Biology subjects

Erban, A.

Publications and source records attributed to Erban, A..

10 recordsLinked to original sources

bioRxiv2025

Insights into CPSFL1 Induced Membrane Dynamics: A Multifaceted Regulator Linking Vesicle Formation to Thylakoid Biogenesis

Light drives plant life through photosynthesis, a process that takes place in the thylakoid membrane of the chloroplast, an organelle of cyanobacterial origin. The formation of thylakoid membranes within the chloroplast involves the eukaryote-specific factor CHLOROPLAST SEC14 LIKE PROTEIN 1 (CPSFL1), which shares strong sequence homology with the vesicle trafficking regulator SEC14. CSPFL1 is essential for vesicle formation, yet its specific molecular function in this process has remained unclear. In this study, we characterized CSPFL1 functions both in vitro and in vivo. Using a minimal membrane system of giant unilamellar vesicles (GUVs), we show that CPSFL1 alone can induce vesiculation. This process is mediated by lipid binding and membrane deformation, driven by curvature sensing and lipid-protein electrostatics. When expressed in the prokaryote E. coli, the eukaryote-specific CSPFL1 induces membrane curvature and vesicle formation. Plastid CPSFL1 co-purifies with vesicular structures. Lipid compositional analysis of CPSFL1-induced vesicles from bacteria reveals the presence of quinone precursors as cargo, linking CSPFL-mediated vesicle formation to prenylquinone transport. Together, our data suggest that during plant evolution, the eukaryotic vesicle formation system was co-opted for the transport of membrane integral metabolites from the inner envelope to the thylakoid membrane.

plant biology↗

bioRxiv2024

Unveiling the potato cultivars with microbiome interactive traits for sustainable agricultural production

Root traits significantly shape rhizosphere microbiomes, yet their interaction with microbes is often overlooked in plant breeding programs. Here, we propose that selecting modern cultivars based on microbiome interactive traits (MITs), such as root biomass, exudate patterns and the rhizosphere microbiome, can enhance agricultural sustainability by interacting effectively with soil microbiomes, which in turn, promotes plant growth and resistance to stress, thereby reducing reliance on synthetic crop protectants. Through a stepwise selection process (in silico and in vitro) that started with approximately 1000 potato genotypes, we chose 51 potato cultivars based on known phenotypical properties and distinct root exudate patterns. We conducted a greenhouse experiment to evaluate their capacity to interact with the soil microbiome and to assess their MITs. Our findings revealed that cultivars significantly influence plant growth, metabolite profiles, and rhizosphere fungal community composition. Moreover, we observed a positive correlation between microbial community diversity and root biomass. Additionally, leaf metabolites were correlated with rhizosphere bacterial composition, supporting the plant holobiont framework. Utilising z-scores, we aggregated all data related to plant growth, metabolomes, and microbiomes, creating a classification of 51 cultivars based on a gradient of MITs. By examining the distribution of low, medium, and high MITs, we identified a group of 11 potato cultivars suitable for further studies to assess their resilience and productivity under low-input production systems. This study provides an in-depth correlation between microbiome and several plant traits across 51 cultivars, offering tools to facilitate and expedite the incorporation of microbiome traits into breeding goals to support sustainable agriculture.

microbiology↗

bioRxiv2024

Morphological, physiological and metabolic responses of diverse barley inbreds to dry down and moderate drought stress

Drought stress alters the metabolic activity, physiological processes, and plant growth and such responses might differ with the intensity of stress. We evaluated the genotypic diversity on plant morphology, photosynthetic responses, metabolite shift and their relationship in diverse barley inbreds under dry down (DD) and moderate drought (MD) stress using 23 genetically diverse parental inbreds of genetic mapping population of barley. MD stress caused a strong growth reduction, while DD stress triggered inhibition of photosynthetic health. We observed that the induced changes occurred in a genotype-dependent manner. Compared to control conditions, the metabolism of simple sugars and polyhydroxy acids increased in MD and DD, while the maximum accumulation of amino acid, lipids and phosphates occurred in DD stress. Accumulation of hexose and metabolites with unknown classification was the metabolic signature of drought tolerant inbreds. The inbreds tolerant to MD originated from the temperate regions while those tolerant to both MD and DD came from semi-arid regions. Low stomata density, reduced water loss and retarded growth under drought stress were the key features of inbreds with better survival capacity under severe dehydration. We identified drought tolerant barley inbreds and our study offers resources for future genetic research on various drought tolerance strategies.

plant biology↗

bioRxiv2024

Positional 13C Enrichment Analysis of Aspartate by GC-MS to Determine PEPC Activity In Vivo

