Biology subjects

Ranf, S.

Publications and source records attributed to Ranf, S..

9 recordsLinked to original sources

bioRxiv2026

The plant immune receptor LORE binds agonistic and antagonistic 3-hydroxy fatty acid ligands via a dynamic loop in its G-type lectin domain

The Arabidopsis thaliana S-domain receptor kinase LORE senses bacterial medium-chain 3-hydroxy fatty acids (mc-3-OH-FAs) as microbe-associated molecular patterns to activate pattern-triggered immunity. How LORE recognises these fatty acid ligands at the molecular level remains unknown. Here, we combined protein structure prediction, protein-ligand interaction modelling and molecular dynamics (MD) simulations with ligand-binding assays using chimeric and mutant receptor ectodomains, and functional analysis of receptor activation to characterise the mc-3-OH-FA binding mechanism. Domain-swap experiments between LORE and its non-binding paralog AtSD1-23 identify the lectin 2 (L2) domain as the ligand-binding domain. Mutational analysis and reverse engineering confirm a hydrophobic pocket in the L2 core as the primary ligand-binding site. Multiple walker Supervised MD (mwSuMD) simulations reveal that the acyl tail enters the pocket first, whilst polar interactions between the headgroup and a flexible L2 loop guide and stabilise the bound state. In support of this model, 3-OH-C10:0 analogues with bulky headgroup modifications dock into the pocket but act as antagonists, presumably by preventing the loop from adopting the conformation required for signalling. Together, these data suggest that the flexible L2 loop has multiple functions: it acts as a dynamic gate regulating pocket access, provides essential anchoring points once the ligand is bound, and contributes to receptor activation. These findings provide a mechanistic framework for immunogenic mc-3-OH-FA sensing by LORE.

plant biology↗

bioRxiv2026

Diversification of ligand preference in conserved Brassicaceae LORE immune receptors matches proteobacterial 3-hydroxy fatty acid profiles

Pattern recognition receptors are often conserved across plant lineages, yet how ligand sensing diversifies during evolution remains poorly understood. The receptor kinase LORE mediates Arabidopsis thaliana immune responses to bacterial medium-chain 3-hydroxy fatty acids (mc-3-OH-FAs), but its distribution and functional variation across Brassicaceae remain largely unexplored. We combined phylogenetic analysis, species-wide immune profiling, ligand-binding assays, heterologous functional testing, and A. thaliana lore-1 complementation to define LORE diversification across Brassicaceae. LORE orthologs with conserved 3-OH-C10:0 binding are present in all major Brassicaceae lineages, including the basal lineage. Functional complementation confirmed mc-3-OH-FA-sensing capacity in orthologs spanning all four lineages. By contrast, chain-length preference profiles among mc-3-OH-FAs varied between species and orthologs, ranging from narrow 3-OH-C10:0-dominated profiles to broader profiles with comparable sensitivity to 3-OH-C10:0 and 3-OH-C12:0. 3-OH-FA profiling of a diverse collection of plant-associated bacteria revealed that high levels of mc-3-OH-FAs are prevalent in Proteobacteria, and that the range of mc-3-OH-FA chain lengths produced corresponds to the chain-length preferences of Brassicaceae LORE receptors. Our results establish LORE as a Brassicaceae-restricted pattern recognition receptor with conserved mc-3-OH-FA sensing but diversified chain-length preferences, and provide an evolutionary framework for studying how ligand preference can diversify within a biologically relevant ligand spectrum while core receptor function is maintained.

plant biology↗

bioRxiv2025

Pantoea ananatis-triggered systemic resistance requires root sensing through the LORE receptor kinase in Arabidopsis

