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Rhizobial Nod factors modulate reactive oxygen species, jasmonates, and pattern-recognizing receptors to suppress immune response

ABSTRACT: Fine-tuning of the immune response plays a key role in legume-rhizobial symbiosis. Rhizobial Nod factors can suppress the defense responses during symbiosis, but the possible mechanisms of such regulation remain poorly understood. Here, we observe that Nod factors effectively suppress the expression of genes encoding defense markers (WRKYs, PRs, PALs), the reactive oxygen species (ROS) formation, and reduce the content of pattern recognition receptor (PRR) LYK9 induced by treatment with deacetylated chitooligosaccharide CO8-DA in pea roots. Since PRR LYK9 may recognize both chitin/COs and peptidoglycan, it likely plays an important role in the activation of defense responses during rhizobial inoculation. To identify potential regulators through which Nod factors suppress the immune response in plants during symbiosis with rhizobia, proteome and transcriptome analyses were performed. This allowed identifying several potential candidates activated by Nod factors, such as superoxide dismutase and catalase enzymes, which prevent excessive ROS accumulation and the development of oxidative stress. We also found ubiquitin ligases and ubiquitin-conjugating enzymes that may target PRRs activated in response to rhizobial inoculation. LYK9 degradation via ubiquitinylation was shown to prevent a hypersensitive response in plants. Nod factors activate enzymes involved in jasmonic acid biosynthesis, which in turn activates the transcription factor ABR1, suppressing the abscisic acid-induced responses and decreasing the immune response. Finally, we showed that LysM-receptor-like kinases PsLYK11/MtLYK11, probable homologs of Arabidopsis AtLYK3 in pea and Medicago, are involved in regulation of the immune response.

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ORGANISM(S): Pisum Sativum (garden Pea)

TISSUE(S): Root

SUBMITTER: Tatiana Leonova

LAB HEAD: Dr. Elena Dolgikh

PROVIDER: PXD069618 | Pride | 2026-07-10

REPOSITORIES: Pride

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Project description:Palmitoylethanolamide (PEA) is an endogenous lipid mediator with recognized immunomodulatory actions, yet its effects in skeletal muscle remain poorly defined. We investigated whether PEA influences myogenesis and profiled the acute transcriptomic response of differentiated C2C12 myotubes. Dose-tolerability assays identified reduced metabolic activity and viability at 100 µM PEA, so 10 µM was used for subsequent experiments. Across differentiation, PEA decreased myotube number (90.3 ± 10.6 vs 112.6 ± 10.1 in control) while increasing nuclear fusion index (37.8 ± 5.7% vs 30.7 ± 3.2%); myotube area was unchanged. In myoblasts, 24 h PEA exposure increased the proportion of cells in G0/G1 (48.2 ± 1.2% vs 42.3 ± 1.9%) with a concomitant reduction in S-phase (21.7 ± 1.2% vs 25.5 ± 1.2%), consistent with G1 arrest and reduced proliferative drive. RNA sequencing revealed a distinct transcriptional signature after 24 h PEA treatment, with 1,952 differentially expressed genes (1,028 upregulated; 924 downregulated, q < 0.05). Pathway-level changes encompassed cytokine–receptor interactions, chemokine, JAK–STAT, NF-κB, NOD-like and Toll-like receptor signalling. PEA downregulated NF-κB target/pro-inflammatory cytokine genes (Il1a, Il1b, Il12a, Il33, Tnfrsf19) while upregulating interferon-related and chemokine genes (e.g., Stat2, Oas1b, Gbp7, Cxcl14, Ccl5), indicating an anti-inflammatory/immune-priming profile. Notably, N-acylethanolamine acid amidase (Naaa), a lysosomal PEA hydrolase, was highly expressed and significantly induced, whereas fatty acid amide hydrolase (Faah) was low and unchanged, suggesting a muscle-specific bias toward NAAA-dependent PEA metabolism. Peroxisome proliferator-activated receptor-α (PPARα) was expressed at low levels and unaffected by PEA, indicating that the observed responses occur largely independent of canonical PPARα signalling. Together, these data show that PEA biases skeletal muscle toward a less proliferative but more fused and inflammation-resolving phenotype, accompanied by transcriptional reprogramming of immune pathways and preferential engagement of NAAA. While in vitro concentrations exceed reported plasma levels and mRNA changes may not fully predict protein/function, these findings provide the first comprehensive transcriptomic characterisation of PEA in myotubes and motivate in vivo studies to test whether PEA’s dual actions translate into benefits for muscle regeneration, adaptation, or anti-atrophy interventions.

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Rhizobial Nod factors modulate reactive oxygen species, jasmonates, and pattern-recognizing receptors to suppress immune response

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