Memory Processes in the Response of Plants to Environmental Signals

Plants are sensitive to stimuli from the environment (e.g., wind, rain, contact, pricking, wounding). They usually respond to such stimuli by metabolic or morphogenetic changes. Sometimes the information corresponding to a stimulus may be “stored” in the plant where it remains inactive until a second stimulus “recalls” this information and finally allows it to take effect. Two experimental systems have proved especially useful in unravelling the main features of these memory-like processes.In the system based on Bidens seedlings, an asymmetrical treatment (e.g., pricking, or gently rubbing one of the seedling cotyledons) causes the cotyledonary buds to grow asymmetrically after release of apical dominance by decapitation of the seedlings. This information may be stored within the seedlings, without taking effect, for at least two weeks; then the information may be recalled by subjecting the seedlings to a second, appropriate, treatment that permits transduction of the signal into the final response (differential growth of the buds). Whilst storage is an irreversible, all-or-nothing process, recall is sensitive to a number of factors, including the intensity of these factors, and can readily be enabled or disabled. In consequence, it is possible to recall the stored message several times successively.In the system based on flax seedlings, stimulation such as manipulation stimulus, drought, wind, cold shock and radiation from a GSM telephone or from a 105 GHz Gunn oscillator, has no apparent effect. If, however, the seedlings are subjected at the same time to transient calcium depletion, numerous epidermal meristems form in their hypocotyls. When the calcium depletion treatment is applied a few days after the mechanical treatment, the time taken for the meristems to appear is increased by a number of days exactly equal to that between the application of the mechanical treatment and the beginning of the calcium depletion treatment. This means that a meristem-production information corresponding to the stimulation treatment has been stored in the plants, without any apparent effect, until the calcium depletion treatment recalls this information to allow it to take effect. Gel electrophoresis has shown that a few protein spots are changed (pI shift, appearance or disappearance of a spot) as a consequence of the application of the treatments that store or recall a meristem-production signal in flax seedlings. A SIMS investigation has revealed that the pI shift of one of these spots is probably due to protein phosphorylation. Modifications of the proteome have also been observed in Arabidopsis seedlings subjected to stimuli such as cold shock or radiation from a GSM telephone.Key Words: memory, environmental signals, meristems, mobile telephone, bud growth, proteome, plants     

The role of calcium in the recall of stored morphogenetic information by plants

Flax seedlings grown in the absence of environmental stimuli, stresses and injuries do not form epidermal meristems in their hypocotyls. Such meristems do form when the stimuli are combined with a transient depletion of calcium. These stimuli include the "manipulation stimulus" resulting from transferring the seedlings from germination to growth conditions. If, after a stimulus, calcium depletion is delayed, meristem production is also delayed; in other words, the meristem-production instruction can be memorised. Memorisation includes both storage and recall of information. Here, we focus on information recall. We show that if the first transient calcium depletion is followed by a second transient depletion there is a new round of meristem production. We also show that if an excess of calcium follows calcium depletion, meristem production is blocked; but if the excess of calcium is in turn followed by another calcium depletion, again there is a new round of meristem production. The same stored information can thus be recalled repeatedly (at least twice). We describe a conceptual model that takes into account these findings.     

Plant sensitivity to low intensity 105 GHz electromagnetic radiation

Exposing seedlings of the flax, Linum usitatissimum L., to a variety of weak environmental stresses followed by a 2 day calcium deprivation, triggers the common response of production of epidermal meristems (actively dividing groups of cells) in the hypocotyl, which is the part of the stem between the root and the cotyledons (the pre-existing leaves in the embryo). This production reaches a plateau of 10-20 meristems after a month in the case of mechanical stimulation and cold shock. Recently, we have shown that radiation from a global system for mobile communication (GSM) telephone also triggers production of meristems with a plateau of around six meristems. Here, we show that a single 2 h exposure to radiation emitted at 105 GHz at non-thermal levels by a Gunn oscillator induces meristem production with kinetics similar to that induced by weak environmental stimuli and radiation from GSM telephone.     

