Biology subjects

Huckelhoven, R.

Publications and source records attributed to Huckelhoven, R..

12 recordsLinked to original sources

bioRxiv2025

Barley C2-Domain Abscisic Acid-Related protein CARa supports susceptibility to Blumeria hordei and localizes to the extrahaustorial membrane

Rho GTPases are key regulators of cellular signalling processes in eukaryotes. In plants, Rho of plants (ROP) proteins function in cell polarization, hormone signalling, and plant immunity or susceptibility to diseases. The barley (Hordeum vulgare) ROP protein RACB is involved in susceptibility towards the biotrophic ascomycete fungus Blumeria hordei (Bh) and supports the accommodation of fungal haustoria. In healthy barley, RACB plays a role in cell development, a function that the pathogen may co-opt for cellular ingrowth of its haustorium into intact epidermal cells of barley. The majority of identified functions of GTP-bound activated RACB appear to be mediated by scaffold proteins. One of them, the ROP Interactive Partner b (RIPb, synonym: Interactor of Constitutive Active ROP b, ICRb) binds to RACB-GTP and co-localizes with RACB at the site of fungal host cell entry. RIPb interacts with RACB-GTP via its C-terminal coiled-coil CC2 domain (RIPbCC2) at the plasma membrane. Here, we show that a barley Calcium-Dependent Phospholipid Binding 2 (C2)-Domain Abscisic Acid-Related protein (CARa) interacts with RIPb and its RIPbCC2 domain in planta. Transient knockdown of CARa renders barley epidermal cells less susceptible to invasion by Bh, whereas overexpression supports fungal haustorium accomodation. Upon fungal attack, CARa is recruited to the site of fungal attack and localizes around the haustorial neck and at the extrahaustorial membrane. These findings further support that Bh profits from host ROP signalling components and CARa. CARa might connect RACB-RIPb-signalling to membrane organization for support of fungal infection structures in barley epidermal cells.

plant biology↗

bioRxiv2025

Multi-omics of barley Fusarium Head Blight converge on pathogen-triggered biosynthesis of aromatic amino acid derived chemical defense compounds

Fusarium Head Blight (FHB) is a devastating fungal disease of small grain cereals like wheat and barley, causing substantial yield and quality losses each year worldwide. FHB is caused by Fusarium species that produce mycotoxins such as deoxynivalenol (DON) that impairs protein biosynthesis. Although defense responses in barley to Fusarium infection have been described at the transcriptional level, it remains unclear to what extent these responses are translated into functional changes at the protein and metabolite levels. In this study, we employed comprehensive transcriptomics, proteomics, and metabolomics to dissect the defense responses of barley heads during infection with Fusarium culmorum. Our integrated analyses revealed a set of significantly regulated gene-protein pairs linked to biosynthetic pathways that consistently correspond to upregulated defense-related metabolites. These include tryptophan-derived stress metabolites such as tryptamine and serotonin, as well as barley-specific hydroxycinnamylamides, including conjugates from the trypthophan metabolism, hordatines, and their biosynthetic precursors. Integrating data across multiple omics layers identifies the upregulation of aromatic amino acid derived secondary metabolism as the most consistent barley response to FHB infection across diverse barley varieties that share barley-typical type II resistance to fungal spreading in the head rachis.

plant biology↗

bioRxiv2024

Barley resistance and susceptibility to fungal cell entry involve the interplay of ROP signaling with phosphatidylinositol-monophosphates

Rho-of-plant small GTPases (ROPs) are regulators of plant polar growth and of plant-pathogen interactions. The barley ROP, RACB, is involved in susceptibility towards infection by the barley powdery mildew fungus Blumeria hordei (Bh) but little is known about the cellular pathways that connect RACB-signaling to disease susceptibility. Here we identify novel RACB interaction partners of plant or fungal origin by untargeted co-immunoprecipitation of constitutively active (CA) RACB tagged by green-fluorescent protein from Bh-infected barley epidermal layers and subsequent analysis by liquid chromatography-coupled mass spectrometry. Three of immunoprecipitated proteins, a plant phosphoinositide phosphatase, a plant phosphoinositide phospholipase and a putative Bh-effector protein are involved in the barley-Bh-pathosystem and support disease resistance or susceptibility, respectively. RACB and its plant interactors bind to overlapping anionic phospholipid species in vitro, and in case of RACB, this lipid-interaction is mediated by its carboxy-terminal polybasic region (PBR). Fluorescent markers for anionic phospholipids show altered subcellular distribution in barley cells during Bh-attack and under expression of a RACB-binding fungal effector. Phosphatidylinositol 4-phosphate, phosphatidylinositol 3,5-bisphosphate and phosphatidylserine show a distinct enrichment at the haustorial neck region, suggesting a connection to subcellular targeting of RACB at this site. The interplay of ROPs with anionic phospholipids and phospholipid-metabolizing enzymes may thus enable the subcellular enrichment of components pivotal for success or failure of fungal penetration.

