1210 T.-C. Xiong et al. / Biochimica et Biophysica Acta 1763 (2006) 1209–1215in metabolism, growth, development and/or adaptation. Under- NADP from NAD+ for photosynthetic reduction to NADPH, isstanding how a simple and ubiquitous nutrient like calcium is activated by Ca2+ . Interestingly, illumination of isolatedimplicated in a plethora of biological processes is an active field chloroplasts results in the uptake of Ca2+ from the incubationof research in eukaryotic cells and organisms including plants medium and leads to the decrease in the NAD+ levels whereas[1–6]. Due to its pleiotropic effects, a possibility is that calcium inhibition of the light-induced Ca2+ uptake by ruthenium redacts as a general switch responding indistinctly to a large array reduces the light-dependent decrease in NAD+ levels in theof stimuli. As a consequence, the specificity of the biological stroma .outcome would depend strictly upon a complex network of The mechanism by which Ca2+ enters chloroplasts duringdownstream signaling and amplification systems . Indeed, illumination was a matter of controversy. A first line of evidenceexamples suggesting that Ca2+ is a simple switch have been suggests that a H+/Ca2+-antiport process fueled by ATP  isdescribed and critically discussed. However, not all calcium responsible of Ca2+ uptake whereas other experimental datasignals act as a simple binary on–off switch in plants . claim that the driving force is most probably due to changes inRather, as established in animal models , changes in free membrane potentials . Recent data have established thatcalcium do not proceed in a stereotypical manner in plant cells both processes exist in the chloroplast and that they arebut according to a signature depending upon the characteristics specifically located on distinct plastidic compartments.of the stimulus including its intensity, duration, timing and Thus, the initial rate of Ca2+ uptake across inner-envelopesubcellular localization [5,8,9]. The calcium signature triggers vesicles of pea chloroplasts is greater in right-side-out than indownstream events in an ordered manner and manipulating a outside-out vesicles . In right-side out vesicles, the uptake isparticular signature will modify the biological outputs of an stimulated by a pH gradient (high pH inside) or by a potassiuminitial stimulus. diffusion gradient (inside negative) in the absence of a pH Authoritative reviews on calcium signaling in plants are gradient. In either condition, addition of valinomycin in thepublished and updated periodically with the refining of the presence of K+, which dissipates the membrane potentialconceptual frame and the methodological approaches [3–5,10]. gradient (Δψ) but leaves the magnitude of the pH gradientMost of these reviews are concentrated on the regulation of unchanged, inhibits calcium uptake. These data suggest that thechanges in cytosolic calcium [Ca2+]cyt and their effects on the transport of Ca2+ is unidirectional via a potential-stimulatedfunctioning of plant cells. Indeed, the two biggest organelles in transport process at the inner- envelope membrane.size e.g. the vacuole and the cell wall may be considered both as Ca2+ transported to the thylakoid lumen must cross thesources and buffering compartments of calcium mobilized to or envelope membranes, the stroma and the thylakoid membraneexpelled from the cytosol to prevent formation of insoluble salts . Data obtained with isolated thylakoid membranes show(e.g. calcium phosphate) that are deleterious to cell integrity. that transthylakoid Ca2+ transport is stimulated by light or, inChloroplasts, mitochondria and nuclei contain also mM total the dark, by adding ATP in the incubation medium. Addition ofcalcium mainly sequestrated as bound calcium . Conse- an H+-translocating uncoupler which dissipates the thylakoidquently, these organelles surrounded by a double membrane proton gradient generated by ATP hydrolysis inhibits Ca2+constituting either a continuous barrier (chloroplasts and transport. The dependence of the reaction on ΔpH suggests thatmitochondria) or punctuated by pores (nuclei) have to take up a Ca2+/H+ antiporter localizes to the thylakoids.calcium from outside and may also contribute to regulate Therefore, two distinct transport systems with differenthomeostasis of Ca 2+ in the cytosol. Moreover, calcium- spatial localization allow Ca2+ to cross the inner-envelopedependent events take place in chloroplasts, mitochondria and membrane and the thylakoids. The coordinated functioning ofnuclei