Biology subjects

Estevez, J. M.

Publications and source records attributed to Estevez, J. M..

7 recordsLinked to original sources

bioRxiv2026

ABF2 and bZIP2 remodel root system architecture under combined phosphate deficiency and salinity in Arabidopsis

Climate change is degrading arable soils and imposing chronic abiotic stress on plants. As the primary interface with the soil, the root system is especially vulnerable, and plants respond by remodelling their root system architecture (RSA). Most studies have examined single stresses in isolation and so fail to capture the multifactorial environments in which plants actually grow. Rising salinity impairs development by compromising cellular integrity and triggering cytotoxic responses, and it simultaneously restricts uptake of phosphorus, a macronutrient required for nucleic acid and protein synthesis. Combined stresses can amplify these effects, yet how plants adjust RSA to cope with them remains poorly understood. Here we performed a meta-analysis of public transcriptomic datasets for salinity and phosphate deficiency in Arabidopsis thaliana. Integrating these data with protein-protein interaction modelling and DNA-binding (DAP-seq) analyses, we identified two basic leucine zipper transcription factors, ABF2 and bZIP2, as candidate integrators of the combined-stress response. Characterization of abf2, bzip2 and double mutants revealed RSA alterations and allowed us to identify downstream targets of each factor. Our findings offer new insight into root adaptation under combined stress and represent one of the few comprehensive analyses of how multiple abiotic stressors jointly shape root development and plant fitness.

plant biology↗

bioRxiv2026

ER-located Ca2+ ATPase ACA2 regulates Ca2+ cytoplasmic pool linked to root hair growth in Arabidopsis thaliana

Root hairs (RH) are excellent model systems for studying cell size and polarity since they elongate several hundred-fold their original size. Their tip growth is regulated by both intrinsic and environmental signals and is associated with the existence of a highly controlled cytoplasmic tip Ca{superscript 2} gradient, whose disruption impairs RH development. The molecular mechanisms underlying the Ca2+ homeostasis fine tuning and the Ca2+ organellar contributions to the cytoplasmic pool remain unclear. In the model plant Arabidopsis thaliana, many efflux routes are present, including those that employ Ca2+-pumps from the Autoinhibited Ca2+-ATPase (ACA) family. Here, we identified that the ER localized ACA2, and to a lower extent ACA7, are crucial ACAs required to control RH growth. By using genetically encoded Ca2+ biosensors we showed that Ca2+-dynamics are compromised in the aca2-2 mutant, having lower cytosolic Ca2+ concentration [Ca2+]cyt and growth rate, showing an altered homeostatic calcium setpoint compared to Col-0. Accordingly, the ACA2 mutation changed the dynamics of [Ca2+]cyt oscillations coupled to growth rate, inducing longer periods and more regular oscillations in the dominant high-frequency range (around 22 s), and slower oscillations (around 1 min) in the low-frequency range. Finally, expression of ACA2 with changes in four putative Ca2+ binding residues (ACA2{Delta}Ca2+) failed to rescue the RH growth phenotype in the aca2-2 mutant. Collectively, our findings indicate that ER-localized ACA2 and possibly ACA7 are crucial for modulating cytoplasmic Ca2+ signals, possibly composing a critical part of a negative feedback loop, and their absence leads to impairments in RH cell elongation.

plant biology↗

bioRxiv2022

Cell surface receptor kinase FERONIA linked to nutrient sensor TORC1 signaling controls root hair growth at low temperature in Arabidopsis thaliana

