Effects of heat stress in the leaf mitotic cell cycle and chromosomes of four wine-producing grapevine varieties

Grapevine varieties respond differentially to heat stress (HS). HS ultimately reduces the photosynthesis and respiratory performance. However, the HS effects in the leaf nuclei and mitotic cells of grapevine are barely known. This work intends to evaluate the HS effects in the leaf mitotic cell cycle and chromosomes of four wine-producing varieties: Touriga Franca (TF), Touriga Nacional (TN), Rabigato, and Viosinho. In vitro plants with 11 months were used in a stepwise acclimation and recovery (SAR) experimental setup comprising different phases: heat acclimation period (3 h—32 °C), extreme HS (1 h—42 °C), and two recovery periods (3 h—32 °C and 24 h—25 °C), and compared to control plants (maintained in vitro at 25 °C). At the end of each SAR phase, leaves were collected, fixed, and used for cell suspensions and chromosome preparations. Normal and abnormal interphase and mitotic cells were observed, scored, and statistically analyzed in all varieties and treatments (control and SAR phases). Different types of chromosomal anomalies in all mitotic phases, treatments, and varieties were found. In all varieties, the percentage of dividing cells with anomalies (%DCA) after extreme HS increased relative to control. TF and Viosinho were considered the most tolerant to HS. TF showed a gradual MI reduction from heat acclimation to HS and the lowest %DCA after HS and 24 h of recovery. Only Viosinho reached the control values after the long recovery period. Extrapolating these data to the field, we hypothesize that during consecutive hot summer days, the grapevine plants will not have time or capacity to recover from the mitotic anomalies caused by high temperatures.     

Pre-treating paclobutrazol enhanced chilling tolerance of sweetpotato

The objective of this work was to study changes in low molecular weight antioxidants and antioxidative enzymes in chilling-stressed sweetpotato, as affected by paclobutrazol (PBZ) pre-treatment 24 h prior to exposure to chilling conditions. Sweetpotato ‘TN71’ and ‘TN65’ were treated with 300 mg PBZ/5 ml/plant, after which plants were subjected to 7°C/7°C (day/night) for periods of 1, 3 and 5 days, followed by a 3-day recovery period at 24°C/20°C (day/night). A factorial experiment in completely randomized design with four replications was used in this study. Young fully expanded leaves at each temperature and period of time were clipped for antioxidative system measurement. We concluded that different varieties displayed variations in their oxidative system, and the differential expressions of each genotype were associated with chilling stress response. Plants with various antioxidative systems responded differently to chilling stress according to the duration of the chilling period and subsequent re-warming period. ASA, GSH and GSSG contents were enhanced in TN71 prior to chilling stress. Increased APX, GR, ASA and MDA activities accounted for chilling tolerance in TN65. Furthermore, our results indicate that the elevated levels of the antioxidative system observed after PBZ pre-treatments afforded the sweetpotato leaf improved chilling-stress tolerance. The levels of ASA and GSSG of both TN71 and TN65 under chilling were significantly raised by pre-treating with PBZ. PBZ pre-treatment exhibited the important function of enhancing the restoration of leaf oxidative damage under chilling stress and increasing the chilling tolerance of plants to mitigate chilling stress effects.     

Plastic and evolutionary responses to heat stress in a temperate dung fly: negative correlation between basal and induced heat tolerance?

