Biology subjects

Weber, A. P. M.

Publications and source records attributed to Weber, A. P. M..

At least 19 recordsLinked to original sources

bioRxiv2026

Lysine biosynthesis impairment shapes heat-stress acclimation through metabolic and transcriptional reprogramming in Arabidopsis thaliana

Global warming is increasing the frequency and intensity of high-temperature episodes, limiting plant productivity. However, the molecular mechanisms integrating primary metabolism with the heat stress response remains poorly understood. Here, we show that lysine biosynthesis contributes to the coordination of physiological, metabolic and transcriptional responses to heat stress in Arabidopsis thaliana. We compared wild-type, the lysine-biosynthesis mutant dapat, and the salicylic acid (SA)-biosynthesis and signaling mutants sid2-1 and npr1-3 under prolonged warming (6C above control for 7 days) and heat shock (38C for 6 h), followed by recovery. We assessed growth, gas exchange, photosynthetic performance, free SA, salicylic acid glucoside (SAG), salicylic acid glucose ester (SGE), and total SA content, primary metabolite profiles, heat-stress-responsive gene expression and transcriptome-wide changes by RNA sequencing. Before heat stress, dapat mutant presented a distinct metabolic state, marked by amino-acid accumulation, altered organic-acid profiles, reduced soluble sugars and elevated endogenous SA. This metabolic configuration persisted during prolonged warming, whereas WT and SA-pathway mutants underwent more dynamic reprogramming. Heat shock, by contrast, elicited a more convergent response across genotypes. Despite reduced basal PSII efficiency, dapat maintained photosynthetic performance during prolonged warming and recovered. Its transcriptional response, however, differed from that of WT and SA-pathway mutants: selected heat-responsive genes were constitutively or more strongly expressed, whereas some canonical heat-stress regulators showed weaker induction after heat shock. RNA-seq further revealed a largely conserved core heat-shock response but genotype-dependent regulation of defense, hormone and amino-acid-metabolism programs, particularly during recovery. Together, these findings indicate that impaired DAPAT activity establishes a metabolically primed but energetically constrained state that reshapes gas exchange, photosynthetic acclimation and heat-responsive transcription. Lysine homeostasis therefore emerges as a regulatory node linking primary metabolism and SA accumulation with SA-dependent and SA-independent components of heat-stress acclimation.

plant biology↗

bioRxiv2026

Glycogen deficiency impairs diurnal energy metabolism and cell division in Synechocystis

Diurnal changes in light availability are a defining feature of life on Earth. Photoautotrophic organisms therefore store reduced carbon during the day to sustain energy metabolism at night. In cyanobacteria, glycogen is the primary carbon storage compound and supports both energy homeostasis and stress responses. Although glycogen-deficient Synechocystis strains have been studied previously, how these mutants cope with the loss of the major daytime carbon sink and can sustain themselves during the night remains unclear. Using single-cell microfluidics, transcriptomics, and metabolomics, we show that {Delta}glgC mutants exhibit pronounced light sensitivity. At sub-lethal light intensities, daytime transcriptional responses are dominated by downregulation of photosynthesis-related genes, likely preventing NADPH overaccumulation in the absence of a carbon sink. During the night, mutants display severe energy limitation, characterized by reduced ATP levels, altered redox balance, and depletion of central carbon intermediates. In contrast, fumarate and malate accumulate, indicating enhanced respiratory flux through succinate dehydrogenase. These metabolic constraints lead to extended lag phases and delayed cell divisions after the onset of light, demonstrating that glycogen-deficient cells fail to efficiently reinitiate growth after dawn. Overall, our results as a snapshot of the initial response to diurnal regimes highlight glycogen as a central integrator of diurnal physiology in Synechocystis, coordinating energy metabolism, redox balance, and cell division, with implications for metabolic robustness and the evolutionary constraints shaping (endo)symbiosis. Short summaryGlycogen deficiency disrupts day-night energy and redox homeostasis in Synechocystis, revealing constraints on growth, division, and symbiotic potential.

plant biology↗

bioRxiv2026

Metabolic modeling links leaf anatomy to environment-specific benefits on the C3-C4 spectrum

