Concepts in plant stress physiology. Application to plant tissue cultures

Because the term stress is used, most often subjectively, with variousmeanings, this paper first attempts to clarify the physiological definition,andthe appropriate terms as responses in different situations. The flexibility ofnormal metabolism allows the development of responses to environmental changeswhich fluctuate regularly and predictably over daily and seasonal cycles. Thusevery deviation of a factor from its optimum does not necessarily result instress. Stress begins with a constraint or with highly unpredictablefluctuations imposed on regular metabolic patterns that cause bodily injury,disease, or aberrant physiology. Stress is the altered physiological conditioncaused by factors that tend to alter an equilibrium. Strain is any physicaland/or chemical change produced by a stress, i.e. every established condition,which forces a system away from its thermodynamic optimal state. The papersecondly summarises the Strasser's state-change concept which is preciselythat suboptimality is the driving force for acclimation (genotype level) oradaptation (population level) to stress. The paper continues with the actualknowledge on the mechanisms of stress recognition and cell signalling. Briefly:plasma membranes are the sensors of environmental changes; phytohormones andsecond messengers are the transducers of information from membranes tometabolism; carbon balance is the master integrator of plant response; betwixtand between, some genes are expressed more strongly, whereas others arerepressed. Reactive oxygen species play key roles in up- and down-regulation ofmetabolism and structure. The paper shows finally that the above concepts canbeapplied to plant tissue cultures where the accumulating physiological andgenetical deviations (from a normal plant behaviour) are related to thestressing conditions of the in vitro culture media and ofthe confined environment. The hyperhydrated state of shoots and the cancerousstate of cells, both induced under conditions of stress in invitro cultures, are identified and detailed, because they perfectlyillustrate the stress-induced state-change concept. It is concluded that stressresponses include either pathologies or adaptive advantages. Stress may thuscontain both destructive and constructive elements : it is a selection factoraswell as a driving force for improved resistance and adaptive evolution.     

CIDEP observations in photosystem I of green plant and algal photosynthesis.

Flash photolysis experiments with electron paramagnetic resonance detection were carried out between 10 K and 300 K on samples of green plant and algal species. Chemically induced dynamic electron polarization was evident for the signals observed in the g = 2.0 region for 100 KHz modulated detection and also for a system with no magnetic field modulation. The light reversible signals decaying in about 1 ms at low temperatures are interpreted as arising from photosystem I of the green plant and algal samples. Evidence is presented which indicates that the origin of the electron spin polarization is the well established radical-pair mechanism.     

Inclusion of biotic stress (consumer pressure) alters predictions from the stress gradient hypothesis

1.  The stress gradient hypothesis (SGH) predicts a shift from net negative interactions in benign environments towards net positive in harsh environments in ecological communities. While several studies found support for the SGH, others found evidence against it, leading to a debate on how nature and strength of species interactions change along stress gradients, and to calls for new empirical and theoretical work.
2.  In the latest attempt in this journal, it is successfully argued how the SGH should be expanded by considering different life strategies of species (stress tolerance versus competitive ability) and characteristics of abiotic stress (resource versus non-resource based) over wider stress gradients (opposed to low–high contrasts), but the crucial role of biotic stress by consumers is largely ignored in this refinement.
3.  We point out that consumers strongly alter the outcome of species interactions in benign and harsh environments, and show how inclusion of consumer-incurred biotic stress alters the predicted outcome of interactions along resource- and non-resource-based stress gradients for stress-tolerant and competitive benefactors and beneficiaries.
4.   Synthesis. New studies should include stress gradients consisting of both abiotic and biotic components to disentangle their impacts, and to improve our understanding of how species interactions change along environmental gradients.     

Chlorophyll-a fluorescence evaluation of PEG-induced osmotic stress on PSII activity in Arabidopsis plants expressing SIP1

