Biology subjects

Vleeshouwers, V.

Publications and source records attributed to Vleeshouwers, V..

4 recordsLinked to original sources

bioRxiv2026

TurboID-based proximity-dependent labeling using SOBIR1 as a bait in potato leads to the identification of novel defense-related signaling partners

The plant immune system comprises a complex signaling network that is activated upon perceiving molecules derived from invading pathogens. The first line of defense at the plant cell surface is mediated by receptor-like proteins (RLPs) and receptor-like kinases (RLKs). RLPs, which lack a cytoplasmic signalling domain themselves, constitutively interact with the RLK SUPRESSOR OF BIR1-1 (SOBIR1), which is a key component initiating immune signal transduction upon pathogen perception. Therefore, elucidating the composition of the SOBIR1 protein complex will contribute to understanding the basic molecular mechanisms of plant disease resistance. Most of the studies focused on the identification of SOBIR1-interacting proteins are limited to model plants, due to technical challenges and lack of reliable genome and proteome databases in crop plants. Here, we evaluate the application of the biotin ligase TurboID (TbID)-based proximity-dependent labeling (PL) approach by transiently expressing SOBIR1 from Nicotiana benthamiana (NbSOBIR1), fused to TbID in leaves of the wild potato Solanum microdontum. We show that NbSOBIR1-YFP-TbID properly accumulates in potato and that proximal proteins are biotinylated. Quantitative proteomic analysis yielded over 130 candidate proteins to be in the proximity of the cytoplasmic kinase domain of NbSOBIR1, of which some could be linked to disease resistance by KEGG pathway and gene ontology (GO) molecular function analysis. We also studied the dynamics of the proteome in proximity of NbSOBIR1 upon perception of the INF1 elicitin of Phytophthora infestans that was co-expressed in potato with the elicitin receptor ELR, which is an RLP that constitutively interacts with SOBIR1. We found more than 80 proteins, including the NB-LRR REQUIRED FOR HR-ASSOCIATED CELL DEATH 1 (NRC1), putatively interacting with NbSOBIR1. In conclusion, we were able to successfully apply PL in potato and a future roadmap for further research on deciphering the composition of protein complexes involved in immune signaling has been established.

plant biology↗

bioRxiv2023

Stacking of PRRs in potato to achieve enhanced resistance against Phytophthora infestans

Plants employ pattern recognition receptors (PRRs) to sense pathogen-associated molecular patterns (PAMPs) or apoplastic effectors at the plant cell surface, as well as nucleotide-binding domain leucine-rich-repeat-containing receptors (NLRs) to sense effectors inside the plant cell. Breeding for potato resistance to P. infestans has focused on the use of NLRs, however, these genes are typically quickly overcome since the matching avirulence genes evolve exceptionally quickly. Here, we stacked two PRRs, PERU and RLP23, that recognize the rather conserved Phytophthora PAMPs Pep-13/25 and nlp20, respectively, in the potato cultivar Atlantic, and evaluated their effect on P. infestans resistance. We found that PERU and RLP23 cooperate for the early immune responses like the accumulation of reactive oxygen species (ROS) and production of ethylene by recognizing their corresponding PAMPs. Furthermore, we show that potato plants overexpressing these two PRRs are slightly less affected by P. infestans compared to the single transformants. Together, our data suggest that pyramiding of surface receptors can provide additional enhanced resistance against pathogens, however, more effective or synergistic combinations that may include intracellular NLR receptors should be explored.

plant biology↗

bioRxiv2020

RLP/K enrichment sequencing; a novel method to identify receptor-like protein (RLP) and receptor-like kinase (RLK) genes

O_LIThe identification of immune receptors in crop plants is time-consuming but important for disease control. Previously, resistance gene enrichment sequencing (RenSeq) was developed to accelerate mapping of nucleotide-binding domain and leucine-rich repeat containing (NLR) genes. However, resistances mediated by pattern recognition receptors (PRRs) remain less utilised. C_LIO_LIHere, our pipeline shows accelerated mapping of PRRs. Effectoromics leads to precise identification of plants with target PRRs, and subsequent RLP/K enrichment sequencing (RLP/KSeq) leads to detection of informative SNPs that are linked to the trait. C_LIO_LIUsing Phytophthora infestans as a model, we identified Solanum microdontum plants that recognize the apoplastic effectors INF1 or SCR74. RLP/KSeq in a segregating Solanum population confirmed the localization of the INF1 receptor on chromosome 12, and lead to the rapid mapping of the response to SCR74 to chromosome 9. By using markers obtained from RLP/KSeq in conjunction with additional markers, we fine-mapped the SCR74 receptor to a 43-kbp G-LecRK locus. C_LIO_LIOur findings show that RLP/KSeq enables rapid mapping of PRRs and is especially beneficial for crop plants with large and complex genomes. This work will enable the elucidation and characterisation of the non-NLR plant immune receptors and ultimately facilitate informed resistance breeding. C_LI

plant biology↗

bioRxiv2019

Cleavage of a pathogen apoplastic protein by plant subtilases activates immunity

The plant apoplast is a harsh environment in which hydrolytic enzymes, especially proteases, accumulate during pathogen infection. However, the defense functions of most apoplastic proteases remains largely elusive. Here, we show that a newly identified small cysteine-rich secreted protein PC2 from the potato late blight pathogen Phytophthora infestans induces immunity in Solanum plant species only after cleavage by plant apoplastic subtilisin-like proteases, such as tomato P69B. A minimal 61-amino-acid core peptide carrying two key cysteines and widely conserved among most oomycete species is sufficient for PC2 activity. Kazal-like protease inhibitors, such as EPI1 produced by P. infestans can prevent PC2 cleavage and dampen PC2 elicited host immunity. This study reveals that cleavage of pathogen proteins to release immunogenic peptides is an important function of apoplastic proteases but that pathogens interfere with these functions using protease inhibitor effectors.

plant biology↗