Extreme weather events such as heat waves are becoming more frequent and intense. Populations can cope with elevated heat stress by evolving higher basal heat tolerance (evolutionary response) and/or stronger induced heat tolerance (plastic response). However, there is ongoing debate about whether basal and induced heat tolerance are negatively correlated and whether adaptive potential in heat tolerance is sufficient under ongoing climate warming. To evaluate the evolutionary potential of basal and induced heat tolerance, we performed experimental evolution on a temperate source population of the dung fly Sepsis punctum. Offspring of flies adapted to three thermal selection regimes (Hot, Cold and Reference) were subjected to acute heat stress after having been exposed to either a hot‐acclimation or non‐acclimation pretreatment. As different traits may respond differently to temperature stress, several physiological and life history traits were assessed. Condition dependence of the response was evaluated by exposing juveniles to different levels of developmental (food restriction/rearing density) stress. Heat knockdown times were highest, whereas acclimation effects were lowest in the Hot selection regime, indicating a negative association between basal and induced heat tolerance. However, survival, adult longevity, fecundity and fertility did not show such a pattern. Acclimation had positive effects in heat‐shocked flies, but in the absence of heat stress hot‐acclimated flies had reduced life spans relative to non‐acclimated ones, thereby revealing a potential cost of acclimation. Moreover, body size positively affected heat tolerance and unstressed individuals were less prone to heat stress than stressed flies, offering support for energetic costs associated with heat tolerance. Overall, our results indicate that heat tolerance of temperate insects can evolve under rising temperatures, but this response could be limited by a negative relationship between basal and induced thermotolerance, and may involve some but not other fitness‐related traits.     

Time-course for attainment and reversal of acclimation to constant temperature in two Ceratitis species

Acclimation in the thermal tolerance range of insects occurs when they are exposed to novel temperatures in the laboratory. In contrast to the large number of studies that have tested for the ability of insects to acclimate, relatively few have sought to determine the time-course for attainment and reversal of thermal acclimation. In this study the time required for the Mediterranean fruit fly, Ceratitis capitata Wiedemann, and the Natal fruit fly, Ceratitis rosa Karsch, to acclimate to a range of constant temperatures was tested by determining the chill-coma recovery time and heat knock-down time of flies that had been exposed to novel benign temperatures for different durations. The time required for reversal of acclimation for both Ceratitis species was also determined after flies had been returned to the control temperature. Acclimation to 31 °C for only one day significantly improved the heat knock-down time of C. capitata, but also led to slower recovery from chill-coma. Heat knock-down time indicated that acclimation was achieved after only one day in C. rosa, but it took three days for C. rosa to exhibit a significant acclimation response to a novel temperature of 33 °C when measured using chill-coma recovery time. Reversal of acclimation after return to initial temperature conditions was achieved after only one day in both C. capitata and C. rosa. Adult C. capitata held at 31.5 °C initially exhibited improved heat knock-down times but after 9 days the heat knock-down time of these flies had declined to levels not significantly different from that of control flies held at the baseline temperature of 24 °C. In both Ceratitis species, heat knock-down time declined with age whereas chill-coma recovery time increased with age, indicating an increased susceptibility to high and low temperatures, respectively.     

The glutathione system as a stress marker in plant ecophysiology: is a stress-response concept valid?

Environmental stress impacts cause an increased formation of reactive oxygen species (ROS) in the chloroplasts (photo-oxidative stress). The role of glutathione in the antioxidative defence system provides a rationale for its use as a stress marker. However, responses of glutathione concentrations and redox states are not consistent among the large number of available publications. In the present review the hypothesis that stress responses of the glutathione system follow a general ecophysiological stress-response concept is investigated. In this view, an initial response phase would be followed by an acclimation phase where a new steady-state is established. Alternatively, if successful acclimation is not achieved, degradation of the system will follow. Recent publications dealing with responses to photochilling, salinity, and drought are analysed as to whether the results fit the concept. In general, an initial stress response was related to changes in the glutathione redox state, whereas acclimation was marked by increased glutathione concentrations, increased related enzyme activities, and/or a more reduced redox state of glutathione. The latter was interpreted as overcompensation leading to enhanced regeneration of glutathione. Deterioration effects upon strong stress impacts were related to progressive degradation and oxidation of the glutathione pool. A time-course analysis, which has rarely been done in the published literature, showed this sequence of events. When apple trees were subjected to progressing drought, the initial response was a slight oxidation of the glutathione pool, followed by increased glutathione concentrations. When the stress increased, glutathione concentrations dropped and redox state became more oxidized, which marked the degradation of the system. In spite of the general congruency of these results with the suggested stress-response concept, several limitations have to be highlighted: The importance of the glutathione system relative to other components of the photoprotective and antioxidative defence system, as well as relative to stress avoidance strategies, has to be established. It is suggested that a variety of parameters taking into account alternative protection pathways (e.g. photorespiration, light dissipation) and other components of the antioxidative systems should be measured. Within such response patterns the glutathione system is a valuable stress marker in ecophysiological studies.     