In this study, we evaluated the effect of osmotic stress on photosynthetic machinery of Arabidopsis plants expressing a gene encoding small basic intrinsic protein (SIP1) isolated from Solanum tuberosum. Intact leaves of SIP Arabidopsis plants were exposed to 15% polyethylene glycol (PEG) solution and fast Chlorophyll-a (Chl-a) fluorescence induction kinetics was measured. Photosynthetic parameters like ratio of variable and maximum fluorescence (F_V/F_M), absorbance of photons per active reaction center (ABS/RC), trapping of photons per active reaction center (TRo/RC), electron transport per active reaction center (ETo/RC), and performance index (PI) were measured. Furthermore, the energy pipeline model was deduced in response to PEG stress. The membrane model includes a visualization of the average “antenna size”, which follows the value of the ABS/RC. Analysis of SIP Arabidopsis plants under PEG stress through fast Chl-a fluorescence transient showed that the damage caused due to PEG is more prominent at the donor side rather than the acceptor side of PSII. Higher PI in SIP plants under PEG stress indicated a better vitality than control plants. Overall, these results indicate that constitutive expression of SIP1 in Arabidopsis plants induces significant changes in the photosynthetic machinery under PEG-induced osmotic stress.     

A new type of thermoluminometer: a highly sensitive tool in applied photosynthesis research and plant stress physiology

Here we describe a newly developed thermoluminescence measuring device that employs flash excitation, peltier heating, and light detection by channel photomultipliers (CPM). The new thermoluminometer is equipped with four sample holders for simultaneous measurements of thermoinduced light emission in the temperature range from -20 degrees C to +180 degrees C. It allows one to measure leaf samples, chloroplasts, thylakoids, algae, or even bioorganic material lacking chlorophyll by means of naturally induced or artificially applied chemilumigenic probes. The temperature range of the thermoluminometer allows one to analyse the thermoinduced radical pair recombination of photosystem II in the lower temperature region as well as chemiluminescence from lipid peroxidation in the higher temperature region. Hence, plant material can be assessed concerning both its photosynthetic and its oxidative stress status. Since the device is equipped with four sample holders and four CPM channels for simultaneous detection of thermoinduced light emission, it facilitates a high throughput. Therefore, the new device is interesting, not only in ecophysiology, but also in the field of plant breeding, as it can be used to study the stress tolerance of various cultivars of cultural crop plants.     

Role of fructose in the adaptation of plants to cold-induced oxidative stress

This work presents findings, which indicate important role of fructose, fructose 6-phosphate (F6P), and fructose 1,6-bisphosphate (FBP) in preservation of homeostasis in plants under low temperature. Cold combined with light is known to incite increased generation of superoxide in chloroplasts leading to photoinhibition, but also an increased level of soluble sugars. In the present study, oxidative stress in pea leaves provoked by cold/light regime was asserted by the observed decrease of the level of oxidized form of PSI pigment P700 (P700(+)). Alongside, the increased antioxidative status and the accumulation of fructose were observed. The antioxidative properties of fructose and its phosphorylated forms were evaluated to appraise their potential protective role in plants exposed to chilling stress. Fructose, and particularly F6P and FBP exhibited high capacities for scavenging superoxide and showed to be involved in antioxidative protection in pea leaves. These results combined with previously established links implicate that the increase in level of fructose sugars through various pathways intercalated into physiological mechanisms of homeostasis represents important non-enzymatic antioxidative defense in plants under cold-related stress.     

Role of light and photosynthesis on the acclimation process of the cyanobacteriumSpirulina platensis to salinity stress

The response ofSpirulina platensis cells to salinity stress was studied. Once adapted to the higher osmoticum, photosynthetic parameters such as the maximum rate of photosynthesis under saturating irradiance (P_max) and the initial slope of the P-I curve () are reduced by 15% and 25% in 0.5 M NaCl grown cells, respectively. Salt-adapted cells have a modified biochemical composition; reduced protein and chlorophyll content, and an increased level of carbohydrates. The reduction in the photosynthetic capacity of the salt-adaptedSpirulina cells reflects a lower ability to utilize light energy and results in an increase in the susceptibility of the stressed cells to photoinhibition. This conclusion is supported by the finding that cultures exposed to salt stress show not only a decrease in growth rate (), but lose the ability to respond to increased irradiance with an increase in growth. The use of variable fluorescence as a fast and reliable measurement to follow the changes in PSII of salt-stressesSpirulina cells enables following the early events of salinity shock. It indicates that as soon as the cells are exposed to salt, a protection mechanism is induced. This mechanism does not require any protein synthesis and may take place even in the dark, though at somewhat reduced effectiveness. The significance of the result in providing a better understanding of the interaction between two environmental stresses — light and salinity — and their application in the outdoor mass cultivation ofSpirulina are discussed.Author for correspondence     

Incorporation of [35S]methionine in higher plants reveals that stimulation of the D1 reaction centre II protein turnover accompanies tolerance to heavy metal stress

