Root Growth Inhibition Research Articles

Drought Shifts Sorghum Root Metabolite and Microbiome Profiles and Enriches for Pipecolic Acid

Plant-associated microbial communities shift in composition as a result of environmental perturbations, such as drought. It has been shown that Actinobacteria are enriched in plant roots and rhizospheres during drought stress. However, the correlations between microbiome dynamics and plant response to drought are poorly understood. Here we apply a combination of bacterial community composition analysis and plant metabolite profiling in Sorghum bicolor roots, rhizospheres, and soil during drought and drought recovery to investigate potential contributions of host metabolism to shifts in bacterial composition. Our results provide a detailed view of metabolic shifts across the plant root during drought and show that the response to rewatering differs between root and soil; additionally, we identify drought-responsive metabolites that are highly correlated with the observed changes in Actinobacteria abundance. Furthermore, we find that pipecolic acid is a drought-enriched metabolite in sorghum roots, and that exogenous application of pipecolic acid inhibits root growth. Finally, we show that this activity functions independent of the systemic acquired resistance pathway and has the potential to impact Actinobacterial taxa within the root microbiome.

Effect of Waste Cooking Oil-Based Composite Materials on Radish Growth and Biochemical Responses

Waste cooking oil poses a serious threat to human health and the environment, both in households and in larger communities. One of the applications of waste cooking oil is composite materials called vegeblocks, which can be used for construction purposes. These composites are formed by the process of polymerisation, esterification and polyesterification. The resulting materials exhibit mechanical strength in line with the requirements for paving blocks. Composite materials that have been annealed for a minimum of 20 h at 200 °C or higher have the highest tensile strength (above 5 MPa). In contrast, composites with the highest flexural strength were obtained after processing at 210 °C for 16 h. The Saxa 2 variety showed the greatest inhibition of storage root growth (almost 43% compared to the control sample), as well as stimulation of root and leaf blade growth (by a maximum of 61.5% and 53.5%, respectively, compared to the control sample). The composite obtained from the maximum process parameters resulted in significant growth of both the root and the green part of both radish varieties by up to 35%. The study showed that the presence of vegeblocks in the plants causes stress conditions, resulting in increased peroxidase content compared to the control sample. The presence of the oil composite in the soil did not increase the amount of catalase in the radish, and even a reduction was observed compared to the control sample.

Selenite reduced cadmium uptake, interfered signal transduction of endogenous phytohormones, and stimulated secretion of tartaric acid based on a combined analysis of non-invasive micro-test technique, transcriptome and metabolome

Selenium (Se) can reduce uptake and translocation of cadmium (Cd) in plants via plenty of ways, including regulation of root morphology. However, the underlying mechanisms on how Se will regulate root morphology under metal(loid) stresses are not fully illustrated. To fill up this knowledge gap, we investigated the effects of 0.5 mg L−1 selenite (Se(IV)) on root exudates, root morphology, root endogenous hormones, and Cd uptake efficiency of rice under the 1 mg L−1 Cd stress condition. The results showed that Se(IV) significantly reduced shoot and root Cd concentrations, and decreased Cd uptake efficiency via root hairs determined by a non–invasive micro-test (NMT) technology. When compared to the 1 mg L−1 Cd (Cd1) treatment, addition of 0.5 mg L−1 Se(IV) (1) significantly reduced root surface area and tip numbers, and non–significantly reduced root length, but significantly enhanced root diameter and root volume; (2) significantly enhanced concentrations of tartaric acid in the root exudate solution, root auxin (IAA) and root jasmonic acid (JA) via a UHPLC or a HPLC analysis; (3) significantly up-regulated metabolites correlated with synthesis of IAA, JA, gibberellin (GA), and salicylic acid, such as GA53, M–SA, (+/−)7–epi–JA, and derivatives of tryptophan and indole in the metabolome analysis. However, results of transcriptome analysis showed that (1) no upregulated differentially expressed genes (DEGs) were enriched in IAA synthesis; (2) some upregulated DEGs were found to be enriched in JA and GA53 synthesis pathways. In summary, although Se(IV) stimulated the synthesis of IAA, JA, and GA53, it significantly inhibited root growth mainly by 1) affecting signal transduction of IAA and GA; 2) altering IAA polar transport and homeostasis; and 3) regulating DEGs including SAUR32, SAUR36, SAUR76, OsSub33, OsEXPA8, OsEXPA18, and Os6bglu24.

