Biology subjects

Gust, A. A.

Publications and source records attributed to Gust, A. A..

7 recordsLinked to original sources

bioRxiv2025

Enhancing plant broad-spectrum resistance through engineered pattern recognition receptors

Conventional plant resistance breeding has primarily focused on intracellular immune receptors, while cell-surface pattern recognition receptors (PRRs) have been underexplored due to their comparatively modest contributions to resistance. However, PRRs offer significant untapped potential for crop improvement. In this study, we demonstrate that the Arabidopsis receptor-like protein RLP23, which recognizes molecular patterns from three distinct microbial kingdoms, confers broad-spectrum resistance when introduced into the Solanaceae crop tomato. We also identify the intracellular (IC) domain of RLP23 as crucial for ensuring compatibility and efficacy during heterologous expression. Targeted engineering of the IC domain significantly enhances RLP23 potential utility, enabling transfer of robust resistance against bacterial, fungal, and oomycete pathogens to other plants without compromising yield. We extended this RLP engineering strategy to rice and poplar, highlighting its broad applicability. These findings establish a versatile framework for PRR-based engineering, opening new avenues for sustainable crop protection.

plant biology↗

bioRxiv2022

Convergent evolution of plant pattern recognition receptors sensing cysteine-rich patterns from three microbial kingdoms

The Arabidopsis thaliana receptor-like protein RLP30 contributes to immunity against the fungal pathogen Sclerotinia sclerotiorum. Here we identified the RLP30-ligand as a small cysteine-rich protein (SCP) that occurs in many fungi and oomycetes and is also recognized by the Nicotiana benthamiana RLP RE02. However, RLP30 and RE02 share little sequence similarity and respond to different parts of the native/folded protein. Interestingly, some Brassicaceae other than Arabidopsis also respond to a linear SCP peptide, suggesting that SCP is an eminent immune target that led to the convergent evolution of distinct immune receptors in plants. Surprisingly, RLP30 shows a second ligand specificity for a SCP-nonhomologous protein secreted by bacterial Pseudomonads. RLP30 expression in N. tabacum resulted in lower susceptibility to bacterial, fungal and oomycete pathogens, thus demonstrating that detection of immunogenic patterns by Arabidopsis RLP30 is involved in defense against pathogens from three microbial kingdoms.

plant biology↗

bioRxiv2021

Genotyping-by-sequencing-based identification of Arabidopsis pattern recognition receptor RLP32 recognizing proteobacterial translation initiation factor IF1

Pattern-triggered immunity (PTI) is a central component of plant immunity. Activation of PTI relies on the recognition of microbe-derived structures, termed patterns, through plant encoded surface-resident pattern recognition receptors (PRRs). We have identified proteobacterial translation initiation factor 1 (IF1) as an immunogenic pattern that triggers PTI in Arabidopsis thaliana and some related Brassicaceae species. Unlike most other immunogenic patterns identified, IF1 elicitor activity cannot be assigned to a small peptide epitope, suggesting that tertiary fold features are required for IF1 receptor activation. We have deployed natural variation in IF1 sensitivity to identify leucine-rich repeat (LRR) receptor-like protein 32 (RLP32) as the corresponding Arabidopsis receptor using a restriction site-associated DNA sequencing (RAD-seq) approach. Transgenic expression of RLP32 confers IF1 sensitivity to rlp32 mutants, IF1-insensitive Arabidopsis accessions and IF1-insensitive Nicotiana benthamiana. RLP32 binds IF1 specifically and forms complexes with LRR receptor kinases SOBIR1 and BAK1 to mediate signaling. Similar to previously identified PRRs RLP32 confers resistance to Pseudomonas syringae infection, highlighting an unexpectedly complex array of bacterial pattern sensors within a single plant species.

plant biology↗

bioRxiv2020

Arabidopsis cell surface LRR immune receptor signaling through the EDS1-PAD4-ADR1 node

Plants use both cell surface and intracellular immune receptors with leucine rich-repeat (LRRs) to detect pathogens. LRR receptor kinases (LRR-RKs) and LRR receptor-like proteins (LRR-RPs) recognize extracellular microbe-derived molecules to confer pattern-triggered immunity (PTI), while nucleotide-binding LRR (NLR) proteins detect microbial effectors inside the cell to confer effector-triggered immunity (ETI). Despite PTI and ETI signaling being initiated in different compartments, both rely on the transcriptional activation of similar sets of genes, suggesting convergence in signaling upstream of nuclear events. Here we report that two sets of molecules, helper NLRs from the ADR1 (ACTIVATED DISEASE RESISTANCE 1) family as well as lipase-like proteins EDS1 (ENHANCED DISEASE SUSCEPTIBILITY 1) and PAD4 (PHYTOALEXIN DEFICIENT 4), are required not only for ETI, but also for PTI. A further similarity is seen in the evolutionary patterns of some PTI and ETI receptor genes, with both often being highly polymorphic, and with nevertheless distinct roles of LRR-RK and LRR-RP receptors in immunity. We find that the LRR-RK SOBIR1 directly links LRR-RPs with the ADR1 helper NLR as well as EDS1 and PAD4, suggesting the formation of constitutive supramolecular signalosome complexes at the inner side of the plasma membrane. We propose that the EDS1-PAD4-ADR1 node is an essential component and convergence point for immune signaling cascades activated by both surface-resident LRR-RP receptors and intracellular NLR receptors.