Photoautotrophic organisms fix inorganic carbon (Ci) by two enzymes, ribulose-1,5-bisphosphate carboxylase/oxygenase (RUBISCO) and phosphoenolpyruvate carboxylase (PEPC). RUBISCO assimilates Ci (CO2) into the 1-C position of 3-phosphoglycerate (3PGA). The Calvin-Benson-Basham (CBB) cycle redistributes fixed carbon atoms into 2,3-C2 of the same molecule. PEPC uses phosphoenolpyruvate (PEP) derived from 3PGA and assimilates Ci (HCO3-) into 4-C of oxaloacetate (OAA). 1,2,3-C3 of OAA and of its transaminase product aspartate originate directly from 1,2,3-C3 of 3PGA. Positional isotopologue analysis of aspartate, the main downstream metabolite of OAA in the model cyanobacterium Synechocystis sp. PCC 6803 (Synechocystis), allows differentiation between PEPC, RUBISCO, and CBB cycle activities within one molecule. We explored in source fragmentation of gas chromatography-electron impact ionization-mass spectrometry (GC-EI-MS) at nominal mass resolution and GC-atmospheric pressure chemical ionization-MS (GC-APCI-MS) at high mass resolution. This enabled the determination of fractional 13C enrichment (E13C) at each carbon position of aspartate. Two prevailing GC-MS derivatization methods, i.e. trimethylsilylation and tert-butyldimethylsilylation, were evaluated. The method was validated by 13C-isotopomer mixtures of positional labeled aspartic acid. Combination with dynamic 13CO2 labeling of Synechocystis cultures allowed direct measurements of PEPC activity in vivo alongside analyses of RUBISCO and CBB cycle activities. Accurate quantification of aspartate concentration and positional E13C provided molar Ci assimilation rates during the day and night phases of photoautotrophic Synechocystis cultures. The validated method offers several applications to characterize the photosynthetic Ci fixation in different organisms.

biochemistry↗

bioRxiv2022

Remodelled Ribosomes Synthesise a Specific Proteome in Proliferating Plant Tissue during Cold

Plant acclimation to low temperatures occurs through system-wide mechanisms that include proteome shifts, some of which occur at the level of protein synthesis. All proteins are synthesised by ribosomes. Rather than being monolithic, transcript-to-protein translation machines, ribosomes can be selective and cause effective proteome shifts required for successful temperature acclimation. Here, we use apical root meristems of germinating seedlings of the monocotyledonous plant barley as a model to study changes in protein abundance and synthesis rates during cold acclimation. We measure metabolic and physiological parameters that allow us to compare protein synthesis rates in different physiological states, e.g., in cold acclimation compared to the optimal temperature state. We show that ribosomal proteins are independently synthesised and assembled into ribosomal complexes in root proliferative tissue, and assess how the ribo-proteome shifts during cold may be associated with changes in synthesis and accumulation of macromolecular complexes. We demonstrate that translation initiation is the limiting step during cold acclimation and based on our data propose a model of a ribosomal code that depends on a reconfigured ribosome population, where as a mode of cold acclimation, specific ribosomal protein compositions may confer selective association capabilities between 60S subunits and 48S initiation complexes.

plant biology↗

bioRxiv2022

Convergence and molecular evolution of floral fragrance after independent transitions to self fertilization

The study of the independent evolution of similar characters can highlight important ecological and genetic factors that drive phenotypic evolution. The transition from reproduction by outcrossing to self-fertilization has occurred frequently throughout plant evolution. A common trend in this transition is the reduction of flower features in the selfing lineages, including display size, flower signals and pollinators rewards. These changes are believed to evolve because resources invested in building attractive flowers are reallocated to other fitness functions as the pressures to attract pollinators decrease. We investigated the similarities in the evolution of flower fragrance after independent transitions to self-fertilization in Capsella. We identified a large number of compounds that are similarly changed in different selfer lineages, such that the composition of the flower scent can predict the mating system in this genus. We further demonstrate that the emission of some of these compounds convergently evolved based on mutations in different genes. In one of the Capsella selfing lineages, the loss of {beta}-ocimene emission was caused by a mutation altering subcellular localization of the ortholog of TERPENE SYNTHASE 2 without apparent effects on its biosynthetic activity. This mutation appears to have been selected at the early stage of this selfing lineage establishment through the capture of a variant segregating in the ancestral outcrossing population. The large extent of convergence in the independent evolution of flower scent, together with the evolutionary history and molecular consequences of a causal mutation, suggest that the emission of specific volatiles has important fitness consequences in self-fertilizing plants without obvious energetic benefits.

plant biology↗

bioRxiv2021

PEPc-mediated CO2 assimilation provides carbons to gluconeogenesis and the TCA cycle in both dark-exposed and illuminated guard cells

Evidence suggests that guard cells have higher rate of phosphoenolpyruvate carboxylase (PEPc)-mediated dark CO2 assimilation than mesophyll cells. However, it is unknown which metabolic pathways are activated following dark CO2 assimilation in guard cells. Furthermore, it remains unclear how the metabolic fluxes throughout the tricarboxylic acid (TCA) cycle and associated pathways are regulated in illuminated guard cells. Here we used 13C-HCO3 labelling of tobacco guard cells harvested under continuous dark or during the dark-to-light transition to elucidate principles of metabolic dynamics downstream of CO2 assimilation. Most metabolic changes were similar between dark-exposed and illuminated guard cells. However, illumination increased the 13C-enrichment in sugars and metabolites associated to the TCA cycle. Sucrose was labelled in the dark, but light exposure increased the 13C-labelling into this metabolite. Fumarate was strongly labelled under both dark and light conditions, while illumination increased the 13C-enrichment in pyruvate, succinate and glutamate. Only one 13C was incorporated into malate and citrate in either dark or light conditions. Our results collectively suggest that the PEPc-mediated CO2 assimilation provides carbons for gluconeogenesis, the TCA cycle and glutamate synthesis and that previously stored malate and citrate are used to underpin the specific metabolic requirements of illuminated guard cells. HighlightPEPc-mediated CO2 assimilation provides carbons for gluconeogenesis and the TCA cycle, whilst previously stored malate and citrate are used to underpin the specific metabolic requirements of illuminated guard cells.