Structure and functional divergence of PIP peptide family revealed by functional studies on PIP1 and PIP2 in Arabidopsis thaliana

PAMP-induced secreted peptide (PIP), one of the small post-translationally modified peptides (PTMPs), plays a crucial role in plant development and stress tolerance. However, little is known about functional divergence among this peptide family. Here, we studied the evolution of the PIP family in 23 plant species (10 monocotyledons and 13 dicotyledons from 7 families) and their functional divergence in Arabidopsis. A total of 128 putative PIP precursors were identified and classified into two subfamilies through phylogenetic analysis. Functional studies on AtPIP1 which represents Clade I family and AtPIP2 which represents Clade II family have shown that AtPIP2 displayed stronger immunity induction activity but weaker root growth inhibition than AtPIP1 in Arabidopsis. Transcriptome analysis of Arabidopsis seedlings treated with AtPIP1 and AtPIP2 showed that differential genes for both polypeptides were significantly enriched in similar plant defense pathways. However, Co-expression and Protein-protein interaction (PPI) analysis showed that the functions of AtprePIP2 co-expressed genes were more enriched in plant defense pathways than AtprePIP1. Molecular docking results show that AtPIP1 binds to RLK7 receptor with a more stable free energy and less binding area than AtPIP2, while hydrogen bond transfer occurs at the SGP motif position. The above results suggest that the PIP family have undergone functional divergence during evolution. Collectively, this work illustrates the relationship between PIP structure and function using Arabidopsis PIP as an example, and provides new insights into the current understanding between growth inhibition and immune responses which may be correlated but not fully coupled.

Ethylene and ROS mediate root growth inhibition induced by the endocrine disruptor bisphenol A (BPA)

Bisphenol A (BPA) functions as a detrimental substance that disrupts the endocrine system in animals while also impeding the growth and development of plants. In our previous study, we demonstrated that BPA hinders the growth of roots in Arabidopsis by diminishing cell division and elongation, which is ascribed to the increased accumulation and redistribution of auxin. Here, we examined the mediation of ROS and ethylene in BPA-induced auxin accumulation and root growth inhibition. BPA enhanced ROS levels, and ROS increased auxin contents but reduced cell division activity and the expression of EXPA8 involved in root elongation. ROS scavenger treatment reversed BPA-triggered root growth retardation, auxin accumulation, and cell division inhibition. In addition, BPA induced ethylene, and ethylene synthesis inhibitor treatment reversed BPA-triggered root growth retardation and auxin accumulation. Taken together, ROS and ethylene are involved in BPA-inhibited cell elongation and cell division by mediating auxin accumulation and redistribution.

Cytotoxic and genotoxic effects of triphenyl phosphate on root tip cells of Allium cepa L.

Triphenyl phosphate (TPhP) is a tri-ester of phosphoric acid and phenol. It is used in products such as polyvinyl chloride, hydraulic fluids, polymers, photographic films and printed circuit cards as a flame retardant. It is also widely used in consumer products like electric and electronic devices, textiles and building necessaries. TPhP can diffuse into its surrounding environment easily, because it does not form a chemical bond with polymeric materials. Therefore, bio-monitoring of such compounds is needed for possible environmental and human health effects. In this study, we evaluated genotoxicity and cytotoxicity of TPhP on Allium cepa on the cells of root tips. A 10 mg/mL stock solution of TPhP was dissolved in DMSO and dilutions of 2, 4, 6, 8 and 10 mg/mL was made in distilled water. Onion bulbs, rooted in test tubes, were exposed to these concentrations of TPhP for 24 h. Distilled water was used as a negative control, 0.7% hydrogen peroxide was used as positive control, and 0.5% DMSO was used as solvent control. Significant inhibition of onion root growth was observed following treatment with the 6, 8 and 10 mg/mL TPhP ranges in comparison with the negative and solvent control groups (p < 0.05). Furthermore, in the TPhP treatment groups, total chromosome aberration ratios were significantly high in comparison with the controls (p < 0.05). These results suggest that TPhP have cytotoxic and genotoxic effects on A. cepa root tips.

