Auxin Transport Inhibitor Research Articles

Auxin controls the division of root endodermal cells.

The root endodermis forms a selective barrier that prevents the free diffusion of solutes into the vasculature; to make this barrier, endodermal cells deposit hydrophobic compounds in their cell walls, forming the Casparian strip. Here, we showed that, in contrast to vascular and epidermal root cells, endodermal root cells do not divide alongside the root apical meristem in Arabidopsis thaliana. Auxin treatment induced division of endodermal cells in wild-type plants, but not in the auxin signaling mutant auxin resistant3-1. Endodermis-specific activation of auxin responses by expression of truncated AUXIN-RESPONSIVE FACTOR5 (ΔARF5) in root endodermal cells under the control of the ENDODERMIS7 promoter (EN7::ΔARF5) also induced endodermal cell division. We used an auxin transport inhibitor to cause accumulation of auxin in endodermal cells, which induced endodermal cell division. In addition, knockout of P-GLYCOPROTEIN1 (PGP1) and PGP19, which mediate centripetal auxin flow, promoted the division of endodermal cells. Together, these findings reveal a tight link between the endodermal auxin response and endodermal cell division, suggesting that auxin is a key regulator controlling the division of root endodermal cells, and that PGP1 and PGP19 are involved in regulating endodermal cell division.

Peroxisomes Implicated in the Biosynthesis of Siderophores and Biotin, Cell Wall Integrity, Autophagy, and Response to Hydrogen Peroxide in the Citrus Pathogenic Fungus Alternaria alternata.

Little is known about the roles of peroxisomes in the necrotrophic fungal plant pathogens. In the present study, a Pex6 gene encoding an ATPase-associated protein was characterized by analysis of functional mutations in the tangerine pathotype of Alternaria alternata, which produces a host-selective toxin. Peroxisomes were observed in fungal cells by expressing a mCherry fluorescent protein tagging with conserved tripeptides serine-lysing-leucine and transmission electron microscopy. The results indicated that Pex6 plays no roles in peroxisomal biogenesis but impacts protein import into peroxisomes. The number of peroxisomes was affected by nutritional conditions and H2O2, and their degradation was mediated by an autophagy-related machinery termed pexophagy. Pex6 was shown to be required for the formation of Woronin bodies, the biosynthesis of biotin, siderophores, and toxin, the uptake and accumulation of H2O2, growth, and virulence, as well as the Slt2 MAP kinase-mediated maintenance of cell wall integrity. Adding biotin, oleate, and iron in combination fully restored the growth of the pex6-deficient mutant (Δpex6), but failed to restore Δpex6 virulence to citrus. Adding purified toxin could only partially restore Δpex6 virulence even in the presence of biotin, oleate, and iron. Sensitivity assays revealed that Pex6 plays no roles in resistance to H2O2 and superoxide, but plays a negative role in resistance to 2-chloro-5-hydroxypyridine (a hydroxyl radical-generating compound), eosin Y and rose Bengal (singlet oxygen-generating compounds), and 2,3,5-triiodobenzoic acid (an auxin transport inhibitor). The diverse functions of Pex6 underscore the importance of peroxisomes in physiology, pathogenesis, and development in A. alternata.

NITRIC OXIDE ASSOCIATED PROTEIN1 (AtNOA1) is necessary for copper-induced lateral root elongation in Arabidopsis thaliana