Storage of environmental signals in flax. Their morphogenetic effect as enabled by a transient depletion of calcium

It is possible to induce the formation of epidermal meristems in the hypocotyl of non-injured and non-hormone-treated plantlets of flax, by combining various sorts of physical stimulations with a transient depletion of calcium. The characteristic times for the decrease of the tissue concentration of calcium during calcium depletion and for the recovery of the normal tissue concentration of calcium after resupplying the latter ion, are close to 1 day. The stimuli may correspond to wind or drought or even to the manipulation stress occurring when the plantlets are transferred from their germination to their growth vessel. Meristem production is increased by combining several physical stimulations. When calcium depletion is delayed relative to the application of the physical stimulation(s), the production of meristems is delayed accordingly. This means that the signal induced by the physical stimulation(s) may be stored within the plants, without apparent effect, until a calcium depletion finally allows the stored signal to take effect (formation of meristems). For storage periods of up to 8 days no loss of the potency of the stored signal was observed. A few other examples of storage of morphogenetic signals in plants have been described in the literature. The mechanism involved in signal storage is still not clearly understood. However, it seems that the sensing and/or storing of the signals require that the plant tissues are sufficiently rich in calcium, whereas the licensing of the plants for the translation of signals into the final response (meristem production) is done by a transient calcium depletion.     

Calcium ions are involved in the delay of plant cell cycle progression by abiotic stresses

Higher plants respond to environmental stresses by a sequence of reactions which include the reduction of growth by affecting cell division. It has been shown that calcium ions plays a role as a second messenger in mediating various defence responses under environmental stresses. In this study, the role of calcium ions on cell cycle progression under abiotic stresses has been examined in tobacco BY-2 suspension culture cells. Using synchronized BY-2 cells expressing the endogenous calcium sensor aequorin as experimental system, we could show that oxidative and hypoosmotic stress both induce an increase of intracellular calcium and cause a delay of the cell cycle. The inhibitory effect of these abiotic stress stimuli on cell cycle progression could be mimicked by increasing the intracellular calcium concentration via application of an external electrical field. Likewise, depletion of calcium ions in the culture medium suppressed the effect of the stimuli tested. These results demonstrate that calcium signalling is involved in the regulation of cell cycle progression in response to abiotic stress.     

Long-distance transport, storage and recall of morphogenetic information in plants. The existence of a sort of primitive plant 'memory'

An asymmetrical treatment of Bidens seedlings (pricking one of the seedling cotyledons) causes the cotyledonary buds to grow asymmetrically after release of apical dominance by decapitation of the seedlings. The symmetry-breaking signal propagates within the seedlings at a rate of at least a fraction of a millimetre per second. This information may be 'stored' (STO function) within the seedlings, without taking effect, for at least 2 weeks; then the information may be 'recalled' (RCL function), thus permitting transduction of the signal into the final response (differential growth of the buds), as a consequence of subjecting the seedlings to various symmetrical or asymmetrical treatments. A similar behaviour was observed with stimuli other than pricking (including non-traumatic stimuli), with plants other than Bidens (flax, tomato), and with responses other than cotyledonary-bud growth (hypocotyl elongation, induction of meristems, thigmomorphogenesis). There are indications that storage may involve the activation of elements implicated in cell cycle control, and that the last steps of the final response involve genes such as tch1 and hsp70. The adaptive advantage for plants in possessing STO/RCL functions is discussed. Manipulating the STO/RCL functions may have interesting practical applications, e.g. in the resistance of plants to natural stresses. The existence of the STO/RCL functions in plants constitutes an elementary form of 'memory' which may provide an experimental system simpler than the animal brain to test the validity of the theoretical models of interpretation of important features such as memory storage and evocation.     

