Effects of temperature and water availability on light energy utilization in photosynthetic processes of Deschampsia antarctica.

Regional climate change in Antarctica would favor the carbon assimilation of Antarctic vascular plants, since rising temperatures are approaching their photosynthetic optimum (10-19°C). This could be detrimental for photoprotection mechanisms, mainly those associated with thermal dissipation, making plants more susceptible to eventual drought predicted by climate change models. With the purpose to study the effect of temperature and water availability on light energy utilization and putative adjustments in photoprotective mechanisms of Deschampsia antarctica Desv., plants were collected from two Antarctic provenances: King George Island and Lagotellerie Island. Plants were cultivated at 5, 10 and 16°C under well-watered (WW) and water-deficit (WD, at 35% of the field capacity) conditions. Chlorophyll fluorescence, pigment content and de-epoxidation state were evaluated. Regardless of provenances, D. antarctica showed similar morphological, biochemical and functional responses to growth temperature. Higher temperature triggered an increase in photochemical activity (i.e. electron transport rate and photochemical quenching), and a decrease in thermal dissipation capacity (i.e. lower xanthophyll pool, Chl a/b and ß carotene/neoxanthin ratios). Leaf mass per unit area was reduced at higher temperature, and was only affected in plants exposed to WD at 16°C and exhibiting lower electron transport rate and amount of chlorophylls. D. antarctica is adapted to frequent freezing events, which may induce a form of physiological water stress. Photoprotective responses observed under WD contribute to maintain a stable photochemical activity. Thus, it is possible that short-term temperature increases could favor the photochemical activity of this species. However, long-term effects will depend on the magnitude of changes and the plant's ability to adjust to new growth temperature.

Novel antimalarial chloroquine- and primaquine-quinoxaline 1,4-di-N-oxide hybrids: Design, synthesis, Plasmodium life cycle stage profile, and preliminary toxicity studies.

Emergence of drug resistance and targeting all stages of the parasite life cycle are currently the major challenges in antimalarial chemotherapy. Molecular hybridization combining two scaffolds in a single molecule is an innovative strategy for achieving these goals. In this work, a series of novel quinoxaline 1,4-di-N-oxide hybrids containing either chloroquine or primaquine pharmacophores was designed, synthesized and tested against both chloroquine sensitive and multidrug resistant strains of Plasmodium falciparum. Only chloroquine-based compounds exhibited potent blood stage activity with compounds 4b and 4e being the most active and selective hybrids at this parasite stage. Based on their intraerythrocytic activity and selectivity or their chemical nature, seven hybrids were then evaluated against the liver stage of Plasmodium yoelii, Plasmodium berghei and Plasmodium falciparum infections. Compound 4b was the only chloroquine-quinoxaline 1,4-di-N-oxide hybrid with a moderate liver activity, whereas compound 6a and 6b were identified as the most active primaquine-based hybrids against exoerythrocytic stages, displaying enhanced liver activity against P. yoelii and P. berghei, respectively, and better SI values than primaquine. Although both primaquine-quinoxaline 1,4-di-N-oxide hybrids slightly reduced the infection of mosquitoes, they inhibited sporogony of P. berghei and compound 6a showed 92% blocking of transmission. In vivo liver efficacy assays revealed that compound 6a showed causal prophylactic activity affording parasitaemia reduction of up to 95% on day 4. Absence of genotoxicity and in vivo acute toxicity were also determined. These results suggest the approach of primaquine-quinoxaline 1,4-di-N-oxide hybrids as new potential dual-acting antimalarials for further investigation.

Decoding the gene coexpression network underlying the ability of Gevuina avellana to live in diverse light conditions.