plant biology↗

bioRxiv2019

The BIR2/BIR3-interacting Phospholipase D gamma 1 negatively regulates immunity in Arabidopsis

Plants have evolved effective strategies to defend themselves against pathogen invasion. Starting from the plasma membrane with the recognition of microbe-associated molecular patterns (MAMPs) via pattern recognition receptors, internal cellular signaling pathways are induced to ultimately fend off the attack. Phospholipase D (PLD) hydrolyzes membrane phospholipids to produce phosphatidic acid (PA), which has been proposed to play a second messenger role in immunity. The Arabidopsis PLD family consists of 12 members and for some a specific function in resistance towards a subset of pathogens has been shown. We demonstrate here that Arabidopsis PLD{gamma}1, but not its close homologs PLD{gamma}2 and PLD{gamma}3, is specifically involved in plant immunity. Genetic inactivation of PLD{gamma}1 resulted in increased resistance towards the virulent bacterium Pseudomonas syringae pv. tomato DC3000 and the necrotrophic fungus Botrytis cinerea. As pld{gamma}1 mutant plants responded with elevated levels of reactive oxygen species to MAMP-treatment, a negative regulatory function for this PLD isoform is proposed. Importantly, PA levels in pld{gamma}1 mutants were not affected compared to stressed wild-type plants, suggesting that alterations in PA levels are unlikely the cause for the enhanced immunity in the pld{gamma}1 line. Instead, the plasma-membrane-attached PLD{gamma}1 protein colocalized and associated with the receptor-like kinases BIR2 and BIR3, which are known negative regulators of pattern-triggered immunity. Moreover, complex formation of PLD{gamma}1 and BIR2 was further promoted upon MAMP-treatment. Hence, we propose that PLD{gamma}1 acts as a negative regulator of plant immune responses in complex with immunity-related proteins BIR2 and BIR3.\n\nOne-sentence summaryA phospholipase D is a novel negative regulator of plant immunity and forms complexes with regulatory receptor-like kinases.

plant biology↗

bioRxiv2018

Symbiosis-related genes sustain the development of a downy mildew pathogen on Arabidopsis thaliana

AO_SCPLOWBSTRACTC_SCPLOWThe interfaces through which nutrients are transferred from plant cells to arbuscular mycorrhiza fungi and biotrophic hyphal pathogens are structurally similar. We report that in Arabidopsis thaliana, mutations in homologs of common symbiosis genes (CSGs) encoding homologs of the symbiosis receptor kinase SYMRK, the nucleoporins NUP133 and SEC13 or the cation channel POLLUX reduce the reproductive success of Hyaloperonospora arabidopsidis (Hpa). Analysis of the multiplication of extracellular bacterial pathogens, Hpa-induced cell death or callose accumulation, as well as Hpa-or flg22-induced defence marker gene expression, did not reveal any traces of constitutive or exacerbated defence responses. We discovered an age-dependent, possibly senescence-related transition of haustorial shape that occurred significantly earlier and at higher frequency in the CSG mutants. These findings point to a function of the homologs of common symbiosis genes in haustorial maintenance thus revealing an overlapping gene set for the intracellular accommodation of hyphal symbionts and pathogens.

cell biology↗

bioRxiv2018

The fungal ligand chitin directly binds and signals inflammation dependent on oligomer size and TLR2

Chitin is a highly abundant polysaccharide and linked to fungal infection and asthma. Unfortunately, its polymeric structure has hampered the identification of immune receptors directly binding chitin and signaling immune activation and inflammation, because purity, molecular structure and molarity are not well definable for a polymer typically extracted from biomass. Therefore, by using defined chitin (N-acetyl-glucosamine) oligomers, we identified six subunit long chitin chains as the smallest immunologically active motif and the innate immune receptor Toll-like receptor (TLR) 2 as the primary fungal chitin receptor on human and murine immune cells. Chitin oligomers directly bound TLR2 with nanomolar affinity and showed both overlapping and distinct signaling outcomes compared to known mycobacterial TLR2 ligands. Conversely, chitin oligomers shorter than 6 subunits were inactive or showed antagonistic effects on chitin/TLR2-mediated signaling, hinting to a size-dependent sensing/activation system unexpectedly conserved in plants and humans. Since blocking the chitin-TLR2 interaction effectively prevented chitin-mediated inflammation in vitro and in vivo, our study highlights the chitin TLR2 interaction as a potential target for developing novel therapies in chitin-related pathologies and fungal disease.

immunology↗