Microbial volatile organic compounds (mVOCs) enable plants to perceive microbial activity prior to physical contact, yet the contribution of individual bacterial volatiles to immune signalling and disease resistance remains incompletely understood. Here, headspace GC-MS analysis demonstrates that the hemibiotrophic pathogen Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) emits a distinct volatile blend containing the bacterium-specific compounds 2-methylbutanoic acid (2-MBA) and 3-methylbutanoic acid (3-MBA), with 2MBA as a dominant component. Exposure of Arabidopsis thaliana to the complete Pst DC3000 mVOC blend induced extensive transcriptional reprogramming activation of pattern-recognition receptor-associated genes, MAP kinase signalling components, WRKY transcription factors, camalexin biosynthetic genes and early defense responses. The latter included cytosolic Ca2+ ([Ca2+]cyt) elevation, K+ channel activation, hydrogen peroxide accumulation, and nitric oxide production, which culminated in camalexin accumulation in wild-type shoots and roots. In contrast, these early signaling and physiological responses were strongly attenuated in the camalexin-deficient pad3 mutant. Phenotypic priming assays confirmed that mVOC pre-exposure enhances resistance against subsequent Pst DC3000 infection largely through this PAD3-dependent mechanism, while exogenous camalexin administration proved independently sufficient to restore robust pathogen protection, indicating that PAD3-dependent camalexin biosynthesis contributes substantially, but not exclusively, to volatile-induced resistance. Application of synthetic 2-MBA, and to a lesser extent 3-MBA, was sufficient to recreate rapid [Ca2+]cyt elevation and ROS production. Quantitative expression profiling revealed that synthetic 2-MBA modulates gene expression in an organ-specific manner, upregulating shoot volatile perception and salicylic acid pathways, inducing root calcium and immune responses, and systemically activating auxin signaling and tryptophan biosynthetic genes. Together, these findings identify 2-MBA as a primary active component of the Pst DC3000 volatile blend and indicate that full volatile-induced defence depends on PAD3-dependent camalexin accumulation together with integration of multiple volatile signals. Our results reveal how plants integrate distinct bacterial volatiles to trigger early signaling and coordinate localized and systemic camalexin-dependent immunity.

The long-term success of human missions to Mars requires the ability of plants, like maize (Zea mays L.), to thrive under Martian conditions. We assessed the combined effects of the planet’s unique biophysical environment by simulating Martian gravity (sMG, 0.38 g) and Mars hypomagnetic field (hMF, ~ 40 nT) and the chemical toxicity of Mars Global Simulant (MGS-1) regolith. We conducted comprehensive mineralogical characterization of MGS-1 using PXRD, SEM-EDXS, and S/TEM-EDXS. Maize was cultivated in a 30% MGS-1 substrate under sMG and hMF conditions. Analyses included morphological assessments, biochemical measurements, and gene expression. MGS-1 characterization revealed mineral phases (e.g., quartz, mica), enhancing its chemical complexity. Applied alone, hMF and sMG triggered specific compensatory responses in auxin transport and metabolism (e.g., sMG increased leaf area; hMF modulated protein content). In dual-stress scenarios, MGS-1 emerged as the dominant stressor, overriding biophysical cues. The hMF + MGS-1 combination enabled metabolic retention (elevated protein and root PIN plasticity), indicating manageable stress. Conversely, sMG + MGS-1 posed the greatest threat, inducing systemic metabolic depletion (decreased protein and proline) and disrupting root development signals (PIN downregulation). Ultimately, maize survival on Mars is primarily constrained by regolith chemical imbalance and toxicity, rather than reduced gravity or hypomagnetism alone.