The effects of cadmium stress (CdCl₂) on photochemical activity and protein behaviour of photosystem II (PSII) were studied in vivo and in vitro . Treatments of pea ( Pisum sativum ) and broad bean ( Vicia faba ) plants with 0·05–5 m M cadmium (CdCl₂) modified PSII activity with a resulting increase in electron transfer followed by an inhibition and damage to the oxygen-evolving complex. Pulse-chase experiments with [³⁵S]methionine in vivo followed by the separation of the radiolabelled thylakoids into grana and stroma exposed regions indicated that the synthesis, degradation and assembly of the D1 protein were greatly affected by cadmium. Initially D1 synthesis increased, later slowing down when the stress became advanced; at the same time the D1 degradation was increased. Binding studies with radiolabelled [¹⁴C]herbicide revealed that the Q_B pocket activity was also altered. However, the primary consequence of cadmium stress was the disassembly of the stacked regions. The measurements indicated differential tolerance to cadmium stress between the two plant species, which was not caused by either differential metal uptake or binding to the PSII complex. This suggests that the resulting changes in D1 turnover are a consequence of an unknown primary effect of cadmium on the PSII apparatus. However, we show that the higher tolerance to heavy metal stress found in broad bean plants relative to pea is accompanied by stimulation of D1 turnover. These experiments supported by previous data suggest that modulation of D1 turnover under stress is a commonly occurring process.     

Sensitivity of photosynthesis by spinach chloroplast membranes to osmotic stress in vitro: Rapid inhibition of O2 evolution in presence of magnesium

Thylakoids prepared from spinach (Spinacea oleracea L.) chloroplasts were exposed to osmotic stress in vitro in the presence or absence of different inorganic salts. By an hour after incubation in 1.0 M sorbitol and 10 mM (or more) MgCl₂, the thylakoids lost approximately 80% of their photosystem (PS) II activity, but not PS I. The inhibition occurred only in presence of magnesium as indicated by the combinations of several cations/anions. The PS II activity was relatively insensitive to osmotic stress in the presence of diphenyl carbazide. We therefore conclude that under conditions of water stress in the presence of 10 mM or higher Mg²⁺, the oxygen evolving system in chloroplasts is rapidly inactivated.Abbreviations DCMU 3(3,4-dichlorophenyl)-1,1-dimethylurea - DCPIP 2,6-dichlorophenol indophenol - DPC diphenyl carbazide - MV methyl viologen - PS photosystem Part of this work was included in the thesis submitted by the first author of M.Phil.degree.     

Escape of a plant virus from amplicon-mediated RNA silencing is associated with biotic or abiotic stress

Strong RNA silencing was induced in plants transformed with an amplicon consisting of full-length cDNA of potato leafroll virus (PLRV) expressing green fluorescent protein (GFP), as shown by low levels of PLRV-GFP accumulation, lack of symptoms and accumulation of amplicon-specific short interfering RNAs (siRNAs). Inoculation of these plants with various viruses known to encode silencing suppressor proteins induced a striking synergistic effect leading to the enhanced accumulation of PLRV-GFP, suggesting that it had escaped from silencing. However, PLRV-GFP escape also occurred following inoculation with viruses that do not encode known silencing suppressors and treatment of silenced plants with biotic or abiotic stress agents. We propose that viruses can evade host RNA-silencing defences by a previously unrecognized mechanism that may be associated with a host response to some types of abiotic stress such as heat shock.     