Dissecting the proteome dynamics of the early heat stress response leading to plant survival or death in Arabidopsis

In many plant species, an exposure to a sublethal temperature triggers an adaptative response called acclimation. This response involves an extensive molecular reprogramming that allows the plant to further survive to an otherwise lethal increase of temperature. A related response is also launched under an abrupt and lethal heat stress that, in this case, is unable to successfully promote thermotolerance and therefore ends up in plant death. Although these molecular programmes are expected to have common players, the overlapping degree and the specific regulators of each process are currently unknown. We have carried out a high‐throughput comparative proteomics analysis during acclimation and during the early stages of the plant response to a severe heat stress that lead Arabidopsis seedlings either to survival or death. This analysis dissects these responses, unravels the common players and identifies the specific proteins associated with these different fates. Thermotolerance assays of mutants in genes with an uncharacterized role in heat stress demonstrate the relevance of this study to uncover both positive and negative heat regulators and pinpoint a pivotal role of JR1 and BAG6 in heat tolerance.     

Thermotolerance and antioxidant response induced by heat acclimation in <Emphasis Type="Italic">Freesia</Emphasis> seedlings

Followed a heat acclimation pretreatment, seedlings of Freesia hybrida ‘Shangnong Jinghuanghou’ were exposed to heat stress at 38°C for 6 h treatment and then recovered at 22°C for 72 h to study the impact of heat acclimation (30°C) on thermotolerance under heat stress. The results showed that the pretreated seedlings performed better under heat stress than control. Heat acclimation could slow down the decrease of chlorophyll contents under heat stress and recover better. Higher levels of soluble sugar and proline and slight lower level of soluble protein were observed in pretreated seedlings. After recovery, similar levels of proline and soluble protein were maintained in all seedlings. However, a higher level of soluble sugar was maintained in pretreated seedlings. MDA content and EL showed a stable level in pretreated seedlings while a significant increase in control, followed by a significant decrease after recovery. Significant different responses of SOD, POD, CAT, and APX activities were observed in pretreated seedlings and control. Heat acclimation led to higher activities of these enzymes and a significant response of antioxidant enzyme activities occurred in a time-dependent manner under heat stress. Exposure to high temperature caused a significant increase in SOD and APX activity, and much higher levels in SOD and APX activity were observed in pretreated seedlings compared to control during heat stress. A slight difference in change pattern of POD and CAT activity was presented. The highest activities of POD and CAT were observed at 4 and 6 h of heat stress in pretreated seedlings and control, respectively. After 72 h recovery, the activities of all tested enzymes decreased to similar levels in all seedlings.     

Photoprotective responses of Mediterranean and Atlantic trees to the extreme heat-wave of summer 2003 in Southwestern Europe

Summer 2003 was extremely hot in Europe. High light in combination with heat and drought exacerbates the generation of photo-oxidative stress. Under these conditions photoprotective responses can be critical for plant survival. Photoprotection was analysed in 2003 in several Mediterranean and Atlantic woody species. These data were compared with previous summers (1998, 1999 and 2001) to evaluate the potential acclimation for each species. A pattern of changes consisting on a decrease in chlorophyll, ascorbate and Fv/Fm and an increase in tocopherol, xanthophyll cycle pigments (VAZ) and de-epoxidation index was regularly observed. Acclimation potential was measured by the use of the plasticity index for each parameter. Mediterranean species were more plastic than Atlantic ones. The latter were unable to increase antioxidant pools to the same extent or to down-regulate the efficiency of light energy conversion. These results indicate that most Mediterranean species are able to perform an efficient acclimation to heat stress, whilst Atlantic species will be more affected by climate warming.     