Copper (Cu2+) is an essential metal micronutrients required for normal plant growth and development, but is highly toxic when in excess. Inhibition of primary root elongation and stimulation of lateral root (LR) formation are usual reported symptoms when exposure to high concentrations of Cu2+. The mechanism underlying Cu2+-induced LR formation remains to be further elucidated. Here, we reported that Cu2+-induced LR formation is regulated by NITRIC OXIDE ASSOCIATED PROTEIN1 (AtNOA1), mainly through mediating Cu2+-induced LR elongation growth, but not LR primordium initiation. The initiation of LR primordium is inhibited by the mutation of AtNOA1. Transcriptional analysis displayed that the expression of genes involved in Cu2+ homeostasis and tolerance showed significant difference between Atnoa1 and WT. Cu2+-induced auxin redistribution was inhibited in Atnoa1, indicated by auxin response reporter plants (DR5:GUS and DR5:GFP). Furthermore, Cu2+-induced LR formation were more sensitive to exogenous auxin transport inhibitors NOA and NPA in Atnoa1 than in WT. Interestingly, Cu2+-induced H2O2 level was higher in Atnoa1 than WT, but Cu2+-induced LR formation was normal in H2O2-related mutants. These results showed that AtNOA1 regulated the initiation of LR primordium and mediates Cu2+-induced LR elongation in Arabidopsis thaliana. AtNOA1 mediates Cu2+-induced LR elongation maybe through affecting Cu2+ homeostasis and Cu2+-induced auxin redistribution, but not depending on Cu2+-induced H2O2 accumulation.

Silicon induces adventitious root formation in rice under arsenate stress with involvement of nitric oxide and indole-3-acetic acid.

Arsenic (As) negatively affects plant development. This study evaluates how the application of silicon (Si) can favor the formation of adventitious roots in rice under arsenate stress (AsV) as a mechanism to mitigate its negative effects. The simultaneous application of AsV and Si up-regulated the expression of genes involved in nitric oxide (NO) metabolism, cell cycle progression, auxin (IAA, indole-3-acetic acid) biosynthesis and transport, and Si uptake which accompanied adventitious root formation. Furthermore, Si triggered the expression and activity of enzymes involved in ascorbate recycling. Treatment with L-NAME (NG-nitro L-arginine methyl ester), an inhibitor of NO generation, significantly suppressed adventitious root formation, even in the presence of Si; however, supplying NO in the growth media rescued its effects. Our data suggest that both NO and IAA are essential for Si-mediated adventitious root formation under AsV stress. Interestingly, TIBA (2,3,5-triiodobenzoic acid), a polar auxin transport inhibitor, suppressed adventitious root formation even in the presence of Si and SNP (sodium nitroprusside, an NO donor), suggesting that Si is involved in a mechanism whereby a cellular signal is triggered and that first requires NO formation, followed by IAA biosynthesis.

Lupeol Accumulation Correlates with Auxin in the Epidermis of Castor.

Lupeol, a natural lupane-type pentacyclic triterpene, possesses various pharmacological properties, and its production attracts attention. Significant quantities of lupeol are deposited on the castor aerial organ surface and are easily extractable as a predominant wax constituent. Thus, castor might be considered as a potential bioreactor for the production of lupeol. The lupeol biosynthesis pathway is well known, but how it is regulated remains largely unknown. Among large numbers of castor cultivars, we targeted one accession line (337) with high levels of lupeol on its stem surface and low levels thereof on its hypocotyl surface, implicating that lupeol synthesis is differentially regulated in the two organs. To explore the underlying mechanisms, we did comparative transcriptome analysis of the first internode of 337 stem and the upper hypocotyl. Our results show that large amounts of auxin-related genes are differentially expressed in both parts, implying some possible interactions between auxin and lupeol production. We also found that several auxin-responsive cis-elements are present in promoter regions of HMGR and LUS genes encoding two key enzymes involved in lupeol production. Furthermore, auxin treatments apparently induced the expression levels of RcHMGR and RcLUS. Furthermore, we observed that auxin treatment significantly increased lupeol contents, whereas inhibiting auxin transport led to an opposite phenotype. Our study reveals some relationships between hormone activity and lupeol synthesis and might provide a promising way for improving lupeol yields in castor.

Integrated Transcriptome and Metabolome Analysis Reveals an Essential Role for Auxin in Hypocotyl Elongation during End-of-Day Far-Red Treatment of Cucurbita moschata (Duch. Ex Lam.)