Gevuina avellana (Proteaceae) is a typical tree from the South American temperate rainforest. Although this species mostly regenerates in shaded understories, it exhibits an exceptional ecological breadth, being able to live under a wide range of light conditions. Here we studied the genetic basis that underlies physiological acclimation of the photosynthetic responses of G. avellana under contrasting light conditions. We analyzed carbon assimilation and light energy used for photochemical processes in plants acclimated to contrasting light conditions. Also, we used a transcriptional profile of leaf primordia from G. avellana saplings growing under different light environments in their natural habitat, to identify the gene coexpression network underpinning photosynthetic performance and light-related processes. The photosynthetic parameters revealed optimal performance regardless of light conditions. Strikingly, the mechanism involved in dissipation of excess light energy showed no significant differences between high- and low-light-acclimated plants. The gene coexpression network defined a community structure consistent with the photochemical responses, including genes involved mainly in assembly and functioning of photosystems, photoprotection, and retrograde signaling. This ecophysiological genomics approach improves our understanding of the intraspecific variability that allows G. avellana to have optimal photochemical and photoprotective mechanisms in the diverse light habitats it encounters in nature.

In situ warming in the Antarctic: effects on growth and photosynthesis in Antarctic vascular plants.

The Antarctic Peninsula has experienced a rapid warming in the last decades. Although recent climatic evidence supports a new tendency towards stabilization of temperatures, the impacts on the biosphere, and specifically on Antarctic plant species, remain unclear. We evaluated the in situ warming effects on photosynthesis, including the underlying diffusive, biochemical and anatomical determinants, and the relative growth of two Antarctic vascular species, Colobanthus quitensis and Deschampsia antarctica, using open top chambers (OTCs) and gas exchange measurements in the field. In C. quitensis, the photosynthetic response to warming relied on specific adjustments in the anatomical determinants of the leaf CO2 transfer, which enhanced mesophyll conductance and photosynthetic assimilation, thereby promoting higher leaf carbon gain and plant growth. These changes were accompanied by alterations in the leaf chemical composition. By contrast, D. antarctica showed no response to warming, with a lack of significant differences between plants grown inside OTCs and plants grown in the open field. Overall, the present results are the first reporting a contrasting effect of in situ warming on photosynthesis and its underlying determinants, of the two unique Antarctic vascular plant species, which could have direct consequences on their ecological success under future climate conditions.

Physiological differences between root suckers and saplings enlarge the regeneration niche in Eucryphia cordifolia Cav.

Many clonal plants produce vegetative recruits that remain connected to the parent plant. Such connections permit resource sharing among ramets, explaining the high survival rates of vegetative recruits during establishment under suboptimal conditions for sexual regeneration. We propose that differences in the regeneration niches of sexual and vegetative recruits reflect different physiological adjustments caused by parental supply of resources to the ramets. We conducted ecophysiological measurements in saplings and root suckers of Eucryphia cordifolia Cav., a tree species of the temperate rainforest of southern South America. We compared the following traits of saplings and suckers: gas exchange at the leaf level, crown architecture, daily crown carbon balance, biomass allocation to above-ground tissues (leaf-to-stem mass ratio, leaf mass area and leaf area ratio), xylem anatomy traits (lumen vessel fraction, vessel density and size) and stem ring width. We also correlated the growth rates of saplings and suckers with relevant environmental data (light and climate). Saplings showed morphological, architectural and physiological traits that enhance daily crown carbon balance and increase water-use efficiency, in order to supply their growth demands while minimizing water loss per unit of carbon gained. The radial growth of saplings diminished under dry conditions, which suggests a strong stomatal sensitivity to water availability. Suckers have low stomatal conductance, likely because the carbon supplied by the parent plant diminishes the necessity of high rates of photosynthesis. The low responsiveness of sucker growth to temporal changes in water availability also supports the existence of parental supply. The physiological differences between sexual and vegetative recruits satisfactorily explain the ecological niche of E. cordifolia, with saplings restricted to more closed and humid sites.

Two Hymenophyllaceae species from contrasting natural environments exhibit a homoiochlorophyllous strategy in response to desiccation stress.

