Biology subjects

Hirt, H.

Publications and source records attributed to Hirt, H..

17 recordsLinked to original sources

bioRxiv2026

Root endophyte sulfur metabolites enhance redox balance and drought tolerance in Arabidopsis

Drought is a major consequence of climate change and significantly limits crop productivity. Plant growth-promoting bacteria offer a promising solution to mitigate drought stress. The root endophyte Pseudomonas argentinensis SA190 has been shown to enhance plant performance under drought stress conditions, but the mechanistic basis of SA190s beneficial effects remained unclear. Given the pivotal role of sulfur-containing compounds in abiotic stress responses, we investigated multiple sulfur-related Arabidopsis mutants under drought stress. We found that SA190 enhances sulfate uptake and promotes glutathione (GSH) accumulation in shoots under stress conditions. SA190 treatment improved the GSH/GSSG ratio, indicating an enhanced redox balance under drought. Selective inhibition of Arabidopsis GSH biosynthesis using buthionine sulfoximine (BSO) confirmed the essential contribution of bacterial GSH to drought stress. In addition, by generation and use of bacterial mutants deficient in the GSH synthesis pathway, we show that the bacteria directly provide Arabidopsis with either GSH or its precursor {gamma}-EC. In summary, SA190 promotes drought tolerance by supplying the host plant with additional GSH thereby maintaining cellular redox homeostasis and enhancing drought stress resilience.

Plant Biology↗

bioRxiv2026

Host specificity in cereal rust fungi is mediated by a conserved glycoside hydrolase family

Non-host resistance refers to the immunity of plant species to virtually all isolates of a potential pathogen and represents an underexplored avenue for breeding and engineering disease resistance. In domesticated and wild barley, cell surface-localized lectin receptor kinases (LecRKs) contribute to determining the host status to leaf rust fungi, which pose a major threat to global cereal production. Here, we identify a conserved family of leaf rust glycoside hydrolases as ligands for these barley LecRKs and show that direct ligand-receptor binding triggers immune responses. This mechanism of pathogen perception is conserved across multiple cereal species and can be functionally transferred between them. We also uncover previously uncharacterized recognition specificities among distinct LecRK variants, expanding the repertoire of LecRK-mediated rust pathogen detection. Our findings define a molecular mechanism underlying non-host resistance in cereals and provide a basis for harnessing non-host rust resistance across diverse crop-pathogen systems.

plant biology↗

bioRxiv2026

Turnip mosaic virus co-opts host RNA methylation to orchestrate plant infection

N6-methyladenosine (m6A) is a key RNA modification that regulates transcript stability and translation. However, its function in plant viruses remains largely unclear. Here, we show that the positive-sense single-stranded +ssRNA Turnip mosaic virus (TuMV) relies on the host m6A machinery to support efficient infection. Our findings uncover a previously unrecognized nuclear phase in the TuMV life cycle, during which viral RNA undergoes extensive methylation by host enzymes. We identify a complex and non-canonical methylation landscape on the TuMV genome, where canonical DRACH motifs are embedded within clusters of additional virus-specific non-canonical m6A sites. Notably, we also detect the presence of another RNA modification, m5C (5-methylcytosine), in close proximity to m6A-marked regions. This coordinated methylation landscape appears to be critical for efficient viral polyprotein synthesis. In its absence, the virus displays aberrant methylation and reduced infectivity as observed in m6A writer and reader mutants. Based on this, we propose a "seeding" model in which initial m6A deposition at canonical sites nucleates the formation of a broader network of m6A and m5C marks, likely guided by RNA structure rather than sequence motifs alone.

plant biology↗

bioRxiv2025

Darwin21 Genome Database: A Curated Whole-Genome Repository of Endophytic Bacteria from Desert Plants