The Earth's Geomagnetic Field (GMF) is rarely considered a major environmental regulator. We investigated the effects of a reduced GMF, or hypomagnetic field (hMF), on the aromatic plant peppermint (Mentha × piperita L.). We used GC-MS for essential oil analysis and qPCR to assess monoterpene and sesquiterpene gene expression. Photosynthetic efficiency was quantified via OJIP, NPQ, and PAM fluorescence kinetics. We also quantified H2O2 and ROS gene expression (RBOHD, SOD1, CAT, APX2), and photoprotective antioxidant flavonoids (by HPLC-ESI-MS/MS). The hMF significantly reduced menthol content despite the paradoxical upregulation of key menthol biosynthesis genes. Photosynthetic efficiency was impaired, evidenced by lower PSII parameters and reduced photochemical quenching under high light, along with modulated NPQ, indicated potential adjustments in regulated photoprotection. However, maximum PSII quantum yield remained unaltered, suggesting no basal stress. Peppermint adjusted ROS homeostasis by increasing ROS generation (upregulation of RBOHD) and high-efficiency detoxification (upregulation of APX2 and downregulation of CAT), leading to reduced steady-state H2O2. This adjustment was associated with a significant decrease in total photoprotective phenolic compounds. In peppermint, the absence of a normal GMF modulated electron transfer chain activity, leading to a metabolic trade-off where resources were reallocated toward maintaining redox balance at the expense of productivity. These findings establish GMF as a critical, fine-tuning factor regulating peppermint's metabolic homeostasis.

Microbial volatile organic compounds (mVOCs) provide early chemical cues of microbial activity in the rhizosphere, yet how plants translate these signals into coordinated intracellular responses and developmental outcomes remains poorly understood. Here we show that mVOCs from Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) remodel Arabidopsis thaliana root system architecture (RSA) by inhibiting primary root elongation, reducing lateral root formation, and altering the lateral root gravitropic setpoint angle. Live-cell imaging revealed that Pst DC3000 mVOCs trigger rapid Ca²⁺ and K⁺ fluxes accompanied by reactive oxygen species (ROS) and nitric oxide (NO) accumulation, followed by callose deposition at plasmodesmata. Genetic and pharmacological dissection uncoupled distinct signaling modules: fls2 mutants lost the early H2O2 burst and showed delayed NO production yet retained wild-type levels of plasmodesmatal callose. This demonstrates that FLS2 functions as a genetic coordinator of early redox timing rather than a mediator of symplastic gating. In contrast, the pdko3 mutant (pdlp1/2/3) suppressed Ca²⁺, ROS and early NO responses, indicating that plasmodesmal components are essential for early signal propagation. Pharmacological inhibition of K⁺ channels eliminated callose deposition in Col-0 roots, placing K⁺ influx upstream of PDLP–PMR4-dependent plasmodesmal regulation. At the developmental level, Pst DC3000 mVOCs induced expression of PDLP2, PDLP3 and PDLP4 and reconfigured auxin signaling and PIN auxin transporter expression, including AXR1-dependent DR5 activation, transient PIN1 induction and sustained PIN3 repression, ultimately driving root architectural remodeling. Finally, the bacterial volatile 2-methylbutanoic acid partially recapitulated these effects, indicating that full RSA reprogramming depends on the combined action of multiple mVOCs.

Sorghum (Sorghum bicolor (L.) Moench) is a forage crop in agricultural systems resilient to environmental stresses typical of hot periods such as water deficit, increased salinity and elevated temperatures. However, these environmental stress can increase the content of dhurrin, a cyanogenic glycoside that is potentially toxic to livestock. While single stresses are studied, integrated responses to combinations that include physiology, metabolism, and gene expression remain underexplored.Sorghum plants underwent single (water deficit at 40% field capacity, 200 mM NaCl salinity, 40°C heat for 4h) and combined stresses. We assessed photosynthetic pigments, stress biomarkers (proline, H2O2, antioxidant capacity), hormones (ABA, JA), lipid profiles/indices, phenolics/flavonoids, dhurrin (HPLC-MS/MS), and gene expression (qRT-PCR for oxidative, biosynthetic, and hormonal pathways), with multivariate analyses.Results showed that combined stresses reduced chlorophyll with carotenoid shifts; elevated proline, H2O2, ABA/JA, and dhurrin (doubling in combined stress). Lipid saturation lowered nutritional indices through phenolic/flavonoid reprofiling and gene modulation (e.g., UGT85B1 for dhurrin). Our results revealed non-additive stress synergies, trading resilience for quality/safety. Understanding sorghum stress responses might inform breeding and management strategies to mitigate dhurrin risks under climate change.

