Lateral roots (LRs) increase the contact area of the root with the rhizosphere and thereby improve water and nutrient uptake from the soil. LRs are generated either via a developmentally controlled mechanism or through induction by external stimuli, such as water and nutrient availability. Auxin regulates LR organogenesis via transcriptional activation by an auxin complex receptor. Endocytic trafficking to the vacuole positively regulates LR organogenesis independently of the auxin complex receptor in Arabidopsis (Arabidopsis thaliana). Here, we demonstrate that phosphatidylinositol 4-phosphate (PI4P) biosynthesis regulated by the phosphatidylinositol 4-kinases PI4KIIIβ1 and PI4KIIIβ2 is essential for the LR organogenesis driven by endocytic trafficking to the vacuole. Stimulation with Sortin2, a biomodulator that promotes protein targeting to the vacuole, altered PI4P abundance at both the plasma membrane and endosomal compartments, a process dependent on PI4K activity. These findings suggest that endocytic trafficking to the vacuole regulated by the enzymatic activities of PI4KIIIβ1 and PI4KIIIβ2 participates in a mechanism independent of the auxin complex receptor that regulates LR organogenesis in Arabidopsis. Surprisingly, loss-of-function of PI4KIIIβ1 and PI4KIIIβ2 induced both LR primordium formation and endocytic trafficking toward the vacuole. This LR primordium induction was alleviated by exogenous PI4P, suggesting that PI4KIIIβ1 and PI4KIIIβ2 activity constitutively negatively regulates LR primordium formation. Overall, this research demonstrates a dual role of PI4KIIIβ1 and PI4KIIIβ2 in LR primordium formation in Arabidopsis.

Pseudomonas syringae pv. syringae is the main causal agent of bacterial canker in sweet cherry in Chile, causing significant economic losses. Cultivars exhibit diverse susceptibility in the field and the molecular mechanisms underlying the differential responses remain unclear. RNA-seq analysis was performed to characterize the transcriptomic response in cultivars Santina and Bing (less and more susceptible to P. syringae pv. syringae, respectively) after 1 and 7 days post-inoculation (dpi) with the bacterium. Symptoms of bacterial canker became evident from the fifth day. At 1 dpi, cultivar Santina showed a faster response to infection and a larger number of differentially expressed genes (DEGs) than cultivar Bing. At 7 dpi, cultivar Bing almost doubled its DEGs, while cultivar Santina tended to the normal DEG levels. P. syringae pv. syringae infection downregulated the expressions of key genes of the photosynthesis process at 1 dpi in the less susceptible cultivar. The results suggest that the difference in susceptibility to P. syringae pv. syringae is linked to the timeliness of pathogen recognition, limiting the bacteria’s dispersion through modeling its cell wall, and regulation of genes encoding photosynthesis pathway. Through this study, it has been possible to progress the knowledge of relevant factors related to the susceptibility of the two studied cherry cultivars to P. syringae pv. syringae.

Bacterial canker caused by Pseudomonas syringae pv. syringae (Pss) is responsible for substantial loss to the production of sweet cherry in Chile. To date, the molecular mechanisms of the Pss–sweet cherry interaction and the disease-related genes in the plant are poorly understood. In order to gain insight into these aspects, a transcriptomic analysis of the sweet cherry cultivar ‘Lapins’ for differentially expressed genes (DEGs) in response to Pss inoculation was conducted. Three Pss strains, A1M3, A1M197, and 11116_b1, were inoculated in young twigs, and RNA was extracted from tissue samples at the inoculation site and distal sections. RNA sequencing and transcriptomic expression analysis revealed that the three strains induced different patterns of responses in local and distal tissues. In the local tissues, A1M3 triggered a much more extensive response than the other two strains, enriching DEGs especially involved in photosynthesis. In the distal tissues, the three strains triggered a comparable extent of responses, among which 11116_b1 induced a group of DEGs involved in defense responses. Furthermore, tissues from various inoculations exhibited an enrichment of DEGs related to carbohydrate metabolism, terpene metabolism, and cell wall biogenesis. This study opened doors to future research on the Pss–sweet cherry interaction, immunity responses, and disease control.