Long, robust hypocotyls are important for facilitating greenhouse transplant production. The use of far-red light at the end of the day (end-of-day far-red, EOD-FR) is known to prompt hypocotyl elongation, but the mechanism of EOD-FR-mediated hypocotyl elongation in pumpkin remains unclear. Here, we found that hypocotyl length, parenchymal cell size in hypocotyls, and plant IAA levels were significantly greater in pumpkin after EOD-FR treatment. This effect was counteracted by the application of the polar auxin transport inhibitor 1-N-naphthylphthalamic acid. Integrated transcriptomic and metabolomic analysis of pumpkin hypocotyls revealed that the expression of auxin-related genes changed significantly after EOD-FR treatment, and the contents of the auxin biosynthetic precursors tryptophan and indole were also significantly higher. Our results show that auxin plays an essential role in EOD-FR-mediated hypocotyl elongation, shed light on the mechanisms of EOD-FR mediated hypocotyl elongation, and provide a theoretical basis for the use of EOD-FR in facility cultivation.

Auxin regulated metabolic changes underlying sepal retention and development after pollination in spinach

BackgroundPollination accelerate sepal development that enhances plant fitness by protecting seeds in female spinach. This response requires pollination signals that result in the remodeling within the sepal cells for retention and development, but the regulatory mechanism for this response is still unclear. To investigate the early pollination-induced metabolic changes in sepal, we utilize the high-throughput RNA-seq approach.ResultsSpinach variety ‘Cornel 9’ was used for differentially expressed gene analysis followed by experiments of auxin analog and auxin inhibitor treatments. We first compared the candidate transcripts expressed differentially at different time points (12H, 48H, and 96H) after pollination and detected significant difference in Trp-dependent auxin biosynthesis and auxin modulation and transduction process. Furthermore, several auxin regulatory pathways i.e. cell division, cell wall expansion, and biogenesis were activated from pollination to early developmental symptoms in sepals following pollination. To further confirm the role auxin genes play in the sepal development, auxin analog (2, 4-D; IAA) and auxin transport inhibitor (NPA) with different concentrations gradient were sprayed to the spinach unpollinated and pollinated flowers, respectively. NPA treatment resulted in auxin transport weakening that led to inhibition of sepal development at concentration 0.1 and 1 mM after pollination. 2, 4-D and IAA treatment to unpollinated flowers resulted in sepal development at lower concentration but wilting at higher concentration.ConclusionWe hypothesized that sepal retention and development might have associated with auxin homeostasis that regulates the sepal size by modulating associated pathways. These findings advanced the understanding of this unusual phenomenon of sepal growth instead of abscission after pollination in spinach.

Mode of Action of 1-Naphthylphthalamic Acid in Conspicuous Local Stem Swelling of Succulent Plant, Bryophyllum calycinum: Relevance to the Aspects of Its Histological Observation and Comprehensive Analyses of Plant Hormones.

The mode of action of 1-naphthylphthalamic acid (NPA) to induce conspicuous local stem swelling in the area of its application to the growing internode in intact Bryophyllum calycinum was studied based on the aspects of histological observation and comprehensive analyses of plant hormones. Histological analyses revealed that NPA induced an increase in cell size and numerous cell divisions in the cortex and pith, respectively, compared to untreated stem. In the area of NPA application, vascular tissues had significantly wider cambial zones consisting of 5–6 cell layers, whereas phloem and xylem seemed not to be affected. This indicates that stem swelling in the area of NPA application is caused by stimulation of cell division and cell enlargement mainly in the cambial zone, cortex, and pith. Comprehensive analyses of plant hormones revealed that NPA substantially increased endogenous levels of indole-3-acetic acid (IAA) in the swelling area. NPA also increased endogenous levels of cytokinins, jasmonic acid, and its precursor, 12-oxo-phytodienoic acid, but did not increase abscisic acid and gibberellin levels. It was shown, using radiolabeled 14C-IAA, that NPA applied to the middle of internode segments had little effect on polar auxin transport, while 2,3,5-triiodobenzoic acid substantially inhibited it. These results strongly suggest that NPA induces changes in endogenous levels of plant hormones, such as IAA, cytokinins, and jasmonic acid, and their hormonal crosstalk results in a conspicuous local stem swelling. The possible different mode of action of NPA from other polar auxin transport inhibitors in succulent plants is extensively discussed.

