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Plant Stress

Elsevier BV

Preprints posted in the last 90 days, ranked by how well they match Plant Stress's content profile, based on 12 papers previously published here. The average preprint has a 0.02% match score for this journal, so anything above that is already an above-average fit.

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Epigenetic plasticity is associated with enhanced tolerance to low temperature stress in woodland strawberry

Njah, R. G.; Randall, S. K.; Davik, J.; Johansen, W.; Alsheikh, M. K.; Wilson, R. C.; Grini, P. E.

2026-04-28 plant biology 10.64898/2026.04.24.719864 medRxiv
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Low temperature stress causes significant damage to the strawberry plant. During cold stress, plants undergo morphological and physiological changes often regulated at the genetic and/or epigenetic levels. Some strawberry cultivars are more cold-hardy than others. Using the diploid woodland strawberry as a model, we analyzed the effects of cold acclimation on methylome and transcriptome dynamics in the crowns and leaves of three ecotypes with contrasting cold tolerance. Alta, which was the most cold-tolerant ecotype, exhibited the highest genetic and epigenetic plasticity in response to cold. CHH-context methylation dominated the differentially methylated regions (DMRs) with more hypomethylation in crowns and hypermethylation in leaves. CG methylation was enriched in gene bodies, while non-CG methylation was prevalent in upstream and downstream regions. Our study revealed that less than a quarter of differentially methylated genes (DMGs) showed changes in transcript accumulation levels. This finding indicates that universal cold response in Fragaria vesca, as reflected by gene expression, cannot be mechanistically attributed to DNA methylation. The majority of differentially expressed differentially methylated genes (DEDMGs) were ecotype- and tissue-specific. Enrichment analysis revealed that these genes were involved in pathways related to stress tolerance, such as carbohydrate metabolism, lipid metabolism, ATP hydrolysis, and cellular detoxification. Each ecotype responded to cold through mobilization of its own set of differentially expressed genes (DEGs), DMGs, and DEDMGs, and variation in expression and methylation patterns exhibited by Alta, FDP817, and NCGR1363 suggest that cold signaling processes and survival depend on the tissue, ecotype, and geographical origin of the plants exposed to cold stress. Therefore, this study highlights the potential of both genetic markers and epialleles as molecular markers for the development of cold-tolerant octoploid strawberry cultivars that are better suited for propagation in Nordic climates.

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Enhanced photosynthetic efficiency and ROS modulation promote cold stress tolerance of indica rice

Roy, V.; Parveen, R.; Dasgupta, P.; Chaudhuri, S.

2026-05-03 plant biology 10.64898/2026.04.30.721858 medRxiv
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Indica rice, being a tropical crop, is highly sensitive to cold temperature. Cold stress affects vegetative growth, photosynthetic efficiency, along with reproductive features. Genetic resource screening in diverse landraces is an approach for identifying cold-tolerant traits. Here, we have characterised a boro germplasm, CB1, with an efficient germination rate and growth vigour when treated at chilling temperatures. CB1 seedlings show a higher survival rate compared to IR36 when subjected to prolonged chilling stress. Biochemical analyses indicated efficient ROS modulation, higher chlorophyll content, enhanced photosystem II efficiency and unique stomatal traits, leading to higher relative water content in CB1 plants during stress and recovery. Transcriptome analysis supported upregulation of chlorophyll biosynthesis, photosystem, & light harvesting complex and ROS scavenger genes in CB1 seedlings. Interestingly, high D1 protein turnover in CB1 promotes damage-repair of PSII for efficient photosynthesis. Furthermore, key transcription factors for stomatal development and expression of photosynthetic genes were upregulated in CB1 during stress recovery. Notably, higher expression of OsGLK1 and enrichment of GLK1 targets were observed in CB1 plants during chilling stress and recovery. Taken together, our results suggested that CB1 plants exhibit cold tolerance by modulating photosynthesis efficiency and stomatal behavior for better adaptability and survival against chilling temperature. HIGHLIGHTSThe efficient photosynthetic recovery, active ROS scavenging system and maintenance of water content through regulating stomatal traits, enhance the survival of indica germplasm CB1 against chilling stress.

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Comparative Metabolomic Profiling Reveals Salinity Tolerance Mechanisms in a Rice Introgression Line

Chaudhary, C.; Guttula, P.; Agrawal, K.; Subudhi, P. K.; Gartia, M. R.