Hymenophyllaceae is a desiccation tolerant family of Pteridophytes which are poikilohydric epiphytes. Their fronds are composed by a single layer of cells and lack true mesophyll cells and stomata. Although they are associated with humid and shady environments, their vertical distribution varies along the trunk of the host plant with some species inhabiting the drier sides with a higher irradiance. The aim of this work was to compare the structure and function of the photosynthetic apparatus during desiccation and rehydration in two species, Hymenophyllum dentatum and Hymenoglossum cruentum, isolated from a contrasting vertical distribution along the trunk of their hosts. Both species were subjected to desiccation and rehydration kinetics to analyze frond phenotypic plasticity, as well as the structure, composition and function of the photosynthetic apparatus. Minimal differences in photosynthetic pigments were observed upon dehydration. Measurements of ϕPSII (effective quantum yield of PSII), ϕNPQ (quantum yield of the regulated energy dissipation of PSII), ϕNO (quantum yield of non-regulated energy dissipation of PSII), and TL (thermoluminescence) indicate that both species convert a functional photochemical apparatus into a structure which exhibits maximum quenching capacity in the dehydrated state with minimal changes in photosynthetic pigments and polypeptide compositions. This dehydration-induced conversion in the photosynthetic apparatus is completely reversible upon rehydration. We conclude that H. dentatum and H. cruentum are homoiochlorophyllous with respect to desiccation stress and exhibited no correlation between inherent desiccation tolerance and the vertical distribution along the host tree trunk.

Transcriptomic analysis suggests a key role for SQUAMOSA PROMOTER BINDING PROTEIN LIKE, NAC and YUCCA genes in the heteroblastic development of the temperate rainforest tree Gevuina avellana (Proteaceae).

Heteroblasty, the temporal development of the meristem, can produce diverse leaf shapes within a plant. Gevuina avellana, a tree from the South American temperate rainforest shows strong heteroblasty affecting leaf shape, transitioning from juvenile simple leaves to highly pinnate adult leaves. Light availability within the forest canopy also modulates its leaf size and complexity. Here we studied how the interaction between the light environment and the heteroblastic progression of leaves is coordinated in this species. We used RNA-seq on the Illumina platform to compare the range of transcriptional responses in leaf primordia of G. avellana at different heteroblastic stages and growing under different light environments. We found a steady up-regulation of SQUAMOSA PROMOTER BINDING PROTEIN LIKE (SPL), NAC, YUCCA and AGAMOUS-LIKE genes associated with increases in age, leaf complexity, and light availability. In contrast, expression of TCP, TPR and KNOTTED1 homeobox genes showed a sustained down-regulation. Additionally, genes involved in auxin synthesis/transport and jasmonate activity were differentially expressed, indicating an active regulation of processes controlled by these hormones. Our large-scale transcriptional analysis of the leaf primordia of G. avellana sheds light on the integration of internal and external cues during heteroblastic development in this species.

Photosynthetic Light Responses May Explain Vertical Distribution of Hymenophyllaceae Species in a Temperate Rainforest of Southern Chile.

Some epiphytic Hymenophyllaceae are restricted to lower parts of the host (< 60 cm; 10-100 µmol photons m(-2) s(-1)) in a secondary forest of Southern Chile; other species occupy the whole host height (≥ 10 m; max PPFD > 1000 µmol photons m(-2) s(-1)). Our aim was to study the photosynthetic light responses of two Hymenophyllaceae species in relation to their contrasting distribution. We determined light tolerance of Hymenoglossum cruentum and Hymenophyllum dentatum by measuring gas exchange, PSI and PSII light energy partitioning, NPQ components, and pigment contents. H. dentatum showed lower maximum photosynthesis rates (A max) than H. cruentum, but the former species kept its net rates (An) near Amax across a wide light range. In contrast, in the latter one, An declined at PPFDs > 60 µmol photons m(-2) s(-1). H. cruentum, the shadiest plant, showed higher chlorophyll contents than H. dentatum. Differences in energy partitioning at PSI and PSII were consistent with gas exchange results. H. dentatum exhibited a higher light compensation point of the partitioning of absorbed energy between photochemical Y(PSII) and non-photochemical Y(NPQ) processes. Hence, both species allocated energy mainly toward photochemistry instead of heat dissipation at their light saturation points. Above saturation, H. cruentum had higher heat dissipation than H. dentatum. PSI yield (YPSI) remained higher in H. dentatum than H. cruentum in a wider light range. In both species, the main cause of heat dissipation at PSI was a donor side limitation. An early dynamic photo-inhibition of PSII may have caused an over reduction of the Qa+ pool decreasing the efficiency of electron donation to PSI. In H. dentatum, a slight increase in heat dissipation due to acceptor side limitation of PSI was observed above 300 µmol photons m(-2)s(-1). Differences in photosynthetic responses to light suggest that light tolerance and species plasticity could explain their contrasting vertical distribution.