The mechanism of auxin transport in lateral root spacing

The mechanism of auxin transport in lateral root spacing

A Role for Auxin in Triggering Lamina Outgrowth of Unifacial Leaves.

A common morphological feature of typical angiosperms is the patterning of lateral organs along primary axes of asymmetry-a proximodistal, a mediolateral, and an adaxial-abaxial axis. Angiosperm leaves usually have distinct adaxial-abaxial identity, which is required for the development of a flat shape. By contrast, many unifacial leaves, consisting of only the abaxial side, show a flattened morphology. This implicates a unique mechanism that allows leaf flattening independent of adaxial-abaxial identity. In this study, we report a role for auxin in outgrowth of unifacial leaves. In two closely related unifacial-leaved species of Juncaceae, Juncus prismatocarpus with flattened leaves, and Juncus wallichianus with transversally radialized leaves, the auxin-responsive gene GLYCOSIDE HYDROLASE3 displayed spatially different expression patterns within leaf primordia. Treatment of J. prismatocarpus seedlings with exogenous auxin or auxin transport inhibitors, which disturb endogenous auxin distribution, eliminated leaf flatness, resulting in a transversally radialized morphology. These treatments did not affect the radialized morphology of leaves of J. wallichianus. Moreover, elimination of leaf flatness by these treatments accompanied dysregulated expression of genetic factors needed to specify the leaf central-marginal polarity in J. prismatocarpus. The findings imply that lamina outgrowth of unifacial leaves relies on proper placement of auxin, which might induce initial leaf flattening and subsequently act to specify leaf polarity, promoting further flattening growth of leaves.

Plant egg cell fate determination depends on its exact position in female gametophyte

Plant fertilization involves both an egg cell, which fuses with a sperm cell, and synergid cells, which guide pollen tubes for sperm cell delivery. Therefore, egg and synergid cell functional specifications are prerequisites for successful fertilization. However, how the egg and synergid cells, referred to as the "egg apparatus," derived from one mother cell develop into distinct cell types remains an unanswered question. In this report, we show that the final position of the nuclei in female gametophyte determines the cell fate of the egg apparatus. We established a live imaging system to visualize the dynamics of nuclear positioning and cell identity establishment in the female gametophyte. We observed that free nuclei should migrate to a specific position before egg apparatus specialization. Artificial changing in the nuclear position on disturbance of the actin cytoskeleton, either in vitro or in vivo, could reset the cell fate of the egg apparatus. We also found that nuclei of the same origin moved to different positions and then showed different cell identities, whereas nuclei of different origins moved to the same position showed the same cell identity, indicating that the final positions of the nuclei, rather than specific nucleus lineage, play critical roles in the egg apparatus specification. Furthermore, the active auxin level was higher in the egg cell than in synergid cells. Auxin transport inhibitor could decrease the auxin level in egg cells and impair egg cell identity, suggesting that directional and accurate auxin distribution likely acts as a positional cue for egg apparatus specialization.

Crosstalk between auxin and gibberellin during stalk elongation in flowering Chinese cabbage

Plant growth and development are tightly regulated by phytohormones. However, little is known about the interaction between auxin and gibberellin acid (GA) during flower stalk elongation and how it is directly related to organ formation. Therefore, the effects of indole acetic acid (IAA) and GA3 treatments and their interaction on flower stalk elongation in flowering Chinese cabbage were investigated. The growth of flowering Chinese cabbage is regulated by IAA and GA3, and the opposite results were observed after treatments with uniconazole (GA synthesis inhibitor) and N-1-naphthylphthalamic acid (NPA) (auxin transport inhibitor). Anatomical analysis of the pith region in stalks revealed that IAA promoted expansion via signal transduction and transport pathways. GA3 regulated the elongation of flower stalks by controlling GA synthesis and partially controlling the IAA signaling pathway. GA3 also had a stronger effect on stalk elongation than IAA. The results of qRT-PCR and histological analysis revealed that GA3 and IAA induced the expansion of cell walls by activating the expression of genes encoding cell wall structural proteins such as Expansin (EXP). These findings provide new insights into the mechanism of stalk formation regulated by the combination of IAA and GA3.

