Biology subjects

Lacombe, B.

Publications and source records attributed to Lacombe, B..

7 recordsLinked to original sources

bioRxiv2026

Biuret inhibits Arabidopsis root growth through an active, reversible, and genetically tractable developmental response

Biuret, a nitrogen-rich by-product of urea and a common contaminant of urea-based fertilisers, has long been considered a passive phytotoxin, affecting plant performances. Yet its effects on root development and the existence of endogenous mechanisms of perception or tolerance remain largely uncharacterised. Here we combined physiological, developmental, genetic and transcriptomic approaches to investigate the response of Arabidopsis thaliana to biuret. Biuret inhibited primary root growth in a dose-dependent manner by reducing meristematic cell division rather than cell elongation, and concomitantly impaired shoot growth by limiting leaf expansion. This root inhibition was reversible upon biuret removal and was accompanied by increased auxin-responsive (DR5) and decreased cytokinin-responsive (TCS) outputs at the root apex, consistent with a regulated remodelling of meristem activity rather than purely cumulative damage. A forward genetic screen identified the biuret-resistant mutant bir29, which sustained root and inflorescence development under inhibitory concentrations. Using {superscript 1}N-labelled biuret, we showed that resistance occurred without any change in biuret influx or accumulation, uncoupling sensitivity from exposure. Whole-genome transcriptomics revealed that bir29 fails to execute the wild-type response, neither repressing the cell-cycle machinery nor deploying the stress-associated programme induced by biuret. Genetic characterisation linked resistance to multiple genomic loci required for full resistance. Together, the results indicate that biuret triggers an active, reversible and genetically tractable developmental response, suggesting that this xenobiotic compound is integrated into endogenous signalling networks. Significance StatementBiuret, a poorly metabolised contaminant of urea fertilisers, is generally regarded as a passive phytotoxin, yet we show that it inhibits Arabidopsis root growth through a reversible and genetically tractable developmental response, accompanied by reorganised auxin and cytokinin signalling, rather than through cumulative chemical injury. The isolation of the resistant mutant bir29 suggests that plants integrate this xenobiotic molecule into endogenous signalling networks, reframing biuret as an informative probe of root developmental regulation.

Plant Biology↗

bioRxiv2026

ZmSWEET Sucrose transporters expressed in the endosperm adjacent to the maize embryo are necessary for carbon partitioning and embryo growth

In cereals such as maize, the kernel accumulates large quantities of storage compounds, including carbohydrates, lipids, and proteins, a process that requires tight regulation of nutrient transport. Seeds are composed of distinct tissues: the embryo, the endosperm, and maternal tissues that are symplastically isolated (not connected through plasmodesmata), necessitating specialized nutrient transfer mechanisms. In maize, nutrient transfer from maternal tissues to the endosperm via specialized basal endosperm transfer layer (BETL) cells is well characterized. However, nutrient transfer at the endosperm/embryo interface remains poorly understood. Consequently, the routes by which maternal carbon-derived sugars support embryo growth are still unclear. Our previous transcriptomic profiling uncovered a novel Endosperm domain Adjacent to the embryo Scutellum (EAS) with strong enrichment for transporter genes. Notably, genes encoding three sugar transporters from the SWEET (Sugars Will Eventually be Exported Transporters) family are highly and preferentially expressed in the EAS, suggesting the existence of a specialized sugar transfer mechanism at this interface. We show that the ZmSWEET proteins encoded by these genes are membrane-localized sucrose transporters and are functionally important for kernel development. A gene-edited triple zmsweet14a/14b/15a knock-out mutant exhibits reduced kernel weight and embryo size, significantly decreased embryo oil accumulation at maturity, and altered carbon partitioning within the kernel. In addition to these defects, mutant kernels display a significant reduction in primary root length during germination, indicating either lasting physiological consequences of disrupted sucrose transport during seed development or an additional role for these SWEET transporters during germination. Together, our findings demonstrate that sucrose transport at the endosperm/embryo interface is critical for proper carbon allocation, embryo development, and seed vigor, and identify the EAS as a key functional domain and potential target for improving seed composition.

plant biology↗

bioRxiv2025

Physical and functional interactions of the potassium uptake protein HAK5 and the nitrate transporter NPF6.2 is critical for the mineral nutrition of Arabidopsis

Potassium (K+) starvation induces the expression of gene HAK5 encoding a high-affinity K+ uptake protein, but how plants perceive the K+ status and the signaling intermediaries involved in the response remains largely unknown. To identify key regulators of K+ nutrition in Arabidopsis, a genetic screen was performed using an pHAK5:LUC reporter line, and a mutant showing stable induction of the reporter under K+-sufficient conditions was isolated. Mapping-by-sequencing identified two linked mutations affecting genes involved in K+ and nitrate nutrition, namely a loss-of-function in the K+ uptake channel AKT1 and a gain-of-function allele of the nitrate transporter NPF6.2/NRT1.4 (NPF6.2V210M) that doubled the rate of nitrate transport. We report that the physical interaction of NPF6.2 and HAK5 transport proteins resulted in reciprocal interference. Co-expression in Xenopus oocytes of NPF6.2 with the regulatory kinase CIPK23 or the mutant protein NPF6.2V210M alone inhibited HAK5 transport, whereas HAK5 inhibited nitrate transport by NPF6.2 and NPF6.2V210M. We conclude that mutation NPF6.2V210M enhanced the nutritional defects associated to the loss of AKT1 function through the inhibition of HAK5. These findings evidence an intimate molecular crosstalk between transporters involved in the mineral nutrition of plants. The mutual interference when both transport systems are operative may represent a novel integrative regulatory mechanism in mineral nutrition.