Root hairs (RH) are excellent model systems for studying cell size and polarity since they elongate several hundred-fold their original size. Their tip growth is determined both by intrinsic and environmental signals. Although nutrient availability and temperature are key factors for a sustained plant growth, the molecular mechanisms underlying their sensing and downstream signaling pathways remain unclear. Here, we identified that low temperature (10{degrees}C) triggers a strong RH elongation response involving the cell surface receptor kinase FERONIA (FER) and the nutrient sensing TOR Complex 1 (TORC). In this study, we found that FER is required to perceive limited nutrient availability caused by low temperature. FER interacts with and activates TORC downstream components to trigger RH growth. In addition, the small GTPase Rho-related protein from plants 2 (ROP2) is also involved in this RH growth response linking FER and TOR. We also found that limited nitrogen nutrient availability can mimic the RH growth response at 10{degrees}C in a NRT1.1-dependent manner. These results uncover a molecular mechanism by which a central hub composed by FER-ROP2-TORC is involved in the control of RH elongation under low temperature and nitrogen deficiency.

plant biology↗

bioRxiv2021

Ethylene signaling increases reactive oxygen species accumulation to drive root hair initiation in Arabidopsis thaliana

Root hair initiation is a highly regulated aspect of root development. The plant hormone, ethylene, and its precursor, 1-amino-cyclopropane-1-carboxylic acid (ACC), induce formation and elongation of root hairs. Using confocal microscopy paired with redox biosensors and dyes, we demonstrated that treatments that elevate ethylene levels led to increased hydrogen peroxide accumulation in hair cells prior to root hair formation. In two ethylene-insensitive mutants, etr1-3 and ein3/eil1, there was no increase in root hair number or ROS accumulation. Conversely, etr1-7, a constitutive ethylene signaling receptor mutant, has increased root hair formation and ROS accumulation like ethylene-treated Col-0 seedlings. The caprice and werewolf transcription factor mutants have decreased and elevated ROS levels, which are correlated with levels of root hair initiation. The rhd2-6 mutant, with a defect in the gene encoding a ROS synthesizing Respiratory Burst Oxidase Homolog C (RBOHC) and the prx44-2 mutant defective in a class III peroxidase, showed impaired ethylene-dependent ROS synthesis and root hair formation and EIN3/EIL1 dependent transcriptional regulation. Together, these results indicate that ethylene increases ROS accumulation through RBOHC and PRX44 to drive root hair formation. SUMMARY STATEMENTThe gaseous hormone ethylene increases root hair initiation by elevating reactive oxygen species (ROS) in trichoblast cells. Genetic and biochemical approaches identified ethylene-regulated ROS producing enzymes that drive root hair initiation.

plant biology↗

bioRxiv2020

PRX1, PRX44, and PRX73 are Class-III extensin-related peroxidases that modulates root hair growth in Arabidopsis thaliana

O_LIRoot hair cells are important sensors of soil conditions. Expanding several hundred times their original size, root hairs grow towards and absorb water-soluble nutrients. This rapid growth is oscillatory and is mediated by continuous remodelling of the cell wall. Root hair cell walls contain polysaccharides and hydroxyproline-rich glycoproteins including extensins (EXTs). C_LIO_LIClass-III peroxidases (PRXs) are secreted into the apoplastic space and are thought to trigger either cell wall loosening, mediated by oxygen radical species, or polymerization of cell wall components, including the Tyr-mediated assembly of EXT networks (EXT-PRXs). The precise role of these EXT-PRXs is unknown. C_LIO_LIUsing genetic, biochemical, and modeling approaches, we identified and characterized three root hair-specific putative EXT-PRXs, PRX01, PRX44, and PRX73. The triple mutant prx01,44,73 and the PRX44 and PRX73 overexpressors had opposite phenotypes with respect to root hair growth, peroxidase activity and ROS production with a clear impact on cell wall thickness. C_LIO_LIModeling and docking calculations suggested that these three putative EXT-PRXs may interact with non-O-glycosylated sections of EXT peptides that reduce the Tyr-to-Tyr intra-chain distances in EXT aggregates and thereby may enhance Tyr crosslinking. These results suggest that these three putative EXT-PRXs control cell wall properties during the polar expansion of root hair cells. C_LI

plant biology↗

bioRxiv2019

A cell surface O-glycosylated peptide, AGP21, acts on the brassinosteroid pathway and modulates root hair cell fate