and, consequently, free calcium concentrations might be the two systems allows the control of Ca2+ entry into the stromaalso regulated in these organelles, and might, by some manner and the thylakoid and regulates calcium-dependent processes.contribute to specify cellular calcium signature. This short Ca2+ accumulated in the light may be bound in the stroma orreview aims at drawing attention to the spatial component of sequestered in the thylakoids. The thylakoid transporter wouldcalcium signaling in plant cell organelles (essentially mitochon- contribute to remove excess Ca2+ from the stroma, in the light,dria, chloroplasts and nuclei) as part of the calcium toolkit. otherwise high levels of free Ca2+ would inhibit carbon dioxide fixation.2. Isolated plant organelles harbor calcium-dependent In fact, the dynamics of changes in free Ca2+ levels in thebiochemical processes and are able to take up calcium from chloroplast is more complex than expected by measurements intheir internal environment isolated chloroplasts or membrane preparation thereof. Using transgenic Nicotiana plumbaginifolia seedlings harboring the For the sake of clarity, we will consider successively the calcium bioluminescent reporter protein targeted to thechloroplast, the mitochondrion and the plant cell nucleus. chloroplast stroma, it has been shown that darkness provokes a large Ca2+ flux in the stroma . The increase starts a few2.1. Calcium movements and biological effects in isolated and minutes after lights off, proceeds maximally within 20 min andin cellular chloroplasts returns to the background levels. The magnitude of Ca2+ flux (and its return to background level) increases with the duration In vitro measurements of enzyme activities in the chlor- of light exposure, showing that the amount of calcium that mayoplastidic stroma have shown that NAD kinase which provides be mobilized is accumulated in chloroplasts during daytime. If
T.-C. Xiong et al. / Biochimica et Biophysica Acta 1763 (2006) 1209–1215 1211the lights are turned on before calcium elevation, no increase in onset of the Ca2+-induced swelling as well as the rate of thefree Ca2+ is obtained in the stroma. The Ca2+ spike is attenuated process are accelerated and indicate that, in vitro, plantalso if lights are turned on during the increasing phase of Ca2+ mitochondria undergo a faster mitochondrial permeabilityelevation. The expected biological consequence of these large transition [18,19]. It has been shown that in response toCa2+ increases in the stroma is the inhibition of photosynthetic oxidative stress, increases in electron transport in mitochondriaprocesses. The intriguing fact is that inhibiting photosynthetic trigger H2O2 production, depletion of ATP, and opening of PTPelectron transport has only a poor effect on the magnitude of the and cell death .Ca2+ increases in the dark. Therefore, the replenishment of themobilizable Ca2+ pool is not strictly dependent upon photo- 2.3. Calcium movements and biological effects in isolated andsynthesis. Moreover, increases in stromal Ca2+ do not correlate cellular nucleiwith corresponding decreases in cytosolic calcium levelsshowing that Ca2+ flux at lights off comes from the chloroplasts. Early experiments have shown that nuclei isolated fromDarkness induces also Ca2+ increases in the cytosol correla- plants are able to phosphorylate proteins in a calcium dependenttively to the decrease in the chloroplastidic stroma. However, manner . Data accumulated over years have refined thesethe magnitude of the cytosolic burst is 3 to 4-fold lower initial results by characterizing a number of Ca2+-binding orsuggesting that most of the Ca2+ is remobilized by chloroplasts. regulated proteins in the nucleus. These include calmodulin,Collectively, these dark-dependent changes in Ca2+ in both the annexin, transcription factors and calcium-dependent proteincytosol and the chloroplast may signal the end of the day to the kinases and phosphatases [23,24]. Recently, it has been showncell . that a Ca2+-calmodulin-dependent kinase necessary for the establishment of the symbiotic association between nitrogen2.2. Calcium movements and biological effects in isolated and fixing bacteria and plant legumes is located in the nucleus .cellular plant mitochondria The nucleus is separated from other cell compartments by a double membrane punctuated by nuclear pore complexes Mitochondria isolated from different plants are able to take (NPC). NPCs allow trafficking of molecules and ions betweenup calcium (at concentrations as low as