Acclimation of leaf cohorts expanded under light and water stresses: an adaptive mechanism of Eucryphia cordifolia to face changes in climatic conditions?

Eucryphia cordifolia Cav. is a long-lived evergreen tree species, commonly found as a canopy emergent tree in the Chilean temperate rain forest. This species displays successive leaf cohorts throughout the entire growing season. Thus, full leaf expansion occurs under different environmental conditions during growing such as air temperature, vapor pressure deficit and the progress of moderate water stress (WS). These climate variations can be reflected as differences in anatomical and physiological characteristics among leaf cohorts. Thus, we investigated the potential adaptive role of different co-existing leaf cohorts in seedlings grown under shade, drought stress or a combination of the two. Photosynthetic and anatomical traits were measured in the first displayed leaf cohort and in a subsequent leaf cohort generated during the mid-season. Although most anatomical and photosynthetic pigments did not vary between cohorts, photosynthetic acclimation did occur in the leaf cohort and was mainly driven by biochemical processes such as leaf nitrogen content, Rubisco carboxylation capacity and maximal Photosystem II electron transport rather than CO2 diffusion conductance. Cohort acclimation could be relevant in the context of climate change, as this temperate rainforest will likely face some degree of summer WS even under low light conditions. We suggest that the acclimation of the photosynthetic capacity among current leaf cohorts represents a well-tuned mechanism helping E. cordifolia seedlings to face a single stress like shade or drought stress, but is insufficient to cope with simultaneous stresses.

Cold-acclimation limits low temperature induced photoinhibition by promoting a higher photochemical quantum yield and a more effective PSII restoration in darkness in the Antarctic rather than the Andean ecotype of Colobanthus quitensis Kunt Bartl (Cariophyllaceae).

BACKGROUND: Ecotypes of Colobanthus quitensis Kunt Bartl (Cariophyllaceae) from Andes Mountains and Maritime Antarctic grow under contrasting photoinhibitory conditions, reaching differential cold tolerance upon cold acclimation. Photoinhibition depends on the extent of photodamage and recovery capability. We propose that cold acclimation increases resistance to low-temperature-induced photoinhibition, limiting photodamage and promoting recovery under cold. Therefore, the Antarctic ecotype (cold hardiest) should be less photoinhibited and have better recovery from low-temperature-induced photoinhibition than the Andean ecotype. Both ecotypes were exposed to cold induced photoinhibitory treatment (PhT). Photoinhibition and recovery of photosystem II (PSII) was followed by fluorescence, CO2 exchange, and immunoblotting analyses. RESULTS: The same reduction (25%) in maximum PSII efficiency (Fv/Fm) was observed in both cold-acclimated (CA) and non-acclimated (NA) plants under PhT. A full recovery was observed in CA plants of both ecotypes under dark conditions, but CA Antarctic plants recover faster than the Andean ecotype.Under PhT, CA plants maintain their quantum yield of PSII, while NA plants reduced it strongly (50% and 73% for Andean and Antarctic plants respectively). Cold acclimation induced the maintenance of PsaA and Cyt b6/f and reduced a 41% the excitation pressure in Antarctic plants, exhibiting the lowest level under PhT. xCold acclimation decreased significantly NPQs in both ecotypes, and reduced chlorophylls and D1 degradation in Andean plants under PhT.NA and CA plants were able to fully restore their normal photosynthesis, while CA Antarctic plants reached 50% higher photosynthetic rates after recovery, which was associated to electron fluxes maintenance under photoinhibitory conditions. CONCLUSIONS: Cold acclimation has a greater importance on the recovery process than on limiting photodamage. Cold acclimation determined the kinetic and extent of recovery process under darkness in both C. quitensis ecotypes. The greater recovery of PSII at low temperature in the Antarctic ecotype was related with its ability to maintain PsaA, Cyt b6/f and D1 protein after photoinhibitory conditions. This is probably due to either a higher stability of these polypeptides or to the maintenance of their turnover upon cold acclimation. In both cases, it is associated to the maintenance of electron drainage from the intersystem pool, which maintains QA more oxidized and may allow the synthesis of ATP and NADPH necessaries for the regeneration of ribulose 1,5-bisphosphate in the Calvin Cycle. This could be a key factor for C. quitensis success under the harsh conditions and the short growing period in the Maritime Antarctic.

