome during cold acclimation. Using this approach, we were able to identify differentially expressed proteins and to assign them to a class of either ‘temperature-responsive’ or ‘photoperiod-responsive’ proteins. The most significant factor affecting the proteome appeared to be low temperature, while the combination of low temperature and short photoperiod was shown to act either synergistically or additively on the expression of some proteins. Fifty-seven protein spots on gels were identified by mass spectrometry. They included proteins involved in carbohydrate metabolism (e.g., enolase, malate dehydrogenase, etc), defense or protective mechanisms (e.g., dehydrin, HSPs, and PR-proteins), energy production and electron transport (e.g., adenosine triphosphate synthases and lyases), and cytoskeleton organization (e.g., tubulins and actins). The information derived from the analysis of the proteome is discussed as a function of the two treatment factors: low temperature and short photoperiod. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

THE ECOLOGY OF ARCTIC AND ALPINE PLANTS

‘How are plants adapted to the low temperatures and other stresses of arctic and alpine environments ?’ At present it is not possible to answer this question completely. Much work remains to be done, particularly on low-temperature metabolism, frost resistance, and the environmental cues and requirements for flowering, dormancy, regrowth, and germination. However, in brief, we can say that plants are adapted to these severe environments by employing combinations of the following general characteristics: 1. Life form: perennial herb, prostrate shrub, or lichen. Perennial herbs have greatest part of biomass underground. 2. Seed dormancy: generally controlled by environment; seeds can remain dormant for long periods of time at low temperatures since they require temperatures well above freezing for germination. 3. Seedling establishment: rare and very slow; it is often several years before a seedling is safely established. 4. Chlorophyll content: in both alpine and arctic ecosystems not greatly different on a land-area basis from that in temperate herbaceous communities. Within a single species there is more chlorophyll in leaves of arctic populations than in those of alpine populations. 5. Photosynthesis and respiration: (a) These are at high rates for only a few weeks when temperatures and light are favourable. (b) Optimum photosynthesis rates are at lower temperatures than for ordinary plants; rates are both genetically and environmentally controlled with phenotypic plasticity very marked. (c) Dark respiration is higher at all temperatures than for ordinary plants; rate is both genetically and environmentally controlled, with phenotypic plasticity very pronounced, i.e. low-temperature environment increases the rate at all temperatures. (d) Alpine plants have higher light-saturation values in photosynthesis than do arctic or lowland plants; light saturation closely tied to temperature. (e) There is some evidence that alpine plants can carry on photosynthesis at lower carbon dioxide concentrations than can other plants. (f) Annual productivity is low, but daily productivity during growing season can be as high as that of most temperate herbaceous vegetation. Productivity can be increased by temperature, nutrients, or water. 6. Drought resistance: most drought stress in winter in exposed sites is due to frozen soils and dry winds. It is met by decreased water potentials, higher concentrations of soluble carbohydrates, and closed stomates. Little drought resistance in snowbank plants. Alpine plants adapted to summer drought stress can carry on photosynthesis at low water potentials; alpine or arctic plants of moist sites cannot do this. 7. Breaking of dormancy: controlled by mean temperatures near or above 0° C., and in some cases by photoperiod also. 8. Growth: very rapid even at low positive temperatures. Respiration greatly exceeds photosynthesis in early re-growth of perennials. Internal photosynthesis may occur in hollow stems of larger plants during early growth. Nitrogen and phosphorus often limiting in cold soil. 9. Food storage: characteristic of all alpine and arctic plants except annuals. Carbohydrates mostly stored underground in herbaceous perennials. Lipids in old leaves and stems of prostrate evergreen shrubs. Depleted in early growth, and usually restored after flowering. 10. Winter survival: survival and frost resistance are excellent after hardening. Cold resistance closely tied to content of soluble carbohydrates, particularly raffinose. 11. Flowering: flower buds are pre-formed the year before. Complete development and anthesis dependent upon temperature of the flowering year and also, in some cases, upon photoperiod. 12. Pollination: mostly insect-pollinated in alpine regions and even in Arctic, but to a lesser extent. Wind-pollination increasingly more important with increasing latitude. Diptera more important than bees in the Arctic and in the highest mountains. 13. Seed production: opportunistic, and dependent upon temperature during flowering period and latter half of growing season. 14. Vegetative reproduction: by rhizomes, bulbils, or layering. More common and important in Arctic than in alpine areas. 15. Onset of dormancy: triggered by photoperiod, low temperatures, and drought. Dormant plant extremely resistant to low temperatures.     

