Pair Of Guard Cells Research Articles

A network-based modeling framework reveals the core signal transduction network underlying high carbon dioxide-induced stomatal closure in guard cells.

Stomata are pores on plant aerial surfaces, each bordered by a pair of guard cells. They control gas exchange vital for plant survival. Understanding how guard cells respond to environmental signals such as atmospheric carbon dioxide (CO2) levels is not only insightful to fundamental biology but also relevant to real-world issues of crop productivity under global climate change. In the past decade, multiple important signaling elements for stomatal closure induced by elevated CO2 have been identified. Yet, there is no comprehensive understanding of high CO2-induced stomatal closure. In this work, we assemble a cellular signaling network underlying high CO2-induced stomatal closure by integrating evidence from a comprehensive literature analysis. We further construct a Boolean dynamic model of the network, which allows in silico simulation of the stomatal closure response to high CO2 in wild-type Arabidopsis thaliana plants and in cases of pharmacological or genetic manipulation of network nodes. Our model has a 91% accuracy in capturing known experimental observations. We perform network-based logical analysis and reveal a feedback core of the network, which dictates cellular decisions in closure response to high CO2. Based on these analyses, we predict and experimentally confirm that applying nitric oxide (NO) induces stomatal closure in ambient CO2 and causes hypersensitivity to elevated CO2. Moreover, we predict a negative regulatory relationship between NO and the protein phosphatase ABI2 and find experimentally that NO inhibits ABI2 phosphatase activity. The experimental validation of these model predictions demonstrates the effectiveness of network-based modeling and highlights the decision-making role of the feedback core of the network in signal transduction. We further explore the model's potential in predicting targets of signaling elements not yet connected to the CO2 network. Our combination of network science, in silico model simulation, and experimental assays demonstrates an effective interdisciplinary approach to understanding system-level biology.

Surrounded by luxury: The necessities of subsidiary cells.

The evolution of stomata marks one of the key advances that enabled plants to colonise dry land while allowing gas exchange for photosynthesis. In large measure, stomata retain a common design across species that incorporates paired guard cells with little variation in structure. By contrast, the cells of the stomatal complex immediately surrounding the guard cells vary widely in shape, sizeand count. Their origins in development are similarly diverse. Thus, the surrounding cells are likely a luxury that the necessity of stomatal control cannot do without (with apologies to Oscar Wilde). Surrounding cells are thought to support stomatal movements as solute reservoirs and to shape stomatal kinetics through backpressure on the guard cells. Their variety may also reflect a substantial diversity in function. Certainly modelling, kinetic analysis and the few electrophysiological studies to date give hints of much more complex contributions in stomatal physiology. Even so, our knowledge of the cells surrounding the guard cells in the stomatal complex is far from complete.

Ploidy Determination in Banana Hybrids through Stomatal Studies and Chloroplast Count

Stomata are probably a viable taxonomy distinguishing feature. The study of stomata characteristics of synthetic banana hybrids and their parentage has been carried out. Ploidy determination is essential in banana breeding programs to understand the genetic makeup of hybrid plants. The purpose of this study was to determine the ploidy level of hybrids in contrast with the properties of plant stomata in synthetic banana hybrids with those of their parent plants. Therefore, by examining components such as stomata location, number of epidermal cells, number of stomata cells, stomata length, width, area, intensity and density, as well as chloroplast count, it is possible to compare and contrast the traits of synthetic banana hybrids and their parentage which have different genomes. The entire mount approach was used to make an incision for the purpose of observing stomata. By characterising the stomata traits of the synthetic hybrids and their parentage, the results were then descriptively and qualitatively assessed. Three different locations on the abaxial of the leaf were taken for the determination. The samples were collected from close to the petiole (R1), middle (R2), and distal end (R3), stomata were observed using the replica method. Results showed that more number of stomata was observed in R2of leaf abaxial. The chloroplast count in pairs of stomatal guard cells from the accessions was also measured. The chloroplast density was determined in pairs of stomatal guard cells from the accessions. Based on morphological descriptions, it was observed that 16.66% of the accessions were diploid, 50% were triploid and 33.33% were tetraploid. When considering the chloroplast count, the accessions were categorized into groups, certain accessions classified as tetraploids, others as triploids, and diploids.