Systemic immunity induced by beneficial bacteria helps plants to cope with subsequent pathogen attacks. However, how these bacteria are sensed by plants and how this sensing results in a systemic response is not well understood. Recently, the Brassicaceae LIPOOLIGOSACCHARIDE-SPECIFIC REDUCED ELICITATION (LORE) receptor kinase was identified as the direct sensor of bacterial medium-chain-length 3-hydroxy fatty acids (mc-3-OH-FAs) and 3-(3-hydroxyalkanoyloxy)alkanoic acids (HAAs), which activate systemic Brassicaceae immunity. Here, we show that the bacterium Pantoea ananatis BRT175, which produces HAAs, can colonize Arabidopsis thaliana roots independently of the production of these compounds. P. ananatis triggers induced systemic resistance (ISR) to the necrotrophic pathogen Botrytis cinerea, but not to the hemibiotrophic pathogen Pseudomonas syringae pv. tomato. Importantly, this ISR can be mimicked by HAAs or 3-hydroxy-decanoic acid (3-OH-C10). Both P. ananatis-triggered and 3-OH-C10-triggered ISR against B. cinerea are mediated by LORE sensing, involve salicylic acid, jasmonic acid and ethylene signaling pathways, and activate the expression of similar defense genes in infected leaves. Thus, 3-OH-C10 and HAAs are perceived by a pattern recognition receptor (PRR) in the root and activate a local immune response in roots as well as systemic immunity in leaves. Our study demonstrates that these lipidic microbe-associated molecular patterns (MAMPs), produced by beneficial microorganisms, are necessary and sufficient to trigger rhizobacteria-driven ISR. It further highlights the role of these MAMPs in differentially activating systemic resistance against biotrophic and necrotrophic pathogens.

plant biology↗

bioRxiv2024

Cross-family transfer of the Arabidopsis cell-surface immune receptor LORE to tomato confers sensing of 3-hydroxylated fatty acids and enhanced disease resistance

Plant pathogens pose a high risk of yield losses and threaten food security. Technological and scientific advances have improved our understanding of the molecular processes underlying host-pathogen interactions, which paves the way for new strategies in crop disease management beyond the limits of conventional breeding. Cross-family transfer of immune receptor genes is one such strategy that takes advantage of common plant immune signaling pathways to improve disease resistance in crops. Sensing of microbe- or host damage-associated molecular patterns (MAMPs/DAMPs) by plasma membrane-resident pattern recognition receptors (PRR) activates pattern-triggered immunity (PTI) and restricts the spread of a broad spectrum of pathogens in the host plant. In the model plant Arabidopsis thaliana, the S-domain receptor-like kinase LIPOOLIGOSACCHARIDE-SPECIFIC REDUCED ELICITATION (AtLORE, SD1-29) functions as PRR, which senses medium chain-length 3-hydroxylated fatty acids (mc-3-OH-FAs), such as 3-OH-C10:0, and 3-hydroxyalkanoates (HAAs) of microbial origin to activate PTI. In this study, we show that ectopic expression of the Brassicaceae-specific PRR AtLORE in the solanaceous crop species Solanum lycopersicum cv. M82 leads to the gain of 3-OH-C10:0 immune sensing without altering plant development. AtLORE-transgenic tomato shows enhanced resistance against Pseudomonas syringae pv. tomato DC3000 and Alternaria solani NL03003. Applying 3-OH-C10:0 to the soil before infection induces resistance against the oomycete pathogen Phytophthora infestans Pi100 and further enhances resistance to A. solani NL03003. Our study proposes a potential application of AtLORE-transgenic crop plants and mc-3-OH-FAs as resistance-inducing bio-stimulants in disease management.

plant biology↗

bioRxiv2023

Mechanosensing and Sphingolipid-Docking Mediate Lipopetide-Induced Immunity in Arabidopsis

Bacteria-derived lipopeptides are immunogenic triggers of host defenses in metazoans and plants. Root-associated rhizobacteria produce cyclic lipopeptides that activate systemically induced resistance (IR) against microbial infection in various plants. How these molecules are perceived by plant cells remains elusive. Here, we reveal that immunity activation in Arabidopsis thaliana by the lipopeptide elicitor surfactin is mediated by docking into specific sphingolipid-enriched domains and relies on host membrane deformation and subsequent activation of mechanosensitive ion channels. This mechanism leads to host defense potentiation and resistance to the necrotroph B. cinerea but is distinct from host pattern recognition receptor-mediated immune activation and reminiscent of damage-induced plant immunity.