2026-07-07 plant biology 10.64898/2026.07.06.736799 medRxiv
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Rice (Oryza sativa) is highly sensitive to salinity, yet the metabolic mechanisms underlying salt tolerance remains incompletely understood. In this study, we performed leaf tissue-specific untargeted metabolomic profiling of the salt-tolerant introgression line JN100 (JN), its donor parent Nona Bokra (NB), and its recurrent parent Jupiter (JU) to characterize metabolic responses to salt stress. Comparative analysis identified differentially accumulated metabolites (DAMs) spanning diverse chemical classes, including amino acids, sugars and carbohydrates, lipids, organic acids, cofactors, electron carriers, and nucleotides. Under salt stress (SS), 201 DAMs (89 upregulated and 112 downregulated) were detected in JN relative to JU. Notably, metabolites such as allantoin, glycitin, nicotinamide ribotide, D-arabinono-1,4-lactone, violanthin, L-methionine S-oxide, ribitol, lysine, rutin, glutamine, pantothenic acid, and quinic acid, showed significant differential accumulation. Pathway enrichment analysis revealed significant enrichment of arginine biosynthesis, purine metabolism, and alanine, aspartate, and glutamate metabolism, indicating extensive reprogramming of nitrogen and energy-associated metabolic pathways under salinity stress. Integration of transcriptomic and metabolomic datasets from the SS experiments further identified ten differentially expressed genes (DEGs) associated with the metabolite network in the JN vs. JU comparison. Among these, OsDHQDT/SDH, OsFd-GOGAT, phenylalanyl-tRNA synthetase, OsP5CS1, OsP5CS2, and a pyridoxal phosphate-dependent transferase were linked to metabolites involved in shikimate, amino acid, and proline metabolism. Collectively, these results demonstrate that salinity tolerance in rice is associated with coordinated transcriptional and metabolic reprogramming that supports oxidative stress mitigation and adaptive stress responses.

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The lack of peroxisomal Glycolate Oxidases 1 and 2 influences mitochondrial electron transport chain and its redox state under control and cadmium stress

Collado-Arenal, A. M.; Rodriguez-Serrano, M.; Pelaez-Vico, M. A.; Terron-Camero, L. C.; Perez-Gordillo, F. L.; Ranea-Robles, P.; Lopez, L. C.; Sandalio, L.; Romero-Puertas, M. C.

2026-05-08 plant biology 10.64898/2026.05.06.723131 medRxiv
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The production of reactive oxygen species (ROS) in response to cadmium (Cd) has been extensively studied, demonstrating that they play a key role in the plants response to this heavy metal. While the role of enzymes like RBOHs has been thoroughly studied, the function of other ROS-producing enzymes, such as peroxisomal glycolate oxidase (GOX), remains largely overlooked. Peroxisomal GOX is a core metabolic enzyme of the photorespiratory pathway occurring in chloroplasts, mitochondria and peroxisomes. Using Arabidopsis (Arabidopsis thaliana) mutants lacking the main peroxisomal GOX genes, GOX1 (gox1-1) and GOX2 (gox2-1) we explored their function in plant response to Cd. Although photosynthetic capacity appears to be affected to the same extent in both mutants under control and Cd stress conditions, GOX2 seems to play a greater role in ROS production in response to the metal. Transcriptomic analyses on WT and gox2-1 pointed to the mitochondrial electron transport chain (mETC) as a target of Cd stress. We further investigated the individual GOX1 and GOX2 functions in mETC regulation and redox state. Although oxidative ratio of mitochondria was higher in both mutants, it was more pronounced in the absence of GOX1. Furthermore, the mETC is affected in both mutants but the regulation of its components differs in each mutant. These results point out the different functions of the two photorespiratory GOX isoforms in Arabidopsis, leading to a better understanding of the photorespiratory pathway.

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Integrated Evaluation of Osmotic and Antioxidant Defense Mechanisms in Cotton Genotypes Exposed to NaCl Stress

Rakhmatova, N. R.; Imamkhodjayeva, A. S.; Salakhutdinov, I. B.; Kamburova, V. S.; Kadirova, S. B.; Radjapov, F. S.; Norbekov, J. K.; Zakirova, M.; Yuldashova, Z. Z.; Jumaev, R. A.; Buriev, Z. T.