Maize LSC encoding a large subunit of ribonucleotide reductase is required for subsidiary cell development and plant growth.

The four-celled stomatal complex consists of a pair of guard cells (GCs) and two subsidiary cells (SCs) in grasses, which benefits fast adjustment of stomatal pore aperture. The formation and development of SCs are thus important for stomatal functionality. Here, we report the maize loss of subsidiary cells (lsc) mutant with a large number of stomata lacking one or two SCs. The loss of SCs is supposed to be resulted from impeded subsidiary mother cells (SMC) polarization and asymmetrical division. Besides the defect in SCs, the lsc mutant also displays a dwarf morphology and pale and stripped newly-grown leaves. LSC encodes a large subunit of ribonucleotide reductase (RNR), an enzyme involved in deoxyribonucleotides (dNTPs) synthesis. Consistently, the dNTPs levels and expression of genes involved in DNA replication, cell cycle progression, and SC development were significantly reduced in the lsc mutant compared with the wild-type B73 inbred line. Conversely, overexpression of maize LSC increase dNTP synthesis and promotes plant growth in both maize and Arabidopsis. Our data indicate that LSC regulates dNTP production and is required for SMC polarization, SC differentiation, and plant growth.

INDUÇÃO DE POLIPLOIDIA EM GENÓTIPOS DE MELANCIA COM RESISTÊNCIA AO OÍDIO (Podosphaera xanthii)

ABSTRACT Triploid watermelon is highly appreciated by the most demanding markets, and due to its small size, it is ideal for consumption by small families. With the growth in areas cultivated with seedless watermelon worldwide, there is a demand for the development of tetraploid germplasm to obtain triploid hybrids with better agronomic performance. This study performed two tests to induce polyploidy in the powdery mildew–resistant line developed by Embrapa Semi-Arid, LDRO, under different colchicine concentrations and application methods. In Experiment 1, the seeds were treated with colchicine (0.0%, 0.1%, and 0.2%) for 24 h and 48 h. In Experiment 2, 0.2% colchicine was applied by different methods: (a) directly on the seed (MDS) with and without scarification, (b) on seeds with radicle emission (MER), (c) at the insertion point between the hypocotyl and the root (MIHR), (d) at the seedling apex (MAP), and (e) in the inverted hypocotyl (MHI). Chromosome count (cytogenetic analysis), number of chloroplasts per pair of guard cells, number of stomata, seedling height, and hypocotyl diameter were measured. In the LDRO line, chromosomal duplication occurred in some plant cells, but it was not possible to obtain 100% tetraploid plants. Colchicine (0.2%) for 48 h without mechanical scarification induced chromosomal duplication in watermelon. The analysis of the number of chloroplasts identified the level of ploidy early, reducing the number of plants needed to be evaluated by cytogenetics, which allowed us to more accurately identify the different levels of ploidy of the plant.

An anatomical comparison of epidermal cells in wild-type and homeotic mutant flowers of Clarkia tembloriensis Vasek (Onagraceae) 1

Flowers consist of four whorls of organs (sepals, petals, stamens, and carpels), each expressing unique characteristics. In floral homeotic mutants, organs develop abnormally, and floral organs of one whorl express characteristics of another whorl. Much of our current understanding of the development of floral organs has been based on studies of floral homeotic mutants. In the homeotic mutant crinkle-petal (cp) of Clarkia tembloriensis, previous investigations demonstrated that the cp petal is a hybrid organ combining characteristics of sepals and petals. This investigation extends those studies with a comparative anatomical study of trichomes, guard cells, and marginal epidermal cells in wild-type (WT) sepals, WT petals, and cp petals. The aim was to determine if cp petals exhibit a hybrid epidermal anatomy of WT sepals and WT petals. Floral buds 1 day pre-anthesis and flowers 1 day post-anthesis were collected and prepared for light microscopy or scanning electron microscopy. Trichomes and guard cell pairs were described and counted, and marginal cells were observed and described. This anatomical study of trichomes, guard cells, and marginal cells demonstrated that cp petals of C. tembloriensis combine characteristics of both WT sepals and WT petals. These results provide further support for the hybrid nature of cp petals in Clarkia tembloriensis.