a few μM) from the the cytosol and the nucleoplasm. Calcium permeation throughmedium in the presence of respiratory substrates . NPC by simple diffusion fully explains the pattern of nuclear Data obtained with oat mitochondria suggest further that the calcium upon stimulation of cardiac myocyte . However,plant photoreceptor, phytochrome, may regulate Ca2+ fluxes in different lines of research performed mainly on animal systemsa photoreversible manner . Red light irradiation reduces the have shown that calcium channels and transporters localize tonet Ca2+ uptake which is restored back to the dark control level the nuclear envelope [27–29]. These data suggest that theupon far-red irradiation. In the presence of ruthenium red (an nucleus is equipped to generate calcium signals, and theinhibitor of active Ca2+ uptake) in the reaction medium, red diffusion hypothesis may not explain all nuclear calciumlight irradiation provokes a Ca2+ release from the mitochondria patterns.via a passive efflux mechanism, suggesting that mitochondria In plants, nuclei isolated from tobacco cells, harboringare able to take up from and to release Ca2+ into the cytosol. aequorin in the nucleoplasm, respond to chemicals like Recent data show that exposure of plant isolated mitochon- mastoparan , to temperature changes or to mechanicaldria to Pi and mM concentrations of Ca2+ induces a fast stimulation  by large Δ[Ca2+]nuc. Incubation of nuclei in ashrinkage followed by a high amplitude swelling [18,19]. Both medium containing high concentrations of Ca2+ has no effect onPi and Ca2+ are required and replacing Ca2+ by an equivalent nucleoplasmic calcium in the absence of stimulation, ruling outconcentration of Mg2+ promotes shrinkage but not swelling, the possibility of a passive diffusion from the incubationshowing that the process is ion-specific. Swelling of mitochon- medium. Symmetrically, chelating extra nuclear calcium withdria reflects an expansion of the matrix that ends up with the EGTA does not inhibit the increase in free nucleoplasmic Ca2+rupture of the external membrane and the release of proteins and elicited by mechanical or thermal stimuli, establishing that theespecially of cytochrome c. Further work on potato mitochon- signal Ca2+ is mobilized from the nucleus itself . Because ofdria has established that cyclosporin A inhibits Ca2-induced its ability to accumulate calcium, in the lumen, the nuclearswelling even after collapsing the Δψ. Collectively, these data envelope may be the source of the mobilized calcium.suggest the occurrence of permeability transition pores (PTP) Based on the above-mentioned data, calcium homeostasislocated at the contact between inner and outer membranes in and disturbance in plant nucleoplasm can be described andplant mitochondria. This process referred to as mitochondrial simulated by a simple mathematical model . The modelpermeability transition is known to allow mitochondria to be considers the isolated nucleus as a closed system composed ofcellular stress sensors and central players in cell death in two compartments: the nucleoplasm (where Δ[Ca2+]nuc takesanimals . Mitochondrial Ca2+ overload activates PTP place) and a calcium store (corresponding to the nuclearresulting in transient mitochondrial depolarization and envelope). Calcium channels located to the inner nucleardecreased ATP production. PTP gating may also cause release membrane mobilize the accumulated calcium which is thenof mitochondrial proteins (and especially cytochrome c) that released into the nucleoplasm. An elusive calcium transporteractivate apoptotic pathways. Interestingly, under anoxia (lead- located to the inner membrane is predicted to expel calciuming to accelerated ATP depletion and Pi increased levels), the from the nucleoplasm and replenish the lumen .
1212 T.-C. Xiong et al. / Biochimica et Biophysica Acta 1763 (2006) 1209–1215 Patch-clamping nuclear membrane reveals the existence of . Thus, coincident variations in [Ca2+]cyt and [Ca2+]m arenon-selective voltage-dependent Ca2+-channels in beet . induced by cold or osmotic shock with similar temporalMoreover, Ca2+-ATPases pumps localize to the outer membrane kinetics. Δ[Ca2+]m is twice lower than Δ[Ca2+]cyt suggestingof the nuclear envelope isolated from tobacco cells . In that mitochondria may simply buffer Δ[Ca2+ ]cyt throughanimals, the inner membrane of nuclear envelope contains uptake. Touch stimulation induces an immediate elevation ofdifferent ligand operated channels . Patch-clamping of [Ca2+]cyt followed by a return to