2026-06-06 plant biology 10.64898/2026.06.03.729956 medRxiv
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Salinity stress is one of the major abiotic factors limiting cotton productivity worldwide by inducing osmotic imbalance, oxidative stress, and metabolic disturbances in plant tissues. The present study aimed to evaluate the physiological and biochemical responses of different cotton (Gossypium hirsutum L.) genotypes under NaCl-induced salinity stress through analysis of proline accumulation, antioxidant enzyme activities, and lipid peroxidation intensity. The experiment was conducted under controlled conditions using several cotton genotypes exposed to different NaCl concentrations. Proline content, superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) levels were analyzed as major biochemical indicators associated with salinity tolerance and oxidative stress responses. In addition, modern bubble heatmap visualization was applied for comparative assessment of genotype-specific stress response patterns under saline treatments. The obtained results demonstrated that increasing NaCl concentrations generally stimulated proline accumulation and enhanced antioxidant enzyme activities in most investigated cotton genotypes. Increased SOD and CAT activities indicated activation of enzymatic antioxidant defense mechanisms under salinity stress conditions. Simultaneously, elevated MDA accumulation reflected enhanced oxidative membrane damage caused by excessive reactive oxygen species (ROS) production under saline environments. Considerable genotype-dependent variability was observed among the investigated cotton varieties. Genotypes such as "Nasaf", "Gulbahor-2", "Ravnaq-1", "Buxoro-6", "Afsona", "Baraka", "Namangan-77", "Porloq-1", and "C-4727" demonstrated comparatively stronger physiological and antioxidant responses under salinity stress conditions, suggesting relatively higher adaptive capacity to NaCl-induced stress. The heatmap visualization additionally confirmed substantial heterogeneity among cotton genotypes in biochemical stress responses and allowed comprehensive comparative interpretation of salinity-induced physiological variability. Overall, the present findings suggest that proline accumulation, antioxidant enzyme activities (SOD and CAT), and MDA content may serve as important biochemical markers for evaluation of salinity tolerance in cotton. The identified stress-tolerant genotypes may therefore represent valuable genetic resources for future breeding programs aimed at improving cotton productivity under saline environmental conditions.

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Belgian endive-derived biostimulant activity in Arabidopsis, lettuce and sweet pepper at different developmental stages, environmental conditions, and application methods

Ogunsanya, H. Y.; Petit, C.; Audenaert, K.; De Zutter, N.; Geelen, D.

2026-04-17 plant biology 10.64898/2026.04.15.717650 medRxiv
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Belgian endive-derived biostimulant (BEE) was previously shown to enhance root and shoot growth of Arabidopsis thaliana and Plectranthus esculentus in in vitro culturing conditions. In this study, we evaluated the effect of BEE on A. thaliana subjected to abiotic stresses and assessed the translatability of its bioactivity on lettuce (Lactuca sativa) and sweet pepper (Capsicum annuum) cultured in substrate and soil. A first set of experiments tested the impact of BEE on protection during, and restoration after, osmotic or salt (NaCl) stress. BEE treatment had little to no rescuing effect when plants were exposed to osmotic stress. In contrast, BEE strongly promoted shoot development and leaf health both under standard and NaCl stress conditions. Under mild stress, BEE enhanced photosynthetic efficiency and chlorophyll content in Arabidopsis, whereas it did not significantly alleviate osmotic stress induced by sorbitol. To evaluate the effect under ex vitro conditions, BEE was applied via root drenching to substrate-grown A. thaliana, lettuce, and sweet pepper. BEE improved leaf greenness and photosynthesis enhancing Arabidopsis rosette development, but it did not increase lettuce head weight. In sweet pepper, BEE increased fruit yield and promoted fruit maturation. Under drought stress conditions, BEE application did not improve sweet pepper yield.

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Long-term performance of disease-resistant grapevine varieties: insights from an 8-year field monitoring across French vineyards

Pelissier, R.; Marolleau, L.; D Mazet, I.; Delmotte, F.; DELIERE, L.; Miclot, A.-S.; Fabre, F.

2026-04-30 plant biology 10.64898/2026.04.28.721351 medRxiv
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Breeding disease-resistant varieties (DRV) is a central strategy for reducing reliance on phytosanitary products. However, the successful deployment and long-term durability of these cultivars rely on acquiring field data across diverse production conditions, a step that remains frequently neglected, especially in perennial crops. Since 2018, the OSCAR observatory, a network of vineyard plots planted in France with varieties resistant to downy and powdery mildew, the two major pathogens of grapevine, has aimed to close this gap for viticulture. The observatory comprises over 199 commercial plots, covering 127 hectares across diverse agroclimatic conditions, all managed by winegrowers under their own production practices. The observatory currently monitors 30 disease-resistant grapevine varieties, tracking both their agronomic performance and the dynamics of key pathogens. Since 2018, while phytosanitary treatments have been reduced by an average of 79% compared to conventional plots, the incidence of downy and powdery mildew, remain low, even in years highly conducive to these diseases. However, the long-term survey also highlights the decline in efficacy of some resistances to downy mildew and the emergence of black rot, a disease effectively controlled by conventional phytosanitary programs. Beyond acting as a rapid warning system for resistance breakdown, the observatory promotes sustainable disease management in viticulture. It provides valuable insights to winegrowers on effective DRV management. It also delivers actionable feedback to breeders to guide more durable DRV breeding strategies. Highlights- OSCAR observatory monitors 199 plots of grapevine disease resistant varieties (DRV) - Grapevine DRV cuts fungicide uses by 79% while maintaining good disease control - Some resistances efficacy declines against downy mildew, but not powdery mildew - Black rot, a disease usually controlled by fungicide, is rising in OSCAR plots OSCAR provides useful feedback to breeders and winegrowers on DRV management