plant biology↗

bioRxiv2021

LORE homomerization is required for 3-OH-C10:0 induced immune signaling

O_LIThe S-domain-type receptor-like kinase (SD-RLK) LIPOOLIGOSACCHARIDE-SPECIFIC REDUCED ELICITATION (LORE) from Arabidopsis thaliana is a pattern recognition receptor that senses medium-chain 3-hydroxy fatty acids, such as 3-hydroxydecanoic acid (3-OH-C10:0), to activate pattern-triggered immunity. Here, we show that LORE homomerization is required to activate 3-OH-C10:0-induced immune signaling. C_LIO_LIFluorescence lifetime imaging in Nicotiana benthamiana demonstrated that AtLORE homomerizes via the extracellular and transmembrane domains. Co-expression of AtLORE truncations lacking the intracellular domain exerts a dominant negative effect on AtLORE signaling in both N. benthamiana and A. thaliana, highlighting that homomerization is essential for signaling. C_LIO_LIScreening for 3-OH-C10:0-induced reactive oxygen species production revealed natural variation within the Arabidopsis genus. Arabidopsis lyrata and Arabidopsis halleri do not respond to 3-OH-C10:0, although both possess a putative LORE orthologue. Both LORE orthologues have defective extracellular domains that bind 3-OH-C10:0 to a similar level but lack the ability to homomerize. Thus, ligand binding is independent of LORE homomerization. Analysis of AtLORE and AlyrLORE chimera suggests that the loss of AlyrLORE homomerization is caused by several amino acid polymorphisms across the extracellular domain. C_LIO_LIOur findings shed light on the activation mechanism of LORE and the loss of 3-OH-C10:0 perception within the Arabidopsis genus. C_LI

plant biology↗

bioRxiv2021

Low cost, medium throughput depletion-binding assay for screening S-domain-receptor ligand interactions using in planta protein expression

BackgroundPlant cell-surface receptors sense various ligands to regulate physiological processes. Matching ligand-receptor pairs requires evidence of their direct interaction and is often a bottleneck in functional receptor studies. The S-domain-type (SD) pattern recognition receptor LORE senses medium chain-3-hydroxy fatty acids (mc-3-OH-FAs) such as 3-hydroxy decanoic acid (3-OH-C10:0) via its extracellular domain (ECD) They are perceived as signals of danger from Gram-negative bacteria and activate immune responses in Arabidopsis thaliana. LORE is found in low levels in planta and is poorly expressed in heterologous systems. Furthermore, chemical modifications of the mc-3-OH-FA ligand affect its biological activity. Taken together, this makes LORE-mc-3-OH-FA binding studies particularly challenging. ResultsTo investigate the LORE-mc-3-OH-FA interaction, we have developed a sensitive assay system based on protein expression in planta. The ECDs of LORE and other proteins of interest were transiently expressed as soluble, apoplastic mCherry fusion proteins in Nicotiana benthamiana and collected in apoplastic washing fluids. Protein-ligand complexes and unbound ligand were separated according to their molecular weight. In a two-step procedure, we first investigated whether the ECD-mCherry fusion protein depletes 3-OH-C10:0 from the low molecular weight fraction (step depletion). Subsequently, protein-bound 3-OH-C10:0 retained in the high molecular weight fraction in the depletion step is released and detected (step binding). Both the unbound and the released 3-OH-C10:0 ligand are detected by a sensitive bioassay using LORE loss- and gain-of-function Arabidopsis plants. Using the depletion-binding assay, we show that the ECD of AtLORE and its ortholog from Capsella rubella, CrubLORE, bind 3-OH-C10:0. The ECD of AtSD1-23, the closest paralog of AtLORE, and mCherry did not bind 3-OH-C10:0 and are suitable negative controls. ConclusionThe depletion-binding assay is a simple method for reliably detecting interactions between plant-expressed SD-type receptor ectodomains and mc-3-OH-FAs. It does not require special equipment or expensive consumables and is suitable for medium throughput screening. The assay is very flexible and can be easily adapted to investigate ligand interactions of other extracellular receptor domains.

