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Functional genomics screening in Chlamydomonas reinhardtii maps the genetic landscape of tolerance to paraquat and diuron
Tim Godec, Carissa Bleker, Katja Stare, Tjaša Lukan, Valentina Levak, Magda Tušek-Žnidarič, Tina Kosjek, Katarina Petra Van Midden, Marina Klemenčič, Kristina Sepčić, Maruša Kerenčič, Tina Eleršek, Urban Bren, Marko Jukič, Samo Lešnik, Anže Županič, 2026, original scientific article

Abstract: Functional genomics offers a powerful, unbiased approach to elucidating the molecular mechanisms underlying the toxicity of environmental pollutants. In this study, we applied genome-wide screening in Chlamydomonas reinhardtii to investigate two classical herbicides: paraquat and diuron. Our screen successfully uncovered critical nuclear-encoded pathways that govern susceptibility. For both herbicides, we identified genes regulating the assembly and maintenance of the photosynthetic machinery, highlighting the central role of nuclear control over these chloroplast-localized targets. Beyond these target-related factors, we discovered novel nontarget-site resistance mechanisms. For paraquat, we identified intracellular trafficking as the central determinant of toxicity, experimentally characterizing a P5B ATPase transporter and a fatty acid elongation pathway whose disruption, we propose, converges on the same endomembrane delivery route through sphingolipid depletion. In contrast, our screening data suggest that diuron tolerance may be associated with a metabolic strategy focused on energy conservation, where the inactivation of specific NADPH-consuming enzymes could preserve reducing power for essential antioxidant defense. Collectively, these findings demonstrate that functional genomics can reveal novel, complex modes of action even for well-characterized chemicals, providing the mechanistic resolution required to advance modern ecotoxicological risk assessments.
Keywords: paraquat, diuron
Published in DiRROS: 09.06.2026; Views: 321; Downloads: 369
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3.
Jasmonic acid and salicylic acid interact to determine spatial regulation of gene expression responses in potato leaf to herbivory by Colorado potato beetle and mechanical wounding : version v1
Valentina Levak, Tjaša Mahkovec Povalej, Karmen Pogačar, Katja Stare, Maja Zagorščak, Tim Hawkins, Joanne Robson, David Dobnik, Tjaša Lukan, Kristina Gruden, 2025, research data

Abstract: Raw supporting material for article Jasmonic acid and salicylic acid interact to determine spatial regulation of gene expression responses in potato leaf to herbivory by Colorado potato beetle and mechanical wounding.
Keywords: potato, jasmonic acid, salicylic acid, Colorado potato beetle, plant response to wounding
Published in DiRROS: 08.05.2026; Views: 311; Downloads: 442
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Jasmonic acid and salicylic acid interact to determine spatial regulation of gene expression responses in potato leaf to herbivory by Colorado potato beetle and mechanical wounding
Valentina Levak, Tjaša Mahkovec Povalej, Karmen Pogačar, Katja Stare, Maja Zagorščak, Tim Hawkins, Joanne Robson, David Dobnik, Tjaša Lukan, Kristina Gruden, 2025, original scientific article

Abstract: We investigated the spatial dynamics of potato (Solanum tuberosum) responses to herbivory and mechanical wounding. We first followed the spatiotemporal response of jasmonic acid (JA) signaling, known to be involved in the response. We generated two potato sensor lines: a JAZ degradation sensor and a downstream multicystatin (MC) transcriptional reporter. Both sensors revealed concentric, locally restricted responses on wounded leaves. Notably, JAdependent gene expression was absent in cells immediately adjacent to the wound, whereas JAZ degradation spread continuously outward from the wound site. This pattern occurred after both herbivore attack and mechanical injury by the needle. To probe the mechanism, a salicylic acid (SA) reporter showed SA accumulation near the wound. Introducing the MC reporter into SA-depleted NahG plants produced a uniform spread of MC expression, confirming that SA attenuates the JA response in proximal cells. Together, these results show that a locally distinct, spatiotemporal SA–JA crosstalk shapes wound responses in potato, extending principles known from pathogen–plant interactions to herbivory and mechanical damage.
Keywords: Colorado potato beetle, herbivore, hormonal crosstalk, jasmonic acid, potato, salicylic acid, wounding
Published in DiRROS: 28.01.2026; Views: 644; Downloads: 690
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Functionality of potato virus Y coat protein in cell-to-cell movement dynamics is defined by its N-terminal region
Anže Vozelj, Tjaša Mahkovec Povalej, Katja Stare, Magda Tušek-Žnidarič, Katarina Bačnik, Valentina Levak, Ion Gutiérrez-Aguirre, Marjetka Podobnik, Kristina Gruden, Anna Coll Rius, Tjaša Lukan, 2025, original scientific article