Thermal acclimation in Arabidopsis lyrata: genotypic costs and transcriptional changes

Frost and heat events can be challenging for sessile organisms that cannot escape thermal extremes. However, adverse effects of thermal stress on fitness may be reduced by pre‐exposure to cold or heat, a process known as acclimation. To understand the ecological and evolutionary implications of acclimation, we investigated (1) the reduction in performance due to stress pre‐exposure, (2) the magnitude of increased leaf resistance to subsequent stress, (3) the costs of acclimation and (4) the genes differing in expression due to stress pre‐exposure. Plants of Arabidopsis lyrata were raised under three treatments of pre‐exposure: bouts of frost, bouts of heat or constant temperature. Resistance of leaves to subsequent frost and heat stress was then measured by electrolyte leakage. RNA‐seq analysis was performed to examine the genes differentially expressed between stress‐pre‐exposed and control plants. Pre‐exposure to stress during growth decreased plant size and increased leaf resistance to subsequent stress independent of whether pre‐exposure was to frost or heat. But the highest increase in leaf resistance to frost was found after pre‐exposure to frost (as a trend) and in leaf resistance to heat after pre‐exposure to heat. No evidence for costs of acclimation was detected. RNA‐sequencing suggested that acclimation by frost and heat pre‐exposure was caused by distinct mechanisms: modification of the chloroplast membrane and modification of the cell wall and membrane, respectively. Our results suggest that thermal resistance is a labile complex of traits, strongly affected by the previously experienced stress environment, with undetermined costs.     

Hydrogen peroxide pre-treatment induces salt-stress acclimation in maize plants

The effect of exogenously applied H2O2 on salt stress acclimation was studied with regard to plant growth, lipid peroxidation, and activity of antioxidative enzymes in leaves and roots of a salt-sensitive maize genotype. Pre-treatment by addition of 1 microM H2O2 to the hydroponic solution for 2 days induced an increase in salt tolerance during subsequent exposure to salt stress. This was evidenced by plant growth, lipid peroxidation and antioxidative enzymes measurements. In both leaves and roots the variations in lipid peroxidation and antioxidative enzymes (superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, glutathione reductase, and catalase) activities of both acclimated and unacclimated plants, suggest that differences in the antioxidative enzyme activities may, at least in part, explain the increased tolerance of acclimated plants to salt stress, and that H2O2 metabolism is involved as signal in the processes of maize salt acclimation.     

Small RNAs in regulating temperature stress response in plants

Due to global climate change, temperature stress has become one of the primary causes of crop losses worldwide. Much progress has been made in unraveling the complex stress response mechanisms in plants, particularly in the identification of temperature stress responsive protein‐coding genes. Recently discovered microRNAs (miRNAs) and endogenous small‐interfering RNAs (siRN As) have also been demonstrated as important players in plant temperature stress response. Using high‐throughput sequencing, many small RNAs, especially miRNAs, have been identified to be triggered by cold or heat. Subsequently, several studies have shown an important functional role for these small RNAs in cold or heat tolerance. These findings greatly broaden our understanding of endogenous small RNAs in plant stress response control. Here, we highlight new findings regarding the roles of miRNAs and siRNAs in plant temperature stress response and acclimation. We also review the current understanding of the regulatory mechanisms of small RNAs in temperature stress response, and explore the outlook for the use of these small RNAs in molecular breeding for improvement of temperature stress tolerance in plants.     