8
Transcriptomic Insights into Drought Tolerance Enhancement in Bread Wheat Induced by a Microalgae-based Biostimulant

Arvanitidou, C.; Ramos-Gonzalez, M.; Garcia-Gomez, M. E.; Garcia-Gonzalez, M.; Romero-Campero, F. J.

2026-05-18 plant biology 10.64898/2026.05.18.725825 medRxiv
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Bread wheat (Triticum aestivum) is a staple food crucial for global caloric intake and food security. The current climate emergency demands the development of sustainable agricultural practices, particularly in the context of drought-induced yield reductions in bread wheat. Microalgae-based biostimulants have emerged as promising tools to enhance crop tolerance to drought stress while concurrently mitigating atmospheric CO2 accumulation. This study characterizes the transcriptomic responses to the foliar application of the microalgae-based biostimulant LRMTM in drought-stressed and fully irrigated wheat plants unveiling its mode of action. Drought stress at the tillering stage significantly altered gene expression activating key pathways related to phosphate starvation response (PSR), inositol phosphate signaling, and tocopherol biosynthesis. The application of the microalgae-based biostimulant LRMTM in drought-stressed plants further enhanced the expression of drought-responsive genes, particularly those involved in PSR and carbon fixation. Specific responses to LRMTM treatment in drought-stressed plants were also found related to abscisic acid (ABA) signaling activating genes involved in stomata closure, which plays a critical role in drought tolerance. In fully irrigated plants, LRMTM treatment was also beneficial modulating circadian rhythms, shade avoidance and attenuating stress responses. Phenotypic analysis showed that LRMTM-treated plants exhibited enhanced drought tolerance, increased height and spike length even under fully irrigated conditions. These results indicate that the microalgae-based biostimulant LRMTM not only enhances wheat response to drought but also promotes growth and productivity in both stressed and non-stressed conditions which could contribute to the development of sustainable agriculture in the face of the current climate challenges.

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Rice Cultivars Carrying the Semi Dwarfing Allele Enables High Yield without Lodging under Hairy Vetch based Green Manure

Fukuda, H.; Sakamoto, T.; Fukuda, A.; Ogawa, D.

2026-05-27 plant biology 10.64898/2026.05.25.727576 medRxiv
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Green manure is a promising strategy for reducing dependence on chemical fertilizers in crop production. However, vigorous growth due to green manure often leads to high yield accompanied by lodging in rice, hindering its practical use in rice cultivation. Here, we show that rice cultivars carrying a semi-dwarfing sd1/ga20ox2 allele achieve high grain yield without lodging under hairy vetch-based green manure conditions. The leading Japanese cultivar Koshihikari exhibited enhanced vegetative growth, increased panicle number, and consequently higher grain yield and quality under green manure conditions in 2023 and 2024 compared with chemical fertilizer management, although this was accompanied by increased culm length and widespread lodging. Among the four GA20-oxidase genes, green manure significantly upregulated Sd1/GA20ox2 mRNA levels. A temperate japonica cultivar, Nijinokirameki, and an indica cultivar, Hokuriku-193, carrying a non-functional sd1/ga20ox2 allele exhibited no lodging under hairy vetch-based green manure management while achieving improved yield performance. Notably, yields obtained under our hairy vetch-based cultivation system were comparable to or exceeded a recently reported high-yield benchmark observed for Hokuriku-193 under chemical fertilizer management in the same region of Japan. These findings suggest that cultivars harboring non-functional sd1/ga20ox2 alleles enable the practical implementation of annual hairy-vetch-rice rotation for sustainable rice production.

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Bark beetle protein elicitors trigger biphasic immune responses in Norway spruce seedlings

Ramires, M. J.; Netherer, S.; Schebeck, M.; Ertl, R.; Ahmad, M.; Arc, E.; van Loo, M.; Trujillo Moya, C.