C4 photosynthesis evolved from the ancestral C3 pathway through coordinated leaf anatomical and metabolic reorganization that concentrates CO2 to reduce photorespiration. Quantitative understanding of these structure-function relationships remains limited. Here we used anatomy-aware metabolic modeling of a mesophyll-bundle sheath cell system to analyze the interdependence between leaf anatomy and photosynthetic metabolism on the C3-C4 spectrum. Our model faithfully recapitulates the transitory steps from C3 to C4 photosynthesis, reveals a crucial role for plasmodesmata in enabling the C3 to C4 transition, and points at potential pre-C2 metabolic states that provide benefits under conditions that favor elevated photorespiration. Incorporating bundle cell suberisation with our model predicts reduction of PSII activity and dominance of the NADP-ME C4 subtype in leaves with suberized bundle sheath cells and proposes a role for oxygen evolution at PSII as a potential driver for this mechanism. Varying bundle sheath leakage and photorespiratory conditions along the C3-C4 spectrum identify conditions under which C3-C4 intermediate photosynthesis provides energetic benefits and underlines the notion of intermediate photosynthesis as a stable evolutionary state. Overall, our study sheds new light on the quantitative relationship between leaf anatomy and metabolism and its interaction with the environment and suggests targets for climate-adaptation in C3 plants.

plant biology↗

bioRxiv2025

Native catalase expression in Arabidopsis thaliana is more than sufficient to limit excess decarboxylation from photorespiratory intermediates

The hydrogen peroxide (H2O2) scavenging enzyme, catalase, plays a critical role in the photorespiratory pathway by maintaining the balance of H2O2, a reactive oxygen species (ROS), in the peroxisome. H2O2 acts as both a signaling molecule and a potential source of ROS depending on its accumulation in the peroxisome. Additionally, H2O2 can also drive non-enzymatic decarboxylation (NED) reactions as well as other decarboxylation reactions, leading to increased CO2 release that is linked to a severe growth phenotype. However, the exact cause of this stunted growth phenotype is not fully understood, and it remains unclear whether the capacity of catalase is critical for minimizing these decarboxylating reactions. Here we elucidate the mechanism behind the decrease in plant growth due to the accumulation of H2O2 from photorespiration using cat2 knock-out lines of Arabidopsis thaliana rescued with transgenic expression lines of Heliobacter pylori catalase. These experiments demonstrated that while one of the three heterologous lines expressing H. pylori catalase isoform (Hp615) had greater catalase activity than cat2-KO and rescued the severe growth and photosynthetic phenotype, its catalase activity was still far below wild type levels. These findings suggest that catalase plays a crucial role in maintaining H2O2 homeostasis within the peroxisome and minimizing decarboxylation reactions, both of which are linked to plant growth. Moreover, once a threshold capacity is reached, increasing catalase capacity further may offer limited benefits in enhancing net carbon fixation. HighlightWe show that peroxisomal catalase is important for maintaining high rates of net carbon fixation associated with plant growth. Native catalase levels in Arabidopsis thaliana are in excess of that which is required to minimize alternative decarboxylation reactions. Therefore, efforts to optimize catalase-mediated degradation of H2O2 may be of limited benefit.

plant biology↗

bioRxiv2025

Impaired lysine biosynthesis drives constitutive energy stress priming and darkness stress responses

Plant survival under prolonged darkness relies on dynamic metabolic reprogramming that redirects limited carbon resources toward mitochondrial respiration and nutrient remobilization. Although amino acids function as alternative respiratory substrates during carbon starvation, how their biosynthesis integrates into this metabolic adjustment remains poorly understood. Here, we investigated the role of lysine biosynthesis using the dapat mutant, which exhibits photoperiod-dependent hypersensitivity to extended darkness. Under short-day conditions, dapat plants exhibited accelerated senescence. reduced photosystem II maximum quantum efficiency, pronounced protein degradation, and accumulation of amino acid, ultimately failing to recover after nine days of darkness. By contrast, survival was largely restored under neutral-day conditions, indicating that restricted carbohydrate reserves during short-day growth exacerbate the mutant phenotype. Remarkably, transcript profiling further revealed that reduced DAPAT activity constitutively activates a catabolic program, including genes associated with amino acid degradation, alternative respiratory pathways, senescence, and autophagy, even under non-stress conditions. Together, these findings identify DAPAT-mediated lysine biosynthesis as a central metabolic hub linking carbon availability to energy and stress signaling. Disruption of this pathway compromises metabolic flexibility and limits the capacity to withstand prolonged carbon starvation. Our results provide new insights into how primary metabolism coordinates stress responses to extended darkness and highlight amino acid biosynthesis as an important component of plant resilience under energy-deprived darkness conditions.

