The N gene protects tomato plants from tomato brown rugose fruit virus infection.

The tobamovirus tomato brown rugose fruit virus (ToBRFV) has recently emerged, causing significant damage to the tomato industry in various regions worldwide, including the US. ToBRFV evades the widely used Tm-2 resistance gene, which encodes a nucleotide-binding leucine-rich repeat (NLR) class immune receptor with an N-terminal coiled-coil (CC) domain that confers resistance to the tomato mosaic virus (ToMV). In this study, we tested a transgenic tomato line (tomato) expressing the Nicotiana glutinosa N gene, which encodes an NLR with a Toll-Interleukin 1 homology domain (TIR) at the N-terminus, for resistance to ToBRFV.

A phosphorylation-driven ubiquitination switch fine-tunes Alfin-like 7-induced ROS signaling in plant immunity.

Maintaining reactive oxygen species (ROS) homeostasis is essential for balancing growth-defense trade-offs in plants. Although the transcription factors (TFs) that regulate ROS production and scavenging genes have been studied, the regulation of these TFs to control ROS accumulation remains poorly understood. Here, we demonstrate that during N nucleotide-binding leucine-rich repeat-mediated immunity, Alfin-like 7 (AL7) is ubiquitinated by the ubiquitin protein ligase E3 component N-recognin 7 (UBR7).

The proxiome of a plant viral protein with dual targeting to mitochondria and chloroplasts revealed MAPK cascade and splicing components as proviral factors.

The coat protein (CP) of the melon necrotic spot virus (MNSV) is a multifunctional factor localized in the chloroplast, mitochondria, and cytoplasm, playing a critical role in overcoming plant defenses such as RNA silencing (RNAi) and the necrotic hypersensitive response. However, the molecular mechanisms through which CP interferes with plant defenses remain unclear. Identifying viral-host interactors can reveal how viruses exploit fundamental cellular processes and help elucidate viral survival strategies.

Heritable virus-induced germline editing in tomato.

Here, we report the successful implementation of heritable virus-induced genome editing (VIGE) in tomato (Solanum lycopersicum). We generated three transgenic tomato lines expressing Streptococcus pyogenes Cas9 (SpCas9) under the control of Cauliflower mosaic virus 35S (35S), S. lycopersicum ribosomal protein S5A (SlRPS5A), or S.

A cell wall-associated kinase phosphorylates NLR immune receptor to negatively regulate resistosome formation.

Plants deploy intracellular nucleotide-binding leucine-rich repeats (NLRs) to detect pathogen effectors and initiate immune responses. Although the activation mechanism of some plant NLRs forming resistosomes has been elucidated, whether NLR resistosome assembly is regulated to fine-tune immunity remains enigmatic. Here we used an antiviral coiled coil-nucleotide-binding site-leucine rich repeat, Barley Stripe Resistance 1 (BSR1), as a model and demonstrate that BSR1 is phosphorylated.

Directed Evolution of a Plant Immune Receptor for Broad Spectrum Effector Recognition.

Rapid development of immune receptors that protect crops from emerging pathogens is a critical challenge . While novel immune receptors that recognize previously undetected pathogen effectors could provide protection against a wider range of pathogens, engineering such receptors has been constrained by the low throughput and speed of testing. We established yeast surface display as a high throughput platform to recapitulate plant immune receptor-ligand interactions and evolve new binding capabilities.

The XopAE Effector from pv. Targets HSP20-like p23 Cochaperone to Suppress Plant Basal Immunity.

Pathogenic bacteria use Type 3 effector proteins to manipulate host defenses and alter metabolism to favor their survival and spread. The non-model bacterial pathogen pv. () causes devastating disease in cassava.

Orchestrating ROS regulation: coordinated post-translational modification switches in NADPH oxidases.

Reactive oxygen species (ROS) are among the most important signaling molecules, playing a significant role in plant growth, development, and responses to various environmental stresses. Respiratory burst oxidase homologs (RBOHs) are key enzymes in ROS production. Plants tightly regulate the activation and deactivation of RBOHs through various post-translational modifications (PTMs), including phosphorylation, ubiquitination, S-nitrosylation, and persulfidation.

CHUP1 restricts chloroplast movement and effector-triggered immunity in epidermal cells.

Chloroplast Unusual Positioning 1 (CHUP1) plays an important role in the chloroplast avoidance and accumulation responses in mesophyll cells. In epidermal cells, prior research showed silencing CHUP1-induced chloroplast stromules and amplified effector-triggered immunity (ETI); however, the underlying mechanisms remain largely unknown. CHUP1 has a dual function in anchoring chloroplasts and recruiting chloroplast-associated actin (cp-actin) filaments for blue light-induced movement.

Structural insights into strigolactone catabolism by carboxylesterases reveal a conserved conformational regulation.

Phytohormone levels are regulated through specialized enzymes, participating not only in their biosynthesis but also in post-signaling processes for signal inactivation and cue depletion. Arabidopsis thaliana (At) carboxylesterase 15 (CXE15) and carboxylesterase 20 (CXE20) have been shown to deplete strigolactones (SLs) that coordinate various growth and developmental processes and function as signaling molecules in the rhizosphere. Here, we elucidate the X-ray crystal structures of AtCXE15 (both apo and SL intermediate bound) and AtCXE20, revealing insights into the mechanisms of SL binding and catabolism.