Exposure to hypomagnetic fields (hMF, < 40 nT) represents one of the least-characterised environmental stressors for plants. Using Ocimum basilicum L. as a compact, metabolite-rich model for bioregenerative life-support systems, we compared plants grown under hMF versus geomagnetic field (GMF, ∼44 μT) conditions. hMF significantly enhanced essential oil accumulation, with eugenol and methyl eugenol increasing 1.3- and 1.8-fold, respectively. Despite the increased accumulation of these metabolites, transcripts of key phenylpropanoid biosynthetic genes, including the two methyltransferases (ObCOMT and ObEOMT), were significantly downregulated. Total phenolic content decreased while flavonoid content rose, accompanied by a clear shift toward more polar glucuronide- and rhamnoside-conjugated flavonols. Steady-state H2O2 production declined, together with differential regulation of genes involved in ROS detoxification (downregulation of ObCSD and ObCAT; upregulation of ObFSD1). Photosynthetic performance was affected under hMF: chlorophyll b increased, the chlorophyll a/b ratio decreased, and OJIP fluorescence parameters were consistent with partial PSII reaction-centre inactivation, reduced electron transport efficiency, and markedly lower non-photochemical quenching capacity. In parallel, several stress-responsive transcription factors (ObICE1, ObCBF4, ObDREB5) and protective genes were upregulated. These results indicate that hMF exposure induces coordinated changes in secondary metabolism, photosynthetic performance, and oxidative homeostasis in basil, providing a framework for understanding plant responses to near-null magnetic environments relevant to space agriculture.

Spider mites (Tetranychus urticae) are a major threat to economically important crops. Here, we investigated the potential of tetranins, in particular Tet3 and Tet4, as T. urticae protein-type elicitors that stimulate plant defense. Truncated Tet3 and Tet4 proteins showed efficacy in activating the defense gene pathogenesis-related 1 (PR1) and inducing phytohormone production in leaves of Phaseolus vulgaris. In particular, Tet3 caused a drastically higher Ca2+ influx in leaves, but a lower reactive oxygen species (ROS) generation compared to other tetranins, whereas Tet4 caused a low Ca2+ influx and a high ROS generation in the host plants. Such specific and non-specific elicitor activities were examined by knockdown of Tet3 and Tet4 expressions in mites, confirming their respective activities and in particular showing that they function additively or synergistically to induce defense responses. Of great interest is the fact that Tet3 and Tet4 expression levels were higher in mites on their preferred host, P. vulgaris, compared to the levels in mites on the less-preferred host, Cucumis sativus, whereas Tet1 and Tet2 were constitutively expressed regardless of their host. Furthermore, mites that had been hosted on C. sativus induced lower levels of PR1 expression, Ca2+ influx and ROS generation, i.e., Tet3- and Tet4-responsive defense responses, in both P. vulgaris and C. sativus leaves compared to the levels induced by mites that had been hosted on P. vulgaris. Taken together, these findings show that selected tetranins respond to variable host cues that may optimize herbivore fitness by altering the anti-mite response of the host plant.

Lanthanum (La) is often used in industry and agriculture, leading to its accumulation in natural environments and potential ecological risks. The objective of this study was to examine the effects on the growth, metabolism, and nutrient composition of Brassica rapa exposed to at low (1 µM), medium (1 mM), and high (10 mM) La concentrations. We used chemical analytical, molecular, and metabolomic methods and found that high La exposure induced a hormetic effect, triggering both stimulatory and inhibitory responses. La reduced aluminum (Al), cobalt (Co), nickel (Ni), and chromium (Cr) levels at all concentrations, while medium and high doses also decreased phosphorus (P) and iron (Fe). La accumulation in B. rapa increased with La levels, affecting metabolic processes by modulating reactive oxygen species (ROS), increasing proline, and reducing total polyphenol content. Flavonoid levels were altered, chlorophyll and carotenoids declined, and non-photochemical quenching increased. Gene expressions related to flavonoid, carotenoid, and chlorophyll metabolism, as well as ion transport, exhibited a dose-dependent modulation. On the contrary, fatty acid composition remained unaffected. Our results indicate that La accumulates in in B. rapa and disrupts the plant metabolism. Despite an evident effect on plant productivity, our results also raise concerns about the potential health risks of consuming La-enriched B. rapa plants.