plant biology↗

bioRxiv2025

Dissecting the genetic basis of drought escape across multiple traits in colonizing Arabidopsis thaliana lineages

Drought response in plants is complex, involving integration across a range of physiological processes. However, our knowledge of how different aspects of drought response are linked at the genetic level is limited. We investigated multi-trait adaptation in Arabidopsis thaliana from the Cape Verde Islands (CVI). Using a high-throughput phenotyping platform that minimizes spatial heterogeneity, we measured variation in rosette area, growth rate, leaf color, water-use efficiency (WUE), and stomatal patterning under precisely controlled water conditions. Relative to the Moroccan outgroup, CVI populations evolved to be smaller rosette size with faster growth and reduced WUE, consistent with drought escape adaptation. Genome-wide association mapping revealed evidence for pleiotropy involving MPK12 (WUE, rosette area, growth rate, and leaf color), NHL26 (WUE and leaf color), SUVH4 (stomatal patterning, rosette area, and leaf color), and FRI (WUE and leaf color), along with an enrichment of signals in ABA response. This study advances our knowledge of the genetic mechanisms driving plant adaptation to novel precipitation environments. By identifying key genetic components and their contributions to multi-trait adaptation, our findings offer insights into how plants respond to environmental challenges and contribute to predicting plant responses to future climate change.

genomics↗

bioRxiv2024

Alternatives to photorespiration: A systems-level analysis reveals mechanisms of enhanced plant productivity

Photorespiration causes a significant decrease in crop yield due to mitochondrial decarboxylation. Alternative pathways (APs) have been designed to relocate the decarboxylating step or even fix additional carbon. To improve the success of transferring those engineered APs from model species to crops we must understand how they will interact with metabolism and how the plant physiology affects their performance. Here we used multiple mathematical modelling techniques to analyse and compare existing AP designs. We show that carbon-fixing APs are the most promising candidates to replace native photorespiration in major crop species. Our results demonstrate the different metabolic routes that APs employ to increase yield and which plant physiology can profit most from them. We anticipate our results to guide the design of new APs and to help improve existing ones. Codehttps://gitlab.com/gain4crops/2024-paper

plant biology↗

bioRxiv2024

Joint operation of the CO2 concentrating mechanism and photorespiration in green algae during acclimation to limiting CO2

Due to low availability of CO2 in aquatic environment, microalgae have evolved a CO2 concentrating mechanism (CCM). It has long been thought that operation of CCM would suppress photorespiration by increasing the CO2 concentration at the Rubisco active site, but experimental evidence is scarce. To better explore the function of photorespiration in algae, we first characterized a Chlamydomonas reinhardtii mutant defected in low-CO2 inducible 20 (LCI20) and show that LCI20 is a chloroplast-envelope glutamate/malate transporter playing a role in photorespiration. By monitoring growth and glycolate excretion in mutants deficient in either CCM or photorespiration, we conclude that: i. CCM induction does not depend on photorespiration, ii. glycolate excretion protects algal cells from the toxicity of unmetabolized photorespiratory intermediates, iii. photorespiration is active at low CO2 when the CCM is operational. This work provides a foundation for a better understanding of the carbon cycle in the ocean where significant glycolate concentrations have been found.

plant biology↗

bioRxiv2024

Inactivation of mitochondrial complex IV in Physcomitrium patens reveals the essential role of respiration in coordinating plants metabolism