Extracellular nitric oxide sustains root surface redox activity and growth under sudden flooding-induced hypoxic conditions in barley root tips

Main conclusionNitric oxide sustains root tip surface redox activity and restricts lipid peroxidation-triggered cell death in the root tips.In order to gain more insight into the involvement of nitric oxide (NO) in plant response to sudden flooding-induced hypoxic stress, we studied the effect of two NO donors, sodium nitroprusside and S-nitroso-L-glutathione, on short-term partial submergence-induced root growth inhibition, alteration in root surface redox activity, lipid peroxidation and cell death in two barley cultivars (cv.) at their early seedling stage. The short-term hypoxic stress induces root growth arrest in cv. Karmel, accompanied by increased lipid peroxidation and cell death. By contrast, in cv. Slaven, short-term hypoxic conditions cause only reduced root growth rate, associated with elevated extracellular NO level in the root tips. The root tip surface redox activity decreases with the increasing timespan of hypoxic conditions in both cultivars; however, this decrease in redox activity started earlier and was greater in the cv. Karmel in comparison with cv. Slaven. Application of NO donors during hypoxic stress sustains the root redox activity and eliminates the hypoxia-induced lipid peroxidation, accompanied by a partial restoration of root growth after short-term hypoxic stress. These results suggest that extracellular NO plays a key role in maintaining the root tip surface redox activity and in the restriction of lipid peroxidation and cell death under short-term hypoxic stress in the root tips of barley seedlings.

AtIAR1 is a Zn transporter that regulates auxin metabolism in Arabidopsis thaliana.

Root growth in Arabidopsis is inhibited by exogenous auxin-amino acid conjugates, and mutants resistant to one such conjugate [indole-3-acetic acid (IAA)-Ala] map to a gene (AtIAR1) that is a member of a metal transporter family. Here, we test the hypothesis that AtIAR1 controls the hydrolysis of stored conjugated auxin to free auxin through zinc transport. AtIAR1 complements a yeast mutant sensitive to zinc, but not manganese- or iron-sensitive mutants, and the transporter is predicted to be localized to the endoplasmic reticulum/Golgi in plants. A previously identified Atiar1 mutant and a non-expressed T-DNA mutant both exhibit altered auxin metabolism, including decreased IAA-glucose conjugate levels in zinc-deficient conditions and insensitivity to the growth effect of exogenous IAA-Ala conjugates. At a high concentration of zinc, wild-type plants show a novel enhanced response to root growth inhibition by exogenous IAA-Ala which is disrupted in both Atiar1 mutants. Furthermore, both Atiar1 mutants show changes in auxin-related phenotypes, including lateral root density and hypocotyl length. The findings therefore suggest a role for AtIAR1 in controlling zinc release from the secretory system, where zinc homeostasis plays a key role in regulation of auxin metabolism and plant growth regulation.

Enhancing Soybean Yield: The Synergy of Sulfur and Rhizobia Inoculation.

Sulfur deficiency severely limits soybean growth, inhibiting the rhizobia nitrogenase and soybean protein synthesis. This study assessed the impact of sulfur fertilization and rhizobia inoculation on soybean growth and nitrogen fixation through bacterial culture and hydroponic experiments. We selected three rhizobia strains for bacterial cultures and used six sulfur levels. The test demonstrated severe inhibition of Rhizobium USDA110 growth without sulfur. In hydroponic experiment, we employed five sulfur levels with USDA110 as the inoculum strain. Soybean growth, nitrogen fixation, yield, and root morphology-related parameters, and root nodule growth, were significantly inhibited without sulfur. Following Rhizobium inoculation, low sulfur concentrations (0.15-0.60 mM) stimulated early-stage (V9) root growth and increased shoot nitrogen accumulation, but inhibited root growth at R5 stage. Furthermore, Rhizobium inoculation notably enhanced soybean growth, nitrogen fixation, and yield, especially within the recommended low sulfur concentration range (0.15-0.30 mM). The maximum nodule nitrogenase activity at R5 stage and highest yield was recorded at a 0.3 mM sulfur concentration with Rhizobium inoculation, which was 9.51-1222.07% higher than other treatments. These findings highlight that low sulfur concentration and rhizobia inoculation enhance soybean growth, nitrogen fixation, and yield but reduce soybean root efficacy, increasing reliance on root nodules.