plant biology↗

bioRxiv2021

Volatile-mediated signaling induces resistance of barley against infection with the biotrophic fungus Blumeria graminis f.sp. hordei

Plants have evolved a vast variety of secondary metabolites to counteract biotic stress. Volatile organic compounds (VOCs) are carbon-based molecules induced by herbivore attack or pathogen infection. A mixture of plant VOCs is released for direct or indirect plant defense, plant-plant or plant-insect communication. Recent studies suggest that VOCs can also induce biotic stress resistance in distant organs and neighboring plants. Among other VOCs, green leaf volatiles (GLVs) are quickly released by plant tissue after the onset of herbivory or wounding. We analysed VOCs emitted by 13-day old barley plants (Hordeum vulgare L.) after mechanical wounding using passive absorbers and TD-GC/MS detection. We investigated the influence of pure (Z)-3-hexenyl acetate (Z3HAC) as well as complex VOCs from wounded barley plants on the barley - powdery mildew interaction by pre-exposure in a static and a dynamic headspace connected to a powdery mildew susceptibility assay. GLVs dominated the volatile profile of wounded barley plants with Z3HAC as the most prominent compound. Pre-exposure with Z3HAC resulted in induced resistance of barley against fungal infection. Barley complex volatiles emitted after mechanical wounding, similarly, enhanced resistance in receiver plants. We found volatile-induced modification of the interaction towards an enhanced resistance against fungal infection. In addition, Z3HAC triggered a modulation of the alcohol dehydrogenase isoenzyme activity in receiver plants, a physiological response that possibly contributes to induced resistance. Plant-originated volatile metabolites could be a useful supplementation for future agronomic or horticultural practices. HighlightVolatile-induced modification of the barley-powdery mildew interaction towards an enhanced resistance against fungal infection.

plant biology↗

bioRxiv2020

Ramularia leaf spot disease of barley is highly host genotype-dependent and suppressed by continuous drought stress in the field

Since the 1980s, Ramularia leaf spot (RLS) is an emerging barley disease world-wide. The control of RLS is increasingly aggravated by a recent decline in fungicide efficacy and a lack of RLS-resistant cultivars. Furthermore, climate change increases drought periods in Europe, enhances variable weather conditions and thus will have an impact on severity of plant diseases. Hence, identification of RLS-resistant cultivars and understanding of disease progression under abiotic stress are important aims in integrated disease management under climate change. In the present study, we evaluated quantitative RLS resistance of 15 spring barley genotypes under drought, controlled irrigation and field conditions between 2016 and 2019 and monitored microclimatic conditions within the canopy. We identified genotypes that show robust quantitative resistance to RLS in different field environments. Our findings suggest that long-lasting drought periods create unfavourable conditions for the disease and supports that the extent and duration of leaf wetness is a key factor for RLS epidemics.

plant biology↗

bioRxiv2020

Population studies in the wild tomato species Solanum chilense reveal maintenance and loss of resistance at geographically distinct locations