Microbial communities associated with desert plants play a pivotal role in enhancing host survival under extreme environmental stressors, including drought, salinity, and nutrient limitation. The Darwin21 Endophytic Microbial Collection is one of the largest curated repositories of 2,500 cultivable endophytic bacteria isolated from 23 native desert plant species across Saudi Arabia, Jordan, and Pakistan. Representing a broad spectrum of arid microhabitats from inland deserts and mountain wadis to coastal mangroves and date palm oases, the collection supports integrative studies on microbial ecology and plant-microbe interactions in water-limited ecosystems. A central component of this initiative is the Darwin21 Genome Database, which currently hosts whole-genome sequences (WGS) of 534 endophytic bacterial isolates annotated with extensive ecological metadata, assembly statistics, functional traits, and host associations. The database interface provides tools for genome exploration, metadata filtering, and functional gene mining, enabling users to identify taxa and traits of agronomic interest, particularly for applications in sustainable agriculture and sustainable desert revegetation. By combining genomic, ecological, and functional data, the Darwin21 Genome Database serves as a foundational platform for the development of targeted microbial inoculants and fosters data-driven research into desert microbiomes and plant resilience mechanisms. Database URLhttps://www.genomedatabase.org/ O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=156 SRC="FIGDIR/small/672100v1_ufig1.gif" ALT="Figure 1"> View larger version (48K): org.highwire.dtl.DTLVardef@1703432org.highwire.dtl.DTLVardef@172fce7org.highwire.dtl.DTLVardef@39bd1borg.highwire.dtl.DTLVardef@ae3fef_HPS_FORMAT_FIGEXP M_FIG C_FIG

microbiology↗

bioRxiv2025

m6A RNA methylation attenuates thermotolerance in Arabidopsis

Heat stress severely affects plant growth, causing significant crop yield losses. To counteract this, plants employ adaptive strategies, including chromatin and transcriptional regulation of heat stress-related genes. However, the role of RNA methylation in heat stress response remains unclear. This study identifies m6A RNA methylation as a negative regulator of thermotolerance in Arabidopsis. Loss of m6A modification enhances resilience to heat stress by increasing transcript levels of heat-responsive genes and stabilizing their mRNAs. Upon heat shock, plants transiently reduce m6A levels by modulating key methylation machinery genes. This m6A deficiency enriches the H3K4me3 histone mark at heat-responsive gene loci, boosting their transcription. The combined effect of m6A-regulated chromatin changes and mRNA stability facilitates the accumulation of heat shock proteins essential for cellular protection and stress response. These findings reveal an interplay between m6A RNA methylation and histone modifications, forming a regulatory network that fine-tunes gene expression during heat stress. This novel mechanism highlights m6A as a critical regulator of plant thermotolerance, offering a promising target for improving heat stress resilience in crops.

plant biology↗

bioRxiv2025

G3BP1 Phosphorylation Regulates Plant Immunity in Arabidopsis

Mitogen-activated protein kinase (MAPK) cascades play critical roles in plant immunity by phosphorylating downstream effectors that regulate stress responses. While MAPK-mediated transcriptional regulation has been well examined, the involvement of MAPKs in post-transcriptional and post-translational regulation is still poorly understood. In this study, we identify the RNA binding-protein AtG3BP1 as a phosphorylation target of MPK3, MPK4, and MPK6 and demonstrate that phosphorylation of AtG3BP1 at Ser257 modulates key aspects of Arabidopsis immunity. Using phospho-mimic (G3BP1D) and phospho-dead (G3BP1A) mutants, we investigated the functional consequences of AtG3BP1 phosphorylation. Our data indicate that phosphorylation of AtG3BP1 promotes susceptibility to bacterial infection, suppresses accumulation of reactive oxygen species (ROS), and downregulates salicylic acid (SA) biosynthesis. Furthermore, we demonstrate that AtG3BP1 phosphorylation influences stomatal immunity by maintaining stomatal opening, thereby regulating pre-invasive defense mechanisms. Additionally, we provide evidence that phosphorylation stabilizes AtG3BP1 and prevents its degradation via the proteasome, thus leading to sustained immune signaling. These findings validate AtG3BP1 as a central integrator of MAPK signaling during plant immunity and reveal a new level of post-translational control. This study enhances our understanding of plant defense mechanisms and provides potential targets for engineering disease-resistant crops.

plant biology↗

bioRxiv2025

Oxalotrophic bacteria in desert and drylands: Enzymatic pathways and Carbon sequestration