Abstract: Potato virus Y (PVY) is one of the top 10 economically most important plant viruses and responsible for major yield losses. We previously suggested the involvement of the N-terminal region of PVY coat protein (CP) in PVY spread. By constructing different N-terminal deletion mutants of the PVY N605 strain, we here show that deletions of 40 or more amino acid residues from the N-terminal region of the CP resulted in the PVY multiplication limited to primary infected cells in Nicotiana clevelandii plants. Deletion of 26 residues profoundly impaired PVY cell-to-cell movement and prevented systemic PVY spread, while deletions of 19-23 residues allowed delayed systemic PVY spread. Introduced point mutations in the identified region prevent (S21G) or delay (G20P) PVY movement. In summary, this work shows the significance of the CP N-terminus for movement of the PVY.
Keywords: potato virus Y, potato, coat proteins, viral movement, point mutations
Published in DiRROS: 18.12.2025; Views: 1236; Downloads: 654
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Evaluating plant growth–defense trade-offs by modelingthe interaction between primary and secondary metabolism
Jan Zrimec, Sandra Correo, Maja Zagorščak, Marko Petek, Carissa Bleker, Katja Stare, Christian Schuy, Sophia Sonnewald, Kristina Gruden, Zoran Nikoloski, 2025, original scientific article

Abstract: Understanding the molecular mechanisms behind plant response to stress can enhancebreeding strategies and help us design crop varieties with improved stress tolerance,yield, and quality. To investigate resource redistribution from growth- to defense-relatedprocesses in an essential tuber crop, potato, here we generate a large-scale compartmen-talized genome-scale metabolic model (GEM), potato-GEM. Apart from a large-scalereconstruction of primary metabolism, the model includes the full known potato sec-ondary metabolism, spanning over 566 reactions that facilitate the biosynthesis of 182distinct potato secondary metabolites. Constraint-based modeling identifies that theactivation of the largest amount of secondary (defense) pathways occurs at a decreaseof the relative growth rate of potato leaf, due to the costs incurred by defense. We thenobtain transcriptomics data from experiments exposing potato leaves to two bioticstress scenarios, a herbivore and a viral pathogen, and apply them as constraints toproduce condition-specific models. We show that these models recapitulate experimen-tally observed decreases in relative growth rates under treatment as well as changes inmetabolite levels between treatments, enabling us to pinpoint the metabolic rewiringunderlying growth–defense trade-offs. Potato- GEM thus presents a useful resource tostudy and broaden our understanding of potato and general plant defense responsesunder stress conditions.
Keywords: systems biology, constraint-based metabolic modeling, growth-defence trade-offs, secondary metabolism
Published in DiRROS: 24.09.2025; Views: 1315; Downloads: 603
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Dynamics of responses in compatible potato - potato virus Y interaction are modulated by salicylic acid
Špela Baebler, Katja Stare, Maja Kovač, Andrej Blejec, Nina Prezelj, Tjaša Stare, Polona Kogovšek, Maruša Pompe Novak, S. Rosahl, Maja Ravnikar, Kristina Gruden, 2011, original scientific article

Abstract: To investigate the dynamics of the potato – Potato virus Y (PVY) compatible interaction in relation to salicylic acid - controlled pathways we performed experiments using non-transgenic potato cv. Désirée, transgenic NahG-Désirée, cv. Igor and PVYNTN, the most aggressive strain of PVY. The importance of salicylic acid in viral multiplication and symptom development was confirmed by pronounced symptom development in NahG-Désirée, depleted in salicylic acid, and reversion of the effect after spraying with 2,6-dichloroisonicotinic acid (a salicylic acid - analogue). We have employed quantitative PCR for monitoring virus multiplication, as well as plant responses through expression of selected marker genes of photosynthetic activity, carbohydrate metabolism and the defence response. Viral multiplication was the slowest in inoculated potato of cv. Désirée, the only asymptomatic genotype in the study. The intensity of defence-related gene expression was much stronger in both sensitive genotypes (NahG-Désirée and cv. Igor) at the site of inoculation than in asymptomatic plants (cv. Désirée). Photosynthesis and carbohydrate metabolism gene expression differed between the symptomatic and asymptomatic phenotypes. The differential gene expression pattern of the two sensitive genotypes indicates that the outcome of the interaction does not rely simply on one regulatory component, but similar phenotypical features can result from distinct responses at the molecular level.
Keywords: plant viruses, plant diseases
Published in DiRROS: 04.03.2025; Views: 1177; Downloads: 713
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Chloroplast vesiculation and induced chloroplast vesiculation and senescence-associated gene 12 expression during tomato flower pedicel abscission
Magda Tušek-Žnidarič, Maja Zagorščak, Živa Ramšak, Katja Stare, Marko Chersicola, Maruša Pompe Novak, Aleš Kladnik, Marina Dermastia, 2025, original scientific article

