Molecular Biology, Biochemistry, Agricultural and Biological Sciences, Plant Science
25
Scopus Publications
Scopus Publications
Metabolomic profiling provides novel insights into pistil adaptation to heat stress at anthesis in durum wheat lines carrying segmental introgressions from the wild grass Thinopyrum ponticum Ljiljana Kuzmanović, Giuseppina Fanelli, Gloria Giovenali, Sara Rinalducci, Carla Ceoloni Current Plant Biology, 2025 Rising global temperature makes sustainable agricultural production increasingly challenging. This is particularly worrying for staple crops like wheat, mainly in regions like the Mediterranean basin where high temperature extremes and heat waves frequently occur. A strategic commodity there at risk is durum wheat (DW), for which development of heat-tolerant varieties represents a key adaptive strategy with potential short-term effects. In contrast with mostly sensitive cultivars, wild relatives are endowed with numerous stress-adaptive and exploitable traits to reinforce the crop resilience. In this frame, three DW- Thinopyrum ponticum near-isogenic recombinant lines (NIRLs+), containing small alien segments on their 7AL arm, were previously subjected to transient heat stress (HS) at anthesis and their physiological and yield-related response compared to that of DW-only sib (NIRLs−) and non-sib control lines. HS at anthesis is known to greatly impair morphology and function of reproductive structures, yet limited knowledge is available on HS-triggered molecular/metabolic mechanisms in wheat floral organs, particularly the female one (pistil), directly involved in seed development and grain yield. Here, untargeted metabolomics was applied to identify pathways/metabolites in pistils sampled from heat-stressed and control plants of the above materials. Differential metabolic avenues were found to be undertaken by NIRLs+ vs. control lines under HS, including tricarboxylic acid cycle, pentose phosphate pathway, purine and pyrimidine, ascorbate and glutathione metabolisms, and specific metabolites (e.g. allantoin) produced, usable as selection biomarkers. The novel insights not only help explain the genotypes’ differential yield formation and stability but are also instrumental to breeding programs in which various effective metabolic strategies could be profitably combined. • Thinopyrum introgressions boost durum wheat tolerance to heat stress at anthesis. • Study targets heat-sensitive pistils, directly correlated to yield but underexplored. • Untargeted metabolomics shows introgression-specific pistil responses to heat. • TCA energy cycle and PPP were upregulated in tolerant, stably yielding introgression lines. • Stress-induced biomarkers allantoin, glutathione, and ascorbic acid were identified.
In Vitro Antagonistic Activity of Trichoderma gamsii Isolated From Hazelnut Twigs Against Colletotrichum spp. Federico Brugneti, Antonella Cardacino, Giuseppina Fanelli, Sara Rinalducci, Angelo Mazzaglia, Silvia Turco Journal of Phytopathology, 2025 Trichoderma species are widely recognised for their antagonistic activity against phytopathogenic fungi and for their ability to promote plant growth, hence representing promising candidates for sustainable crop protection. In this study, a Trichoderma gamsii isolate was recovered from the phyllosphere of hazelnut ( Corylus avellana ) and characterised through morphological, molecular and phylogenetic approaches. Its antagonistic potential was evaluated in vitro against six Colletotrichum species using dual culture assays, as well as through exposure to volatile and non‐volatile metabolites. To elucidate the chemical basis of its antagonism, volatile organic compounds (VOCs) and non‐volatile metabolites (non‐VOCs) produced in pure culture were profiled by gas chromatography–mass spectrometry (GC–MS) and liquid chromatography–mass spectrometry (LC–MS/MS), respectively. The isolate exhibited strong inhibitory effects, reducing pathogen growth by up to 79%, depending on the Colletotrichum species. These results expand current knowledge of the metabolic repertoire of T. gamsii and support its potential as an effective and eco‐friendly biocontrol agent for the sustainable management of anthracnose diseases.