The widespread use of rare earth elements (REEs) in agriculture, particularly Lanthanum (La), raises concerns about their ecological impact on non-target organisms. We investigated the direct and indirect effects of La on the insect pest Spodoptera littoralis and its host plant, Brassica rapa. Direct exposure to La-supplemented diets reduced larval growth, survival, and egg production. Interestingly, a transgenerational effect was observed, where larvae from La-exposed parents exhibited increased resilience, showing no performance reduction on the same diets. Indirectly, La accumulation in plants mediated a hormetic response in herbivores, increasing larval weight at low concentrations but reducing it at high concentrations, while modulating their oxidative stress and detoxification gene expression. From the plant perspective, La exposure amplified herbivory-induced calcium signalling and altered the expression of key genes related to calcium and reactive oxygen species pathways. These findings reveal the complex ecological risks of La accumulation in agroecosystems, affecting both plants and insects directly and through novel transgenerational effects.

An interesting aspect that links the geomagnetic field (GMF) to the evolution of life lies in how plants respond to the reduction of the GMF, also known as hypomagnetic field (HMF). In this work, tomato plants (Solanum lycopersicum cv Microtom) were exposed either to GMF or HMF and were studied during the development of leaves and fruit set. Changes of expression of genes encoding for primary and secondary metabolites, including Reactive Oxygen Species (ROS), proteins, fatty acids, polyphenols, chlorophylls, carotenoids and phytohormones were assessed by qRT-PCR, while the corresponding metabolite levels were quantified by GC-MS and HPLC-MS. Two tomato homologs of the fruit fly magnetoreceptor MagR, Isca-like 1 and erpA 2, were modulated by HMF, as were numerous tomato genes under investigation. In tomato leaves, positive correlations were observed with most of the genes associated with phytohormones production, ROS scavenging and production, and lipid metabolism, whereas an almost reversed trend was found in flowers and fruits. Interestingly, downregulation of Isca-like 1 and erpA 2 was found to correlate with an upregulation of most unripe fruit genes. Exposure to HMF reduced chlorophyll and carotenoid content, decreased photosynthetic efficiency and increased non-photochemical quenching. Auxins, gibberellins, cytokinins, abscisic acid, jasmonic acid and salicylic acid content and the expression of genes related to their metabolism correlated with tomato ISCA modulation. The results here reported suggest that Isca-like 1 and erpA 2 might be important players in tomato magnetoreception.

Plants are critical for sustaining human life and planetary health. However, their potential to enable humans to survive and thrive beyond Earth remains unrealized. This Viewpoint presents a collective vision outlining priorities associated with plant science to support a new frontier of human existence. These priorities are drawn from the International Space Life Sciences Working Group (ISLSWG) Plants for Space Exploration and Earth Applications workshop, held at the European Low Gravity Research Association (ELGRA) conference in September 2024. First, we highlight transformative advances gained from using the 'laboratory of space' in understanding how plants respond to gravity and other stressors. Second, we introduce a new crop Bioregenerative Life Support System (BLSS) readiness level (BRL) framework - extending the existing Crop Readiness Level (CRL) - to assist in overcoming challenges to establish resilient, sustainable crop production. Materializing the vision of plants as enablers of space exploration will require innovative approaches, including predictive modeling, synthetic biology, robust Earth-based analogue systems, and reliable space-based instruments to monitor biological processes. Success relies upon a unified international community to promote sharing of resources, facilities, expertise, and data to accelerate progress. Ultimately, this work will both advance human space exploration and provide solutions to enhance sustainable plant production on Earth.

Cerium (Ce), the most abundant of the rare Earth elements (REEs), is increasingly recognized as an environmental contaminant due to its growing applications in various industrial and agricultural sectors. This study investigates the physiological, biochemical, and molecular responses of Brassica rapa L. plants to varying concentrations of Ce exposure to elucidate its effects on plant growth, metabolism, and stress responses. Through chemical analytical, biochemical, and gene expression methods, we revealed a biphasic (hormetic) effect of Ce on B. rapa. Low-level Ce exposure (1 µM) stimulated plant growth, evidenced by increased leaf area and fresh biomass. Conversely, elevated Ce concentrations (1 mM and 10 mM) induced significant photosynthetic dysfunction, characterized by diminished chlorophyll a and b content, impaired photosystem II (PSII) efficiency, and altered chlorophyll fluorescence. Ce exposure also modulated oxidative stress responses, exhibiting a hormetic pattern in reactive oxygen species (ROS) accumulation, alongside a general increase in proline. Secondary metabolism was selectively impacted, with higher Ce levels specifically promoting the accumulation of kaempferol derivatives. Mineral nutrient analysis revealed substantial Ce accumulation in leaves and a concomitant decrease in essential elements (Al, Se, Na). Gene expression analysis further elucidated that Ce exposure triggered differential expression of genes involved in carotenoid and flavonoid biosynthesis, chlorophyll metabolism, and ion transport. These comprehensive findings offer novel insights into the multifaceted physiological, biochemical, and molecular responses of B. rapa to Ce, underscoring both the potential ecological risks of Ce contamination and the intricate adaptive strategies employed by plants under REE stress.