Ants on plants: a meta-analysis of the role of ants as plant biotic defenses

We reviewed the evidence on the role of ants as plant biotic defenses, by conducting meta-analyses for the effects of experimental removal of ants on plant herbivory and fitness with data pooled from 81 studies. Effects reviewed were plant herbivory, herbivore abundance, hemipteran abundance, predator abundance, plant biomass and reproduction in studies where ants were experimentally removed (n = 273 independent comparisons). Ant removal exhibited strong effects on herbivory rates, as plants without ants suffered almost twice as much damage and exhibited 50% more herbivores than plants with ants. Ants also influenced several parameters of plant fitness, as plants without ants suffered a reduction in biomass (−23.7%), leaf production (−51.8%), and reproduction (−24.3%). Effects were much stronger in tropical regions compared to temperate ones. Tropical plants suffered almost threefold higher herbivore damage than plants from temperate regions and exhibited three times more herbivores. Ant removal in tropical plants resulted in a decrease in plant fitness of about 59%, whereas in temperate plants this reduction was not statistically significant. Ant removal effects were also more important in obligate ant–plants (=myrmecophytes) compared to plants exhibiting facultative relationships with hemiptera or those plants with extrafloral nectaries and food bodies. When only tropical plants were considered and the strength of the association between ants and plants taken into account, plants with obligate association with ants exhibited almost four times higher herbivory compared to plants with facultative associations with ants, but similar reductions in plant reproduction. The removal of a single ant species increased plant herbivory by almost three times compared to the removal of several ant species. Altogether, these results suggest that ants do act as plant biotic defenses, but the effects of their presence are more pronounced in tropical systems, especially in myrmecophytic plants. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. N. P. de U. Barbosa, L. Diniz, Y. Oki and F. Pezzini contributed equally to this work and are listed in alphabetical order.     

短时间热胁迫对疏叶骆驼刺光系统II、Rubisco活性和活性氧化剂的影响

生长在温带沙漠地区的植物在夏季时常遭受正午短时间的高温胁迫, 频繁和骤然的热胁迫在很大程度上限制了荒漠植物的光合作用。以塔克拉玛干沙漠南缘防风固沙的优势植物疏叶骆驼刺(Alhagi sparsifolia)为材料, 分别用叶绿素荧光诱导动力学和CO₂响应方法分析热胁迫后光系统II (PSII)和RuBP羧化酶的热稳定性。结果表明: (1)在叶片温度超过43 ℃后PSII最大光化学量子产量、有活性反应中心数目、活力指数均出现明显的降低; 中高温度下PSII的电子供体侧比电子受体侧组分更容易受到热胁迫的伤害; 在58 ℃出现明显的K点(300 μs), 说明放氧复合体放氧结构受到破坏而失去活性。(2)随着叶片温度的上升, Rubisco活性先升高后降低, 在34 ℃时具有最高的活性水平。(3)叶片受到高温胁迫时, 细胞内氨态氮和活性氧分子等大量积累。(4)疏叶骆驼刺叶片处于短时间的高温环境时, 光合作用的光反应和暗反应阶段均表现出功能的不稳定性, 其中PSII和Rubisco是主要的热敏感位点。     

Molecular insights into the transgenerational inheritance of stress memory

There is accumulating evidence to show that environmental stressors can regulate a variety of phenotypes in descendants through germline-mediated epigenetic inheritance. Studies of model organisms exposed to environmental cues (e.g., diet, heat stress, toxins) indicate that altered DNA methylations, histone modifications, or non-coding RNAs in the germ cells are responsible for the transgenerational effects. In addition, it has also become evident that maternal provision could provide a mechanism for the transgenerational inheritance of stress adaptations that result from ancestral environmental cues. However, how the signal of environmentally-induced stress response transmits from the soma to the germline, which may influence offspring fitness, remains largely elusive. Small RNAs could serve as signaling molecules that transmit between tissues and even across generations. Furthermore, a recent study revealed that neuronal mitochondrial perturbations induce a transgenerational induction of the mitochondrial unfolded protein response mediated by a Wnt-dependent increase in mitochondrial DNA levels. Here, we review recent work on the molecular mechanism by which parental experience can affect future generations and the importance of soma-to-germline signaling for transgenerational inheritance.     

Structural changes of the thylakoid membrane network induced by high light stress in plant chloroplasts

Land plants live in a challenging environment dominated by unpredictable changes. A particular problem is fluctuation in sunlight intensity that can cause irreversible damage of components of the photosynthetic apparatus in thylakoid membranes under high light conditions. Although a battery of photoprotective mechanisms minimize damage, photoinhibition of the photosystem II (PSII) complex occurs. Plants have evolved a multi-step PSII repair cycle that allows efficient recovery from photooxidative PSII damage. An important feature of the repair cycle is its subcompartmentalization to stacked grana thylakoids and unstacked thylakoid regions. Thus, understanding the crosstalk between stacked and unstacked thylakoid membranes is essential to understand the PSII repair cycle. This review summarizes recent progress in our understanding of high-light-induced structural changes of the thylakoid membrane system and correlates these changes to the efficiency of the PSII repair cycle. The role of reversible protein phosphorylation for structural alterations is discussed. It turns out that dynamic changes in thylakoid membrane architecture triggered by high light exposure are central for efficient repair of PSII.     