Biomarker Preservation in Antarctic Sandstones after Prolonged Space Exposure Outside the International Space Station During the ESA EXPOSE-E Lichens and Fungi Experiment Alessia Cassaro, Claudia Pacelli, Giuseppina Fanelli, Mickael Baqué, Alessandro Maturilli, Patrick Leo, Veronica Lelli, Jean-Pierre Paul de Vera, Silvano Onofri, Annmaria Timperio Astrobiology, 2025 A primary aim of current and future space exploration missions is the detection and identification of chemical and biological indicators of life, namely biomarkers, on Mars. The Mars Sample Return NASA-ESA program will bring to Earth samples of martian soil, acquired from up to 7 cm depth. The ESA Rosalind Franklin rover will search for signs of life in the subsurface (down to a depth of 2 meters), given the highly radioactive conditions on Mars' surface, which are not ideal for life as we know it and for the preservation of its traces. In the frame of the Lichens and Fungi Experiment, small fragments of Antarctic sandstones colonized by cryptoendolithic microbial communities were exposed to space and simulated martian conditions in low Earth orbit for 18 months, aboard the EXPOSE-E payload. Through the use of Raman and infrared spectroscopies, as well as a metabolomic approach, we aimed to detect organic compounds in a quartz mineral matrix. The results show that pigments, such as melanin, carotenoids, and chlorophyll, lipids, and amino acids, maintained their stability within minerals under simulated martian conditions in space, which makes them ideal biomarkers for the exploration of putative life on Mars.
Untargeted metabolomics fingerprints in seminal plasma of patients with abnormal sperm morphology using high-performance liquid chromatography and mass spectrometry Serena Correnti, Giuseppina Fanelli, Mariaimmacolata Preianò, Veronica Lelli, Mariagrazia Tarantino, Annalisa Fregola, Massimo Bitonti, Emanuela Chiarella, Anna Maria Timperio, Sara Rinalducci, Rocco Savino, Rosa Terracciano Frontiers in Molecular Biosciences, 2025 Teratozoospermia, a qualitative sperm disorder characterized by abnormal sperm morphology, represents one of the causes of male infertility worldwide. The metabolic analysis of human seminal plasma (SP), can provide insights into the underlying molecular mechanisms of this condition, identifying novel biomarkers and facilitating the development of diagnostic tests. In this study, an untargeted High-Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS) approach was performed to explore SP metabolic alterations associated with teratozoospermia. SP samples from 15 teratozoospermic (TZ) vs. 20 normozoospermic (NZ) subjects were analyzed to identify metabolic pathways linked to sperm morphology dysfunction. Multivariate statistical analysis, including Partial Least Squares Discriminant Analysis (PLS-DA) and Orthogonal PLS-DA, revealed a distinct separation between TZ and NZ, highlighting 14 significantly altered metabolites. Based on Variable Importance in Projection scores, O-acetyl-L-serine showed the highest score. Main findings include alterations in Creatine, Histidine, Adenine, Allantoin and Deoxyuridine levels, suggesting perturbations in inflammation, oxidative stress and sperm DNA damage in teratozoospermia. Correlation and Receiver Operating Characteristic (ROC) analyses identified potential biomarkers, including O-acetyl-L-serine, Creatine, and Histidine, with robust discriminatory power (AUC >0.7). These findings highlight potential metabolic pathways implicated in the pathophysiology of teratozoospermia and provide a foundation for enabling personalized patient management with precision treatment.
Amino Acid Metabolism in Leukocytes Showing In Vitro IgG Memory from SARS-CoV2-Infected Patients Giuseppina Fanelli, Veronica Lelli, Sara Rinalducci, Anna Maria Timperio Diseases, 2024 The immune response to infectious diseases is directly influenced by metabolic activities. COVID-19 is a disease that affects the entire body and can significantly impact cellular metabolism. Recent studies have focused their analysis on the potential connections between post-infection stages of SARS-CoV2 and different metabolic pathways. The spike S1 antigen was found to have in vitro IgG antibody memory for PBMCs when obtaining PBMC cultures 60–90 days post infection, and a significant increase in S-adenosyl homocysteine, sarcosine, and arginine was detected by mass spectrometric analysis. The involvement of these metabolites in physiological recovery from viral infections and immune activity is well documented, and they may provide a new and simple method to better comprehend the impact of SARS-CoV2 on leukocytes. Moreover, there was a significant change in the metabolism of the tryptophan and urea cycle pathways in leukocytes with IgG memory. With these data, together with results from the literature, it seems that leukocyte metabolism is reprogrammed after viral pathogenesis by activating certain amino acid pathways, which may be related to protective immunity against SARS-CoV2.