2026-05-22 plant biology 10.64898/2026.05.22.727111 medRxiv
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Norway spruce (Picea abies) responds to attacks by the spruce bark beetle (Ips typographus) through the rapid activation of local defense mechanisms, but field studies can be difficult to standardize due to variable attack pressure and environmental heterogeneity. Here, we developed a phytotron-based assay that mimics early beetle-associated stress using insect-derived protein extracts, enabling reproducible molecular analyses under controlled conditions. Ten-week-old spruce seedlings were stem-treated with mock buffer or beetle protein extracts, followed by transcriptomic analyses of stem tissues and targeted metabolomic profiling of needles at 2 and 48 h post-inoculation. RT-qPCR analysis revealed rapid transcriptional activation of signaling and defense genes in Norway spruce, with NP-40-based protein extracts producing the most consistent early response. RNA-seq analysis revealed transcriptional dynamics, with 488 differentially expressed genes detected at 2 h and 84 at 48 h post-inoculation relative to mock-treated controls. Early responses at 2 h were characterized by activation of genes associated with immune perception and signal transduction. By 48 h, the response shifted toward accumulation of transcripts encoding defense proteins such as chitinases, defensins, proteinase inhibitors, and pathogenesis-related (PR) proteins. Importantly, a substantial proportion of differentially expressed genes overlapped with those previously identified in mature Norway spruce trees during pioneer bark beetle attack under field conditions, supporting the biological relevance of the assay. In contrast, targeted analyses of secondary metabolites performed in needle tissue revealed limited systemic changes across time points, suggesting that early induced defenses may remain largely localized to the stem. Together, these results demonstrate that beetle-derived proteins trigger a rapid and temporally structured defense response in Norway spruce seedlings and establish a reproducible elicitor-based platform for dissecting conifer immune responses and screening spruce genotypes for bark beetle resistance. HighlightBark beetle protein elicitors trigger temporally structured immune responses in Norway spruce seedlings that overlap with responses observed in mature trees, with rapid immune signaling at 2 h followed by defense protein accumulation at 48 h.

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Transient uncoupling of the Suc-Tre6P-SnRK1 nexus during salt stress associates with biphasic metabolic reprogramming and root plasticity

Barbieri, G.; Parola, R.; Feil, R.; Rodriguez, M. S.

2026-05-12 plant biology 10.64898/2026.05.08.723798 medRxiv
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Soil salinization threatens global agriculture reducing yields, yet the metabolic signals controlling salt-sensitive root plasticity in alfalfa remain unclear. We hypothesize that salinity transiently uncouples the sucrose-trehalose-6-P (Tre6P)- Sucrose non-fermenting kinase 1 (SnRK1) nexus, aligning with a biphasic root metabolic response and altered root architecture. Alfalfa seedlings were grown in a hydroponic system and exposed to 200 mM NaCl, with root samples collected from 1 h to 7 d. While primary root growth and biomass remained unchanged, lateral root development was enhanced under salinity. Early response (1 h-1 d) was characterized by reduced carbon metabolites, low Tre6P, increased malondialdehyde, and SnRK1 activation, with a decline in glycolytic and TCA intermediates. During this phase, sucrose was negatively correlated with both Tre6P and SnRK1. Late response (3-7 d) showed a SnRK1 reactivation, Tre6P recovery, and osmoprotectant accumulation, including increased antioxidant capacity (+75% at 3dpt), proline (+178%), and sucrose (+18%) and starch depletion (-57%) at 7dpt respect to control. These metabolic changes coincided with the enhanced lateral root emergence. These findings indicate a two-phase response: early metabolic downscaling with transient Suc-Tre6P-SnRK1 disruption, followed by recovery with Tre6P restoration, SnRK1 reactivation, osmoprotection, and sustained root plasticity under salinity. HighlightSalinity triggers a temporary metabolic shift in alfalfa roots: plants first conserve energy, then adapt to stress, maintaining lateral root growth and flexible root architecture.

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Peacock Eye Disease Management (PedMan) System: Validation and Implementation in Addressing Key Biological Questions

Gilat, Y.; Ygzao, D.; Shtienberg, D.; Ezra, D.