Comprehensive Analysis and Expression Profiling of PIN, AUX/LAX, and ABCB Auxin Transporter Gene Families in Solanum tuberosum under Phytohormone Stimuli and Abiotic Stresses

Simple SummaryIn this study, we provide comprehensive information on auxin transporter gene families in potato, including basic parameters, chromosomal distribution, phylogeny, co-expression network analysis, gene structure, tissue-specific expression patterns, subcellular localization, transcription analysis under exogenous hormone stimuli and abiotic stresses, and cis-regulatory element prediction. The responsiveness of auxin transporter family genes to auxin and polar auxin transport inhibitors implied their possible roles in auxin homoeostasis and redistribution. Additionally, the differential expression levels of auxin transporter family genes in response to abscisic acid and abiotic stresses suggested their specific adaptive mechanisms on tolerance to various environmental stimuli. Promoter cis-regulatory element description analyses indicated that a number of cis-regulatory elements within the promoters of auxin transporter genes in potato were targeted by relevant transcription factors to respond to diverse stresses. We are confident that our results provide a foundation for a better understanding of auxin transporters in potato, as we have demonstrated the biological significance of this family of genes in hormone signaling and adaption to environmental stresses.Auxin is the only plant hormone that exhibits transport polarity mediated by three families: auxin resistant (AUX) 1/like AUX1 (LAX) influx carriers, pin-formed (PIN) efflux carriers, and ATP-binding cassette B (ABCB) influx/efflux carriers. Extensive studies about the biological functions of auxin transporter genes have been reported in model plants. Information regarding these genes in potato remains scarce. Here, we conducted a comprehensive analysis of auxin transporter gene families in potato to examine genomic distributions, phylogeny, co-expression analysis, gene structure and subcellular localization, and expression profiling using bioinformatics tools and qRT-PCR analysis. From these analyses, 5 StLAXs, 10 StPINs, and 22 StABCBs were identified in the potato genome and distributed in 10 of 18 gene modules correlating to the development of various tissues. Transient expression experiments indicated that three representative auxin transporters showed plasma membrane localizations. The responsiveness to auxin and auxin transport inhibitors implied their possible roles in mediating intercellular auxin homoeostasis and redistribution. The differential expression under abscisic acid and abiotic stresses indicated their specific adaptive mechanisms regulating tolerance to environmental stimuli. A large number of auxin-responsive and stress-related cis-elements within their promoters could account for their responsiveness to diverse stresses. Our study aimed to understand the biological significance of potato auxin transporters in hormone signaling and tolerance to environmental stresses.

N-Glycosylation is Important for Root Growth Under Alkaline Stress by Involving Auxin Transport and Proton Efflux

Summary Asparagine (N)-linked glycosylation is a major modification of glycoproteins, and then class I α-mannosidases (MNS) are involved in early N-glycan processing which play crucial roles in root growth and stress response. However, their roles in alkaline stress are unclear. Using the α-mannosidases deficient Arabidopsis lines, we found that the root growth inhibition in mns1 mns2 mns3 were partially rescued under alkaline condition. N-glycoproteomic analysis revealed that N-glycoproteins associated with auxin action were changed by alkaline stress. Our previous studies have linked the alkaline adaptation of Arabidopsis to auxin transport and proton extrusion. Consistently, conversion of proton flux from inward to outward and inhibition of root growth by auxin transport inhibitor indicated the impacts of proton extrusion and auxin transport on alkaline-maintained root growth in mns1 mns2 mns3. Therefore, we suggested that N-glycosylation is important for root growth under alkaline stress probably by involving auxin transport and proton efflux.