A novel PGPR strain homologous to Beijerinckia fluminensis induces biochemical and molecular changes involved in Arabidopsis thaliana salt tolerance

The use of PGPR is widely accepted as a promising tool for a more sustainable agricultural production and improved plant abiotic stress resistance. This study tested the ability of PVr_9, a novel bacterial strain, homologous to Beijerinckia fluminensis, to increase salt stress tolerance in A. thaliana. In vitro plantlets inoculated with PVr_9 and treated with 150 mM NaCl showed a reduction in primary root growth inhibition compared to uninoculated ones, and a leaf area significantly less affected by salt. Furthermore, salt-stressed PVr_9-inoculated plants had low ROS and 8-oxo-dG, osmolytes, and ABA content along with a modulation in antioxidant enzymatic activities. A significant decrease in Na+ in the leaves and a corresponding increase in the roots were also observed in salt-stressed inoculated plants. SOS1, NHX1 genes involved in plant salt tolerance, were up-regulated in PVr_9-inoculated plants, while different MYB genes involved in salt stress signal response were down-regulated in both roots and shoots. Thus, PVr_9 was able to increase salt tolerance in A. thaliana, thereby suggesting a role in ion homeostasis by reducing salt stress rather than inhibiting total Na+ uptake. These results showed a possible molecular mechanism of crosstalk between PVr_9 and plant roots to enhance salt tolerance, and highlighted this bacterium as a promising PGPR for field applications on agronomical crops.

Integrated transcriptomic and metabolomic analysis reveals the potential mechanisms underlying indium-induced inhibition of root elongation in wheat plants

Soil contamination by indium, an emerging contaminant from electronics, has a negative impact on crop growth. Inhibition of root growth serves as a valuable biomarker for predicting indium phytotoxicity. Therefore, elucidating the molecular mechanisms underlying indium-induced root damage is essential for developing strategies to mitigate its harmful effects. Our transcriptomic findings revealed that indium affects the expression of numerous genes related to cell wall composition and metabolism in wheat roots. Morphological and compositional analysis revealed that indium induced a 2.9-fold thickening and a 17.5 % increase in the content of cell walls in wheat roots. Untargeted metabolomics indicated a substantial upregulation of the phenylpropanoid biosynthesis pathway. As the major end product of phenylpropanoid metabolism, lignin significantly accumulated in root cell walls after indium exposure. Together with increased lignin precursors, enhanced activity of lignin biosynthesis-related enzymes was observed. Moreover, analysis of the monomeric content and composition of lignin revealed a significant enrichment of p-hydroxyphenyl (H) and syringyl (S) units in root cell walls under indium stress. The present study contributes to the existing knowledge of indium toxicity. It provides valuable insights for developing sustainable solutions to address the challenges posed by electronic waste and indium contamination on agroecosystems.

Uncovering root compaction response mechanisms: new insights and opportunities.

Compaction disrupts soil structure, reducing root growth, nutrient and water uptake, gas exchange, and microbial growth. Root growth inhibition by soil compaction was originally thought to reflect the impact of mechanical impedance and reduced water availability. However, using a novel gas diffusion-based mechanism employing the hormone ethylene, recent research has revealed that plant roots sense soil compaction. Non-compacted soil features highly interconnected pore spaces that facilitate diffusion of gases such as ethylene which are released by root tips. In contrast, soil compaction stress disrupts the pore network, causing ethylene to accumulate around root tips and trigger growth arrest. Genetically disrupting ethylene signalling causes roots to become much less sensitive to compaction stress. Such new understanding about the molecular sensing mechanism and emerging root anatomical traits provides novel opportunities to develop crops resistant to soil compaction by targeting key genes and their signalling pathways. This expert view discusses these recent advances and the molecular mechanisms associated with root-soil compaction responses.

