Biology subjects

Rodrigo, I.

Publications and source records attributed to Rodrigo, I..

6 recordsLinked to original sources

bioRxiv2026

Hydroxylated monoterpenes mimic bacterial attack to trigger jasmonate-dependent self-amplifying immunity in tomato

Hydroxylated monoterpenes (HMTPs) are emitted during resistant tomato-Pseudomonas syringae interactions and confer antibacterial resistance, yet their integration into immune signalling remains poorly understood. Here we show that HMTPs act as endogenous mimics of pathogen attack that engage canonical defence pathways in tomato. Using -terpineol as a representative HMTP, we demonstrate that this volatile activates MPK kinase-, calcium- and reactive oxygen species (ROS)-dependent signalling, promotes jasmonate and salicylic acid accumulation, and induces pathogen-like stomatal immunity independently of abscisic acid. Functional analyses revealed that HMTPs biosynthesis depends on ROS and jasmonate signalling, and HMTPs further promote their own accumulation, establishing a self-reinforcing feed-forward mechanism. Moreover, in vitro oxidative conditions drive chemical remodelling and selective interconversion among HMTPs, contributing to volatile diversification and favouring the accumulation of highly bioactive hydroxylated forms. Consistently, deuterium-labelled linalool is incorporated into plant metabolism and converted into deuterated -terpineol in planta, providing direct evidence for volatile interconversion. Together, our findings establish HMTPs as dynamic amplifiers of tomato antibacterial immunity and key actors in pathogen-associated signalling.

plant biology↗

bioRxiv2025

Mitophagy-related prohibitin 2 is an orthoflavivirus restriction factor targeted for degradation during infection

1.Orthoflaviviruses (OFVs) are the primary cause of arboviral disease worldwide, leading to large numbers of hospitalisations, sequelae and deaths associated. Viral genome replication is catalysed by non-structural protein 5 (NS5), the largest and most conserved protein in these viruses. Here, we used quantitative proteomics to identify host cell interactors of Zika virus (ZIKV), Usutu virus (USUV), and West Nile virus (WNV) NS5 proteins. A total of 141 proteins were detected in the analysis, being 26 of them associated with all three NS5s. Downregulation of myoferlin and LGALS3BP led to decreases in the virus yields, indicating that their presence is needed for efficient virus replication. Conversely, silencing of mitophagy-related prohibitin 2 (PHB2) resulted in significant increases in ZIKV and USUV titres and genomic RNA. Supporting a connection between this antiviral behaviour and mitophagy, an inhibitor of this pathway also led to larger virus titres and viral RNA copies. During infection, PHB2 protein levels gradually decreased, becoming transiently undetectable, and suggesting that it is specifically targeted by the virus for degradation. Ectopic expression of NS5 alone did not affect PHB2 intracellular abundance, however a significant decrease was observed in cells expressing the viral protease NS2B/NS3. Its elimination, was more pronounced in cells where both viral enzymes were co-expressed, supporting that NS5 could be assisting PHB2 recognition by NS2B/NS3. Overall, our results support that PHB2 is an orthoflaviviral restriction factor which is targeted for degradation to favour viral replication. 2. IMPORTANCEOrthoflaviviruses (OFVs) are one of the main causes of disease transmitted by mosquitoes, with millions of infections occurring annually, and large numbers of deaths associated. Some important members include viruses causing dengue, yellow fever, and Zika-associated disease (e.g. microcephaly). OFVs contain a genome which is copied by a highly conserved viral protein termed NS5. Prohibitin 2 (PHB2) is a host cellular protein involved in different cellular processes including mitophagy, a mechanism to remove damaged mitochondria. Here, we show that human PHB2 interacts with orthoflaviviral NS5. Depletion of PHB2 results in increased viral yields, suggesting that this protein restricts orthoflaviviral infection. We found that PHB2 is targeted for degradation by the orthoflaviviral protease in a process aided by NS5, supporting that several viral proteins act concertedly to eliminate PHB2, counteracting its antiviral activity, and thus favouring infection.

microbiology↗

bioRxiv2024

TOR Inhibition Enhances Autophagic Flux and Immune Response in Tomato Plants Against PSTVd Infection

Viroids are small, non-coding RNA pathogens known for their ability to cause severe plant diseases. Despite their simple structure, viroids like Potato Spindle Tuber Viroid (PSTVd) can interfere plant cellular processes, including both transcriptional and post-transcriptional mechanisms, thereby impacting plant growth and yield. In this study, we have investigated the role of the Target Of Rapamycin (TOR) signalling pathway in modulating viroid pathogenesis in tomato plants infected with PSTVd. Our findings reveal that PSTVd infection induces the accumulation of the selective autophagy receptor NBR1, potentially inhibiting autophagic flux. Pharmacological inhibition of TOR with AZD8055 mitigated PSTVd symptomatology by reducing viroid accumulation. Furthermore, TOR inhibition promoted the recovery of autophagic flux through NBR1 and primed the plant defence response, as evidenced by enhanced expression of both the defence-related gene PR1b and S5H, a gene involved in the salicylic acid catabolism. These results suggest a novel role for TOR in regulating viroid-induced pathogenesis and highlight the potential of TOR inhibitors as tools for enhancing plant resistance against viroid infections.

plant biology↗

bioRxiv2023

Signaling mechanisms and agricultural applications of (Z)-3-Hexenyl Butyrate-mediated stomatal closure