Root hairs (RHs) develop from specialized epidermal cells called trichoblasts, whereas epidermal cells that lack RHs are known as atrichoblasts. The mechanism controlling root epidermal cell fate is only partially understood. Root epidermis cell fate is regulated by a transcription factor complex that promotes the expression of the homeodomain protein GLABRA 2 (GL2), which blocks RH development by inhibiting ROOT HAIR DEFECTIVE 6 (RHD6). Suppression of GL2 expression activates RHD6, a series of downstream TFs including ROOT HAIR DEFECTIVE 6 LIKE-4 (RSL4 [Yi et al. 2010]) and their target genes, and causes epidermal cells to develop into RHs. Brassinosteroids (BRs) influence root epidermis cell fate. In the absence of BRs, phosphorylated BIN2 (a Type-II GSK3-like kinase) inhibits a protein complex that directly downregulates GL2 [Chen et al. 2014]. Here, we show that the genetic and pharmacological perturbation of the arabinogalactan peptide (AG) AGP21 in Arabidopsis thaliana, triggers aberrant RH development, similar to that observed in plants with defective BR signaling. We reveal that an O-glycosylated AGP21 peptide, which is positively regulated by BZR1, a transcription factor activated by BR signaling, affects RH cell fate by altering GL2 expression in a BIN2-dependent manner. These results suggest that perturbation of a cell surface AGP disrupts BR responses and inhibits the downstream effect of BIN2 on the RH repressor GL2 in root epidermal cells. In addition, AGP21 also acts in a BR-independent, AGP-dependent mode that together with BIN2 signalling cascade controls RH cell fate. SignificanceIn the plant Arabidopsis thaliana, the root epidermis forms in an alternating pattern atrichoblasts with trichoblast cells that end up developing root hairs (RHs). Atrichoblast cell fate is directly promoted by the transcription factor GLABRA2 (GL2) while the lack of GL2 allows RH formation. The loss of AGP21 peptide triggers an abnormal RH cell fate in two contiguous cells in a similar manner as brassinosteroid (BRs) mutants. In the absence of BR signaling, BIN2 (a GSK3 like-kinase) in a phosphorylated state, downregulate GL2 expression to trigger RH cell fate. The absence of AGP21 is able to repress GL2 expression and activates the expression of RSL4 and EXP7 root hair proteins.

plant biology↗

bioRxiv2017

The Molecular Link Between Auxin And ROS-Mediated Polar Growth

Root hair polar growth is endogenously controlled by auxin and sustained by oscillating levels of reactive oxygen species (ROS). These cells extend several hundred-fold their original size toward signals important for plant survival. Although their final cell size is of fundamental importance, the molecular mechanisms that control it remain largely unknown. Here, we show that ROS production is controlled by the transcription factors RSL4, which in turn is transcriptionally regulated by auxin through several Auxin Responsive Factors (ARFs). In this manner, auxin controls ROS-mediated polar growth by activating RSL4, which then upregulates the expression of genes encoding NADPH oxidases (also known as RBOHs, RESPIRATORY BURST OXIDASE HOMOLOG proteins) and Class-III Peroxidases (PER), which catalyse ROS production. Chemical or genetic interference with the ROS balance or peroxidase activity affect root hair final cell size. Overall, our findings establish a molecular link between auxin regulated ARFs-RSL4 and ROS-mediated polar root hair growth.\n\nSignificance StatementTip-growing root hairs are excellent model systems to decipher the molecular mechanism underlying reactive oxygen species (ROS)-mediated cell elongation. Root hairs are able to expand in response to external signals, increasing several hundred-fold their original size, which is important for survival of the plant. Although their final cell size is of fundamental importance, the molecular mechanisms that control it remain largely unknown. In this study, we propose a molecular mechanism that links the auxin-Auxin Response Factors (ARFs) module to activation of RSL4, which directly targets genes encoding ROS-producing enzymes, such as NADPH oxidases (or RBOHs) and secreted type-III peroxidases (PERs). Activation of these genes impacts apoplastic ROS homeostasis, thereby stimulating root hair cell elongation.

plant biology↗