the background level withinnuclei isolated from osteoblastic-like MC3T3-E1 cells allows less than 30 s. Under the same conditions, after an elevation ascharacterizing mechano-sensitive Ca2+-channels responsible for rapid as in the cytosol, Δ[Ca2+]m is maintained at 37% of itscalcium elevation . In animals, TRP-like channels are measured maximum for at least 1 min. Lastly, challengingknown to sense either temperature or pH [37,38] but their plants with hydrogen peroxide induces Δ[Ca2+]m faster andpresence in the nuclear compartment has not been established. longer than Δ[Ca2+]cyt. Collectively, these data show thatIn nuclei isolated from tobacco cells, Δ[Ca2+]nuc elicited by mitochondria have the potential machinery to discriminate andeither pH and or temperature changes are inhibited by drugs generate specific Ca2+ signaling pathways in response to anknown to inhibit TRP-like channels . array of stimuli. Studying the biological effect of two distinct stress2.4. The dynamics of cytosolic and organelle calcium are conditions acting both of Δ[Ca2+]nuc and Δ[Ca2+]cyt shedsdifferentially regulated light on the subtlety of cellular Ca2+ signaling. Thus, wind signals induce Δ[Ca2+]cyt and Δ[Ca2+]nuc to peak at 0.3 s and The data described in the preceding paragraphs are indicative 0.6 s, respectively in tobacco seedlings . In response to coldof the ability of chloroplasts, mitochondria and nuclei to shock, Δ[Ca2+]cyt peaks at about 4 s whereas the nucleus reactsgenerate intra organelle changes in free Ca2+ concentrations. maximally after 7 s. Consequently, wind stimuli and cold shockThe availability of bioluminescent or fluorescent Ca2+-probes implicate distinct calcium signaling pathways. Interestinglythat may be targeted to a particular compartment allows both wind and cold shock induce a particular isoform ofdetermining relationships between the organelles . calmodulin gene referred to as NpCaM-1 (for Nicotiana Thus, pulses of blue light induce Δ[Ca2+]cyt in Arabidopsis plumbaginifolia Calmodulin gene 1). Comparison of Ca2+and tobacco seedlings . The spectral response of the Ca2+ dynamics with NpCaM-1 expression after stimulation suggeststransient is similar to phototropism and suggests the involve- that Δ[Ca2+]nuc are the preferential transducers of windment of NPH1 as the photoreceptor. Use of organelle-targeted stimulation and Δ[Ca2+]cyt of cold shock .aequorin either in the nucleus or the chloroplasts shows that Ca2+ In line with these data, other abiotic and biotic stimuli elicitincreases occur only in the cytosol. These observations suggest specific cytosolic and nuclear calcium patterns. Thus, in responsethat physiological responses implicating NPH1 may be to osmotic constraints of identical intensity (150 mosM) butspecifically transduced through [Ca2+]cyt. A more complex sensed by the cell as either “tension” or “pressure”, distinctsituation has been described depending upon the physiological calcium responses are recorded . Hypo-osmotic constraintsconditions experienced by plants. elicit large Δ[Ca2+]cyt and Δ[Ca2+]nuc. In contrast, hyper-osmotic Thus, under continuous illumination of tobacco plantlet free constraints with the same intensity induce only a cytosolicCa2+ concentrations vary rhythmically in the cytosol ; upon response. Moreover, the cell suspensions responded withtransfer to darkness the periodic fluctuations stop and resume characteristic nuclear and cytosolic Δ[Ca2+] patterns as a functionupon turning lights on. The oscillations may be phase-shifted by of the nature (non-ionic/ionic osmoticum) and the intensitydark–light transitions and present the characteristics of a (osmolarity) of the stimulus.circadian rhythm. Interestingly, in contrast to the cytosolic Cryptogein, a polypeptide secreted by the oomycete Phy-compartment, there is no circadian variation in Δ[Ca2+]chl under tophthora infestans, triggers defense reaction to pathogen attackcontinuous illumination. Rather, on transfer to darkness, a large in tobacco . Cryptogein induces calcium transients in bothincrease followed by circadian oscillations in Δ[Ca2+]chl is the cytosol and the nucleus of tobacco cell suspension culturesobserved. Therefore, Ca2+ oscillations are regulated differen- [47,48]. Interestingly, Δ[Ca2+]nuc is maximal 15 min after thetially in the two considered compartments. cytosolic peak and other elicitors provoking equivalent Ca2+ Cooperation between mitochondria and the cytosol is clearly changes in the cytosol have no effect on Δ[Ca2+]nuc. Theoreticalshown when plants experience