Boron deficiency responses in maize (Zea mays L.) roots

AbstractBackgroundBoron (B) is an essential micronutrient for plants. Dicot plants respond to insufficient B supply by altering root architecture and root hair growth. How root systems of rather low‐B demanding monocot species such as maize (Zea mays L.) respond to B deficiency in terra has not been experimentally resolved, yet.AimsThe study aims to investigate root responses and their physiological consequences under B deficiency during the vegetative growth of maize.MethodsB73 wild‐type (WT) maize and its root hairless rth3 mutant were grown under varying B supply conditions in soil columns and in an automated root phenotyping facility. Biomass data, root system architecture traits, the mineral elemental composition and molecular B‐deficiency responses were quantified.ResultsThough having very low leaf B concentrations, no major growth deficit, apart from chlorotic stripes on leaves, was recorded on maize root and shoot development, with or without root hairs, on B‐deficient conditions. Although leaf B concentration of the rth3 mutant is significantly lower under B‐deficient and under B‐surplus conditions compared to the WT, the rth3 mutant neither developed a larger total root length, more fine roots nor displayed a higher expression of B uptake transporters as compensatory adaptations.ConclusionsStrikingly, maize plants did neither react with an inhibited root growth nor by a compensatory root foraging behaviour to severe B‐deficient in terra growth conditions. This is rather atypical for plants. The performance and altered leaf B concentrations of rth3 mutants may be biased by secondary effects, such as an overall reduced root growth.

Receptor Plants Alleviated Allelopathic Stress from Invasive Chenopodium ambrosioides L. by Upregulating the Production and Autophagy of Their Root Border Cells.

Chenopodium ambrosioides L. is an invasive plant native to the Neotropics that has seriously threatened the ecological security of China, and allelopathy is one of the mechanisms underlying its successful invasion. Maize (Zea mays L.) and soybean (Glycine max (L.) Merr.), as the main food crops, are usually affected by C. ambrosioides in their planting areas. The purpose of this study was to investigate the ultrastructure, autophagy, and release-related gene expression of receptor plant root border cells (RBCs) after exposure to volatile oil from C. ambrosioides and its main component α-terpene, which were studied using maize and soybean as receptor plants. The volatiles inhibited root growth and promoted a brief increase in the number of RBCs. As the volatile concentration increased, the organelles in RBCs were gradually destroyed, and intracellular autophagosomes were produced and continuously increased in number. Transcriptomic analysis revealed that genes involved in the synthesis of the plasma membrane and cell wall components in receptor root cells were significantly up-regulated, particularly those related to cell wall polysaccharide synthesis. Meanwhile, polygalacturonase and pectin methylesterases (PME) exhibited up-regulated expression, and PME activity also increased. The contribution of α-terpene to this allelopathic effect of C. ambrosioides volatile oil exceeded 70%. Based on these results, receptor plant root tips may increase the synthesis of cell wall substances while degrading the intercellular layer, accelerating the generation and release of RBCs. Meanwhile, their cells survived through autophagy of RBCs, indicating the key role of RBCs in alleviating allelopathic stress from C. ambrosioides volatiles.

The allelochemical potential of an invasive ornamental plant, the Indian blanket flower (Gaillardia pulchella Foug.)

AbstractGlobal spread of invasive plant species threatens biodiversity significantly, with a particularly high presence of invasives in the Asteraceae family. This is partly due to their wide use as ornamental plants and their rapid reproduction and allelopathy. The Indian blanket flower (Gaillardia pulchella) is a native North American species widely used as an ornamental plant and has become invasive in other countries. Although it contains bioactive compounds, its allelopathic impact on other plants has been largely unexplored. We conducted in vitro tests on oilseed rape (Brassica napus) germination and early growth to assess blanket flower's allelopathy. Seeds were exposed to aqueous extracts at three different concentrations (10%, 20%, and 40%). The 40% extract significantly inhibited germination during the treatment. All three concentrations inhibited root growth, with the 40% treatment showing this effect from the second day. In contrast, the extracts promoted significantly shoot elongation. Fresh weight of seedlings was not affected by the extract, but the 10% treatment resulted in higher mass. These findings provide evidence of blanket flower's allelopathic effects, shedding light on its invasive potential.