Oxalotrophy refers to the ability of bacteria to utilize oxalate as a carbon and energy source. This is a critical process with significant implications for the global carbon cycle. Oxalate-degrading bacteria play a key role in carbon sequestration through the oxalate-carbonate pathway (OCP), contributing to stable inorganic carbon pools. In this study, we identified and cataloged 20 enzymes associated with various facets of oxalate metabolism to characterize the oxalotrophic potential of bacteria. Within this group, sets of enzymes were grouped into two functional categories in the context of carbon sequestration: a biomineralization toolkit for converting oxalate to inorganic carbon and an assimilation toolkit for incorporating oxalate into metabolic pathways such as amino acid biosynthesis and energy production. Using bioinformatic approaches, we analyzed a collection of 536 bacterial genomes from desert and dryland strains spanning 81 genera to identify oxalotrophs. To validate our findings, we tested several bacterial strains for growth on media supplemented with exogenous oxalate. Notably, while multiple bacterial strains grew on oxalate media, two Pseudomonas species, namely JZ043 and JZ097, failed to grow despite genomic predictions suggesting otherwise. Further investigation of these strains revealed several non-conservative amino acid substitutions in the glyoxylate carboligase enzyme (EC 4.1.1.47), a key player in oxalate metabolism, suggesting a potential link between these mutations and their inability to metabolize oxalate. Our findings highlight the significance of our approach for identifying oxalotrophic bacteria (OxB) and offer valuable insights into the molecular basis of oxalate metabolism. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=123 SRC="FIGDIR/small/637404v1_ufig1.gif" ALT="Figure 1"> View larger version (61K): org.highwire.dtl.DTLVardef@13d1408org.highwire.dtl.DTLVardef@ee2e5eorg.highwire.dtl.DTLVardef@1265323org.highwire.dtl.DTLVardef@11c56d0_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioinformatics↗

bioRxiv2025

USP1 acts as a chaperone for HSFA2 and plays a crucial role in thermopriming in Arabidopsis

Plants employ diverse strategies to cope with different types of heat stress. The response to short-term acute heat stress differs significantly from that to moderate heat stress followed by severe stress events. After experiencing moderate heat stress, plants exhibit a more robust response to subsequent severe stress, a phenomenon known as thermopriming or acquired thermotolerance. Thermopriming creates a memory by maintaining the heat stress (HS) memory-related genes in an alert state. In this work, we investigated the role of Arabidopsis Universal Stress Protein 1 (USP1) in plant heat stress responses. CRISPR-Cas9 generated knockout usp1 mutant lines showed no morphological changes during development and normal growth conditions. However, usp1 mutant plants showed enhanced levels of apoplast hydrogen peroxide and superoxide reactive oxygen species accumulation upon heat stress. Transcriptome analyses revealed that genes related to protein folding, electron transport, and oxidative phosphorylation are strongly upregulated in usp1 mutant plants. USP1 is essential for acquired thermotolerance, as usp1 mutants are compromised in heat stress memory but show normal responses to acute heat stress similar to hsfa2 mutants. Biochemical assays showed that USP1 functions as a molecular chaperone, protecting the transcription factor HSFA2 from heat-induced denaturation. Moreover, usp1 mutant plants show decreased transcript levels of heat stress response genes and reduced H3K4me3 enrichment at memory gene loci. These data show that USP1 plays an important role as a chaperone of HSFA2 in mediating plant heat stress memory.

biochemistry↗

bioRxiv2024

Mangrove endophytes enhance yield and convey flooding and salt tolerance to Nipponbare rice

Global climate change increasingly challenges agriculture with flooding and salinity. Among strategies to enhance crop resilience to these stresses, we tested several endophytic bacterial strains from mangroves, which are permanently exposed to flooding and high salinity. We show several strains that can enhance flooding and salinity tolerance in Arabidopsis and rice plants. Two strains and their combination massively enhanced the growth and yield of Oryza sativa cv. Nipponbare under both soil and hydroponic growth conditions with and without salt treatment. The bacteria-induced transcriptome changes in O. sativa roots related to ABA-signaling with lignin and suberin deposition in root tissues explain the altered responses of colonized rice plants to hypoxic and saline stress conditions. While enhancing yield and grain quality, bacterially colonized rice plants also show much earlier flowering, thereby massively shortening the life cycle of rice plants and opening the possibility for an additional harvest per year. These results show that microbes can be a powerful tool for enhancing the yield and resilience of rice to hypoxic and saline stress conditions.