Natural plant populations encounter strong pathogen pressure and defense-associated genes are known to be under different selection pressure dependent on the pressure by the pathogens. Here we use wild tomato Solanum chilense populations to investigate natural resistance against Cladosporium fulvum, a well-known pathogenic fungus of domesticated tomatoes. We show that populations of S. chilense differ in resistance against the pathogen. Next, we explored the underlying molecular processes in a species wide-context. Then, focusing on recognition of the two prominent avirulence factors secreted by C. fulvum (Avr4 and Avr9) in central and northern populations of S. chilense we observed high complexity in the cognate homologues of Cladosporium resistance (Hcr9) locus underlying the recognition of these effectors. Presence of canonical genomic regions coding for Cf-4 and Cf-9, two major dominant resistance genes in the Hcr9 locus recognizing Avr4 and Avr9, respectively, does not meet prediction from Avr response phenotypes. We find both genes in varying fractions of the plant populations and we show possible co-existence of two functionally active resistance genes, previously thought to be allelic. Additionally, we observed the complete local absence of recognition of additional Avr proteins of C. fulvum. In the southern populations we attribute this to changes in the coregulatory network. As a result of loss of pathogen pressure or adaptation to extreme climatic conditions. This may ultimately explain the observed pathogen susceptibility in the southern populations. This work puts major gene mediated disease resistance in an ecological context.

ecology↗

bioRxiv2020

Posttranslational modification of the RHO of plants protein RACB by phosphorylation and cross-kingdom conserved ubiquitination

Small RHO-type G-proteins act as signaling hubs and master regulators of polarity in eukaryotic cells. Their activity is tightly controlled, as defective RHO signaling leads to aberrant growth and developmental defects. Two major processes regulate G-protein activity: canonical shuttling between different nucleotide bound states and posttranslational modification (PTM), of which the latter can support or suppress RHO signaling, depending on the individual PTM. In plants, regulation of Rho of plants (ROPs) signaling activity has been shown to act through nucleotide exchange and GTP hydrolysis, as well as through lipid modification, but there is little data available on phosphorylation or ubiquitination of ROPs. Hence, we applied proteomic analyses to identify PTMs of the barley ROP RACB. We observed in vitro phosphorylation by barley ROP binding kinase 1 and in vivo ubiquitination of RACB. Comparative analyses of the newly identified RACB phosphosites and human RHO protein phosphosites revealed conservation of modified amino acid residues, but no overlap of actual phosphorylation patterns. However, the identified RACB ubiquitination site is conserved in all ROPs from Hordeum vulgare, Arabidopsis thaliana and Oryza sativa and in mammalian Rac1 and Rac3. Point mutation of this ubiquitination site leads to stabilization of RACB. Hence, this highly conserved lysine residue may regulate protein stability across different kingdoms.

plant biology↗

bioRxiv2019

Barley RIC157 is involved in RACB-mediated susceptibility to powdery mildew

Successful obligate pathogens benefit from host cellular processes. For the biotrophic ascomycete fungus Blumeria graminis f.sp. hordei (Bgh) it has been shown that barley RACB, a small monomeric G-protein (ROP, RHO of plants), is required for full susceptibility to fungal penetration. The susceptibility function of RACB probably lies in its role in cell polarisation, which may be co-opted by the pathogen for invasive ingrowth of its haustorium. However, the actual mechanism of how RACB supports the fungal penetration success is little understood. RIC proteins (ROP-Interactive and CRIB-(Cdc42/Rac Interactive Binding) motif-containing) are considered scaffold proteins which can interact directly with ROPs via a conserved CRIB motif. Here we describe a yet uncharacterised RIC protein, RIC157, which can interact directly with RACB in planta. We show that RIC157 undergoes a recruitment from the cytoplasm to the cell periphery in the presence of activated RACB. During fungal infection, RIC157 and activated RACB colocalise at the penetration site, particularly at the haustorial neck. In a RACB-dependent manner, transiently overexpressed RIC157 renders barley epidermal cells more susceptible to fungal penetration. This suggests that RIC157 promotes fungal penetration into barley epidermal cells via its function downstream of RACB.

plant biology↗

bioRxiv2019

ROP INTERACTIVE PARTNER b interacts with the ROP GTPase RACB and supports fungal penetration into barley epidermal cells