Acclimation to singlet oxygen stress in Chlamydomonas reinhardtii

In an aerobic environment, responding to oxidative cues is critical for physiological adaptation (acclimation) to changing environmental conditions. The unicellular alga Chlamydomonas reinhardtii was tested for the ability to acclimate to specific forms of oxidative stress. Acclimation was defined as the ability of a sublethal pretreatment with a reactive oxygen species to activate defense responses that subsequently enhance survival of that stress. C. reinhardtii exhibited a strong acclimation response to rose bengal, a photosensitizing dye that produces singlet oxygen. This acclimation was dependent upon photosensitization and occurred only when pretreatment was administered in the light. Shifting cells from low light to high light also enhanced resistance to singlet oxygen, suggesting an overlap in high-light and singlet oxygen response pathways. Microarray analysis of RNA levels indicated that a relatively small number of genes respond to sublethal levels of singlet oxygen. Constitutive overexpression of either of two such genes, a glutathione peroxidase gene and a glutathione S-transferase gene, was sufficient to enhance singlet oxygen resistance. Escherichia coli and Saccharomyces cerevisiae exhibit well-defined responses to reactive oxygen but did not acclimate to singlet oxygen, possibly reflecting the relative importance of singlet oxygen stress for photosynthetic organisms.     

Two Rubisco activase isoforms may play different roles in photosynthetic heat acclimation in the rice plant

Studies on some plant species have shown that increasing the growth temperature gradually or pretreating with high temperature can lead to obvious photosynthetic acclimation to high temperature. To test whether this acclimation arises from heat adaptation of ribulose 1,5‐bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) activation mediated by Rubisco activase (RCA), gene expression of RCA large isoform (RCA_L) and RCA small isoform (RCA_S) in rice was determined using a 4‐day heat stress treatment [40/30°C (day/night)] followed by a 3‐day recovery under control conditions [30/22°C (day/night)]. The heat stress significantly induced the expression of RCA_L as determined by both mRNA and protein levels. Correlative analysis indicated that RCA_S protein content was extremely significantly related to Rubisco initial activity and net photosynthetic rate (Pn) under both heat stress and normal conditions. Immunoblot analysis of the Rubisco–RCA complex revealed that the ratio of RCA_L to Rubisco increased markedly in heat‐acclimated rice leaves. Furthermore, transgenic rice plants expressing enhanced amounts of RCA_L exhibited higher thermotolerance in Pn and Rubisco initial activity and grew better at high temperature than wild‐type (WT) plants and transgenic rice plants expressing enhanced amounts of RCA_S. Under normal conditions, the transgenic rice plants expressing enhanced amounts of RCA_S showed higher Pn and produced more biomass than transgenic rice plants expressing enhanced amounts of RCA_L and wild‐type plants. Together, these suggest that the heat‐induced RCA_L may play an important role in photosynthetic acclimation to moderate heat stress in vivo, while RCA_S plays a major role in maintaining Rubisco initial activity under normal conditions.     

Heat acclimation: phenotypic plasticity and cues to the underlying molecular mechanisms

Acclimation, in contrast to evolutionary adaptation, is a “within life time phenotypic adaptation” resulting in a widening of the dynamic regulatory range of body temperature. Increased efficiency and capacity of the thermoregulatory effectors, and delayed onset of the temperature threshold for thermal injury, contribute to the beneficial effects of acclimation. Reprogrammed gene expression and changes in cellular signaling underlie these responses. Constitutive elevation of the inducible heat shock protein (HSP) 72 kDa provides cytoprotection and delays thermal injury without the need for de novo HSP synthesis upon thermal stress. The time window for evocation of heat acclimation is the early phase of acclimation, the short-term heat acclimation (STHA), with accelerated sympathetic excitability and a drop in plasma thyroxin playing an essential role. An important consequence of thermal acclimation is the development of cross-tolerance between heat acclimation and ischemia/reperfusion insults. The beneficial implications of this feature are discussed.     