An untargeted metabolomic approach to investigate antiviral defence mechanisms in memory leukocytes secreting anti-SARS-CoV-2 IgG in vitro Gevi Federica, Fanelli Giuseppina, Lelli Veronica, Zarletti Gianpaolo, Tiberi Massimo, De Molfetta Veronica, Scapigliati Giuseppe, Timperio Anna Maria Scientific Reports, 2023 Evidence shows that individuals infected by SARS-CoV-2 experience an altered metabolic state in multiple organs. Metabolic activities are directly involved in modulating immune responses against infectious diseases, yet our understanding of how host metabolism relates to inflammatory responses remains limited. To better elucidate the underlying biochemistry of the leukocyte response, we focused our analysis on possible relationships between SARS-CoV-2 post-infection stages and distinct metabolic pathways. Indeed, we observed a significant altered metabolism of tryptophan and urea cycle pathways in cultures of peripheral blood mononuclear cells obtained 60–90 days after infection and showing in vitro IgG antibody memory for spike-S1 antigen (n = 17). This work, for the first time, identifies metabolic routes in cell metabolism possibly related to later stages of immune defence against SARS-CoV-2 infection, namely, when circulating antibodies may be absent but an antibody memory is present. The results suggest reprogramming of leukocyte metabolism after viral pathogenesis through activation of specific amino acid pathways possibly related to protective immunity against SARS-CoV-2.
Untargeted Metabolomics Reveals a Multi-Faceted Resistance Response to Fusarium Head Blight Mediated by the Thinopyrum elongatum Fhb7E Locus Transferred via Chromosome Engineering into Wheat Giuseppina Fanelli, Ljiljana Kuzmanović, Gloria Giovenali, Silvio Tundo, Giulia Mandalà, Sara Rinalducci, Carla Ceoloni Cells, 2023 The Thinopyrum elongatum Fhb7E locus has been proven to confer outstanding resistance to Fusarium Head Blight (FHB) when transferred into wheat, minimizing yield loss and mycotoxin accumulation in grains. Despite their biological relevance and breeding implications, the molecular mechanisms underlying the resistant phenotype associated with Fhb7E have not been fully uncovered. To gain a broader understanding of processes involved in this complex plant–pathogen interaction, we analysed via untargeted metabolomics durum wheat (DW) rachises and grains upon spike inoculation with Fusarium graminearum (Fg) and water. The employment of DW near-isogenic recombinant lines carrying or lacking the Th. elongatum chromosome 7E region including Fhb7E on their 7AL arm, allowed clear-cut distinction between differentially accumulated disease-related metabolites. Besides confirming the rachis as key site of the main metabolic shift in plant response to FHB, and the upregulation of defence pathways (aromatic amino acid, phenylpropanoid, terpenoid) leading to antioxidants and lignin accumulation, novel insights were revealed. Fhb7E conferred constitutive and early-induced defence response, in which specific importance of polyamine biosynthesis, glutathione and vitamin B6 metabolisms, along with presence of multiple routes for deoxynivalenol detoxification, was highlighted. The results suggested Fhb7E to correspond to a compound locus, triggering a multi-faceted plant response to Fg, effectively limiting Fg growth and mycotoxin production.
Mechano-induced cell metabolism disrupts the oxidative stress homeostasis of SAOS-2 osteosarcoma cells Giuseppina Fanelli, Giulia Alloisio, Veronica Lelli, Stefano Marini, Sara Rinalducci, Magda Gioia Frontiers in Molecular Biosciences, 2023 There has been an increasing focus on cancer mechanobiology, determining the underlying-induced changes to unlock new avenues in the modulation of cell malignancy. Our study used LC-MS untargeted metabolomic approaches and real-time polymerase chain reaction (PCR) to characterize the molecular changes induced by a specific moderate uniaxial stretch regimen (i.e., 24 h-1 Hz, cyclic stretch 0,5% elongation) on SAOS-2 osteosarcoma cells. Differential metabolic pathway analysis revealed that the mechanical stimulation induces a downregulation of both glycolysis and the tricarboxylic acid (TCA) cycle. At the same time, the amino acid metabolism was found to be dysregulated, with the mechanical stimulation enhancing glutaminolysis and reducing the methionine cycle. Our findings showed that cell metabolism and oxidative defense are tightly intertwined in mechanically stimulated cells. On the one hand, the mechano-induced disruption of the energy cell metabolism was found correlated with an antioxidant glutathione (GSH) depletion and an accumulation of reactive oxygen species (ROS). On the other hand, we showed that a moderate stretch regimen could disrupt the cytoprotective gene transcription by altering the expression levels of manganese superoxide dismutase (SOD1), Sirtuin 1 (SIRT1), and NF-E2-related factor 2 (Nrf2) genes. Interestingly, the cyclic applied strain could induce a cytotoxic sensitization (to the doxorubicin-induced cell death), suggesting that mechanical signals are integral regulators of cell cytoprotection. Hence, focusing on the mechanosensitive system as a therapeutic approach could potentially result in more effective treatments for osteosarcoma in the future.