RHO of Plants (ROP) G-proteins are key components of cell polarization processes in plant development. The barley (Hordeum vulgare) ROP protein RACB, is a susceptibility factor in the interaction of barley with the barley powdery mildew fungus Blumeria graminis f.sp. hordei (Bgh). RACB also drives polar cell development, and this function might be coopted during formation of fungal haustoria in barley epidermal cells. In order to understand RACB signaling during the interaction of barley with Bgh, we searched for potential downstream interactors of RACB. Here, we show that ROP INTERACTIVE PARTNER b (RIPb, synonym: INTERACTOR OF CONSTITUTIVE ACTIVE ROP b; ICRb) directly interacts with RACB in yeast and in planta. Over-expression of RIPb supports susceptibility of barley to Bgh. RIPb further interacts with itself at microtubules. However, the interaction with activated RACB takes place at the plasma membrane. Both, RIPb and RACB are recruited to the site of fungal attack around the neck of developing haustoria suggesting locally enhanced ROP activity. We further assigned different functions to different domains of the RIPb protein. The N-terminal coiled-coil CC1 domain is required for microtubule localization, while the C-terminal coiled-coil CC2 domain is sufficient to interact with RACB and to fulfill a function in susceptibility at the plasma membrane. Hence, RIPb appears to be localized at microtubules and is then recruited by activated RACB for a function at the plasma membrane during formation of the haustorial complex. One Sentence summaryRIPb acts downstream of the powdery mildew susceptibility factor RACB of barley and influences susceptibility

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↗

bioRxiv2019

Barley ROP-INTERACTIVE PARTNER-a organizes into RAC1- and MICROTUBULE-ASSOCIATED ROP-GTPASE ACTIVATING PROTEIN 1-dependent membrane domains

Small ROP (also called RAC) GTPases are key factors in polar cell development and in interaction with the environment. ROP-Interactive Partner (RIP) proteins are predicted scaffold or ROP-effector proteins, which function downstream of activated GTP-loaded ROP proteins in establishing membrane heterogeneity and cellular organization. Grass ROP proteins function in cell polarity, resistance and susceptibility to fungal pathogens but grass RIP proteins are little understood.\n\nWe found that the barley (Hordeum vulgare L.) RIPa protein can interact with barley ROPs in yeast. Fluorescent-tagged RIPa, when co-expressed with the constitutively activated ROP protein CA RAC1, accumulates at the cell periphery or plasma membrane. Additionally, RIPa, locates into membrane domains, which are laterally restricted by microtubules, when co-expressed with RAC1 and MICROTUBULE-ASSOCIATED ROP-GTPASE ACTIVATING PROTEIN 1. Both structural integrity of MICROTUBULE-ASSOCIATED ROP-GTPASE ACTIVATING PROTEIN 1 and microtubule stability are key to maintenance of RIPa-labeled membrane domains. In this context, RIPa also accumulates at the interface of barley and invading hyphae of the powdery mildew fungus Blumeria graminis f.sp. hordei.\n\nData suggest that barley RIPa interacts with barley ROPs and specifies RAC1 activity-associated membrane domains with potential signaling capacity. Lateral diffusion of this RAC1 signaling capacity is restricted the resulting membrane heterogeneity requires intact microtubules and MICROTUBULE-ASSOCIATED ROP-GTPASE ACTIVATING PROTEIN 1. Focal accumulation of RIPa at sites of fungal attack may indicate locally restricted ROP activity at sites of fungal invasion.

plant biology↗

bioRxiv2018

A Barley Powdery Mildew Fungus Non-Autonomous Retrotransposon Encodes a Peptide that Supports Penetration Success on Barley

Plant immunity is overcome by pathogens by the means of secreted effectors. Host effector targets might be proteins acting in pathogen defense or serve demands of the pathogen. The barley ROP GTPase HvRACB is involved in entry of the powdery mildew fungus Blumeria graminis f.sp. hordei (Bgh) into barley epidermal cells. We found that HvRACB interacts with the ROP-interactive peptide 1 (ROPIP1) that is encoded on the active non-long terminal repeat retroelement Eg-R1 of Bgh. Over-expression of ROPIP1 in barley epidermal cells and host-induced post-transcriptional gene silencing (HIGS) of ROPIP1 suggested that ROPIP1 is involved in virulence of Bgh. Bimolecular fluorescence complementation and co-localization supported that ROPIP1 can interact with activated HvRACB in planta. We show that ROPIP1 is expressed by Bgh on barley and translocated into the cytoplasm of infected barley cells. ROPIP1 is recruited to microtubules upon co-expression of MICROTUBULE ASSOCIATED ROP GTPase ACTIVATING PROTEIN (HvMAGAP1) and can destabilize cortical microtubules. Bgh ROPIP might target HvRACB and manipulate host cell microtubule organization for facilitated host cell entry. Data suggest a possible neo-functionalization of retroelement-derived transcripts for the evolution of a pathogen virulence effector.

plant biology↗