Deceleration of the cell cycle underpins a switch from proliferative to terminal divisions in plant stomatal lineage

SummaryDifferentiation of specialized cell types requires precise cell-cycle control. Plant stomata are generated through asymmetric divisions of a stem-cell-like precursor followed by a single symmetric division that creates paired guard cells surrounding a pore. The stomatal-lineage-specific transcription factor MUTE terminates the asymmetric divisions and commits to differentiation. However, the role of cell-cycle machineries in this transition remains unknown. We discover that the symmetric division is slower than the asymmetric division in Arabidopsis. We identify a plant-specific cyclin-dependent kinase inhibitor, SIAMESE-RELATED4 (SMR4), as a MUTE-induced molecular brake that decelerates the cell cycle. SMR4 physically and functionally associates with CYCD3;1 and extends the G1 phase of asymmetric divisions. By contrast, SMR4 fails to interact with CYCD5;1, a MUTE-induced G1 cyclin, and permits the symmetric division. Our work unravels a molecular framework of the proliferation-to-differentiation switch within the stomatal lineage and suggests that a timely proliferative cell cycle is critical for stomatal-lineage identity.

OsBC1L1 and OsBC1L8 function in stomatal development in rice

Stomata that are bordered by pairs of guard cells are specialized for regulating gas exchange and transpiration in plants. The stomatal morphology of grass is unique, characterized by two dumbbell-shaped guard cells flanked by two lateral subsidiary cells. This morphology and developmental pattern enable grass stomata to respond to environmental signals efficiently. In this study, we demonstrated that knockout either OsBC1L1 or OsBC1L8, two close homologs of OsBC1L family causes no discernible defects in rice stomatal development, however, the double knockout mutant osbc1l1 osbc1l8 exhibits excess stomatal production and stomatal clustering. OsBC1L1 overexpression also causes abnormal stomatal patterning in rice. Moreover, osbc1l1 osbc1l8 has many defective stomata complexes with only one subsidiary cell. The expression of OsSPCH2 and OsFAMA, two genes key to stomatal development is both down-regulated in osbc1l1 osbc1l8. In contrast, overexpressing OsBC1L1 suppresses only the expression of OsSPCH2. Both OsBC1L1 and OsBC1L8 could be detected to be localized at the cell plate and plasma membrane during cell division of guard mother cells and subsidiary mother cells. Taken together, these results suggest that OsBC1L1 and OsBC1L8 play essential roles in the development of rice stomatal complex likely through their involvement in cell reproduction.

Autotetraploid Emergence via Somatic Embryogenesis in Vitis vinifera Induces Marked Morphological Changes in Shoots, Mature Leaves, and Stomata.

Polyploidy plays an important role in plant adaptation to biotic and abiotic stresses. Alterations of the ploidy in grapevine plants regenerated via somatic embryogenesis (SE) may provide a source of genetic variability useful for the improvement of agronomic characteristics of crops. In the grapevine, the SE induction process may cause ploidy changes without alterations in DNA profile. In the present research, tetraploid plants were observed for 9.3% of ‘Frappato’ grapevine somatic embryos regenerated in medium supplemented with the growth regulators β-naphthoxyacetic acid (10 µM) and N6-benzylaminopurine (4.4 µM). Autotetraploid plants regenerated via SE without detectable changes in the DNA profiles were transferred in field conditions to analyze the effect of polyploidization. Different ploidy levels induced several anatomical and morphological changes of the shoots and mature leaves. Alterations have been also observed in stomata. The length and width of stomata of tetraploid leaves were 39.9 and 18.6% higher than diploids, respectively. The chloroplast number per guard cell pair was higher (5.2%) in tetraploid leaves. On the contrary, the stomatal index was markedly decreased (12%) in tetraploid leaves. The observed morphological alterations might be useful traits for breeding of grapevine varieties in a changing environment.