Photosynthetic organisms use sunlight as energy source but rely on respiration during the night and in non-photosynthetic tissues. Respiration is also active in photosynthetically active cells, where its role is still unclear due to a lack of viable mutants. Plants lacking cytochrome c oxidase (complex IV) activity are generally lethal but were here isolated exploiting the possibility of generating knockout lines through vegetative propagation in the moss Physcomitrium patens. The mutants showed severely impaired growth, with an altered composition of the respiratory apparatus and increased electron transfer through the alternative oxidase. The light phase of photosynthesis remained largely unaffected while the efficiency of carbon fixation was moderately reduced. Transcriptomic and metabolomic analyses showed that the disruption of the cytochrome pathway had broad consequences for carbon and nitrogen metabolism. A major alteration in nitrogen assimilation was observed with a general reduction in amino acid abundance. A partial rescue of the growth could be obtained by growing the plants with an external supply of amino acids but not with sugars, demonstrating that respiration in plant photosynthetic cells plays an essential role at the interface between carbon and nitrogen metabolism and a key role in providing carbon skeletons for amino acid biosynthesis.

plant biology↗

bioRxiv2024

The host genotype actively shapes its microbiome across generations

BackgroundThe microbiome greatly affects health and wellbeing. Evolutionarily, it is doubtful that a host would rely on chance alone to pass on microbial colonization to its offspring. However, the literature currently offers only limited evidence regarding two alternative hypotheses: active microbial shaping by host genetic factors or transmission of a microbial maternal legacy. ResultsTo further dissect the influence of host genetics and maternal inheritance, we collected 2-cell stage embryos from two representative wildtypes, C57BL6/J and BALB/c, and transferred a mixture of both genotype embryos into hybrid recipient mice to be inoculated by an identical microbiome at birth. ConclusionsObserving the offspring for six generations unequivocally emphasizes the impact of host genetic factors over maternal legacy in constant environments, akin to murine laboratory experiments. Interestingly, maternal legacy solely controlled the microbiome in the first offspring generation. However, current evidence supporting maternal legacy has not extended beyond this initial generation, resolving the aforementioned debate. O_FIG O_LINKSMALLFIG WIDTH=197 HEIGHT=200 SRC="FIGDIR/small/584950v2_ufig1.gif" ALT="Figure 5"> View larger version (48K): org.highwire.dtl.DTLVardef@790822org.highwire.dtl.DTLVardef@4f486org.highwire.dtl.DTLVardef@151ab46org.highwire.dtl.DTLVardef@c3ab66_HPS_FORMAT_FIGEXP M_FIG graphical abstract C_FIG

ecology↗

bioRxiv2023

Mitochondrial Apolipoprotein MIC26 is a metabolic rheostat regulating central cellular fuel pathways

Mitochondria play central roles in metabolism and metabolic disorders such as type 2 diabetes. MIC26, a MICOS complex subunit, was linked to diabetes and modulation of lipid metabolism. Yet, the functional role of MIC26 in regulating metabolism under hyperglycemia is not understood. We employed a multi-omics approach combined with functional assays using WT and MIC26 KO cells cultured in normoglycemia or hyperglycemia, mimicking altered nutrient availability. We show that MIC26 has an inhibitory role in glycolysis and cholesterol/lipid metabolism under normoglycemic conditions. Under hyperglycemia, this inhibitory role is reversed demonstrating that MIC26 is critical for metabolic adaptations. This is partially mediated by alterations of mitochondrial metabolite transporters. Furthermore, MIC26 deletion led to a major metabolic rewiring of glutamine utilization as well as oxidative phosphorylation. We propose that MIC26 acts as a metabolic rheostat, that modulates mitochondrial metabolite exchange via regulating mitochondrial cristae, allowing cells to cope with nutrient overload.

cell biology↗

bioRxiv2023

A transcription factor module mediating C2 photosynthesis

C4 photosynthesis has arisen from the ancestral C3 state in over sixty lineages of angio-sperms. It is widely accepted that an early step in C4 evolution is restriction of glycine decarboxylase activity to bundle sheath cells to generate the so-called C2 pathway. In C2 Moricandia species, changes to the cis-regulatory region controlling expression of the P-subunit of GLYCINE DECARBOXYLASE (GLDP) in mesophyll cells enables this trait, but the mechanism underpinning GLDP expression in the bundle sheath is not known. We identify a MYC-MYB transcription factor module previously associated with the control of glucosinolate bio-synthesis as the basis of GLDP expression in bundle sheath cells. In C3 Arabidopsis thaliana this module drives GLDP expression in bundle sheath cells along with as yet unidentified factors driving expression in mesophyll cells. In the C2 species Moricandia arvensis, GLDP expression is lost from mesophyll cells and the MYC-MYB dependent expression in the bundle sheath is revealed. Evolution of C2 photosynthesis is thus associated with a MYC-MYB based transcriptional network already present in the C3 state. This work identifies a molecular genetic mechanism underlying the bundle sheath accumulation of glycine decarboxylase required for C2 photosynthesis and thus a foundational step in the evolution of C4 photosynthesis.