Tree size and light availability increase photochemical instead of non-photochemical capacities of Nothofagus nitida trees growing in an evergreen temperate rain forest.

Nothofagus nitida (Phil.) Krasser (Nothofagaceae) regenerates under the canopy in microsites protected from high light. Nonetheless, it is common to find older saplings in clear areas and adults as emergent trees of the Chilean evergreen forest. We hypothesized that this shade to sun transition in N. nitida is supported by an increase in photochemical and non-photochemical energy dissipation capacities of both photosystems in parallel with the increase in plant size and light availability. To dissect the relative contribution of light environment and plant developmental stage to these physiological responses, the photosynthetic performance of both photosystems was studied from the morpho-anatomical to the biochemical level in current-year leaves of N. nitida plants of different heights (ranging from 0.1 to 7 m) growing under contrasting light environments (integrated quantum flux (IQF) 5-40 mol m(-2). Tree height (TH) and light environment (IQF) independently increased the saturated electron transport rates of both photosystems, as well as leaf and palisade thickness, but non-photochemical energy flux, photoinhibition susceptibility, state transition capacity, and the contents of D1 and PsbS proteins were not affected by IQF and TH. Spongy mesophyll thickness and palisade cell diameter decreased with IQF and TH. A(max), light compensation and saturation points, Rubisco and nitrogen content (area basis) only increased with light environment (IQF), whereas dark respiration (R(d)) decreased slightly and relative chlorophyll content was higher in taller trees. Overall, the independent effects of more illuminated environment and tree height mainly increased the photochemical instead of the non-photochemical energy flux. Regardless of the photochemical increase with TH, carbon assimilation only significantly improved with higher IQF. Therefore it seems that mainly acclimation to the light environment supports the phenotypic transition of N. nitida from shade to sun.

Genotypic variation in morphology and freezing resistance of Eucalyptus globulus seedlings subjected to drought hardening in nursery / Variación genotípica en la morfología y resistencia a la congelación del Eucalyptus globulus a la sequía sometido al endurecimiento en vivero

Eucalyptus globulus Labill is one of the most planted species in Chile, because of its fast growth and superior pulp qualities. Nevertheless, the incidence of drought and frost damage immediately after planting is frequent. The purpose of this work was to study the effect of drought hardening on frost resistance and on variations in morphological traits that may increase drought resistance at nursery phase in four genotypes of E. globulus Labill. Drought hardening treatments consisted in induced water stress by watering restriction, until pre-dawn stem xylem water potentials (Psi pd) reached -0.2, -1.8 and -2.6 MPa. Two water stress-rewatering cycles were applied during 54 days of hardening. Plant and root biomasses were affected by the interaction of drought hardening and genotypes. The rest of morphological and alometrical traits were affected independently by drought or genotype. Plant height, leaf area, specific leaf area (SLA), stem, and leaf biomasses decreased with drought hardening, while collar diameter was not affected. Genotypes responded differentially to drought hardening in plant height, leaf area, SLA, and stem, and leaf biomasses. Ice nucleation temperature (INT), and freezing temperatures (FRT), and 50 percent freezing damage index of leaves (LT50) were affected by the interaction between drought hardening and genotypes. EG-13, EG-23 and EG-22 genotypes became freezing tolerant with drought hardening (-2.6 MPa). Additionally, EG-14 genotype increased its freezing resistance at -1.8 MPa. Therefore, freezing resistance levels and mechanism depend on genotype and drought hardening treatment. The success in tree breeding by genetic selection should be facilitated by improved understanding of the physiology of stress resistance development and survival during water supply limitations. The knowledge of morphological and freezing resistance dependency on the interaction between genotype and drought hardening may be useful...

Thermal energy dissipation and its components in two developmental stages of a shade-tolerant species, Nothofagus nitida, and a shade-intolerant species, Nothofagus dombeyi.