Maize transcription factor ZmEREB20 enhanced salt tolerance in transgenic Arabidopsis

Soil salinity severely limits agricultural crop production worldwide. As one of the biggest plant specific transcription factor families, AP2/ERF members have been extensively studied to regulate plant growth, development and stress responses. However, the role of AP2/ERF family in maize salt tolerance remains largely unknown. In this study, we identified a maize AP2-ERF family member ZmEREB20 as a positive salinity responsive gene. Overexpression of ZmEREB20in Arabidopsis enhanced ABA sensitivity and resulted in delayed seed germination under salt stress through regulating ABA and GA related genes. ZmEREB20 overexpression lines also showed higher survival rates with elevated ROS scavenging toward high salinity. Furthermore, root hair growth inhibition by salt stress was markedly rescued in ZmEREB20 overexpression lines. Auxin transport inhibitor TIBA drastically enhanced root hair growth in ZmEREB20 overexpression Arabidopsis under salt stress, together with the increased expression of auxin-related genes, ion transporter genes and root hair growth genes by RNA-seq analysis. ZmEREB20 positively regulated salt tolerance through the molecular mechanism associated with hormone signaling, ROS scavenging and root hair plasticity, proving the potential target for crop breeding to improve salt resistance.

Naphthylphthalamic acid associates with and inhibits PIN auxin transporters

N-1-naphthylphthalamic acid (NPA) is a key inhibitor of directional (polar) transport of the hormone auxin in plants. For decades, it has been a pivotal tool in elucidating the unique polar auxin transport-based processes underlying plant growth and development. Its exact mode of action has long been sought after and is still being debated, with prevailing mechanistic schemes describing only indirect connections between NPA and the main transporters responsible for directional transport, namely PIN auxin exporters. Here we present data supporting a model in which NPA associates with PINs in a more direct manner than hitherto postulated. We show that NPA inhibits PIN activity in a heterologous oocyte system and that expression of NPA-sensitive PINs in plant, yeast, and oocyte membranes leads to specific saturable NPA binding. We thus propose that PINs are a bona fide NPA target. This offers a straightforward molecular basis for NPA inhibition of PIN-dependent auxin transport and a logical parsimonious explanation for the known physiological effects of NPA on plant growth, as well as an alternative hypothesis to interpret past and future results. We also introduce PIN dimerization and describe an effect of NPA on this, suggesting that NPA binding could be exploited to gain insights into structural aspects of PINs related to their transport mechanism.

Loss of Arabidopsis matrix metalloproteinase-5 affects root development and root bacterial communities during drought stress.

Matrix metalloproteinases (MMPs) are zinc‐dependent endo‐peptidases that in mammals are known to be involved in remodeling the extracellular matrix (ECM) in developmental and pathological processes. In this study, we report At5‐MMP of Arabidopsis thaliana to be important for root development and root bacterial communities. At5‐MMP is mainly localized in the root vasculature and lateral root, an At5‐MMP T‐DNA insertion mutant (mmp5 KO) showed reduced root growth and a lower number of root apexes, causing reduced water uptake from the soil. Subsequently, mmp5 KO is sensitive to drought stress. Inhibited auxin transport was accompanied with resistance to indole‐3‐acetic acid (IAA), 2, 4‐dichlorophenoxyacetic acid (2, 4‐D), and 1‐naphthaleneacetic acid (NAA). The content of endogenous abscisic acid (ABA) was lower in roots of mmp5 KO than in wild type. Genes responsive to ABA as well as genes encoding enzymes of the proline biosynthesis were expressed to a lower extent in mmp5 KO than in wild type. Moreover, drought stress modulated root‐associated bacterial communities of mmp5 KO: the number of Actinobacteria increased. Therefore, At5‐MMP modulates auxin/ABA signaling rendering the plant sensitive to drought stress and recruiting differential root bacterial communities.