anoxia. In these conditions, considerations lead to the conclusion that if calcium signalmitochondrial Ca2+ varies rapidly and reversibly in response to kinetics in the cytosol and the nucleoplasm differ each other bychanges in O2 availability . The Δ[Ca2+]m is inversely at least 1 s, then a simple diffusion of calcium from the cytosolrelated to Δ[Ca2+]cyt. Moreover, Δ[Ca2+]cyt colocalize essen- to the nucleus is ruled out . In animals cells differenttially with mitochondria showing that mitochondria contribute regulation of cytosolic and nuclear calcium has been reportedmost probably to Δ[Ca2+]cyt under anoxia and also to its [50,51]. As stated above, isolated plant nuclei are able torestoration upon returning back to normoxia. Interestingly, convert physical constraints into Δ[Ca2+]nuc . Conversely,when challenged with different stress conditions, tobacco when challenged with cryptogein, isolated nuclei do notplantlets respond by a rapid and large variation (up to 1 μM) respond by Δ[Ca2+]nuc. Δ[Ca2+]nuc triggered by cryptogeinin mitochondrial free Ca2+. Interestingly, Δ[Ca2+]m depends are only detectable in intact cells and needs the initialupon the nature of the stress and generates differential responses recognition of the elicitor by its receptors localizing to the
T.-C. Xiong et al. / Biochimica et Biophysica Acta 1763 (2006) 1209–1215 1213plasma membrane . Therefore only stimuli whose putative Another example of intracellular movements of proteinsensors are located onto the nucleus may be converted into related to calcium signaling is illustrated by the case of OsERG1Δ[Ca2+]nuc. protein, a small protein containing a Ca2+-dependent mem- These two chosen examples clearly show that Δ[Ca2+]nuc brane-binding motif (C2- domain) . OsERG1 protein whichmay be directly generated in response to stimuli but may be also is mainly located in the cytosol accumulates at the plasmaa relatively late event that needs the activation of signaling steps membrane upon challenging rice cells with either a fungallocated in the cytosol or/and the integrity of a functional elicitor from Magnaportha grisea or a calcium ionophore .continuum between the plasma membrane and the nucleus. Collectively, these data show that a particular calcium- binding protein (effector) may localize to a particular compart-3. Discussion and prospects ment as a function of the physiological status of the cell (metabolic status, defense responses). Such versatility may have Plant cell organelles surrounded by a double membrane (e.g. a profound influence on the coordination of calcium-dependentmitochondria, chloroplasts and nuclei) have multiple functions events in the plasma membrane and the nucleus (CaM53) or inin Ca2+ signaling, regardless of the needs for high Ca2+ level for the cytosol and the plasma membrane (C2-proteins) usingmaintaining membrane structure and intactness. Firstly, they identical effector protein.contribute to the regulation of Δ[Ca2+]cyt by taking up Ca2+ Ca2+ by itself may be involved in protein translocation/from the cytosol and storing the cation as a mobilizable form import into organelles and secretion.that may be released again in the cytosol. Secondly, they harbor For example, chloroplast imports of a subset of proteins thatcalcium-dependent processes needing the fine-tuning of free need a cleavable transit peptide like the small subunit of RibuloseCa2+ in the considered organelle. Thirdly, in each organelle 1, 5-Bisphosphate carboxylase/oxygenase are inhibited either bysubtle compartmentation of mobilizable and free Ca2+ exists calmodulin inhibitors or calcium ionophore. Addition of externalmost probably (e.g. in chloroplasts, free Ca2+ concentrations calmodulin or calcium restores the import process . Inhave to be regulated in the stroma and the thylakoid lumen). growing pollen tube, Ca2+-dependent protein kinase activity isCa2+ release in discrete and highly localized region of the highly concentrated in the apical region . According tonucleus may also be generated by the existence of a nucleo- Moutinho et al.  the modification of growth direction orplasmic reticulum identified in animal epithelial cell . This localized Δ[Ca2+]cyt leading to reorientation greatly increased thebranching intranuclear network forming a continuum with the kinase activity presumably in connection with Ca2+ -mediatednuclear envelope and the endoplasmic reticulum functions as a exocytosis.store from which Ca2+ may be mobilized at precise location in Local changes in Δ[Ca2+] in a given compartment maythe