Genotoxic Potential, Total Flavonoid and Phenolic Contents of Solanum anomalum Thonn. Ex Schumach Leaf

Solanum anomalum, a medicinal plant, used in ethnomedicine for their antimalarial, antidiabetic, and antiphlogistic activities was investigated for cytotoxic and genotoxic effects on the root meristem cells of Allium cepa. The extract and fractions was also screened for total flavonoid and phenol content using standard methods. Onion bulbs were exposed to 2.5 mg/mL, 5mg/mL, and 10 mg/mL concentrations of the leaf extract for macroscopic and microscopic analysis. Tap water was used as a negative control and Methotrexate (0.1 mg/mL) as a positive control. There was statistically significant (P &lt; 0.05) inhibition of root growth depending on concentration by the extract when compared with the negative control group. All the tested concentrations of the extract were observed to have cytotoxic effects on cell division in A. cepa. The extract- induced chromosomal aberrations and micronuclei (MNC) formations in A. cepa root tip cells were significant (p&lt;0.05) when compared with control group. The extract treatment further induced cell death, ghost cells, cells membrane damage, and binucleated cells. The dichloromethane fraction was found to contain the highest level of total flavonoid and phenol followed by the crude extract. These results suggest that Solanum anomalum leaf extract possess cytotoxic and genotoxic effects on A. cepa which is as a results of its phytochemical constituents.

The Brassica napus boron deficient inflorescence transcriptome resembles a wounding and infection response.

Oilseed rape and other crops of Brassica napus have a high demand for boron (B). Boron deficiencies result in the inhibition of root growth, and eventually premature flower abortion. Understanding the genetic mechanisms underlying flower abortion in B-limiting conditions could provide the basis to enhance B-efficiency and prevent B-deficiency-related yield losses. In this study, we assessed transcriptomic responses to B-deficiency in diverse inflorescence tissues at multiple time points of soil-grown plants that were phenotypically unaffected by B-deficiency until early flowering. Whilst transcript levels of known B transporters were higher in B-deficient samples, these remained remarkably stable as the duration of B-deficiency increased. Meanwhile, GO-term enrichment analysis indicated a growing response resembling that of a pathogen or pest attack, escalating to a huge transcriptome response in shoot heads at mid-flowering. Grouping differentially expressed genes within this tissue into MapMan functional bins indicated enrichment of genes related to wounding, jasmonic acid and WRKY transcription factors. Individual candidate genes for controlling the "flowering-without-seed-setting" phenotype from within MapMan biotic stress bins include those of the metacaspase family, which have been implicated in orchestrating programmed cell death. Overall temporal expression patterns observed here imply a dynamic response to B-deficiency, first increasing expression of B transporters before recruiting various biotic stress-related pathways to coordinate targeted cell death, likely in response to as yet unidentified B-deficiency induced damage-associated molecular patterns (DAMPs). This response indicates new pathways to target and dissect to control B-deficiency-induced flower abortion and to develop more B-efficient crops.

Jarin-1, an inhibitor of JA-Ile biosynthesis in Arabidopsis thaliana, acts differently in other plant species