Flavonol‐mediated stabilization of PIN efflux complexes regulates polar auxin transport

The transport of auxin controls the rate, direction and localization of plant growth and development. The course of auxin transport is defined by the polar subcellular localization of the PIN proteins, a family of auxin efflux transporters. However, little is known about the composition and regulation of the PIN protein complex. Here, using blue‐native PAGE and quantitative mass spectrometry, we identify native PIN core transport units as homo‐ and heteromers assembled from PIN1, PIN2, PIN3, PIN4 and PIN7 subunits only. Furthermore, we show that endogenous flavonols stabilize PIN dimers to regulate auxin efflux in the same way as does the auxin transport inhibitor 1‐naphthylphthalamic acid (NPA). This inhibitory mechanism is counteracted both by the natural auxin indole‐3‐acetic acid and by phosphomimetic amino acids introduced into the PIN1 cytoplasmic domain. Our results lend mechanistic insights into an endogenous control mechanism which regulates PIN function and opens the way for a deeper understanding of the protein environment and regulation of the polar auxin transport complex.

Plant Growth Alterations in Halophyte Prosopis strombulifera (Lam.) Benth. and Natrophile Glycine max (L.) Merril in Response to Salinity and Changes in Auxin Transport

The effects of changes in auxin transport in response to different osmotic potentials (Ψo) were analyzed in the roots and leaves of two leguminous species with different degrees of salt tolerance: the halophyte Prosopis strombulifera (Lam.) Benth. and the glycophyte Glycine max L. The potentials were generated with sodium chloride (NaCl), sodium sulfate (Na2SO4), and the iso-osmotic mixture of both salts (NaCl + Na2SO4). The values evaluated were −1, −1.8, and −2.6 MPa for P. strombulifera, and −0.47, −0.69, and −0.87 MPa for G. max. In addition, the plants were sprayed with 2,3,5-triiodobenzoic acid (TIBA), an auxin transport inhibitor. The parameters measured included root length, shoot height, number of leaves, relative membrane permeability in roots, and endogenous levels of abscisic acid (ABA), indole-3-acetic acid (IAA), and zeatin (Z). Significant responses were observed at the lowest potentials evaluated (−2.6 MPa for P. strombulifera and −0.89 MPa for G. max). Treatment with Na2SO4 inhibited plant growth more than the others, increased the relative membrane permeability, and enhanced ABA production to its highest levels. These toxic effects were slightly reversed in the presence of iso-osmotic mixture. TIBA brought about impaired development in shoots and roots, the highest values for membrane permeability, alterations in the distribution of endogenous ABA, IAA, and Z, as well as harmful effects on the growth response of both species. In general, the alterations in auxin transport intensified the effects of salinity, which confirms the essential role of this mechanism in plants under salt stress or in normal, non-stressful conditions.

Intrinsic Cell Polarity Coupled to Growth Axis Formation in Tobacco BY-2 Cells

SummarySeveral plant proteins are preferentially localized to one end of a cell, allowing a polarity to be assigned to the cell. These cell polarity proteins often exhibit coordinated patterns between neighboring cells, termed tissue cell polarity. Tissue cell polarity is widespread in plants and can influence how cells grow, divide, and differentiate [1, 2, 3, 4, 5]. However, it is unclear whether cell polarity is established through cell-intrinsic or -extrinsic mechanisms and how polarity is coupled to growth. To address these issues, we analyzed the behavior of a tissue cell polarity protein BASL (BREAKING OF ASYMMETRY IN THE STOMATAL LINEAGE) in the simplifying context of cultured cell filaments and in protoplasts before and during regeneration. We show that BASL is polarly localized when ectopically expressed in tobacco BY-2 cell cultures. Ectopic BASL is found preferentially at the developing tips of cell filaments, likely marking a polarized molecular address. Polarity can shift during the cell cycle and is resistant to treatment with microtubule, actin or auxin transport inhibitors. BASL also exhibits polar localization in spherical protoplasts, in contrast to other polarity proteins so far tested. BASL polarity within protoplasts is dynamic and resistant to auxin transport inhibitors. As protoplasts regenerate, polarity remains dynamic in isotropically growing cells but becomes fixed in anisotropic cells and aligns with the axis of cell growth. Our findings suggest that plant cells have an intrinsic ability to polarize and that environmental or developmental cues may act by biasing the direction of this polarity and thus the orientation of anisotropic growth.