Structure and activity of SLAC1 channels for stomatal signaling in leaves

Stomata in leaves regulate gas exchange between the plant and its atmosphere. Various environmental stimuli elicit abscisic acid (ABA); ABA leads to phosphoactivation of slow anion channel 1 (SLAC1); SLAC1 activity reduces turgor pressure in aperture-defining guard cells; and stomatal closure ensues. We used electrophysiology for functional characterizations of Arabidopsis thaliana SLAC1 (AtSLAC1) and cryoelectron microscopy (cryo-EM) for structural analysis of Brachypodium distachyon SLAC1 (BdSLAC1), at 2.97-Å resolution. We identified 14 phosphorylation sites in AtSLAC1 and showed nearly 330-fold channel-activity enhancement with 4 to 6 of these phosphorylated. Seven SLAC1-conserved arginines are poised in BdSLAC1 for regulatory interaction with the N-terminal extension. This BdSLAC1 structure has its pores closed, in a basal state, spring loaded by phenylalanyl residues in high-energy conformations. SLAC1 phosphorylation fine-tunes an equilibrium between basal and activated SLAC1 trimers, thereby controlling the degree of stomatal opening.

SPEECHLESS and MUTE Mediate Feedback Regulation of Signal Transduction during Stomatal Development.

Stomatal density, spacing, and patterning greatly influence the efficiency of gas exchange, photosynthesis, and water economy. They are regulated by a complex of extracellular and intracellular factors through the signaling pathways. After binding the extracellular epidermal patterning factor 1 (EPF1) and 2 (EPF2) as ligands, the receptor-ligand complexes activate by phosphorylation through the MAP-kinase cascades, regulating basic helix-loop-helix (bHLH) transcription factors SPEECHLESS (SPCH), MUTE, and FAMA. In this review, we summarize the molecular mechanisms and signal transduction pathways running within the transition of the protodermal cell into a pair of guard cells with a space (aperture) between them, called a stoma, comprising asymmetric and symmetric cell divisions and draw several functional models. The feedback mechanisms involving the bHLH factors SPCH and MUTE are not fully recognized yet. We show the feedback mechanisms driven by SPCH and MUTE in the regulation of EPF2 and the ERECTA family. Intersections of the molecular mechanisms for fate determination of stomatal lineage cells with the role of core cell cycle-related genes and stabilization of SPCH and MUTE are also reported.

Influence of Environmental Factors Light, CO2, Temperature, and Relative Humidity on Stomatal Opening and Development: A Review

Stomata, the microscopic pores surrounded by a pair of guard cells on the surfaces of leaves and stems, play an essential role in regulating the gas exchange between a plant and the surrounding atmosphere. Stomatal development and opening are significantly influenced by environmental conditions, both in the short and long term. The rapid rate of current climate change has been affecting stomatal responses, as a new balance between photosynthesis and water-use efficiency has to be found. Understanding the mechanisms involved in stomatal regulation and adjustment provides us with new insights into the ability of stomata to process information and evolve over time. In this review, we summarize the recent advances in research on the underlying mechanisms of the interaction between environmental factors and stomatal development and opening. Specific emphasis is placed on the environmental factors including light, CO2 concentration, ambient temperature, and relative humidity, as these factors play a significant role in understanding the impact of global climate change on plant development.

A Guard Cell Abscisic Acid (ABA) Network Model That Captures the Stomatal Resting State.