Attenuation of phytofungal pathogenicity of Ascomycota by autophagy modulators.

Autophagy in eukaryotes functions to maintain homeostasis by degradation and recycling of long-lived and unwanted cellular materials. Autophagy plays important roles in pathogenicity of various fungal pathogens, suggesting that autophagy is a novel target for development of antifungal compounds. Here, we describe bioluminescence resonance energy transfer (BRET)-based high-throughput screening (HTS) strategy to identify compounds that inhibit fungal ATG4 cysteine protease-mediated cleavage of ATG8 that is critical for autophagosome formation.

Conserved transcription factors NRZ1 and NRM1 regulate NLR receptor-mediated immunity.

Plant innate immunity mediated by the nucleotide-binding leucine-rich repeat (NLR) class of immune receptors plays an important role in defense against various pathogens. Although key biochemical events involving NLR activation and signaling have been recently uncovered, we know very little about the transcriptional regulation of NLRs and their downstream signaling components. Here, we show that the Toll-Interleukin 1 receptor homology domain containing NLR (TNL) gene N (Necrosis), which confers resistance to Tobacco mosaic virus, is transcriptionally induced upon immune activation.

High-quality assembled and annotated genomes of Nicotiana tabacum and Nicotiana benthamiana reveal chromosome evolution and changes in defense arsenals.

Nicotiana tabacum and Nicotiana benthamiana are widely used models in plant biology research. However, genomic studies of these species have lagged. Here we report the chromosome-level reference genome assemblies for N.

Virus-Induced Heritable Gene Editing in Plants.

Gene editing using clustered, regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) nuclease is an excellent tool for assessing gene function in plants. However, delivery of CRISPR/Cas-editing components into plant cells is still a major bottleneck and requires tissue culture-based approaches and regeneration of plants. To overcome this limitation, several plant viral vectors have recently been engineered to deliver single-guide RNA (sgRNA) targets into SpCas9-expressing plants.

TurboID-Based Proximity Labeling: A Method to Decipher Protein-Protein Interactions in Plants.

Proteins form complex networks through interaction to drive biological processes. Thus, dissecting protein-protein interactions (PPIs) is essential for interpreting cellular processes. To overcome the drawbacks of traditional approaches for analyzing PPIs, enzyme-catalyzed proximity labeling (PL) techniques based on peroxidases or biotin ligases have been developed and successfully utilized in mammalian systems.

Calponin homology domain containing kinesin, KIS1, regulates chloroplast stromule formation and immunity.

Chloroplast morphology changes during immunity, giving rise to tubule-like structures known as stromules. Stromules extend along microtubules and anchor to actin filaments along nuclei to promote perinuclear chloroplast clustering. This facilitates the transport of defense molecules/proteins from chloroplasts to the nucleus.

The NLR Immune Receptor Induces Early Transcriptional Changes in Response to Thrips and Mechanical Modes of Inoculation of .

The NLR (nucleotide-binding leucine-rich repeat) class immune receptor confers resistance to (TSWV). Although is known to activate immunity upon recognition of the TSWV movement protein NSm, we know very little about the downstream events that lead to resistance. Here, we investigated the -mediated early transcriptomic changes that occur in response to mechanical and thrips-mediated inoculation of TSWV, using near-isogenic tomato lines CNPH-LAM 147 () and Santa Clara ().

NLRs derepress MED10b- and MED7-mediated repression of jasmonate-dependent transcription to activate immunity.

Plant intracellular nucleotide-binding domain, leucine-rich repeat-containing receptors (NLRs) activate a robust immune response upon detection of pathogen effectors. How NLRs induce downstream immune defense genes remains poorly understood. The Mediator complex plays a central role in transducing signals from gene-specific transcription factors to the transcription machinery for gene transcription/activation.

Breaking Boundaries: The Perpetual Interplay Between Tobamoviruses and Plant Immunity.

Plant viruses of the genus cause significant economic losses in various crops. The emergence of new tobamoviruses such as the tomato brown rugose fruit virus (ToBRFV) poses a major threat to global agriculture. Upon infection, plants mount a complex immune response to restrict virus replication and spread, involving a multilayered defense system that includes defense hormones, RNA silencing, and immune receptors.

Activation of Tm-2 resistance is mediated by a conserved cysteine essential for tobacco mosaic virus movement.

The tomato Tm-2 gene was considered to be one of the most durable resistance genes in agriculture, protecting against viruses of the Tobamovirus genus, such as tomato mosaic virus (ToMV) and tobacco mosaic virus (TMV). However, an emerging tobamovirus, tomato brown rugose fruit virus (ToBRFV), has overcome Tm-2 , damaging tomato production worldwide. Tm-2 encodes a nucleotide-binding leucine-rich repeat (NLR) class immune receptor that recognizes its effector, the tobamovirus movement protein (MP).