Disease severity and drought due to climate change present significant challenges to orchard productivity. This study examines the effects of spring inoculation with Pseudomonas syringae pv. syringae (Pss) on sweet cherry plants, cvs. Bing and Santina with varying defense responses, assessing plant growth, physiological variables (water potential, gas exchange, and plant hydraulic conductance), and the levels of abscisic acid (ABA) and salicylic acid (SA) under two summer irrigation levels. Pss inoculation elicited a more pronounced response in ‘Santina’ compared to ‘Bing’ at 14 days post-inoculation (dpi), and those plants inoculated with Pss exhibited a slower leaf growth and reduced transpiration compared to control plants during 60 dpi. During differential irrigations, leaf area was reduced 14% and 44% in Pss inoculated plants of ‘Bing’ and ‘Santina’ respectively, under well-watered (WW) conditions, without changes in plant water status or gas exchange. Conversely, water-deficit (WD) conditions led to gas exchange limitations and a 43% decrease in plant biomass compared to that under WW conditions, with no differences between inoculation treatments. ABA levels were lower under WW than under WD at 90 dpi, while SA levels were significantly higher in Pss-inoculated plants under WW conditions. These findings underscore the influence on plant growth during summer in sweet cherry cultivars that showed a differential response to Pss inoculations and how the relationship between ABA and SA changes in plant drought level responses.

Bacterial canker is an important disease of sweet cherry plants mainly caused by Pseudomonas syringae pv. syringae (Pss). Water deficit profoundly impairs the yield of this crop. Nitric oxide (NO) is a molecule that plays an important role in the plant defense mechanisms. To evaluate the protection exerted by NO against Pss infection under normal or water-restricted conditions, sodium nitroprusside (SNP), a NO donor, was applied to sweet cherry plants cv. Lapins, before they were exposed to Pss infection under normal or water-restricted conditions throughout two seasons. Well-watered plants treated with exogenous NO presented a lower susceptibility to Pss. A lower susceptibility to Pss was also induced in plants by water stress and this effect was increased when water stress was accompanied by exogenous NO. The lower susceptibility to Pss induced either by exogenous NO or water stress was accompanied by a decrease in the internal bacterial population. In well-watered plants, exogenous NO increased the stomatal conductance and the net CO₂ assimilation. In water-stressed plants, NO induced an increase in the leaf membranes stability and proline content, but not an increase in the CO₂ assimilation or the stomatal conductance.

Climate change is intensifying the simultaneous occurrence of biotic and abiotic stresses in fruit crops, but yet the molecular mechanisms underlying plant responses remain poorly understood. The physiological and transcriptomic responses of two sweet cherry (Prunus avium L.) cultivars, Santina and Bing, grafted onto Gisela 12, were investigated under single and combined stresses imposed by Pseudomonas syringae pv. syringae and water deficit. Although biomass, gas exchange, and hormone accumulation showed only minor changes, combined stress triggered distinct cultivar-dependent transcriptional reprogramming. The cultivar Bing exhibited a pronounced response with 4261 differentially expressed genes (DEGs), characterized by strong repression of photosynthetic processes and activation of defense- and hormone-related pathways. In contrast, the cultivar Santina showed a moderate response with 674 DEGs, primarily reinforcing structural and secondary metabolism. Cultivar-specific modulation of abscisic acid sensitivity was associated with the contrasting regulation of WRKY40 and Sin3-like repressors, despite comparable ABA levels. Strikingly, both cultivars upregulated the GIGANTEA gene, underscoring its role as a central regulatory hub linking circadian rhythm, stomatal function, and hormonal crosstalk under dual stress. Collectively, these results reveal non-additive, genotype-specific transcriptional strategies in sweet cherry trees, providing insights into stress integration in fruit trees and identifying regulatory genes that may inform breeding and management strategies for resilience under climate change.