Stomatal pores play a central role in the control of carbon assimilation and plant water status. The guard cell pair that borders each pore integrates information from environmental and endogenous signals and accordingly swells or deflates, thereby increasing or decreasing the stomatal aperture. Prior research shows that there is a complex cellular network underlying this process. We have previously constructed a signal transduction network and a Boolean dynamic model describing stomatal closure in response to signals including the plant hormone abscisic acid (ABA), calcium or reactive oxygen species (ROS). Here, we improve the Boolean network model such that it captures the biologically expected response of the guard cell in the absence or following the removal of a closure-inducing signal such as ABA or external Ca2+. The expectation from the biological system is reversibility, i.e., the stomata should reopen after the closing signal is removed. We find that the model’s reversibility is obstructed by the previously assumed persistent activity of four nodes. By introducing time-dependent Boolean functions for these nodes, the model recapitulates stomatal reopening following the removal of a signal. The previous version of the model predicts ∼20% closure in the absence of any signal due to uncertainty regarding the initial conditions of multiple network nodes. We systematically test and adjust these initial conditions to find the minimally restrictive combinations that appropriately result in open stomata in the absence of a closure signal. We support these results by an analysis of the successive stabilization of feedback motifs in the network, illuminating the system’s dynamic progression toward the open or closed stomata state. This analysis particularly highlights the role of cytosolic calcium oscillations in causing and maintaining stomatal closure. Overall, we illustrate the strength of the Boolean network modeling framework to efficiently capture cellular phenotypes as emergent outcomes of intracellular biological processes.

Secreted Peptide PIP1 Induces Stomatal Closure by Activation of Guard Cell Anion Channels in Arabidopsis.

Plant stomata which consist of a pair of guard cells, are not only finely controlled to balance water loss as transpiration and CO2 absorption for photosynthesis, but also serve as the major sites to defend against pathogen attack, thus allowing plants to respond appropriately to abiotic and biotic stress conditions. The regulatory signaling network for stomatal movement is complex in nature, and plant peptides have been shown to be involved in signaling processes. Arabidopsis secreted peptide PIP1 was previously identified as an endogenous elicitor, which induced immune response through its receptor, RLK7. PIP1-RLK7 can activate stomatal immunity against the bacterial strain Pst DC3118. However, the molecular mechanism of PIP1 in stomatal regulation is still unclear and additional new factors need to be discovered. In this study, we further clarified that PIP1 could function as an important regulator in the induction of stomatal closure. The results showed that PIP1 could promote stomata to close in a certain range of concentrations and response time. In addition, we uncovered that PIP1-RLK7 signaling regulated stomatal response by activating S-type anion channel SLAC1. PIP1-induced stomatal closure was impaired in bak1, mpk3, and mpk6 mutants, indicating that BAK1 and MPK3/MPK6 were required for PIP1-regulated stomatal movement. Our research further deciphered that OST1 which acts as an essential ABA-signaling component, also played a role in PIP1-induced stomatal closure. In addition, ROS participated in PIP1-induced stomatal closure and PIP1 could activate Ca2+ permeable channels. In conclusion, we reveal the role of peptide PIP1 in triggering stomatal closure and the possible mechanism of PIP1 in the regulation of stomatal apertures. Our findings improve the understanding of the role of PIP1 in stomatal regulation and immune response.

H2O2, Ca2+, and K+ in subsidiary cells of maize leaves are involved in regulatory signaling of stomatal movement