nucleus. Although its biological role has not been addressed, impact on the protein (and other metabolites) repertoire of thecomparable structure has been described in plant cell nuclei others. In the end, the dynamics of the exchanges that allow. redistribution of key components of the signaling pathway All these experimental evidence show that each organelle has (enzymes, substrates, transcription factors) determines thethe ability to control its own Δ[Ca2+] and, in this way, is biological output and the mounting of an adaptive response.autonomous with respect to the cytosol and is not a simple Not only macromolecules are redistributed but the intracel-passive sensor of local [Ca2+]cyt (once the calcium pool is lular architecture may be reorganized also, in response toaccumulated in the organelle). However, cross-talk between stimuli. Due to the large number of examples, we will justdifferent cellular organelles is crucial in Ca2+ signaling in quote: migration of the nucleus at the site of application of arelation to the dynamic localization of Ca2+ effectors, the pathogen most probably driven by cytoskeleton reorganization,Ca2+-dependent translocation and/or post-translational modifi- local contacts between the endoplasmic reticulum and mito-cations of proteins and the dynamic intracellular reorganization chondria or nuclei. Because the diffusion rate of Ca2+ is veryin response to stimuli. slow, the formation of clusters of organelles facilitates contacts Thus, a particular calmodulin referred to as CaM53 is post- between Ca2+ stores, calcium channels and transporters andtranslationally isoprenylated at its C-terminus domain in reduces the transit time of information, and increases thePetunia . Blockade of the isoprenoid biosynthesis results efficiency of the overall system.in the localization of the protein in the nucleus, showing that An exciting challenge in plant Ca 2+ signaling is anisoprenylation may drive the subcellular localization of CaM53. integrative approach allowing quantifying and putting in aIn leaves exposed to light for several days, CaM53 localizes to logical order different steps that link an initial stimulus to timethe plasma membrane whereas the protein accumulates in the and space changes in calcium and ending with a biologicalnuclear compartment in samples maintained in the dark during quantifiable output. Importantly, time-lapse changes in localthe same period. Interestingly, dark exposure on a medium Ca2+ and other second messengers (the proton, active oxygensupplemented with sucrose prevents nuclear translocation. species, and lipid-derived compounds) should be carefullyRecombinant CaM53 activates glutamate decarboxylase a established. Opportune models to cope with such difficult tasksplant calmodulin dependent enzyme and CaM53 gene rescues are already available in plant biology and include physiology ofyeast cmd1Δ mutant defective in the single gene for CaM by stomata, cell growth (pollen tube) and symbiotic associationcomplementation . Therefore, the unusual CaM isoform is between nitrogen-fixing bacteria and legumes . Becausefunctionally active. genomic as well as genetic and cell biology resources are
1214 T.-C. Xiong et al. / Biochimica et Biophysica Acta 1763 (2006) 1209–1215becoming available, other systems might be of value to consider Mitochondria and cell death. Mechanistic aspects and methodologicalless specialized but specific aspects of plant physiology [46,57]. issues, Eur. J. Biochem. 264 (1999) 687–701.  B.S. Tiwari, B. Belenghi, A. Levine, Oxidative stress increased respirationWe believe that an integrated and multifaceted approach aiming and generation of reactive oxygen species, resulting in ATP depletion,at understanding how plant specific metabolisms are reoriented opening of mitochondrial permeability transition, and programmed cellin response to quantifiable stimuli is worth studying. The topic death, Plant Physiol. 128 (2002) 1271–1281.should give clues on the coupling between Ca2+ signaling, gene  N. Datta, Y. Chen, S. Roux, Phytochrome and calcium stimulation of protein phosphorylation in isolated pea nuclei, Biochem. Biophys. Res.expression, metabolite production/use and transport in space Commun. 128 (1985) 1403–1408.and time, and how the metabolite phenotype might be  N. Bouché, A. Yellin, W.A. Snedden, H. Fromm, Plant-specificcontrolled by acting on signaling processes. calmodulin-binding proteins, Ann. Rev. Plant Biol. 56 (2005) 435–466.  J.F. Harper, G. Breton, A. Harmon, Decoding Ca(2+) signals through plantAcknowledgements protein kinases, Annu. Rev. Plant Biol. 55 (2004) 263–288.  