plant biology↗

bioRxiv2021

Distinct metabolic flux modes through the tricarboxylic acid cycle in mesophyll and guard cells revealed by GC-MS-based 13C-positional isotopomer analysis

O_LI13C-Metabolic flux analysis (13C-MFA) have greatly contributed to revealing the regulation of plant metabolism. However, mass spectrometry (MS) approaches have hitherto been limited in their power to deduce flux information due to lack of positional information. C_LIO_LIHere we established an MS-based 13C-positional isotopomer labelling approach and performed a multi-species/cell-types analysis based on previous 13C-MFA to compare flux modes through the tricarboxylic acid (TCA) cycle and associated pathways in mesophyll (MCs) and guard cells (GCs). C_LIO_LIBoth cell types showed high 13C-enrichment in pyruvate. However, GCs and sink MCs, but not source MCs showed high 13C-incorporation into Glu/Gln following provision of 13C-sucrose. Only GCs showed higher 13C-enrichment in the carbon 1 atom of Gln, which is derived from PEPc-mediated CO2 fixation. Increased 13C-enrichment in the carbon 1 of Glu was also observed in both trxo1 and ntra ntrb mutants, but not in wild type Arabidopsis plants, following provision of 13C-glucose. C_LIO_LIOur results suggest that the mitochondrial thioredoxin system restricts the fluxes from PEPc and glycolysis to Glu in illuminated MCs and reveal that fluxes throughout the TCA cycle of GCs resemble those of sink MCs but operate different non-cyclic flux modes to support Gln synthesis in the light. C_LI

plant biology↗

bioRxiv2019

The fungal endophyte Fusarium solani provokes differential effects on the energy balance of two Lotus species

Background and aimsThe interactions established between plants and endophytic fungi span a continuum from beneficial to pathogenic associations. The aim of this work was to explore the mechanisms underlying the potentially beneficial effects provoked by a fungal strain in legume species of the genus Lotus.\n\nMethodsThe ability to solubilise phosphorous was evaluated in nine fungal strains isolated from roots of L. tenuis. A selected strain was further assessed for its ability to colonize plant roots in different Lotus species. The effects of the two interactions were assessed by analysis of the photosynthesis, sugar amount, and macronutrient status of leaves and roots.\n\nResultsA fungal isolate identified as Fusarium solani shows the highest phosphate-solubilisation activity and grows endophytically in roots of L. japonicus and L. tenuis. Fungal invasion enhances plant growth in L. japonicus but provokes a contrasting effect in L. tenuis. Photosynthesis, sugars and K content showed a differential effect in both plant species.\n\nConclusionsOur results indicate neither of the plant species evaluated in this work were significantly stressed by F. solani. Thus, the differential responses observed are due to distinct mechanisms involving photosynthesis, potassium homeostasis, and carbohydrate metabolism that are employed by plants to maintain fitness during the endophytic interaction.

microbiology↗

bioRxiv2018

Transcriptome and metabolome analysis provide insights into root and root released organic anion responses to phosphorus deficiency in oat

Root and root-released organic anions play important roles in uptake of phosphorus (P), an essential macronutrient for food production. Oat, ranking sixth in the worlds cereal production, contains valuable nutritional compounds and can withstand poor soil conditions. The aim of this research was to investigate root transcriptional and metabolic responses of oat grown under P-deficient and P-sufficient conditions. We conducted a hydroponic experiment and measured root morphology, organic anions exudation, and analysed changes in the transcriptome and metabolome, to understand oat root adaptation to P deficiency. We found that oat roots showed enhanced citrate and malate exudation after four weeks of P-deficiency. After 10 days of P-deficiency, we identified 9371 differentially expressed transcripts with a two-fold or greater change (p < 0.05): forty-eight sequences predicted to be involved in organic anion biosynthesis and efflux were consistently up-regulated; twenty-four up-regulated transcripts in oat were also found up-regulated upon P starvation in rice and wheat under similar conditions. Phosphorylated metabolites (i.e. glucose-6-phosphate, myo-inositol-phosphate) reduced dramatically, while citrate and malate, some sugars and amino acids increased slightly in P-deficient oat roots. Our data provide new insights into the root responses to P deficiency and root-released organic anions in oat.\n\nHighlightWe found oat- a monocot food crop, showed high exudation rate of citrate under phosphorus deficiency; root transcriptome and metabolome were then investigated to understand oat adaptation to P deficiency.

plant biology↗