Nothofagus dombeyi (Mirb.) Blume and Nothofagus nitida (Phil.) Krasser, two evergreens in the South Chilean forest, regenerate in open habitats and under the canopy, respectively. Both overtop the forest canopy when they are in the adult stage, suggesting that their photoprotective mechanisms differ in ontogenetic dynamics. We postulated that N. nitida, a shade-tolerant species increases its capacity to tolerate photoinhibitory conditions (low temperature and high irradiance) by thermal energy dissipation of excess energy during its transition from the seedling to the adult stage, whereas N. dombeyi, a shade-intolerant species, maintains a high capacity for photoprotection by thermal energy dissipation from the seedling to the adult stage. To test this hypothesis, the main photoprotective mechanisms in plants - the fast- and slow-relaxing components of thermal energy dissipation (NPQ, non-photochemical quenching) NPQ(F) and NPQ(S), respectively, and state transitions - were studied in seedlings and adults of both species grown in their natural habitats and in a common garden. In adults, NPQ(F) and NPQ(S) did not differ between species and seasons. The greatest differences in these parameters were observed in seedlings. The xanthophyll cycle was more active in N. dombeyi seedlings than in N. nitida seedlings at low temperature and high irradiance, consistent with a higher NPQ(F) in N. dombeyi. Under all study conditions, N. nitida seedlings had higher NPQ(S) than N. dombeyi seedlings. The state transition capability was higher in N. nitida seedlings than in N. dombeyi seedlings. Therefore, although (shade-intolerant) N. dombeyi was able to thermally dissipate the excess absorbed energy, under natural conditions its photochemical energy quenching was efficient in both developmental stages, decreasing its need for thermal dissipation. In contrast, the seedlings of N. nitida were more sensitive to photoinhibition than the adult trees, suggesting a change from shade-grown to sun-exposed phenotype from the seedling to the adult stage. These results help to explain the differences in the regeneration patterns of N. nitida and N. dombeyi and the presence of N. nitida adult stage in the upper canopy.

Changes during early development in photosynthetic light acclimation capacity explain the shade to sun transition in Nothofagus nitida.

Nothofagus nitida (Phil.) Krasser, an emergent tree of the Chilean evergreen forest, regenerates under the canopy. Nonetheless, it is common to find older saplings in clear areas. We hypothesized that this transition from shade to sun during the early developmental stages is made possible by an ontogenetic increase in the light acclimation capacity of photosynthesis. To test our hypothesis, we studied photosynthetic performance and photoprotection in N. nitida saplings at different developmental stages corresponding with three different height classes (short: 16.2 cm; medium-height: 48.0 cm; and tall: 73.7 cm) grown under contrasting light conditions (photosynthetic photon flux (PPF) of 20, 300 or 600 micromol m(-2) s(-1)) until newly expanded leaves had developed. Light-saturated CO(2) assimilation rate and light compensation and saturation points increased with increasing PPF. Medium-height and tall saplings acclimated to high light had higher electron transport rates and higher proportions of open Photosystem II reaction centers than shorter plants acclimated to high light. Short saplings showed higher thermal dissipation and contents of xanthophylls than taller saplings. Only medium-height and tall saplings acclimated to high light recovered after photoinhibition. State transitions were higher in short plants growing in low light, and decreased with plant height and growth irradiance. Thus, during development, N. nitida changes the balance of light energy utilization and photoprotective mechanisms, supporting a phenotypic transition from shade to sun during its early ontogeny.

Effect of cold acclimation on the photosynthetic performance of two ecotypes of Colobanthus quitensis (Kunth) Bartl.