plant biology↗

bioRxiv2023

Arabidopsis actin-binding protein WLIM2A links PAMP-triggered immunity and cytoskeletal organization

MAPKs are a family of highly conserved serine/threonine protein kinases that link upstream receptors to their downstream targets which can be localized in the cytoplasm or the nucleus. Pathogens produce pathogen-associated molecular patterns (PAMPs) that trigger the activation of MAPK cascades in plants. Phosphoproteomic analysis of PAMP-induced Arabidopsis plants led to the identification of several putative MAPK targets, WLIM2A. Here, we investigated the role of WLIM2A in plant immunity via a reverse-genetics approach generating wlim2a knockout lines using CRISPR-Cas9, as well as complementation and phosphosite mutated WLIM2A expression lines in the wlim2a background. The wlim2a lines were compromised in their response to Pst DC3000 but showed enhanced resistance to fungal infection by Botrytis cinereae. Transcriptome analyses revealed that immune hormone signaling and biosynthesis genes of salicylic acid (SA), jasmonic acid (JA), and ethylene (ET) are differentially regulated in the wlim2a knockout lines. Pathogen assays with Pst DC3000 showed altered stomatal phenotypes in wlim2a mutants. Importantly, WLIM2A phosphomutants had opposing stomatal behaviour and resistance phenotypes in response to Pst DC3000 infection. Overall, these data show that phosphorylation of WLIM2A by MAPKs regulates Arabidopsis stomatal immunity.

plant biology↗

bioRxiv2023

Microbe-induced plant drought tolerance by ABA-mediated root morphogenesis and epigenetic reprogramming of gene expression

The use of beneficial microbes to mitigate drought stress tolerance of plants is of great potential albeit little understood. We show here that a root endophytic desert bacterium, Pseudomonas argentinensis sp. SA190, enhances drought stress tolerance in Arabidopsis. Transcriptome and genetic analysis demonstrate that SA190-induced root morphogenesis and gene expression is mediated via the plant abscisic acid (ABA) pathway. Moreover, we demonstrate that SA190 primes the promoters of target genes in an epigenetic manner which is ABA-dependent. Application of the SA190 priming technology on crops is demonstrated for alfalfa in field trials, showing enhanced performance under desert agriculture conditions. In summary, a single beneficial root bacterial strain can help to perform agriculture under drought and water limiting conditions. Synopsis O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=161 SRC="FIGDIR/small/522604v1_ufig1.gif" ALT="Figure 1"> View larger version (39K): org.highwire.dtl.DTLVardef@b0bd17org.highwire.dtl.DTLVardef@14e3262org.highwire.dtl.DTLVardef@cdd103org.highwire.dtl.DTLVardef@5526a0_HPS_FORMAT_FIGEXP M_FIG C_FIG O_LIBeneficial root endophyte Pseudomonas argentinensis sp. SA190 confers drought tolerance in plants C_LIO_LISA190 modulates the expression of genes under drought stress in an ABA-dependent manner C_LIO_LISA190 primes genes via H3K4me3 histone mark enrichment C_LIO_LISA190 alters host plant physiology by improving the plant water status C_LIO_LISA190 enhances crop performance in open field conditions with limited irrigation C_LI

plant biology↗

bioRxiv2022

Molecular insights into the Darwin paradox of coral reefs from the sea anemone Aiptasia