plant biology↗

bioRxiv2021

Chlorhexidine reduced susceptibility associated to tetracycline resistance in clinical isolates of Escherichia coli

Chlorhexidine is a widely used antiseptic in hospital and community healthcare. Decreased susceptibility to this compound has been recently described in Klebsiella pneumoniae and Pseudomonas aeruginosa, together with cross-resistance to colistin. Surprisingly, few data are available for Escherichia coli, the main species responsible for community and healthcare-associated infections. In order to decipher chlorhexidine resistance mechanisms in E. coli, we studied both in vitro derived and clinical isolates through whole-genome sequence analysis. Comparison of strains grown in vitro under chlorhexidine pressure identified mutations in the gene mlaA coding for a phospholipid transport system. Phenotypic analyses of single-gene mutant from the Keio collection confirmed the role of this mutation in the decreased susceptibility to chlorhexidine. However, mutations in mlaA were not found in isolates from large clinical collections. In contrast, genome wide association studies (GWAS) showed that, in clinical strains, chlorhexidine reduced susceptibility was associated with the presence of tetA genes of class B coding for efflux pumps and located in a Tn10 transposon. Construction of recombinant strains in E. coli K-12 confirmed the role of tetA determinant in acquired resistance to both chlorhexidine and tetracycline. Our results reveal two different evolutionary paths leading to chlorhexidine decreased susceptibility: one restricted to in vitro evolution conditions and involving a retrograde phospholipid transport system; the other observed in clinical isolates associated with efflux pump TetA. None of these mechanisms provides cross-resistance to colistin or to the cationic surfactant octenidine. This work demonstrates the GWAS power to identify new resistance mechanisms in bacterial species.

microbiology↗

bioRxiv2018

Revisiting the functional properties of NPF6.3/NRT1.1/CHL1 in xenopus oocytes

Within the Arabidopsis NPF proteins, most of the characterized nitrate transporters are low-affinity transporters, whereas the functional characterization of NPF6.3/NRT1.1 has revealed interesting transport properties: the transport of nitrate and auxin, the eletrogenicity of the nitrate transport and a dual-affinity transport behavior for nitrate depending on external nitrate concentration. However, some of these properties remained controversial and were challenged here. We functionally express WT NPF6.3/NRT1.1 and some of its mutant in Xenopus oocytes and used a combination of uptake experiments using 15N-labelled nitrate and two-electrode voltage-clamp. In our experimental conditions in xenopus oocytes, in the presence or in the absence of external chloride, NPF6.3/NRT1.1 behaves as a non-electrogenic and pure low-affinity transporter. Moreover, further functional characterization of a NPF6.3/NRT1.1 point mutant, P492L, allowed us to hypothesize that NPF6.3/NRT1.1 is regulated by internal nitrate concentration and that the internal perception site involves the P492 residue.

plant biology↗

bioRxiv2018

HRS1/HHOs GARP transcription factors and reactive oxygen species are regulators of Arabidopsis nitrogen starvation response.

Plants need to cope with strong variations in the nitrogen content of the soil solution. Although many molecular actors are being discovered concerning how plants perceive NO3- provision, it is less clear how plants recognize a lack of Nitrogen. Indeed, following N removal plants activate their Nitrogen Starvation Response (NSR) being characterized in particular by the activation of very high affinity nitrate transport systems (NRT2.4, NRT2.5) and other sentinel genes such as GDH3. Here we show using a combination of functional genomics (via TF perturbation) and molecular physiology studies, that the GARP Transcription Factors (TFs) belonging the HHO sub-family are important regulators of the NSR through two potential mechanisms. First, HHOs directly repress NRT2.4 and NRT2.5 high-affinity nitrate transporters. Genotypes affected in HHO genes (mutants and overexpressors) display modified high-affinity nitrate transport activities opening interesting perspectives in biotechnology applications. Second, we show that Reactive Oxygen Species (ROS) are important to control NSR in wild type plants and that HRS1 and HHO1 overexpressors are affected in their ROS content, defining a potential feedforward branch of the signaling pathway. Taken together our results define two new classes of molecular actors in the control of NSR including ROS and the first transcription factors to date. This work (i) opens perspectives on a poorly understood nutrient related signaling pathway, and (ii) defines targets for molecular breeding of plants with enhanced NO3- uptake.

plant biology↗

bioRxiv2018

Root Responses to Heterogeneous Nitrate Availability are Mediated by trans-Zeatin in Arabidopsis Shoots

Plants are subjected to variable nitrogen (N) availability including frequent spatial nitrate (NO3-) heterogeneity in soil. Thus, plants constantly adapt their genome expression and root physiology in order to optimize N acquisition from this heterogeneous source. These adaptations rely on a complex and long-distance root-shoot-root signaling network that is still largely unknown. Here, we used a combination of reverse genetics, transcriptomic analysis, NO3- uptake experiments and hormone profiling under conditions of homogeneous or heterogeneous NO3- availability to characterize the systemic signaling involved. We demonstrate the important role of the trans-zeatin form of cytokinin (CK) in shoots, in particular using a mutant altered for ABCG14-mediated trans-zeatin-translocation from the root to the shoot, in mediating: (i) rapid long distance N-demand signaling and (ii) long term functional adaptations to heterogeneous NO3- supply, including changes in NO3- transport capacity and root growth modifications. We also provide insights into the potential CK-dependent and independent shoot-to-root signals involved in root adaptation to heterogeneous N availability.

plant biology↗