The stomata of maize (Zea mays) contain a pair of guard cells and a pair of subsidiary cells. To determine whether H2O2, Ca2+, and K+ in subsidiary cells were involved in stomatal movement, we treated four-week-old maize (Zhengdan 958) leaves with H2O2, diphenylene iodonium (DPI), CaCl2, and LaCl3. Changes in content and distribution of H2O2, Ca2+, and K+ during stomatal movement were observed. When exogenous H2O2 was applied, Ca2+ increased and K+ decreased in guard cells, while both ions increased in subsidiary cells, leading to stomatal closure. After DPI treatment, Ca2+ decreased and K+ increased in guard cells, but both Ca2+ and K+ decreased in subsidiary cells, resulting in open stomata. Exogenous CaCl2 increased H2O2 and reduced K+ in guard cells, while significantly increasing them in subsidiary cells and causing stomatal closure. After LaCl3 treatment, H2O2 decreased and K+ increased in guard cells, whereas both decreased in subsidiary cells and stomata became open. Results indicate that H2O2 and Ca2+ correlate positively with each other and with K+ in subsidiary cells during stomatal movement. Both H2O2 and Ca2+ in subsidiary cells promote an inflow of K+, indirectly regulating stomatal closure.

TAXONOMIC IMPORTANCE OF ANTICLINAL WALLS AND STOMATA PATTERNING IN SOME MELASTOMA L. SPECIES FROM FRASER HILL

The epidermal characteristics of five selected Melastoma L. species in Fraser Hill, Pahang that belongs to Melastomataceae family had been investigated. These five species namely M. muticum Ridl., M. decemfidum Roxb., M. perakense Ridl., M. sanguineum x malabathricum and M. malabathricum var. normale. The objective of this study is to determine whether epidermal characteristics in Melastoma could be taxonomic value in systematic and diagnostic investigations. Methods of the investigation involved epidermal peel and scanning electron microscopy. Results obtained revealed that the presence of hypostomatic stomata and guard cells pairs were elliptic in shaped for all species studied. Apart, the pattern of anticlinal walls on adaxial and abaxial surfaces was straight to wavy for all species studied except for wavy to sinuous anticlinal walls on abaxial surface of M. sanguineum x malabathricum. Furthermore, two types of stomata were observed among species studied such as anomocytic and diacytic stomata. Results showed that diacytic type was only present in M. sanguineum x malabathricum therefore could be a criterion to diagnose the species. Lastly, this present study was also reported on the presence of two types of guard cell pairs such as raised or slightly raised and sunken guard cell pairs. In conclusion, the present study revealed that the anticlinal walls and stomata patterning possess as taxonomic importance in identification and classification of Melastoma either at genus or species level.

The Role of Grass MUTE Orthologues During Stomatal Development.

Gas exchange between the plant and the atmosphere takes place through stomatal pores formed by paired guard cells. Grasses develop a unique stomatal structure that consists of two dumbbell-shaped guard cells flanked by lateral subsidiary cells. These structures confer a very efficient gas exchange capacity, which may have contributed to the evolutionary success of grasses. Recent works have identified orthologues of Arabidopsis MUTE in three grass species: BdMUTE in Brachypodium distachyon, BZU2/ZmMUTE in maize, and OsMUTE in rice. These genes induce the recruitment of subsidiary cells, and it appears to rely upon the ability of intercellular movement, from the guard mother cell to subsidiary mother cells, of the proteins encoded by them. Unexpectedly, this function of these grass MUTE genes contrasts with that of Arabidopsis MUTE, which promotes guard mother cell identity. These MUTE orthologues also appear to control guard mother cell fate progression, with the action of BdMUTE being less severe than those of BZU2/ZmMUTE and OsMUTE. The emerging picture unravels that grass MUTE genes have not only diverged, due to neo-functionalization, from Arabidopsis MUTE, but also among them.

Metabolomics of red-light-induced stomatal opening in Arabidopsis thaliana: Coupling with abscisic acid and jasmonic acid metabolism.