plant biology↗

bioRxiv2023

PLANT UNCOUPLING MITOCHONDRIAL PROTEIN 2 localizes to the Golgi

Mitochondria act as cellular hubs of energy transformation and metabolite conversion in most eukaryotes. Plant mitochondrial electron transport chains are particularly flexible, featuring alternative components, such as ALTERNATIVE NAD(P)H DEHYDROGENASES and ALTERNATIVE OXIDASES (AOXs), that can bypass proton translocation steps. PLANT UNCOUPLING MITOCHONDRIAL PROTEINS (named PUMPs or plant UCPs) have been identified in plants as homologues of mammalian Uncoupling Proteins (UCPs), and their biochemical and physiological roles have been investigated in the context of mitochondrial energy metabolism. To dissect UCP function in Arabidopsis, the two most conserved (UCP1 and UCP2) have been targeted in recent work by combining mutant lines to circumvent potential functional redundancy in vivo. Such approaches rely on the assumption that both proteins reside in the inner mitochondrial membrane as a prerequisite for functional redundancy. Yet, contradicting results have been reported on UCP2 localization in plants. Here we provide evidence that, conversely to UCP1, which is an abundant inner mitochondrial membrane protein, UCP2 localizes to the Golgi rather than to mitochondria. Based on multiple lines of new and prior evidence, we summarize the consensus view that we have reached and provide an example of how open, critical exchange within the research community is able to constructively address ambiguities. Our observations and considerations provide direction to the ongoing discussion about the functions of UCP proteins. They further offer new perspectives for the study of Golgi membrane transport and subcellular targeting principles of membrane proteins. Since 20 to 30 % of genes in plant genomes are predicted to encode transmembrane proteins and the function of most of those proteins has not been experimentally investigated, we highlight the importance of using independent evidence for localization as a prerequisite for understanding physiological function of membrane proteins.

plant biology↗

bioRxiv2023

The genome of Talinum fruticosum

Research on crassulacean acid metabolism (CAM) has in recent years focused on obligate CAM species, such as Kalanchoe fedtschenkoi and pineapple (Ananas comosus). To fully understand the plasticity of the CAM pathway, its evolutionary trajectory and regulation, genomic resources of additional species, including facultative CAM species are desirable. To this end, we sequenced the genome and full-length transcripts (Iso-Seq) of the facultative CAM dicot Talinum fruticosum. The provided resources may aid in CAM engineering as an approach to improving crop water-use efficiency.

plant biology↗

bioRxiv2023

Helixer--de novo Prediction of Primary Eukaryotic Gene Models Combining Deep Learning and a Hidden Markov Model.

AO_SCPLOWBSTRACTC_SCPLOWGene structural annotation is a critical step in obtaining biological knowledge from genome sequences yet remains a major challenge in genomics projects. Current de novo Hidden Markov Models are limited in their capacity to model biological complexity; while current pipelines are resource-intensive and their results vary in quality with the available extrinsic data. Here, we build on our previous work in applying Deep Learning to gene calling to make a fully applicable, fast and user friendly tool for predicting primary gene models from DNA sequence alone. The quality is state-of-the-art, with predictions scoring closer by most measures to the references than to predictions from other de novo tools. Helixers predictions can be used as is or could be integrated in pipelines to boost quality further. Moreover, there is substantial potential for further improvements and advancements in gene calling with Deep Learning. Helixer is open source and available at https://github.com/weberlab-hhu/Helixer A web interface is available at https://www.plabipd.de/helixer_main.html

bioinformatics↗

bioRxiv2023

A genomic panel for studying C3-C4 intermediate photosynthesis in the Brassiceae tribe