Biotic and abiotic stresses can severely limit crop productivity. In response to drought, plants close stomata to prevent water loss. Besides, stomata are considered the main entrance of several pathogens. Therefore, the development of natural products to control stomata closure can be considered a sustainable strategy to cope with stresses in agriculture. Plants respond to different stresses by releasing volatile organic compounds (VOCs). Green leaf volatiles (GLVs), which are commonly produced across different plant species after tissue damage, comprise an important group within VOCs. Among them, (Z)-3-hexenyl butyrate (HB) was described as a natural inducer of stomatal closure, playing an important role in stomatal immunity, although its mechanism of action is still unknown. Here, through different genetic, pharmacological, and biochemical approaches, we uncover that HB perception initiates various defense signaling events such as activation of Ca2+ permeable channels, mitogen-activated protein kinases (MPKs) and production of NADPH oxidase-mediated reactive oxygen species (ROS). Furthermore, HB-mediated stomata closure resulted to be independent of abscisic acid (ABA) biosynthesis and signaling. Additionally, exogenous treatments with HB alleviate water stress and improve fruit productivity in tomato plants. The efficacy of HB was also tested under open field conditions, leading to enhanced resistance against Phytophthora spp. and Pseudomonas syringae infection in potato and tomato plants, respectively. Taken together, our results provide insights into HB signaling transduction pathway, confirming its role in stomatal closure and plant immune system activation, and proposing HB as a new phytoprotectant for the sustainable control of biotic and abiotic stresses in agriculture.

molecular biology↗

bioRxiv2023

Metabolic crosstalk between hydroxylated monoterpenes and salicylic acid in tomato defence response against Pseudomonas syringae pv tomato

Hydroxylated monoterpenes (HMTPs) are differentially emitted by tomato plants efficiently resisting a bacterial infection. We have studied the defensive role of these volatiles in the tomato response to bacteria, whose main entrance are stomata apertures. Treatments with some HMTPs resulted in stomatal closure and PR1 induction. Particularly, -terpineol induced stomatal closure in a salicylic (SA) and abscisic acid-independent manner, and conferred resistance to bacteria. Interestingly, transgenic tomato plants overexpressing or silencing the monoterpene synthase MTS1, which displayed alterations in the emission of HMTPs, exhibited changes in the stomatal aperture but not in plant resistance. Measures of both 2-C-methyl-D-erythritol-2,4-cyclopyrophosphate (MEcPP) and SA levels, revealed a competition for MEcPP by the methylerythritol phosphate (MEP) pathway and the SA biosynthesis activation, thus explaining the absence of phenotype in transgenic plants. These results were confirmed by chemical inhibition or activation of the MEP pathway. Besides, treatments with BTH, a SA functional analogue, conferred enhanced resistance in transgenic tomato plants overexpressing MTS1. Finally, plants overexpressing MTS1 induced PR1 and stomata closure in neighbouring plants. Our results confirm the role of HMTPs in both intra and inter-plant immune signalling, and reveal a metabolic crosstalk between the MEP and SA pathways in tomato plants. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=124 SRC="FIGDIR/small/539605v2_ufig1.gif" ALT="Figure 1"> View larger version (19K): org.highwire.dtl.DTLVardef@1048cddorg.highwire.dtl.DTLVardef@1bc1948org.highwire.dtl.DTLVardef@1ff0eaorg.highwire.dtl.DTLVardef@101c4fa_HPS_FORMAT_FIGEXP M_FIG Metabolic crosstalk between hydroxylated monoterpenes and salicylic acid in tomato defence response against Pseudomonas syringae pv tomato. Created with BioRender.com. C_FIG

plant biology↗

bioRxiv2022

SlS5H silencing reveals specific pathogen-triggered salicylic acid metabolism in tomato

The phytohormone salicylic acid (SA or 2-hydroxybenzoic acid) plays an important role in plant biotic and abiotic responses. Gentisic acid (GA or 2,5-Dihydroxybenzoic acid, 2,5-DHBA) is the product of the SA 5-hydroxylation which is catalysed by the S5H enzyme, also known as DMR6. GA has been described to accumulate at high levels in compatible plant-pathogen interactions such as tomato plants infected by Citrus Exocortis Viroid (CEVd), and to a much lesser extend upon Pseudomonas syringae DC3000 pv. tomato (Pst) infection. Here we describe the specific effect that tomato SlS5H impairment produces on both plant-pathogen interactions. The induction of SlS5H in tomato plants by different pathogens was corroborated by qRT-PCR and correlated with previously described 2,5-DHBA accumulations. Transient SlS5H over-expression assays in Nicotiana benthamiana confirmed that SA is a substrate for SlS5H in vivo. RNAi_SlS5H tomato transgenic plants were generated and characterized upon CEVd and Pst infections. Transgenic tomato plants displayed an activation of defences and therefore a loss of susceptibility against both pathogens, and alternative SA homeostasis seems to occur for each specific interaction. Metabolomic assays revealed that whilst the glycosylated form of SA was the most discriminant metabolite found in CEVd infected RNAi_SlS5H transgenic plants, trans-feruloyldopamine, feruloylquinic acid, feruloylgalactarate and 2-hydroxyglutarate were the most accumulated compounds in the Pst-infected transgenic tomato leaves. Transgenic lines also displayed hyper susceptibility to Botrytis cinerea, as well as a smaller size and early senescence. Collectively, our results reveal a novel mechanism by which tomato plants specifically set SA homeostasis upon different pathogen attacks. One sentence summaryThe impairment of SA hydroxylation in tomato plants uncovers specific SA homeostasis upon CEVd or Pseudomonas syringae infections.

biochemistry↗