Lectins of oil-seed flax plants exposed to abiotic stress

The seedlings of six cultivars of oil-seed flax (Linum humile Mill.) differing in the extent of adaptation to abiotic stresses (hypo- and hyperthermia, osmotic stress, and salinity) were used to assess hemag-glutination activity and carbohydrate specificity of total lectin preparations extracted from various cell compartments. In the course of adaptation of plants resistant to hyperthermia, osmotic stress and salinity, we observed a considerable rise in the coefficient of activity of membrane lectins, whereas the adaptation to hypothermia elevated the coefficient of activity of cell wall lectins. As to total soluble lectins, the adaptation of flax plants was associated with the changes in the range of their carbohydrate specificity. For instance, following the adaptation to hyperthermia, they were found to bind glucose and glucosamine, to osmotic stress—mannose and xylose, to salinity—galactose, glucose, and glucosamine; after cold resistance was developed, total soluble lectins were found to recognize lactose and fructose. It was concluded that lectins may participate in specific adaptation of flax plants to various abiotic stress factors.     

Barium-induced electro-mechanical uncoupling in myocardial cells

The heart of the adult moth Hyalophora cecropia requires extracellular calcium to maintain electrogenesis as well as tension development. In this study we ask whether the processes of autorhythmicity, driven electrogenesis and tension development require calcium specifically or whether the divalent cation Ba2+ can be substituted for calcium to support these activities. Ba2+ substituted for Ca2+ in equimolar amounts caused a marked (25 mV) hyperpolarization, suppression both of pacemaker activity and of tension development in spontaneously beating semi-isolated heart cells. Heart cells bathed in Ba2+ saline and paced by action potentials (produced by external stimuli) of greatly increased amplitude, prolonged phase 2 (plateau) and increased latency, and after 30 min, no mechanical activity was observed. These changes were completely reversible when calcium was reintroduced. We conclude that Ba2+ substitution for Ca2+ is an effective electromechanical uncoupler in moth heart cells. Although Ba2+ can support electrogenesis, it cannot replace 'trigger'-Ca2+ needed to release calcium from sarcoplasmic stores to effect tension development.     

The induction of soluble peroxidase activity in bean leaves by wind-induced mechanical perturbation

The induction of defense-related peroxidase (POD) activity in plants occurs in response to many biotic and abiotic stimuli. This controlled greenhouse study was an attempt to provide insight into the nature of the induction of soluble POD activity by noninjurious wind-induced mechanical perturbation (MP). In a time course study, exposure of common bean (Phaseolus vulgaris) seedlings to daily periods of fan-produced wind induced a significant and sustained increase in soluble POD activity in primary leaves of 7-9-d-old seedlings. In a wind-gradient study, wind-induced MP led to increases in soluble POD activity in leaves that were proportionally related to the wind speed experienced by individual seedlings. Wind-induced MP enhanced soluble POD activity to a degree similar to treatment with 5 mmol/L HgCl(2), a potent oxidizing elicitor of POD activity in plants. However, no further increases in POD activity were induced by HgCl(2) on plants that were preconditioned with wind-induced MP. Finally, short periods of brushing-induced MP enhanced soluble POD activity to the same degree as longer periods of wind-induced MS, suggesting a greater sensitivity to thigmic stimuli than to seismic stimuli in leaves of bean seedlings. This study illustrates the potential importance of wind and other mechanical stimuli as inducers of POD activity and interacting factors in the elicitation of POD activity by other environmental stimuli.     

Numbing the senses: role of TRPA1 in mechanical and cold sensation

In this issue of Neuron, Kwan et al. demonstrate that TRPA1 is critical for the transduction of noxious cold and mechanical stimuli, as well as in mediating the activation of nociceptors by endogenous and natural irritants. Differences between the present report and a previous study indicate that further study is needed to reach a consensus on the role of TRPA1 in the transduction of mechanical and noxious cold stimuli.     