Symbiotic cnidarians such as corals and anemones form highly productive and biodiverse coral-reef ecosystems in nutrient-poor ocean environments, a phenomenon known as Darwins Paradox. Resolving this paradox requires elucidating the molecular bases of efficient nutrient distribution and recycling in the cnidarian-dinoflagellate symbiosis. Using the sea anemone Aiptasia, we show that during symbiosis, the increased availability of glucose and the presence of the algae jointly induce the coordinated upregulation and re-localization of glucose and ammonium transporters. These molecular responses are critical to support symbiont functioning and organism-wide nitrogen assimilation through GS/GOGAT-mediated amino-acid biosynthesis. Our results reveal crucial aspects of the molecular mechanisms underlying nitrogen conservation and recycling in these organisms that allow them to thrive in the nitrogen-poor ocean environments. One-sentence summaryWhole-organism nitrogen assimilation fueled by glucose from symbiotic algae enables corals to flourish in oligotrophic waters.

molecular biology↗

bioRxiv2022

Linker histone H1 regulates defense priming and immunity in plants

Linker H1 histones play an important role in animal and human pathogenesis, but their function in plant immunity is poorly understood. Here, we analyzed mutants of the three canonical variants of Arabidopsis H1 histones, namely H1.1, H1.2 and H1.3. We observed that double h1.1h1.2 and triple h1.1h1.2h1.3 (3h1) mutants were resistant to Pseudomonas syringae and Botrytis cinerea infections. Transcriptome analysis of 3h1 mutant plants showed that histone H1s play a key role in regulating the expression of early and late defense genes upon pathogen challenge. Moreover, 3h1 mutant plants showed enhanced production of reactive oxygen species and activation of mitogen activated protein kinases upon pathogen-associated molecular pattern (PAMP) treatment. However, 3h1 mutant plants were insensitive to priming with flg22, a well-known bacterial PAMP (pathogen-associated molecular pattern) which induces enhanced resistance in WT plants. The defective defense response in 3h1 was correlated with the enhanced DNA methylation and reduced H3K56ac levels upon priming. Our data place H1 as a molecular gatekeeper in governing dynamic changes in the chromatin landscape of defense genes during plant pathogen interaction.

plant biology↗

bioRxiv2020

Enterococcal PrgA extends far outside the cell and provides surface exclusion to protect against unwanted conjugation

Horizontal gene transfer between Gram-positive bacteria leads to a rapid spread of virulence factors and antibiotic resistance. This transfer is often facilitated via Type 4 Secretion Systems (T4SS), which frequently are encoded on conjugative plasmids. However, donor cells that already contain a particular conjugative plasmid resist acquisition of a 2nd copy of said plasmid. They utilize different mechanisms, including surface exclusion for this purpose. Enterococcus faecalis PrgA, encoded by the conjugative plasmid pCF10, is a surface protein that has been implicated to play a role in both virulence and surface exclusion, but the mechanism by which this is achieved has not been fully explained. Here, we report the structure of full-length PrgA, which shows that PrgA protrudes far out from the cell wall (approximately 40 nm), where it presents a protease domain. In vivo experiments show that PrgA provides a physical barrier to cellular adhesion, thereby reducing cellular aggregation. This function of PrgA contributes to surface exclusion, reducing the uptake of its cognate plasmid by approximately one order of magnitude. Using variants of PrgA with mutations in the catalytic site we show that the surface exclusion effect is dependent on the activity of the protease domain of PrgA. In silico analysis suggest that PrgA can interact with another enterococcal adhesin, PrgB, and that these two proteins have co-evolved. PrgB is a strong virulence factor, and PrgA is involved in post-translational processing of PrgB. Finally, competition mating experiments show that PrgA provides a significant fitness advantage to plasmid-carrying cells.

biochemistry↗

bioRxiv2020

Enterobacter sp. SA187 mediates plant thermotolerance by chromatin modification of heat stress genes

Global warming has become a critical challenge to food safety, causing severe yield losses of major crops worldwide. Heat acclimation empowers plants to survive under extreme temperature conditions but the potential of beneficial microbes to make plants thermotolerant has not been considered so far. Here, we report that the endophytic bacterium Enterobacter sp. SA187 induces heat tolerance in Arabidopsis thaliana by reprogramming the plant transcriptome to a similar extent as acclimation. Acclimation induces priming of heat stress memory genes such as APX2 and HSP18.2 via the transcription factors HSFA1A, B, D, and E and the downstream master regulator HSFA2. hsfa1a,b,d,e and hsfa2 mutants compromised both acclimation and bacterial priming through the same pathway of HSF transcription factors. However, while acclimation transiently modifies H3K4me3 levels at heat stress memory gene loci, SA187 induces the constitutive priming of these loci. In summary, we demonstrate the molecular mechanism by which SA187 imparts thermotolerance in A. thaliana, suggesting that beneficial microbes might be a promising way to enhance crop production under global warming conditions.