2026-06-04 plant biology 10.64898/2026.06.02.729514 medRxiv
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Peacock eye disease, caused by Venturia oleaginea, is a major foliar disease of olive (Olea europaea) in Mediterranean regions, resulting in defoliation, reduced tree vigor, and yield losses. Effective disease management relies on precise fungicide timing. In this study, we validated a decision support system (DSS) named PedMan (Peacock eye disease Manager), designed to predict infection events and optimize fungicide application timing in highly susceptible olive cultivars based on rainfall and temperature conditions. The system was validated in seven independent grove experiments conducted during the 2023/24 and 2024/25 growing seasons in commercial olive orchards in Israel. In addition, simulation analyses were performed using weather data from 11 meteorological stations representing diverse climatic regions. Field validation showed that fungicide applications timed according to PedMan significantly reduced leaf abscission by approximately 60% compared with untreated controls. Applications made contrary to system recommendations did not improve disease control, confirming the reliability of both positive and negative predictions. Multi-season analyses indicated cumulative disease suppression, with up to 85% reduction in leaf abscission after three consecutive years of correctly timed applications. The system was implemented by olive growers in 2025/6 with applicable success. Simulation results showed that most infection events occurred in autumn and early winter, with rainfall as the primary driver in autumn and temperature as the main limiting factor in winter and spring. Across all regions and seasons, PedMan recommended 0-3 fungicide applications per season, comparable to or fewer than conventional spray programs. These findings demonstrate that PedMan is a robust, field-validated DSS that improves fungicide timing, enhances disease control efficiency, and supports sustainable management of peacock eye disease without increasing spray frequency under Mediterranean conditions.

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Cytokinin N-conjugate Form Activity, Metabolism, and Signaling During Leaf Senescence

Hasannin, O.; Petrik, I.; Strnad, M.; Novak, O.; Cerny, M.; Rashotte, A. M.

2026-05-13 plant biology 10.64898/2026.05.08.723873 medRxiv
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Cytokinin (CK) N-glucosides are the most abundant CK metabolites in Arabidopsis and most angiosperms, yet their role in cytokinin activity and response is unclear. Here, we examined metabolomic, transcriptomic, and proteomic profiles of seven CK N-glucoside conjugates in detached Arabidopsis leaves across a 144-hour dark-induced senescence (DIS) timecourse. All tested N-glucosides were found to undergo a slow conversion to their corresponding base forms at position-dependent rates, with N9-glucosides releasing base faster than their corresponding N7-glucosides. Conversion during DIS was strictly isoform-specific and not accompanied by coordinated induction of CK biosynthesis genes, arguing against de novo synthesis as the source of accumulated base. Despite progressive base accumulation, N-glucoside-treated leaves produced substantially fewer Differentially Expressed Genes than direct base application at comparable base concentrations, revealing a disconnect between hormone presence and transcriptional output. Unbiased model comparison identified the base:glucoside ratio as a stronger predictor of CK-Two Component Signaling (TCS) gene expression than absolute base concentration, though modulated by base-type-specific receptor affinities. Early proteomic profiling further revealed a coordinated response shared across N-glucosides but largely absent from base treatments. Together, these findings support that CK N-glucosides as kinetically slow, position-dependent reservoirs whose presence in abundance modulate activation of CK-TCS elicited by bioactive forms. HighlightsPhysiology, metabolomic, transcriptomic, and proteomic findings here support CK N-glucosides as kinetically slow, position-dependent reservoirs whose presence in abundance modulate activation of CK-TCS elicited by bioactive forms.

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Dual Knockout of StAMY23 and StVINV Improves Postharvest Storage Traits in Potato

Teper-Bamnolker, P.; Steinberg, T.; Shtein, C.; Peer, R.; Doron-Faigenboim, A.; Belausov, E.; Sherman, A.; Eshel, D.

2026-06-10 plant biology 10.64898/2026.06.08.730856 medRxiv
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Starch is the primary carbohydrate reserve in potato (Solanum tuberosum L.) tubers and a critical determinant of their industrial value. The rate of starch degradation during postharvest storage directly influences key traits such as endodormancy (ED) duration and cold-induced sweetening (CIS), which affect sprouting behavior. In this study, we used CRISPR/Cas9 genome editing to knockout StAMY23, a gene encoding -amylase involved in starch breakdown. stamy23 plants exhibited higher yield and extended tuber ED postharvest, without significantly altering CIS or starch granule content. To further reduce CIS, we knockout StAMY23 in VACUOLAR INVERTASE knockout (stvinv) backgrounds, generating stamy23/stvinv double-knockouts plants. These lines showed significantly reduced CIS, prolonged ED, and elevated starch content, along with altered starch granule content. Collectively, our findings demonstrate that simultaneous downregulation of StAMY23 and StVINV can additively enhance desirable postharvest traits, providing a promising strategy for improving potato storage quality through precision genome editing.

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Non-plastic gene expression underlies root phenotypes involved in drought adaptation in Vitis spp.

Chedid, E.; Patin, E. R.; Tran, J.; de Miguel, M.