The effects of cold acclimation of two ecotypes (Antarctic and Andes) of Colobanthus quitensis (Kunth) Bartl. Caryophyllaceae on their photosynthetic characteristics and performance under high light (HL) were compared. Non-acclimated plants of the Antarctic ecotype exhibited a higher (34%) maximal rate of photosynthesis than the Andes ecotype. In cold-acclimated plants the light compensation point was increased. Dark respiration was significantly increased during the exposure to 4 degrees C in both ecotypes. Cold-acclimated Antarctic plants showed higher Phi(PSII) and qP compared with the Andes ecotype. In addition, the Antarctic ecotype exhibited higher heat dissipation (NPQ), especially in the cold-acclimated state, which was mainly associated with the fast relaxing component of non-photochemical quenching (NPQ(F)). By contrast, the Andes ecotype exhibited a lower NPQ(F) and a significant increase in the slowly relaxing component (NPQ(s)) at low temperature and HL, indicating higher sensitivity to low temperature-induced photoinhibition. Although the xanthophyll cycle was fully operational in both ecotypes, cold-acclimated Antarctic plants exposed to HL exhibited higher epoxidation state of the xanthophyll cycle pigments (EPS) compared with the cold-acclimated Andes ecotype. Thus, the photosynthetic apparatus of the Antarctic ecotype operates more efficiently than that of the Andes one, under a combination of low temperature and HL. The ecotype differences are discussed in relation to the different climatic conditions of the two Colobanthus.

Arabidopsis thaliana avoids freezing by supercooling.

Arabidopsis thaliana (L.) Heynh. has been described as a freezing-tolerant species based on freezing-resistance assays. Nonetheless, this type of experiment does not discriminate between freezing-tolerance and freezing-avoidance mechanisms. The purpose of this paper was to determine which of these two freezing-resistance mechanisms is responsible for freezing resistance in A. thaliana. This was achieved by comparing the thermal properties (ice-nucleation temperature and the freezing temperature) of leaves and the lethal temperature to 10, 50 and 90% of the plants (LT10, LT50, and LT90, respectively). Two wild-type genotypes were used (Columbia and Ler) and their mutants (esk-1 and frs-1, respectively), which differ in their freezing resistance. This study's results indicated that the mutant esk-1, described as a freezing-tolerant species showed freezing tolerance only after a cold-acclimation period. The mutant frs-1, described as freezing sensitive, presented freezing avoidance. Both wild genotypes presented LT50 similar to or higher than the ice-nucleation temperature. Thus, the main freezing-resistance mechanism for A. thaliana is avoidance of freezing by supercooling. No injury of the photosynthetic apparatus was shown by measuring the maximal photochemical efficiency (Fv/Fm) and pigments (chlorophyll and carotenoid) during cold acclimation in all genotypes. During cold acclimation, Columbia and esk-1 increased total soluble carbohydrates in leaves. esk-1 was the only genotype that presented freezing tolerance after cold acclimation. This feature could be related to an increase in sugar accumulation in the apoplast.

Light regulation of sucrose-phosphate synthase activity in the freezing-tolerant grass Deschampsia antarctica.

Deschampsia antarctica, a freezing-tolerant grass that has colonized the Maritime Antarctic, has an unusually high content of sucrose (Suc) in leaves, reaching up to 36% of dry weight. Suc accumulation has often been linked with increased activity of sucrose phosphate synthase (SPS; EC: 2.4.1.1.14). SPS, a key enzyme in sucrose biosynthesis, is controlled by an intricate hierarchy of regulatory mechanisms including allosteric modulators, reversible covalent modification in response to illumination, and transcriptional regulation. We hypothesized that during long day conditions in the Antarctic summer D. antarctica can maintain high SPS activity longer by indirect light regulation, thereby leading to a high sucrose accumulation in the leaves. The objectives of this study were to investigate a possible indirect light regulation of SPS activity and the effect of cold and day length on transcriptional and protein level of SPS in D. antarctica. Although SPS activity did not display an endogenous rhythm of activity in continuous light, activation of SPS at the end of the dark period was observed in D. antarctica. This activation of SPS is possibly controlled by covalent modification, because it was inhibited by okadaic acid while the SPS protein level did not significantly change. The highest SPS activity increase was observed after 21 days of cold-acclimation under long day conditions. This increased activity was not related to an increase in SPS gene expression or protein content. High SPS activity in cold long days leading to hyper accumulation of Suc appears to be among the features that permit D. antarctica to survive in the harsh Antarctic conditions.

Phosphate deficiency regulates phosphoenolpyruvate carboxylase expression in proteoid root clusters of white lupin.

