Ojha, B. | bioRxiv Science

bioRxiv2026

AtNHR2A and AtNHR2B participate in unconventional protein secretion in response to environmental stress

The Arabidopsis thaliana nonhost resistance proteins 2A (AtNHR2A) and 2B (AtNHR2B) play crucial roles in plant immunity as the single mutants Atnhr2a and Atnhr2b and the double mutant Atnhr2bAtnhr2a are susceptible to the non-adapted pathogen Pseudomonas syringae pv. tabaci that is unable to infect wild-type Col-0 plants. The localization of fluorescent versions of AtNHR2A and AtNHR2B to compartments of the endomembrane system together with their interaction with secreted proteins suggested a function in endomembrane-mediated secretory processes participating in plant immunity. Comparative apoplastic proteomics analysis between wild type Col-0 and the double mutant Atnhr2bAtnhr2a after treatment P. syringae pv. tabaci, revealed that AtNHR2A and AtNHR2B are indeed required for the secretion of proteins containing N-terminal signal peptides that occurs through the conventional protein secretion pathway. In this work, we leveraged these apoplastic proteomics datasets to identify proteins lacking N-terminal signal peptide and expected to be secreted through unconventional secretion pathway(s). We discovered that AtNHR2A and AtNHR2B are also required for the secretion of proteins through an unconventional secretion pathway that, intriguingly, included proteins previously associated with abiotic stress. These findings led us to define the subcellular dynamics of AtNHR2A and AtNHR2B, and through co-localization analyses and the use of vesicle trafficking inhibitors, we uncovered their trafficking pathways transitioning through Golgi-dependent and Golgi-independent pathways to ultimately reach the central vacuole. Our findings suggest that AtNHR2A and AtNHR2B participate in a multivesicular bodies-vacuole-mediated unconventional secretion pathway that results in the release of proteins involved in plant responses to environmental stresses.

plant biology↗

bioRxiv2025

Azospirillum brasilense Sp7 and tomato seedling exudate or cytidine reduced disease symptoms on yellow pear tomatoes caused by Pseudomonas syringae pv. tomato DC3000

Tomatoes are an important crop worldwide and phytopathogens can cause devastating losses. This study describes a treatment, consisting of Azospirillum brasilense Sp7 and either tomato seedling exudate or the exudate compound cytidine. The combination of A. brasilense Sp7 with cytidine showed a remarkable reduction of 83.4% in disease severity of tomatoes challenged with Pseudomonas syringae pv. tomato DC3000. Replacing cytidine with exudate was less effective at 71%, but the reduction in disease severity was still larger than by A. brasilense Sp7 alone at 55%. This reduction in disease severity was not paralleled by a decrease in P. syringae in leaf homogenates. Cytidine caused a 6.7 fold increase in A. brasilense Sp7 16S rDNA in root homogenates. In phyllosphere and rhizosphere, treatments modulated the microbial composition. In the phyllosphere, specific associations between treatment groups and bacterial orders could be computed. In the rhizosphere, principal component analysis revealed that variation along PC1 was dominated by the presence or absence of A. brasilense. Intriguingly, the inoculant caused an increase in the abundance of other Azospirillales species. Les tomates sont une culture dimportance mondiale, et les phytopathogenes peuvent entrainer des pertes devastatrices. Notre etude decrit un traitement compose dAzospirillum brasilense Sp7 et soit dun exsudat de plantules de tomate, soit de la cytidine, un compose particulier provenant des exsudats et suppose avoir un effet chimiotactique. Lassociation de A. brasilense Sp7 avec la cytidine a entraine une reduction remarquable de 83,4 % de la severite de la maladie parmi les plantes de tomate infectees par Pseudomonas syringae pv. tomato DC3000. Le remplacement de la cytidine par des exsudats etait moins efficace (71 %), bien que la reduction de la severite de la maladie ait demeure superieure a celle obtenue avec A. brasilense Sp7 seul (55 %). Cette diminution de la severite de la maladie na toutefois pas ete accompagnee dune reduction de la concentration de P. syringae dans les homogenats foliaires. La cytidine a provoque une augmentation de 6,7 fois de lADNr 16S de A. brasilense Sp7 dans les homogenats racinaires. Dans la phyllosphere et la rhizosphere, les traitements ont module la composition microbienne. Dans la phyllosphere, des associations specifiques entre les groupes du traitement et les ordres bacteriens ont pu etre calculees. Dans la rhizosphere, lanalyse en composantes principales a montre que la variation de laxe PC1 etait dominee par la presence ou labsence dA. brasilense. De maniere interessante, linoculum a entraine une augmentation de labondance dautres especes dAzospirillales.

microbiology↗