plant biology↗

bioRxiv2019

Wounding and insect feeding trigger two independent MAPK pathways with distinct regulation and kinetics

Wounding is caused by abiotic and biotic factors and triggers complex short- and long-term responses at the local and systemic level. These responses are under the control of complex signaling pathways, which are still poorly understood. Here, we show that the rapid activation of MKK4/5-MPK3/6 by wounding is independent of jasmonic acid (JA) signaling and that, contrary to what happens in tobacco, this fast module does not control wound-triggered JA accumulation in Arabidopsis. We also demonstrate that a second MAPK module, constituted by MKK3 and the clade-C MAPKs MPK1/2/7, is activated by wounding in an independent manner. We provide evidence that the activation of this MKK3-MPK1/2/7 module occurs mainly through wound-induced JA production via the transcriptional regulation of upstream clade-III MAP3Ks and particularly MAP3K14. We show that mkk3 mutant plants are more susceptible to the larvae of the generalist lepidopteran herbivore Spodoptera littoralis, indicating that the MKK3-MPK1/2/7 module is involved in counteracting insect feeding. One sentence summaryWounding induces the parallel activation of a rapid signaling module (MKK4/5-MPK3/6) and a JA-dependent slow one (MAP3K14-MKK3-MPK1/2/7/14) to restrict insect feeding.

plant biology↗

bioRxiv2019

Role of MPK4 in pathogen-associated molecular pattern-triggered alternative splicing in Arabidopsis

Alternative splicing (AS) of pre-mRNAs in plants is an important mechanism of gene regulation in environmental stress tolerance but plant signals involved are essentially unknown. Pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) is mediated by mitogen-activated protein kinases and the majority of PTI defense genes are regulated by MPK3, MPK4 and MPK6. These responses have been mainly analyzed at the transcriptional level, however many splicing factors are direct targets of MAPKs. Here, we studied alternative splicing induced by the PAMP flagellin in Arabidopsis. We identified 506 PAMP-induced differentially alternatively spliced (DAS) genes. Although many DAS genes are targets of nonsense-mediated degradation (NMD), only 19% are potential NMD targets. Importantly, of the 506 PAMP-induced DAS genes, only 89 overlap with the set of 1849 PAMP-induced differentially expressed genes (DEG), indicating that transcriptome analysis does not identify most DASevents. Global DAS analysis of mpk3, mpk4, and mpk6 mutants revealed that MPK4 is a key regulator of PAMP-induced differential splicing, regulating AS of a number of splicing factors and immunity-related protein kinases, such as the calcium-dependent protein kinase CPK28, the cysteine-rich receptor like kinases CRK13 and CRK29 or the FLS2 co-receptor SERK4/BKK1. These data suggest that MAP kinase regulation of splicing factors is a key mechanism in PAMP-induced AS regulation of PTI.\n\nSignificance statementAlternative pre-mRNA splicing (AS) affects plant responses to environmental stresses. So far, however, the regulation of AS is little understood. Here, we studied AS induced by the pathogen-associated molecular pattern (PAMP) flagellin in Arabidopsis. We identified 506 differentially alternatively spliced (DAS) genes, 89 of which overlap with the 1849 DEG, indicating that the majority of DAS events go undetected by common transcriptome analysis. PAMP-triggered immunity is mediated by mitogen-activated protein kinases. Global DAS analysis of MAPK mutants revealed that MPK4 is a key regulator of AS by affecting splicing factors and a number of important protein kinases involved in immunity. Since PAMP-triggered phosphorylation of several splicing factors is directly mediated by MAPKs, we discovered a key mechanism of AS regulation.

plant biology↗