Environmental stimuli-triggered stomatal movement is a key physiological process that regulates CO2 uptake and water loss in plants. Stomata are defined by pairs of guard cells that perceive and transduce external signals, leading to cellular volume changes and consequent stomatal aperture change. Within the visible light spectrum, red light induces stomatal opening in intact leaves. However, there has been debate regarding the extent to which red-light-induced stomatal opening arises from direct guard cell sensing of red light versus indirect responses as a result of red light influences on mesophyll photosynthesis. Here we identify conditions that result in red-light-stimulated stomatal opening in isolated epidermal peels and enlargement of protoplasts, firmly establishing a direct guard cell response to red light. We then employ metabolomics workflows utilizing gas chromatography mass spectrometry and liquid chromatography mass spectrometry for metabolite profiling and identification of Arabidopsis guard cell metabolic signatures in response to red light in the absence of the mesophyll. We quantified 223 metabolites in Arabidopsis guard cells, with 104 found to be red light responsive. These red-light-modulated metabolites participate in the tricarboxylic acid cycle, carbon balance, phytohormone biosynthesis and redox homeostasis. We next analyzed selected Arabidopsis mutants, and discovered that stomatal opening response to red light is correlated with a decrease in guard cell abscisic acid content and an increase in jasmonic acid content. The red-light-modulated guard cell metabolome reported here provides fundamental information concerning autonomous red light signaling pathways in guard cells.

Salt acclimation induced salt tolerance in wild-type and abscisic acid-deficient mutant barley

Salt acclimation is a process to enhance salt tolerance in plants. The salt acclimation induced salt tolerance was investigated in a spring barley (Hordeum vulgare L.) cv. Steptoe (wild type, WT) and its abscisic acid (ABA)-deficient mutant Az34. Endogenesis ABA concentration in leaf was significantly increased by salt stress in WT, while it was not affected in Az34. Under salt stress, the salt acclimated Az34 plants had 14.8% lower total soluble sugar concentration and 93.7% higher sodium (Na) concentration in leaf, compared with salt acclimated WT plants. The acclimated plants had significantly higher leaf water potential and osmotic potential than non-acclimated plants in both WT and Az34 under salt stress. The salt acclimation enhanced the net photosynthetic rate (by 22.9% and 12.3%) and the maximum quantum yield of PS II (22.7% and 22.0%) in WT and Az34 under salt stress. However, the stomatal conductance in salt acclimated Az34 plants was 28.9% lower than WT under salt stress. Besides, the guard cell pair width was significantly higher in salt acclimated Az34 plants than that in WT plants. The results indicated that the salt acclimated WT plants showed a higher salt tolerance than Az34 plants, suggesting that ABA deficiency has a negative effect on the salt acclimation induced salt tolerance in barley.

Arabidopsis thaliana Leaf Epidermal Guard Cells: A Model for Studying Chloroplast Proliferation and Partitioning in Plants.

The existence of numerous chloroplasts in photosynthetic cells is a general feature of plants. Chloroplast biogenesis and inheritance involve two distinct mechanisms: proliferation of chloroplasts by binary fission and partitioning of chloroplasts into daughter cells during cell division. The mechanism of chloroplast number coordination in a given cell type is a fundamental question. Stomatal guard cells (GCs) in the plant shoot epidermis generally contain several to tens of chloroplasts per cell. Thus far, chloroplast number at the stomatal (GC pair) level has generally been used as a convenient marker for identifying hybrid species or estimating the ploidy level of a given plant tissue. Here, we report that Arabidopsis thaliana leaf GCs represent a useful system for investigating the unexploited aspects of chloroplast number control in plant cells. In contrast to a general notion based on analyses of leaf mesophyll chloroplasts, a small difference was detected in the GC chloroplast number among three Arabidopsis ecotypes (Columbia, Landsberg erecta, and Wassilewskija). Fluorescence microscopy often detected dividing GC chloroplasts with the FtsZ1 ring not only at the early stage of leaf expansion but also at the late stage. Compensatory chloroplast expansion, a phenomenon well documented in leaf mesophyll cells of chloroplast division mutants and transgenic plants, could take place between paired GCs in wild-type leaves. Furthermore, modest chloroplast number per GC as well as symmetric division of guard mother cells for GC formation suggests that Arabidopsis GCs would facilitate the analysis of chloroplast partitioning, based on chloroplast counting at the individual cell level.