非生物逆境胁迫下植物钙信号转导的分子机制

Ca2+作为植物细胞中最重要的第二信使, 参与植物对许多逆境信号的转导。在非生物逆境条件下, 植物细胞质内的钙离子在时间、空间及浓度上会出现特异性变化, 即诱发产生钙信号。钙信号再通过其下游的钙结合蛋白进行感受和转导, 进而在细胞内引起一系列的生物化学反应以适应或抵制各种逆境胁迫。目前在植物细胞中发现Ca2+/CDPK、Ca2+/CaM和Ca2+/CBL 3类钙信号系统, 研究表明它们与非生物逆境胁迫信号转导密切相关。本文通过从植物在非生物逆境条件下钙信号的感受、转导到产生适应性和抗性等方面, 介绍钙信号转导分子机制的一些研究进展。     

Cell-type-specific calcium responses to drought, salt and cold in the Arabidopsis root

Little is known about the signalling processes involved in the response of roots to abiotic stresses. The Arabidopsis root is a model system of root anatomy with a simple architecture and is amenable to genetic manipulation. Although it is known that the root responds to cold, drought and salt stress with increases in cytoplasmic free calcium, there is currently no information about the role(s) of the functionally diverse cell types that comprise the root. Transgenic Arabidopsis with enhancer-trapped GAL4 expression in specific cell types was used to target the calcium reporting protein, aequorin, fused to a modified yellow fluorescent protein (YFP). The luminescence output of targeted aequorin enabled in vivo measurement of changes in cytosolic free calcium concentrations ([Ca2+]cyt) in specific cell types during acute cold, osmotic and salt stresses. In response to an acute cold stress, all cell types tested as well as plants constitutively expressing aequorin displayed rapid [Ca2+]cyt peaks. However, there were significant quantitative differences between different cell types in terms of their response to cold stress, osmotic stress (440 mM mannitol) and salt stress (220 mM NaCl), implying specific roles for certain cell types in the detection and/or response to these stimuli. In response to osmotic and salt stress, the endodermis and pericycle displayed prolonged oscillations in cytosolic calcium that were distinct from the responses of the other cell types tested. Targeted expression of aequorin circumvented the technical difficulties involved in fluorescent dye injection as well as the lack of cell specificity of constitutively expressed aequorin, and revealed a new level of complexity in root calcium signalling.     

冷激诱导高温胁迫下番茄幼苗矮化机理

为了探明冷激诱导高温胁迫下番茄幼苗矮化的机理,育苗期间,每天8:00对幼苗分别进行5、10、15 ℃持续时间依次为10、20、30 min的冷激处理,测试了不同冷激强度下番茄幼苗乙烯释放速率,研究了冷激处理T10 ℃ D10 min(10 ℃持续10 min)结合不同生长调节物质对番茄幼苗乙烯释放速率、赤霉素(GA₃)含量和幼苗生长特性的影响.结果表明： 冷激处理刺激了番茄幼苗乙烯的产生,随着冷激温度的降低和冷激时间的延长,冷激诱导乙烯释放的效应显著增强.5 ℃持续30 min的冷激处理番茄幼苗乙烯产生速率最大,达到60.3 nL·h^-1·g^-1,为对照的6.5倍;乙烯利(ETH)、硫代硫酸银(STS)、GA₃和多效唑(PP333)均不能完全阻止冷激处理T10 ℃D10 min诱发的高乙烯产生率.冷激处理T10 ℃D10 min番茄幼苗茎叶GA₃含量为80.8 μg·g^-1,与对照(130.6 μg·g^-1)相比降低了38.1%.喷施ETH、STS对冷激诱发的幼苗矮化效应无显著影响,而GA₃显著减弱了冷激的矮化效应,PP333显著增强了冷激的矮化效应.以株高作为衡量指标,浓度为4.0 mg·L^-1的PP333处理,相当于10 ℃冷激处理.冷激诱导的番茄幼苗矮化效应主要原因在于冷激降低了番茄幼苗茎叶GA₃的含量.T10 ℃ D10 min可以在降低幼苗株高的同时不降低幼苗干物质的积累.     