P. Kalo, C. Gleason, A. Edwards, J. Marsh, R.M. Mitra, S. Hirsch, J. Jakab, S. Sims, S.R. Long, J. Rogers, G.B. Kiss, J.A. Downie, G.E. Oldroyd, Due to space limitation, we apologize for not having quoted Nodulation signaling in legumes requires NSP2, a member of the GRASmany contributions to the field of calcium signaling in plants. family of transcriptional regulators, Science 308 (2005) 1786–1789.  C. Genka, H. Ishida, K. Ichimori, Y. Hirota, T. Tanaami, H. Nakazawa,References Visualization of biphasic Ca2+ diffusion from cytosol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging  P.J. White, M.R. Broadley, Calcium in plants, Ann. Bot. (London) 92 system, Cell Calcium 25 (1999) 199–208. (2003) 487–511.  L. Santella, K. Kyozuka, Effects of 1-methyladenine on nuclear Ca2+  K.D. Hirschi, The calcium conundrum. Both versatile nutrient and specific transients and meiosis resumption in starfish oocytes are mimicked by the signal, Plant Physiol. 136 (2004) 2438–2442. nuclear injection of inositol 1,4,5-trisphosphate and cADP-ribose, Cell  D. Sanders, C. Brownlee, J.F. Harper, Communicating with calcium, Plant Calcium 22 (1997) 11–20. Cell 11 (1999) 691–706.  M.D. Bootman, D. Thomas, S.C. Tovey, M.J. Berridge, P. Lipp, Nuclear  D. Sanders, J. Pelloux, C. Brownlee, J.F. Harper, Calcium at the crossroads calcium signalling, Cell Mol. Life Sci. 57 (2000) 371–378. of signaling, Plant Cell 14 (2002) S401–S417.  A.N. Malviya, The nuclear inositol 1,4,5-trisphosphate and inositol  A.M. Hetherington, C. Brownlee, The generation of Ca(2+) signals in 1,3,4,5-tetrakisphosphate receptors, Cell Calcium 16 (1994) 301–313. plants, Annu. Rev. Plant Biol. 55 (2004) 401–427.  N. Pauly, M.R. Knight, P. Thuleau, A.H. Van der Luit, M. Moreau, A.J.  M.J. Berridge, M.D. Bootman, H.L. Roderick, Calcium signalling: Trewavas, R. Ranjeva, C. Mazars, Control of free calcium in plant cell dynamics, homeostasis and remodelling, Nat. Rev., Mol. Cell Biol. 4 nuclei, Nature 405 (2000) 754–755. (2003) 517–529.  T.C. Xiong, A. Jauneau, R. Ranjeva, C. Mazars, Isolated plant nuclei as  S.A. Scrase-Field, M.R. Knight, Calcium: just a chemical switch? Curr. mechanical and thermal sensors involved in calcium signalling, Plant J. 40 Opin. Plant Biol. 6 (2003) 500–506. (2004) 12–21.  R. Malho, A. Moutinho, A. Vanderluit, A.J. Trewavas, Spatial character-  C. Brière, T.C. Xiong, C. Mazars, R. Ranjeva, Autonomous regulation of istics of calcium signalling: the calcium wave as a basic unit in plant cell free Ca(2+) concentrations in isolated plant cell nuclei: a mathematical calcium signalling, Philos. Trans. R. Soc. Lond., B Biol. Sci. 353 (1998) analysis, Cell Calcium 39 (2006) 293–303. 1463–1473.  C. Grygorczyk, R. Grygorczyk, A Ca2+- and voltage-dependent cation  M.R. McAinsh, A.M. Hetherington, Encoding specificity in Ca2+ channel in the nuclear envelope of red beet, Biochim. Biophys. Acta 1375 signalling systems, Trends Plant Sci. 3 (1998) 32–36. (1998) 117–130. D.S. Bush, Calcium regulation in plant cells and its role in signaling, Annu.  T.D. Bunney, P.J. Shaw, P.A. Watkins, J.P. Taylor, A.F. Beven, B. Wells, G. Rev. Plant Physiol., Plant Mol. Biol. 46 (1995) 95–122. M. Calder, B.K. Drobak, ATP-dependent regulation of nuclear Ca(2+) G. Kreimer, M. Melkonian, J.A.M. Holtum, E. Latzko, Characterization of levels in plant cells, FEBS Lett. 476 (2000) 145–149. calcium fluxes across the envelope of intact spinach chloroplasts, Planta  J.P. Humbert, N. Matter, J.C. Artault, P. Koppler, A.N. Malviya, Inositol 166 (1985) 515–523. 1,4,5-trisphosphate receptor is located to the inner nuclear membrane S. Muto, S. Izawa, S. Miyachi, Light-induced Ca2+ uptake by intact vindicating regulation of nuclear calcium signaling by inositol 1,4,5- chloroplasts, FEBS Lett. 139 (1982) 250–254. trisphosphate. Discrete distribution of inositol phosphate receptors to inner M.H. Roh, R. Shingles, M.J. Cleveland, R.E. McCarty, Direct measure- and outer nuclear membranes, J. Biol. Chem. 271 (1996) 478–485. ment of calcium transport across chloroplast inner-envelope vesicles, Plant  N. Itano, S. Okamoto, D. Zhang, S.A. Lipton, E. Ruoslahti, Cell spreading Physiol. 118 (1998) 1447–1454. controls endoplasmic and nuclear calcium: a physical gene regulation W.F. Ettinger, A.M. Clear, K.J. Fanning, M. Lou Peck, Identification of a pathway from the cell surface to the nucleus, Proc. Natl. Acad. Sci. U. S. A. Ca2+/H+ antiport in the plant chloroplast thylakoid membrane, Plant 100 (2003) 5181–5186. Physiol. 