Anthocyanin accumulation in the illuminated surface of maize leaves enhances protection from photo-inhibitory risks at low temperature, without further limitation to photosynthesis

At suboptimal temperatures, anthocyanins accumulate in the illuminated leaf surface of some maize genotypes and, if the anthocyanins shade chloroplasts, they can effectively reduce the risk of photo‐inhibition but also photo‐synthesis. To investigate this phenomenon, gas exchange, fluorescence, superoxide dismutase activity and xantho‐phyll composition of anthocyanin‐containing HOPI and anthocyanin‐deficient W22 maize genotypes were measured in either white or red light, where the latter is not absorbed by anthocyanins. Despite differences in light absorption in chloroplasts, photosynthesis did not differ between HOPI and W22 under either light source, suggesting that neither CO₂ supply nor photochemistry were more limiting in red leaves than in green leaves. In fact, no major differences in transpiration were detected. The ΔF/F_m (photosystem II quantum yield) of HOPI in white light was higher than in red light and higher than ΔF/F_m of W22 with either light source. This probably compensated for the lower white light absorption of HOPI chloroplasts compared with W22 because of the presence of anthocyanins and led to similar rates of calculated electron transport for both genotypes. After exposure to high white light at 5 °C, xanthophyll de‐epoxidation and superoxide dismutase activity were lower in HOPI than in W22. Further, HOPI could be exposed to a much higher irradiance than W22 before F_v/F_m was reduced to that of W22.     

The effect of leaffolder Cnaphalocrocis medinalis (Guenee) [Lepidoptera: Pyralidae] injury on the plant physiology and yield loss in rice

Influence of leaffolder feeding on chlorophyll, PS II activity and plant–water relations, effect of larval density on leaf damage and time course studies on larval feeding behaviour on altered physiological changes in TN1 rice culture were studied. Quantification of yield losses in the field caused by leaffolder was also assessed. Leaffolder damage resulted in 57% reduction in chlorophyll content, 23% reduction in PS II activity and 23% reduction in relative water content in comparison with control. Rice leaffolder larva folds the leaf and scrapes the green tissue from within the fold resulting in scorching and drying of the leaves. Larval density had differentially influenced effective leaf area of rice crop. Larval densities of more than 3 larvae per hill at maximum tillering stage resulted up to 20% unfilled grains, 28–57% reduction in PS II activity and 23% reduction in relative water content in comparison with the control. At flowering stage, flag leaf area damage of above 25% resulted in more than 50% unfilled grains over control, indicating direct effect of yield reduction in rice. Thus, a cumulative effect of loss in chlorophyll, reduced photosynthate availability and altered water relations caused by the leaffolder injury to flag leaf lead to greater yield loss in rice.     

Water stress in Eucalyptus pauciflora: comparison of effects on stomatal conductance with effects on the mesophyll capacity for photosynthesis,and investigation of a possible involvement of photoinhibition

Seedlings of Eucalyptus pauciflora Sieb. ex Spreng., grown in 4-1 pots, were stressed by withholding water while relationships between net assimilation rate (A) and intercellular partial pressure of CO₂ (p_i) in selected leaves were obtained repeatedly throughout the stress cycle. Water stress at first caused stomatal closure without any decline in the A(p_i) relationship. As stress became more severe, the A(p_i) relationship was affected as well. This always affected assimilation rate at both high and low intercellular partial pressures of CO₂. It was then tested whether water-stressed leaves were more prone to photoinhibition than unstressed ones. Plants were water-stressed while at the same time subjected to strong photon flux area density (2000 mol quanta·m^-2·s^-1). A possible light-induced inhibition was assessed by comparing quantum yields of photosynthesis with light directed onto one or the other surface of the leaf. A decline in quantum yield was observed, and the decline on the previously irradiated side was more pronounced than on the previously shaded side, but the effect was small and disappeared entirely within 1 d of rewatering the plants. It is concluded that photoinhibition can play a role, but not an important one, in the effect of water stress on the A(p_i) relationship in leaves of E. pauciflora.Abbreviations and symbols RuBP ribulose-1,5-bisphosphate - A net assimilation rate - p_i intercellular partial pressure of CO₂ - quantum yield of photosynthesis (net assimilation or RuBP-regeneration rate) - w difference in water content between air saturated at leaf temperature and the actual vapor content of the air, expressed as mole fraction