plant biology↗

bioRxiv2020

Rhamnolipids and their 3-(3-hydroxyalkanoyloxy)alkanoic acid precursors activate Arabidopsis innate immunity through two independent mechanisms

Plant innate immunity is activated upon perception of invasion pattern molecules by plant cell-surface immune receptors. Several bacteria of the genera Pseudomonas and Burkholderia produce rhamnolipids (RLs) from L- rhamnose and (R)-3-hydroxyalkanoate precursors (HAAs). RL and HAA secretion is required to modulate bacterial surface motility, biofilm development, and thus successful colonization of hosts. Here, we show that the lipidic secretome from the opportunistic pathogen Pseudomonas aeruginosa mostly comprising RLs and HAAs stimulates Arabidopsis immunity. We demonstrate that HAAs are sensed by the bulb-type lectin receptor kinase LIPOOLIGOSACCHARIDE-SPECIFIC REDUCED ELICITATION/S-DOMAIN-1-29 (LORE/SD1-29) that also mediates medium-chain 3-hydroxy fatty acid (mc-3-OH-FA) perception in the plant Arabidopsis thaliana. HAA sensing induces canonical immune signaling and local resistance to plant pathogenic Pseudomonas infection. By contrast, RLs trigger an atypical immune response and resistance to Pseudomonas infection independent of LORE. Thus, the glycosyl moieties of RLs, albeit abolishing sensing by LORE, do not impair their ability to trigger plant defense. In addition, our results show that RL-triggered immune response is affected by the sphingolipid composition of the plasma membrane. In conclusion, RLs and their precursors released by bacteria can both be perceived by plants but through distinct mechanisms. SignificanceActivation of plant innate immunity relies on the perception of microorganisms through self and nonself elicitors. Rhamnolipids and their precursor HAAs are exoproducts produced by beneficial and pathogenic bacteria. They are involved in bacterial surface dissemination and biofilm development. As these compounds are released in the extracellular milieu, they have the potential to be perceived by the plant immune system. Our work shows that both compounds independently activate plant immunity. We demonstrate that HAAs are perceived by the receptor protein kinase LORE. By contrast, rhamnolipids are not senses by LORE but activate a non-canonical immune response affected by the sphingolipid composition of the plant plasma membrane. Thus, plants are able to sense bacterial molecules as well as their direct precursors to trigger a distinct immune response.

plant biology↗

bioRxiv2019

The Arabidopsis leucine-rich repeat receptor kinase MIK2 is a crucial component of pattern-triggered immunity responses to Fusarium fungi

Fusarium is a genus of fungi causing severe economic damage in many crop species exemplified by Fusarium Head Blight of wheat or Panama Disease of banana. Plants sense immunogenic patterns (termed elicitors) at the cell surface contributing to disease resistance via the activation of pattern-triggered immunity (PTI). Knowledge of such elicitors or corresponding plant immunity components is largely lacking for Fusarium species. We describe a new peptide elicitor fraction present in several Fusarium spp. which elicits canonical PTI responses in Arabidopsis thaliana but depends on a currently unknown perception mechanism. We therefore employed a forward-genetics screen using Arabidopsis plants containing a cytosolic calcium reporter (apoaequorin) to isolate fere (Fusarium Elicitor Reduced Elicitation) mutants. The fere1 mutant showed impaired PTI marker responses to an enriched elicitor fraction derived from Fusarium oxysporum but normal responses to other fungal elicitors. We mapped the causal mutation to the receptor-like kinase MIK2 (MALE DISCOVERER1-INTERACTING RECEPTOR LIKE KINASE 2) with a hitherto undescribed role in PTI pathways but documented functions in other cell surface signalling pathways. The strength of the phenotype in fere1 and independent mik2 mutants supports that MIK2 is a new key component in sensing Fusarium. Fusarium elicitor responses also partially depend on PTI signalling components known for other cell surface elicitor responses such as BAK1, BIK1, PBL1, FERONIA, LLG1 and RBOHD. This shows that Arabidopsis senses Fusarium by a novel receptor complex at the cell surface that feeds into common PTI pathways and positions MIK2 as a central player that potentially integrates plant endogenous signals with biotic and abiotic stress responses.

plant biology↗