Proteoid roots play a major role in enabling white lupin (Lupinus albus L.) to adapt to phosphate (Pi) deficiency. Such roots release citrate from proteoid rootlets, which allows this species to mobilize Pi from sparingly soluble Pi sources. Release of citrate is preceded by a significant accumulation of organic acids, in which a Pi deficiency-inducible phosphoenolpyruvate carboxylase (PEPC) activity has been involved. To gain an insight into this adaptive mechanism, the expression of three different transcripts coding for PEPC was examined in proteoid rootlets of Pi-starved and Pi-starved-and-rescued white lupin. Semi-quantitative reverse transcriptase (RT)-PCR experiments performed with gene-specific primers targeted to the 3'-end region of the corresponding cDNAs revealed that the transcripts for these three PEPCs differentially accumulate in both Pi-starved and Pi-starved-and-rescued proteoid rootlets. Semi-quantitative RT-PCR analysis in Pi-starved proteoid rootlets sampled at different times after being rescued from Pi deficiency showed that Pi levels differentially down-regulated the three PEPC transcripts. RT-PCR experiments were further extended to Pi-starved and Pi-fed whole roots, cotyledons, and leaves on which a tissue-specific, Pi-dependent PEPC expression was observed. These results indicate that there exists at least three different transcripts coding for PEPC in proteoid root clusters of white lupin, whose expression are differentially regulated by Pi.

Abscisic acid and jasmonic acid affect proteinase inhibitor activities in barley leaves.

Proteinase inhibitor (PI) accumulation has been described as a plant defense response against insects and pathogens. The induction of PIs is known to be regulated by endogenous chemical factors including phytohormones. We studied the induction of barley chymotrypsin and trypsin inhibitory activities by aphid infestation, mechanical wounding, abscisic acid (ABA) and jasmonic acid (JA). Wounding experiments led to a minimal accumulation of PI activity (16% over controls) compared to that found in barley seedlings infested by aphids, where chymotrypsin inhibitor activity showed a two-fold increment. No systemic induction could be detected in healthy leaves of an infested or mechanically injured plant. Exogenous ABA applied on barley leaves increased the chymotrypsin inhibitory activity, while JA only increased trypsin inhibitory activity locally and systemically when applied exogenously. Our data suggest that two different mechanisms may be regulating the induction of these two types of inhibitors.

The role of photochemical quenching and antioxidants in photoprotection of Deschampsia antarctica.

Deschampsia antarctica Desv. (Poaceae) is the only grass that grows in the maritime Antarctic. Constant low temperatures and episodes of high light are typical conditions during the growing season at this latitude. These factors enhance the formation of active oxygen species and may cause photoinhibition. Therefore, an efficient mechanism of energy dissipation and / or scavenging of reactive oxygen species (ROS) would contribute to survival in this harsh environment. In this paper, non-acclimated and cold-acclimated D. antarctica were subjected to high light and / or low temperature for 24 h. The contribution of non-photochemical dissipation of excitation light energy and the activities of detoxifying enzymes in the development of resistance to chilling induced photoinhibition were studied by monitoring PSII fluorescence, total soluble antioxidants, and pigments contents and measuring variations in activity of superoxide dismutase (SOD; EC 1.15.1.1), ascorbate peroxidase (APX; EC 1.11.1.11), and glutathione reductase (GR; EC 1.6.4.2). The photochemical efficiency of PSII, measured as Fv / F m, and the yield of PSII electron transport (ΦPSII) both decreased under high light and low temperatures. In contrast, photochemical quenching (qP) in both non-acclimated and cold-acclimated plants remained relatively constant (approximately 0.8) in high-light-treated plants. Unexpectedly, qP was lower (0.55) in cold-acclimated plants exposed to 4°C and low light intensity. Activity of SOD in cold-acclimated plants treated with high light at low temperature showed a sharp peak 2-4 h after the beginning of the experiment. In cold-acclimated plants APX remained high with all treatments. Activity of GR decreased in cold-acclimated plants. Compared with other plants, D. antarctica exhibited high levels of SOD and APX activity. Pigment analyses show that the xanthophyll cycle is operative in this plant. We propose that photochemical quenching and particularly the high level of antioxidants help D. antarctica to resist photoinhibitory conditions. The relatively high antioxidant capacity of D. antarctica may be a determinant for its survival in the harsh Antarctic environment.