2026-07-10 plant biology 10.64898/2026.07.09.737455 medRxiv
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Drought is a major abiotic stress threatening plant productivity and agricultural sustainability, yet the molecular mechanisms underlying adaptive root responses to water deficit in the water use strategies continuum remain insufficiently understood, particularly in perennial crops. In this study, we explored drought responses in nine accessions belonging to three wild Vitis species (V. acerifolia, V. candicans, and V. doaniana) displaying varying drought-response strategies. Plants were subjected to moderate drought stress (40% soil water content) for three weeks under greenhouse conditions. By integrating physiological, metabolic, and transcriptomic analyses, we aimed to identify both conserved and species-specific mechanisms associated with drought adaptation. Differential expression analyses revealed a conserved core set of drought-responsive genes shared among species, including genes involved in abscisic acid signaling, reactive oxygen species detoxification, solute transport, and plant defense. In parallel, each species exhibited distinct transcriptional and metabolic signatures reflecting alternative adaptive strategies related to osmoregulation, and oxidative stress mitigation. Weighted gene co-expression network analysis (WGCNA) further revealed significant associations between constitutive, non-plastic gene expression and root phenotypic traits. Overall, our findings demonstrate that wild Vitis species rely on both conserved stress-responsive pathways and species-specific constitutive regulation to cope with drought stress. These results highlight the importance of root-associated traits and intrinsic regulatory networks in shaping drought adaptation and provide new targets for the development of drought-resilient grapevine rootstocks.

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Assessment of EMS-induced mutagenesis in Fagopyrum esculentum Moench (Buckwheat)

BADAR, A.; Siddique, I.; Mubeen, H.

2026-04-29 plant biology 10.64898/2026.04.25.720850 medRxiv
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Global demand for pseudocereals, including buckwheat, has surged in recent years due to their higher nutritional and pharmaceutical value than cereals and also due to them being a climate-resilient, gluten-free, and potential crop for combating cancer, type ll diabetes, and overcoming micronutrients hidden hunger problems that lack in cereals. Major efforts are needed to make its cultivation more popular by improving its quantitative and qualitative traits through crop genetics by adopting modern genetic, molecular, and mutational approaches, which also necessitate the induction of genetic variation for better yielding and improved varieties. In this experimental study, the induced mutant populations of widely recommended VL-7 and PRB-1 varieties of buckwheat were generated using different concentrations treatments of ethyl methane sulfonate (EMS). Investigation on induced phenotypical and genotypical variations in individual plants of M1 population of different treatments resulted in morphological and cytological mutant types affecting plant germination, survival, height and morphology, leaf morphology, flower morphology, growth period, chlorophyll and pigments abnormalities in leaves, leaf growth pattern, plant fertility, yield, and cytological aberrations. This experiment showed that plant survival decreased with the concentration of the mutagen doses. The lower doses resulted in dwarf varieties suitable for cultivation as they increased yield by having higher breaking force and lower lodging index over the tall plants. Studies on various quantitative parameters revealed the general effectiveness of intermediate doses and stimulatory effectiveness of lower and higher concentrations in M1 generation.

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Genomic Prediction Enables Same-Season Selection for Reduced Glycosidic Nitrile in Eastern U.S. Winter Barley

Perry, A. D.; Sabadin, F.; Brooks, W.; Brown-Guedira, G.; Uhlmann, H.; Bettenhausen, H.; Santantonio, N.

2026-06-06 plant biology 10.64898/2026.06.03.729884 medRxiv
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Glycosidic nitriles (GN) in barley are precursors to carcinogens formed during distillation, making GN reduction a critical breeding objective for malting and distilling industries. Measurement of GN is time-consuming. Grain must first be malted before GN can be quantified, and generally cannot be completed before selections must be made in a winter barley breeding program. Here, feasibility of same-season genomic selection against GN content was evaluated in elite Virginia Tech winter barley germplasm. In 2023, all 176 elite breeding lines screened for presence of GN were shown to be GN producers. A subset of 95 lines was then quantitatively measured for GN concentration to determine the genetic variability for the trait. Efficacy of genomic selection for GN was first assessed using a divergent selection approach on the remaining 81 predicted lines. The highest 16 and lowest 16 of the predicted lines were chosen for GN quantification. A significant phenotypic difference was found between the predicted high and low group means (0.8 ppm; P = 0.003). An additional 120 lines were quantified the following year to determine repeatability. GN exhibited moderate narrow-sense heritability (h2 = 0.42) and a high genetic correlation (r = 0.79) across years. Moderate predictive ability as was observed in cross-validation (range 0.38 - 0.61), and forward prediction using 2023 to predict 2024 (r = 0.39). A genome-wide scan did not identify any major-effect loci, suggesting GN content is polygenic, thus enabling same-season genomic selection to reduce GN content in this germplasm.