119 (1999) 1379–1386.  D.E. Clapham, TRP channels as cellular sensors, Nature 426 (2003) J. Sai, C.H. Johnson, Dark-stimulated calcium ion fluxes in the chloroplast 517–524. stroma and cytosol, Plant Cell 14 (2002) 1279–1291.  C. Montell, The TRP superfamily of cation channels, Sci. STKE 2005 C. Chen, A. Lehninger, Ca2+ transport activity in mitochondria from some (2005) re3. plant tissues, Arch. Biochem. Biophys. 157 (1973) 183–196.  H. Knight, M.R. Knight, Recombinant aequorin methods for intracellular S.J. Roux, K. McEntire, R.D. Slocum, T.E. Cedel, C.C. Hale, Phytochrome calcium measurement in plants, Methods Cell Biol. 49 (1995) 201–216. induces photoreversible calcium fluxes in a purified mitochondrial fraction  G. Baum, J. Long, G. Jenkins, A. Trewavas, Stimulation of the blue light from oats, Proc. Natl. Acad. Sci. U. S. A. 78 (1981) 283–287. phototropic receptor NPH1 causes a transient increase in cytosolic Ca2+, S. Arpagaus, A. Rawyler, R. Braendle, Occurrence and characteristics of Proc. Natl. Acad. Sci. 96 (1999) 13554–13559. the mitochondrial permeability transition in plants, J. Biol. Chem. 277  C. Johnson, M. Knight, T. Kondo, P. Masson, J. Sedbrook, A. Haley, A. (2002) 1780–1787. Trewavas, Circadian oscillations of cytosolic and chloroplastic free E. Virolainen, O. Blokhina, K. Fagersedt, Ca2+-induced high amplitude calcium in plants, Science 269 (1995) 1863. swelling and cytochrome c release from wheat (Triticum aestivum L.)  C.C. Subbaiah, D.S. Bush, M.M. Sachs, Mitochondrial contribution to the mitochondria under anoxic stress, Ann. Bot. 90 (2002) 509–516. anoxic Ca2+ signal in maize suspension-cultured cells, Plant Physiol. 118 P. Bernardi, L. Scorrano, R. Colonna, V. Petronilli, F. Di Lisa, (1998) 759–771.
T.-C. Xiong et al. / Biochimica et Biophysica Acta 1763 (2006) 1209–1215 1215 D.C. Logan, M.R. Knight, Mitochondrial and cytosolic calcium dynamics  M.F. Leite, E.C. Thrower, W. Echevarria, P. Koulen, K. Hirata, A.M. are differentially regulated in plants, Plant Physiol. 133 (2003) 21–24. Bennett, B.E. Ehrlich, M.H. Nathanson, Nuclear and cytosolic calcium are A. van der Luit, C. Olivari, A. Haley, M. Knight, A. Trewavas, Distinct regulated independently, Proc. Natl. Acad. Sci. U. S. A. 100 (2003) calcium signaling pathways regulate calmodulin gene expression in 2975–2980. tobacco, Plant Physiol. 121 (1999) 705–714.  D.A. Collings, C.N. Carter, J.C. Rink, A.C. Scott, S.E. Wyatt, N.S. Allen, N. Pauly, M.R. Knight, P. Thuleau, A. Graziana, S. Muto, R. Ranjeva, C. Plant nuclei can contain extensive grooves and invaginations, Plant Cell 12 Mazars, The nucleus together with the cytosol generates patterns of (2000) 2425–2440. specific cellular calcium signatures in tobacco suspension culture cells,  M. Rodriguez-Concepcion, S. Yalovsky, M. Zik, H. Fromm, W. Gruissem, Cell Calcium 30 (2001) 413–421. The prenylation status of a novel plant calmodulin directs plasma D. Lecourieux, R. Ranjeva, A. Pugin, Calcium in plant defence-signalling membrane or nuclear localization of the protein, EMBO J. 18 (1999) pathways, New Phytol. 171 (2006) 249–269. 1996–2007. D. Lecourieux, C. Mazars, N. Pauly, R. Ranjeva, A. Pugin, Analysis and  C. Kim, Y. Koo, J. Jin, B. Moon, C. Kang, S. Kim, B. Park, S. Lee, M. effects of cytosolic free calcium increases in response to elicitors in Ni- Kim, I. Hwang, Rice C2-domain proteins are induced and translocated to cotiana plumbaginifolia cells, Plant Cell 14 (2002) 2627–2641. the plasma membrane in response to a fungal elicitor, Biochemistry 42 D. Lecourieux, O. Lamotte, S. Bourque, D. Vendehenne, C. Mazars, R. (2003) 11625–11633. Ranjeva, A. Pugin, Proteinaceous and oligosaccharidic elicitors induce  F. Chigri, J. Soll, U. Vothknecht, Calcium regulation of chloroplast protein different calcium signatures in the nucleus of tobacco cells, Cell Calcium import, Plant J. 42 (2005) 821. 38 (2005) 527–538.  A. Moutinho, A. Trewavas, R. Malhó, Relocation of a Ca2+-dependent T. Meyer, E. Oancea, N. Allbritton, Nuclear calcium signals, Ciba Found. protein kinase activity during pollen tube reorientation, Plant Cell 10 Symp. 188 (1995) 252–266. (1998) 1499–1510. W. Echevarria, M.F. Leite, M.T. Guerra, W.R. Zipfel, M.H. Nathanson,  T.C. Xiong, S. Bourque, C. Mazars, A. Pugin, R. Ranjeva, Signalisation Regulation of calcium signals in the nucleus by a nucleoplasmic reticulum, calcique cytosolique et nucléaire et réponses des plantes aux stimuli Nat. Cell Biol. 5 (2003) 440–446. biotiques et abiotiques, Medecine-Science, in press.