Abiotic stresses activate a MAPkinase in the model grass species Lolium temulentum

Forage and turf grasses are utilized in diverse environments that expose them to a variety of abiotic stresses, however very little is known concerning the perception or molecular responses to these various stresses. In the model grass species Lolium temulentum, a 46kDa mitogen-activated protein kinase (MAPK) was activated in the leaf within 10min of exposing the roots to salt stress. When plants were subjected cold stress, no significant activation of the MAPK was observed. However, the 46kDa MAPK was rapidly activated in the leaves of plants within 3min of exposure to heat stress. Previously, mechanical wounding has been shown to rapidly activate a 46kDa and a 44kDa MAPK in L. temulentum. The wound activation of the MAPKs was delayed and diminished in plants undergoing cold treatment. In plants subjected simultaneously to 40°C and wounding, the activation of the 46kDa MAPK was enhanced. However if plants were subjected to heat and cold stress for more than 2h or exposed to 300mM NaCl for 24h prior to wounding, the wound activation of the 46kDa and a 44kDa MAPKs were significantly inhibited. These results suggest that the 46kDa MAPK plays a role in the response to various environmental stimuli.     

Somatic Chromosome Counts from Leaf Meristems in the Tribe Triticeae

The Feulgen procedure was applied to chromosome preparations of leaf meristems from growing plants of wheat, barley, and wheat-wheatgrass hybrids. Leaf primordia from the base of secondary tillers were pretreated in cold water (overnight, 2 C), fixed in glacial acetic acid (20 min, 2 C), hydrolyzed in 1 N HCl (14 min), stained in leuco-basic fuchsin (about 15 min) and squashed in 1% acetocarmine. The chromosome spreads from leaf meristems were generally superior to those of the root meristems of the same plant. Mitotic index in leaf meristems was higher than in root meristems in some species. The method appears useful for counting the chromosome number of growing plants, detecting chimeras, and verifying root tip chromosome counts.     

Water Deficit in Pea Root Tips: Effects on the Cell Cycle and on the Production of Dehydrin-like Proteins

Dehydrin-like proteins have been detected in nuclei and cytoplasmof meristematic root tip cells from pea seedlings subjectedto slow dehydration at 90% relative humidity for 48 h or more.Evidence was gained from Western blotting and immunocytochemicalexperiments using an antibody raised against the conserved domainof dehydrin proteins. Flow cytometer analysis has shown thatcycling cells of root tip meristems from dehydrated seedlingsare mostly arrested in G2 phase. Other stress treatments thoughtto involve water depletion (osmotic stress, cold treatment)or to modulate cell response to water deficit (abscisic acid)gave less clear-cut results with all treatments lowering theproportion of cells entering the S phase, but without a definiteand persistent arrest in any preferential phase of the cycle.Possible interrelationships between G2 arrest and dehydrin productionare discussed. Cell cycle; dehydrins; flow cytometry; nuclei; pea; Pisum sativum L.; water stress     

沙冬青CBL1蛋白纯化及性质研究

CBL是近年来发现的一类钙信号转导蛋白,CBL-CIPK组成的信号通路在植物应答生物和非生物刺激中发挥重要作用。其中CBL1和相应的CIPK在低钾,渗透压,干旱,机械损伤,及冻伤等环境胁迫中发挥重要作用。通过对沙冬青CBL1表面带电氨基酸定点突变,表面赖氨酸甲基化后电荷消除证明了沙冬青CBL1(AmCBL1)在钙离子存在下的非特异性聚集是由于分子间的电荷相互作用引起,三体蛋白很可能是沙冬青CBL1蛋白发挥功能的单位。通过甲基化可以得到聚合状态均一的蛋白,为CBL1晶体生长奠定了基础。