ABSTRACT Jasmonates (JAs), including jasmonic acid (JA) and its biologically active derivative JA-Ile, are lipid-derived plant signaling molecules. They govern plant responses to stresses, such as wounding and insect herbivory. Wounding elicits a rapid increase of JA and JA-Ile levels as well as the expression of JAR1, coding for the enzyme involved in JA-Ile biosynthesis. Endogenous increase and application of JAs, such as MeJA, a JA methylester, result in increased defense levels, often accompanied by diminished growth. A JA-Ile biosynthesis inhibitor, jarin-1, was shown to exclusively inhibit the JA-conjugating enzyme JAR1 in Arabidopsis thaliana. To investigate whether jarin-1 does function similarly in other plants, we tested this in Medicago truncatula, Solanum lycopersicum, and Brassica nigra seedlings in a root growth inhibition assay. Application of jarin-1 alleviated the inhibition of root growth after MeJA application in M. truncatula seedlings, proving that jarin-1 is biologically active in M. truncatula. Jarin-1 did not show, however, a similar effect in S. lycopersicum and B. nigra seedlings treated with MeJA. Even JA-Ile levels were not affected by application of jarin-1 in wounded leaf disks from S. lycopersicum. Based on these results, we conclude that the effect of jarin-1 is highly species-specific. Researchers intending to use jarin-1 for studying the function of JAR1 or JA-Ile in their model plants, must test its functionality before use.

The response of Arabidopsis to the apocarotenoid β-cyclocitric acid reveals a role for SIAMESE-RELATED 5 in root development and drought tolerance

Abstract New regulatory functions in plant development and environmental stress responses have recently emerged for a number of apocarotenoids produced by enzymatic or nonenzymatic oxidation of carotenoids. β-Cyclocitric acid (β-CCA) is one such compound derived from β-carotene, which triggers defense mechanisms leading to a marked enhancement of plant tolerance to drought stress. We show here that this response is associated with an inhibition of root growth affecting both root cell elongation and division. Remarkably, β-CCA selectively induced cell cycle inhibitors of the SIAMESE-RELATED (SMR) family, especially SMR5, in root tip cells. Overexpression of the SMR5 gene in Arabidopsis induced molecular and physiological changes that mimicked in large part the effects of β-CCA. In particular, the SMR5 overexpressors exhibited an inhibition of root development and a marked increase in drought tolerance which is not related to stomatal closure. SMR5 up-regulation induced changes in gene expression that strongly overlapped with the β-CCA–induced transcriptomic changes. Both β-CCA and SMR5 led to a down-regulation of many cell cycle activators (cyclins, cyclin-dependent kinases) and a concomitant up-regulation of genes related to water deprivation, cellular detoxification, and biosynthesis of lipid biopolymers such as suberin and lignin. This was correlated with an accumulation of suberin lipid polyesters in the roots and a decrease in nonstomatal leaf transpiration. Taken together, our results identify the β-CCA–inducible and drought-inducible SMR5 gene as a key component of a stress-signaling pathway that reorients root metabolism from growth to multiple defense mechanisms leading to drought tolerance.

The Cytokinins BAP and 2-iP Modulate Different Molecular Mechanisms on Shoot Proliferation and Root Development in Lemongrass (Cymbopogon citratus).

The known activities of cytokinins (CKs) are promoting shoot multiplication, root growth inhibition, and delaying senescence. 6-Benzylaminopurine (BAP) has been the most effective CK to induce shoot proliferation in cereal and grasses. Previously, we reported that in lemongrass (Cymbopogon citratus) micropropagation, BAP 10 µM induces high shoot proliferation, while the natural CK 6-(γ,γ-Dimethylallylamino)purine (2-iP) 10 µM shows less pronounced effects and developed rooting. To understand the molecular mechanisms involved, we perform a protein-protein interaction (PPI) network based on the genes of Brachypodium distachyon involved in shoot proliferation/repression, cell cycle, stem cell maintenance, auxin response factors, and CK signaling to analyze the molecular mechanisms in BAP versus 2-iP plants. A different pattern of gene expression was observed between BAP- versus 2-iP-treated plants. In shoots derived from BAP, we found upregulated genes that have already been demonstrated to be involved in de novo shoot proliferation development in several plant species; CK receptors (AHK3, ARR1), stem cell maintenance (STM, REV and CLV3), cell cycle regulation (CDKA-CYCD3 complex), as well as the auxin response factor (ARF5) and CK metabolism (CKX1). In contrast, in the 2-iP culture medium, there was an upregulation of genes involved in shoot repression (BRC1, MAX3), ARR4, a type A-response regulator (RR), and auxin metabolism (SHY2).