Calcium chloride enhances the antioxidative system of sweet potato (Ipomoea batatas) under flooding stress

Three sweet potato varieties, Taoyuan 2, Simon 1 and Sushu 18, were pretreated with four levels of CaCl₂ (0, 60, 120 and 180 kg ha^?1) weekly for 50 days from planting before being subjected to non‐flooding (control) and flooding conditions. The experiment used a randomised complete block design with a split‐split plot arrangement of treatments. Young, fully expanded leaves from each plant were clipped for measuring enzyme activities and antioxidant contents. The three genotypes exhibited unique abilities and specificities through the antioxidative systems in response to flooding stress. The level of activity of the antioxidative system in sweet potato leaves was related to CaCl₂ pretreatment during flooding. The ascorbate peroxidase, superoxide dismutase, glutathione reductase, reduced ascorbate, total ascorbate, reduced glutathione and malondialdehyde contents of the three sweet potato varieties under flooding stress significantly increased because of pretreatment with 60 and 120 kg ha^?1 of CaCl₂. Moreover, pretreatment with 60 and 120 kg ha^?1 CaCl₂ enhanced the flooding tolerance of all three sweet potato varieties and mitigated the effects of flooding stress. However, pretreatment with 180 kg ha^?1 CaCl₂ markedly decreased some enzyme activities and antioxidant contents under a flooded condition. Calcium most likely played a role in the antioxidative system in the leaves of these three sweet potato varieties under flooding stress, as an optimum amount strengthened their oxidative systems.     

热胁迫对豌豆下胚轴生理的一些影响

通过测定热驯和热胁迫下3个豌豆品种幼苗下胚轴生长、细胞膜损伤、抗坏血酸（AsA）和丙二醛（佃A）含量的变化及热激蛋白70（HSP70）表达,探讨热胁迫对豌豆生理的影响。结果表明,在48℃高温胁迫下豌豆种子萌发率下降,幼苗下胚轴生长受抑制,细胞膜受损,AsA含量下降,MDA含量升高;经37℃热驯再48℃热激处理的下胚轴长度和ASA明显高于直接热胁迫的,细胞膜受损程度和MDA含量则低于后者。HSP70测定表明,除台湾品种外,37℃热驯1h不足以诱导HSP70表达;而37℃热驯后常温恢复再48℃热激和直接48℃热激均能诱导HSP70表达,其中蒙自品种经热驯后再热激的HSP70表达量高于直接热激的。     

Thermal response of juvenile Hawaiian mullet Mugil cephalus (L.) to acclimation time and fluctuating low temperatures*

Rates of acclimation and the response to fluctuating low temperatures of juvenile striped mullet Mugil cephalus (L.) have been described in terms of changes in heat resistance over a period of time. Fish changed from 25.5 to 27 or 29° C were acclimated within seven days. Acclimation to 23 or 15° C required a maximum of 11 days. Thermal responses to fluctuating low temperatures were lower than responses to constant temperature levels. Cyclic variations in heat resistance were present in all experimental and control tests.     

Rate of acclimation of the capacity for isoprene emission in response to light and temperature

Isoprene emission from plants accounts for nearly half of all non‐methane hydrocarbons entering the atmosphere. Light and temperature regulate the instantaneous rate of isoprene emission but there is increasing evidence that they also affect the capacity for isoprene emission (i.e. the rate measured under standard conditions). We tested the rate of acclimation of the capacity for isoprene emission following step changes in growth conditions. Acclimation to new growth temperatures was very rapid, with most of the change occurring within a few hours and complete adjustment occurring within a day. Acclimation to new light levels was more complicated. Following a switch from low‐light growth conditions to standard assay conditions (30 °C and 1000 µmol photons m^?2 s^?1), there was a rapid (5–10 min) and a slightly slower (10–50 min) acclimation of the capacity for isoprene emission. After accounting for these short‐term changes, there was also a small, long‐term (4–6 d) acclimation of the isoprene emission capacity to the light level of growth conditions. We found no effect of growth conditions on the coefficients used to describe the instantaneous light and temperature response of isoprene emission. Therefore, current models of isoprene emission will only need to be altered to account for changes in the capacity for isoprene emission.