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LC-MS Metabolomics Reveals No Significant Impact of Microbial Inoculation with Bacillus velezensis and Lachnum sp. on Cranberry Metabolome

Ali, E. T.; Findlay, B.

2026-06-06 plant biology 10.64898/2026.06.02.729675 medRxiv
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Sustainable agriculture has driven increased exploration of microbial inoculants as a promising strategy to boost plant growth for higher yield and enhance secondary metabolism to increase crop nutritional value. However, their influence on fruit-specific metabolites under field conditions remains understudied. This study investigated the impact of inoculating cranberry (Vaccinium macrocarpon) plants with Bacillus velezensis EB37 and Lachnum sp. EC5, applied individually and in combination, on cranberry fruit phytochemistry. Over two growing seasons (2019 and 2021), cranberries were collected from treated and control plots and analysed using untargeted and targeted LC-MS-based metabolomics. Multivariate analysis revealed no significant metabolomic differences due to treatments. However, samples clustered strongly by year of harvest, highlighting a pronounced environmental effect. Quantitative analysis of six representative phenolic compounds: chlorogenic acid, catechin, p-coumaric acid, phloridzin, myricetin, and quercetin, showed no statistically significant differences between treated and control cranberries. These findings indicate that microbial inoculation alone does not alter cranberry fruit metabolome, including phenolic levels, at field conditions. This study underscores how multiple factors, such as environmental conditions, can affect the outcome of microbial inoculation under field conditions and suggests that additional interventions may be required to achieve microbiome-based improvements in cranberry fruit quality.

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Herbivory-induced alterations in cytosolic proteins of pigeon pea (Cajanus cajan) leaves

S, A.; Kalita, P. J.; Meshram, S. K.; Das, A.; Patil, R. I.; Das, S.; Jaba, J.; Das, D.; Acharjee, S.

2026-05-08 plant biology 10.64898/2026.05.07.723431 medRxiv
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Insect herbivory triggers cytosolic proteome reprogramming by activating defense pathways and modulating key metabolic processes. We found that simulated herbivory in pigeon pea (Cajanus cajan) induced reactive oxygen species (ROS) production and molecular alterations within 12 hours (h) of post treatment. We compared the leaf proteome profiles of two cultivated genotypes, ICPL 332 (moderately resistant) and ICPL 87 (susceptible), using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) coupled with mass spectrometry (MS). More than 220 protein spots were detected in ICPL 332 and over 200 in ICPL 87. Comparative analysis revealed 75 differentially accumulated proteins (DAPs), of which 40 were consistently reproducible across biological replicates. These included 11 unique to ICPL 87, 9 unique to ICPL 332, and 10 common to both genotypes. Among the shared DAPs, ICPL 332 showed five upregulated and five downregulated, whereas ICPL 87 exhibited only two upregulated and eight downregulated. Functional categorization grouped DAPs into primary metabolism, stress response, and growth and development. Proteins related to primary metabolism were largely downregulated in both genotypes, while stress-associated proteins exhibited substantial downregulation in ICPL 87 compared to ICPL 332. Overall, the results demonstrate proteomic adjustments underlying defense responses in pigeon pea genotypes.

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Effects of an increase in water temperature on inter- and transgenerational plasticity reveal a short-term metabolic and phenotypic memory in an aquatic plant species

Loupit, G.; Sancharme, M.; Petriacq, P.; Valls Fonayet, J.; Bittebiere, A.-K.

2026-07-07 plant biology 10.64898/2026.07.06.736556 medRxiv
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Transgenerational plasticity can shape plant phenotype and influence plant response to environmental changes in interaction with the current conditions. While how past stress interact with either current optimal or stress conditions is increasingly documented within a single plant, transgenerational plasticity remains particularly poorly understood especially at the metabolome level. In our study, we investigated whether heat stress induces transgenerational metabolic and phenotypic modifications along two successive clonal ramet generations of the sub-Antarctic aquatic plant Limosella australis. We performed untargeted metabolomic approaches and measured morphologic and performance traits, to assess both transgenerational plasticity of the metabolome and the phenotype. We found that heat stress remodelled the metabolic profile and influenced the foraging strategy of our clonal plant, and that some of these metabolic changes persisted into the first clonal generation. This one therefore adopted an intermediate growth strategy, even though culture conditions were optimal. By comparing differentially accumulated features between daughter ramets from heat stressed mother ramets and from unstressed mother ramets, we identified common and specific metabolites accumulation to heat stress response, belonging to diverse compound families. However, we did not observe any adaptative advantage and any metabolic imprint during another heat stress applied on the second clonal generation. This work provides especially new clues into how plant metabolome integrates and transfers previous stressed clonal generation's information.