Abstract: Abscission is a tightly regulated process in which plants shed unnecessary, infected, damaged, or aging organs, as well as ripe fruits, through predetermined abscission zones in response to developmental, hormonal, and environmental signals. Despite its importance, the underlying mechanisms remain incompletely understood. This study highlights the deleterious effects of abscission on chloroplast ultrastructure in the cells of the tomato flower pedicel abscission zone, revealing spatiotemporal differential gene expression and key transcriptional networks involved in chloroplast vesiculation during abscission. Significant changes in chloroplast structure and vesicle formation were observed 8 and 14 h after abscission induction, coinciding with the differential expression of vesiculation-related genes, particularly with upregulation of Senescence-Associated Gene 12 (SAG12) and Chloroplast Vesiculation (CV). This suggests a possible vesicle transport of chloroplast degrading material for recycling by autophagy-independent senescence-associated vacuoles (SAVs) and CV-containing vesicles (CCVs). Ethylene signaling appears to be involved in the regulation of these processes, as treatment with a competitive inhibitor of ethylene action, 1-methylcyclopropene, delayed vesiculation, reduced the expression of SAG12, and increased expression of Curvature Thylakoid 1A (CURT1A). In addition, chloroplast vesiculation during abscission was associated with differential expression of photosynthesis-related genes, particularly those involved in light reactions, underscoring the possible functional impact of the observed structural changes. This work provides new insights into the molecular and ultrastructural mechanisms underlying abscission and offers potential new targets for agricultural or biotechnological applications.
Keywords: abscission, chloroplast vesiculation, CURT1A, CV-containing vesicle, senescence-associated vacuole, ethylene, gene expression, tomato flower pedicel
Published in DiRROS: 10.01.2025; Views: 1534; Downloads: 1062
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10.
The StPti5 ethylene response factor acts as a susceptibility factorby negatively regulating the potato immune responseto pathogens
Anna Coll Rius, Tjaša Lukan, Katja Stare, Maja Zagorščak, Tjaša Mahkovec Povalej, Špela Baebler, Salomé Prat, Núria Sánchez Coll, Marc Valls, Marko Petek, Kristina Gruden, 2024, original scientific article

Abstract: Ethylene response factors (ERFs) have been associated with biotic stress in Arabidopsis, while their function in non-model plants is still poorly understood. Here we investigated the role of potato ERF StPti5 in plant immunity. We show that StPti5 acts as a susceptibility factor. It negatively regulates potato immunity against potato virus Y and Ralstonia solanacearum, pathogens with completely different modes of action, and thereby has a different role than its orthologue in tomato. Remarkably, StPti5 is destabilised in healthy plants via the autophagy pathway and accumulates exclusively in the nucleus upon infection. We demonstrate that StEIN3 and StEIL1 directly bind the StPti5 promoter and activate its expression, while synergistic activity of the ethylene and salicylic acid pathways is required for regulated StPti expression. To gain further insight into the mode of StPti5 action in attenuating potato defence responses, we investigated transcriptional changes in salicylic acid deficient potato lines with silenced StPti5 expression. We show that StPti5 regulates the expression of other ERFs and downregulates the ubiquitin-proteasome pathway as well as several proteases involved in directed proteolysis. This study adds a novel element to the complex puzzle of immune regulation, by deciphering a two-level regulation of ERF transcription factor activity in response to pathogens.
Keywords: ethylene response factor, immune signalling, potato virus Y, Pti5, Ralstonia solanacearum, Solanum tuberosum, susceptibility factor
Published in DiRROS: 29.08.2024; Views: 1640; Downloads: 1543
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