Research on C4 and C3-C4 photosynthesis has attracted significant attention because the understanding of the genetic underpinnings of this trait will support the introduction of its characteristics into commercially relevant crop species. We used a panel of 19 taxa of 18 Brassiceae species with different photosynthesis characteristics (C3 and C3-C4) with the following objectives: (i) create draft genome assemblies and annotations, (ii) quantify the level of orthology using synteny maps between all pairs of taxa, (iii) describe the phylogenetic relatedness across all the species, and (iv) track the evolution of C3-C4 intermediate photosynthesis in the Brassiceae tribe. Our results indicate that the draft de novo genome assemblies are of high quality and cover at least 90% of the gene space. Therewith we more than doubled the sampling depth of genomes of the Brassiceae tribe that comprises commercially important as well as biologically interesting species. The gene annotation generated high-quality gene models, and for most genes extensive upstream sequences are available for all taxa, yielding potential to explore variants in regulatory sequences. The genome-based phylogenetic tree of the Brassiceae contained two main clades and indicated that the C3-C4 intermediate photosynthesis has evolved five times independently. Furthermore, our study provides the first genomic support of the hypothesis that Diplotaxis muralis is a natural hybrid of D. tenuifolia and D. viminea. Altogether, the de novo genome assemblies and the annotations reported in this study are a valuable resource for research on the evolution of C3-C4 intermediate photosynthesis.

genomics↗

bioRxiv2022

Brassicaceae display diverse photorespiratory carbon recapturing mechanisms

Carbon concentrating mechanisms enhance the carboxylase efficiency of the central photosynthetic enzyme rubisco by providing supra-atmospheric concentrations of CO2 in its surrounding. In the C4 photosynthesis pathway, this is achieved by combinatory changes to leaf biochemistry and anatomy. Carbon concentration by the photorespiratory glycine shuttle requires fewer and less complex modifications. It could represent an early step during evolution from C3 to C4 photosynthesis and an inspiration for engineering approaches. Plants displaying CO2 compensation points between 10 to 40 ppm are therefore often termed C3-C4 intermediates. In the present study, we perform a physiological, biochemical and anatomical survey of a large number of Brassicaceae species to better understand the C3-C4 intermediate phenotype. Our phylogenetic analysis suggested that C3-C4 metabolism evolved up to five times independently in the Brassicaceae. The efficiency of the pathways showed considerable variation between the species but also within species. Centripetal accumulation of organelles in the bundle sheath was consistently observed in all C3-C4 classified accessions indicating a crucial role of anatomical features for CO2 concentrating pathways. Leaf metabolite patterns were strongly influenced by the individual plant accessions, but accumulation of photorespiratory shuttle metabolites glycine and serine was generally observed. Analysis of PEPC activities suggests that C4-like shuttles have not evolve in the investigated Brassicaceae. HighlightOur physiological, biochemical and anatomical survey of Brassicaceae revels multiple evolution of C3-C4 intermediacy connected to variation in photorespiratory carbon recapturing efficiency and a distinct C3-C4 bundle sheath anatomy.

plant biology↗

bioRxiv2022

Transposable elements contribute to the establishment of the glycine shuttle in Brassicaceae species

C3-C4 intermediate photosynthesis has evolved at least five times convergently in the Brassicaceae, despite this family lacking bona fide C4 species. The establishment of this carbon concentrating mechanism is known to require a complex suite of ultrastructural modifications as well as changes in spatial expression patterns, which are both thought to be underpinned by a reconfiguration of existing gene-regulatory networks. However, to date, the mechanisms which underpin the reconfiguration of these gene networks are largely unknown. In this study, we used a pan-genomic association approach to identify genomic features that could confer differential gene expression toward the C3-C4 intermediate state by analysing eight C3 species and seven C3-C4 species from five independent origins in the Brassicaceae. We found a strong correlation between transposable element (TE) insertions in cis-regulatory regions and the C3-C4 intermediacy. Specifically, our study revealed 113 gene models in which presence of a TE within a gene correlates with C3-C4 intermediate photosynthesis. In this set, genes involved in the photorespiratory glycine shuttle are enriched, including the glycine decarboxylase P-protein whose expression domain undergoes a spatial shift during the transition to C3-C4 photosynthesis. When further interrogating this gene, we discovered independent TE insertions in its upstream region which we conclude to be responsible for causing the spatial shift in GLDP1 gene expression. Our findings hint at a pivotal role of TEs in the evolution of C3-C4 intermediacy, especially in mediating differential spatial gene expression.

plant biology↗