Plastic and Placenta: Identification of Polyethylene Glycol (PEG) Compounds in the Human Placenta by HPLC-MS/MS System Antonio Ragusa, Veronica Lelli, Giuseppina Fanelli, Alessandro Svelato, Sara D’Avino, Federica Gevi, Criselda Santacroce, Piera Catalano, Mauro Ciro Antonio Rongioletti, Caterina De Luca, Alessandra Gulotta, Sara Rinalducci, Anna Maria Timperio International Journal of Molecular Sciences, 2022 The placenta is a crucial interface between the fetus and the maternal environment. It allows for nutrient absorption, thermal regulation, waste elimination, and gas exchange through the mother’s blood supply. Furthermore, the placenta determines important adjustments and epigenetic modifications that can change the phenotypic expression of the individual even long after birth. Polyethylene glycol (PEG) is a polyether compound derived from petroleum with many applications, from medicine to industrial manufacturing. In this study, for the first time, an integration of ultra-high-performance liquid chromatography (UHPLC) coupled with mass spectrometry (MS) was used to detect suites of PEG compounds in human placenta samples, collected from 12 placentas, originating from physiological pregnancy. In 10 placentas, we identified fragments of PEG in both chorioamniotic membranes and placental cotyledons, for a total of 36 samples.
Supplementation with uric and ascorbic acid protects stored red blood cells through enhancement of non-enzymatic antioxidant activity and metabolic rewiring Vassilis L. Tzounakas, Alkmini T. Anastasiadi, Vasiliki-Zoi Arvaniti, Veronica Lelli, Giuseppina Fanelli, Efthymios C. Paronis, Anastasia C. Apostolidou, Evangelos G. Balafas, Nikolaos G. Kostomitsopoulos, Effie G. Papageorgiou, Issidora S. Papassideri, Konstantinos Stamoulis, Anastasios G. Kriebardis, Sara Rinalducci, Marianna H. Antonelou Redox Biology, 2022 Redox imbalance and oxidative stress have emerged as generative causes of the structural and functional degradation of red blood cells (RBC) that happens during their hypothermic storage at blood banks. The aim of the present study was to examine whether the antioxidant enhancement of stored RBC units following uric (UA) and/or ascorbic acid (AA) supplementation can improve their storability as well as post-transfusion phenotypes and recovery by using in vitro and animal models, respectively. For this purpose, 34 leukoreduced CPD/SAGM RBC units were aseptically split in 4 satellite units each. UA, AA or their mixture were added in the three of them, while the fourth was used as control. Hemolysis as well as redox and metabolic parameters were studied in RBC units throughout storage. The addition of antioxidants maintained the quality parameters of stored RBCs, (e.g., hemolysis, calcium homeostasis) and furthermore, shielded them against oxidative defects by boosting extracellular and intracellular (e.g., reduced glutathione; GSH) antioxidant powers. Higher levels of GSH seemed to be obtained through distinct metabolic rewiring in the modified units: methionine-cysteine metabolism in UA samples and glutamine production in the other two groups. Oxidatively-induced hemolysis, reactive oxygen species accumulation and membrane lipid peroxidation were lower in all modifications compared to controls. Moreover, denatured/oxidized Hb binding to the membrane was minor, especially in the AA and mix treatments during middle storage. The treated RBC were able to cope against pro-oxidant triggers when found in a recipient mimicking environment in vitro, and retain control levels of 24h recovery in mice circulation. The currently presented study provides (a) a detailed picture of the effect of UA/AA administration upon stored RBCs and (b) insight into the differential metabolic rewiring when distinct antioxidant "enhancers" are used.