Heat and cold shock protein synthesis in arctic and temperate strains of rhizobia.

We compared heat shock proteins (HSPs) and cold shock proteins (CSPs) produced by different species of Rhizobium having different growth temperature ranges. Several HSPs and CSPs were induced when cells of three arctic (psychrotrophic) and three temperate (mesophilic) strains of rhizobia were shifted from their optimal growth temperatures (arctic, 25 degrees C; temperate, 30 degrees C) to shock temperatures outside their growth temperature ranges. At heat shock temperatures, three major HSPs of high molecular weight (106,900, 83,100, and 59,500) were present in all strains for all shock treatments (29, 32, 36.4, 38.4, 40.7, 41.4, and 46.4 degrees C), with the exception of temperate strains exposed to 46.4 degrees C, in which no protein synthesis was detected. Cell survival of arctic and temperate strains decreased markedly with the increase of shock temperature and was only 1% at 46.4 degrees C. Under cold shock conditions, five proteins (52.0, 38.0, 23.4, 22.7, and 11.1 kDa) were always present for all treatments (-2, -5, and -10 degrees C) in arctic strains. Among temperate strains, five CSPs (56.1, 37.1, 34.4, 17.3, and 11.1 kDa) were present at temperatures down to 0 degrees C. The 34.4- and the 11.1-kDa components were present in all temperate strains at -5 degrees C and in one strain at -10 degrees C. Survival of all strains decreased with cold shock temperatures but was always higher than 50%. These results show that rhizobia can synthesize proteins at temperatures not permissive for growth. In all shock treatments, no correspondence between the number of HSPs or CSPs produced and rhizobial survival was found.(ABSTRACT TRUNCATED AT 250 WORDS)     

Heat and cold shock protein synthesis in arctic and temperate strains of rhizobia.

We compared heat shock proteins (HSPs) and cold shock proteins (CSPs) produced by different species of Rhizobium having different growth temperature ranges. Several HSPs and CSPs were induced when cells of three arctic (psychrotrophic) and three temperate (mesophilic) strains of rhizobia were shifted from their optimal growth temperatures (arctic, 25 degrees C; temperate, 30 degrees C) to shock temperatures outside their growth temperature ranges. At heat shock temperatures, three major HSPs of high molecular weight (106,900, 83,100, and 59,500) were present in all strains for all shock treatments (29, 32, 36.4, 38.4, 40.7, 41.4, and 46.4 degrees C), with the exception of temperate strains exposed to 46.4 degrees C, in which no protein synthesis was detected. Cell survival of arctic and temperate strains decreased markedly with the increase of shock temperature and was only 1% at 46.4 degrees C. Under cold shock conditions, five proteins (52.0, 38.0, 23.4, 22.7, and 11.1 kDa) were always present for all treatments (-2, -5, and -10 degrees C) in arctic strains. Among temperate strains, five CSPs (56.1, 37.1, 34.4, 17.3, and 11.1 kDa) were present at temperatures down to 0 degrees C. The 34.4- and the 11.1-kDa components were present in all temperate strains at -5 degrees C and in one strain at -10 degrees C. Survival of all strains decreased with cold shock temperatures but was always higher than 50%. These results show that rhizobia can synthesize proteins at temperatures not permissive for growth. In all shock treatments, no correspondence between the number of HSPs or CSPs produced and rhizobial survival was found.(ABSTRACT TRUNCATED AT 250 WORDS)     

Observations on Net Assimilation Rates in Arctic Environments: With two Figures in the Text

Examination of the net assimilation rate (E) during the growingseason in arctic regions by a detached-leaf method revealedno differences between species or with soil richness, but showeda reduction of E with exposure to wind–probably resultingfrom cooling–and a tendency for E to fall towards thelater part of the growing season. E generally lay in the range0·5 to o·8 g./dm.²/week. E for detached leaves ignores respiratory losses in other partsof the plant and is not comparable with E for whole plants;failure to appreciate this confused a previous comparison ofE under arctic and temperate environments. E for detached leavesin temperate summer conditions is normally around 1·1to 1·5 g./dm.²/week. Thus E is reduced in arctic environmentsto about half the value in temperate conditions. This reductionis due mainly to the cold climate.