The intricate interplay of quantum coherence, entanglement, radical pair mechanisms, and tunneling, suggests that plants operate at a level of sophistication beyond classical expectations. The potential to harness these quantum principles for agricultural innovation and environmental sustainability is immense. This review provides a comprehensive overview of plant quantum biology, extending beyond photosynthesis and magnetosensitivity the exploration of enzyme catalysis and stress responses. The quantum coherence and entanglement in photosynthetic light harvesting and energy transfer, examining their role in efficient energy transduction is evaluated. Plant magnetosensitivity, mediated by cryptochromes and iron–sulfur clusters, is discussed as a potential quantum sensing mechanism. The radical pair mechanism influence on plant growth, development, and circadian rhythms via magnetic field perception is analyzed. Quantum tunnelling impact on enzyme reaction rates and substrate specificity is also discussed. The critical intersection of quantum biology and plant stress responses, encompassing light, oxidative stress, temperature, and biotic stress, is examined. How quantum effects might modulate these responses, offering opportunities for developing stress-tolerant crops reveal that challenges posed by biological complexity, transient quantum phenomena, and experimental limitations, along the need for robust theoretical models are future trends in plant stress biology. Future research should focus on manipulating quantum effects in vivo, bridging fundamental science and agricultural applications for enhanced sustainability.

The Earth’s geomagnetic field (GMF) is a fundamental environmental signal for plants, with its perception rooted in quantum biology. Specifically, the radical pair mechanism (RPM) explains how this weak force influences electron spin states in metabolic pathways, providing a framework for its profound biological impact. Research shows that a hypomagnetic field (hMF) directly reduces the production of reactive oxygen species (ROS), creating a quantum signature in plants. This is a counterintuitive finding, as it suggests the plant perceives less oxidative stress and, in response, downregulates its antioxidant defenses. This multi-level effect, from a quantum trigger to molecular and metabolic changes, ultimately affects the plant’s growth and phenotype. This review suggests a possible link between the GMF and plant health, identifying the GMF as a potential physiological modulator. Manipulating the magnetic field could therefore be a novel strategy for improving crop resilience and growth. However, the fact that some effects cannot be fully explained by the RPM suggests other quantum mechanisms are involved, paving the way for future research into these undiscovered processes and their potential inheritance across generations.

The appearance of new respiratory virus infections in humans with epidemic or pandemic potential has underscored the urgent need for effective broad-spectrum antivirals (BSAs). Bioactive compounds derived from plants may provide a natural source of new BSA candidates. Here, we investigated the novel phytocomplex formulation SP4™ as a candidate direct-acting BSA against major current human respiratory viruses, including coronaviruses and influenza viruses. SP4™ inhibited the in vitro replication of SARS-CoV-2, hCoV-OC43, hCoV-229E, Influenza A and B viruses, and respiratory syncytial virus in the low-microgram range. Using hCoV-OC43 as a representative respiratory virus, most of the antiviral activity of SP4™ was observed to stem primarily from its dimeric A-type proanthocyanidin (PAC-A) component. Further investigations of the mechanistic mode of action showed SP4™ and its PAC-A-rich fraction to prevent hCoV-OC43 from attaching to target cells and exert virucidal activity. This occurred through their interaction with the spike protein of hCoV-OC43 and SARS-CoV-2, thereby interfering with spike functions and leading to the loss of virion infectivity. Overall, these findings support the further development of SP4™ as a candidate BSA of a natural origin for the prevention of human respiratory virus infections.

The stress that the space environment can induce on plant physiology is of both abiotic and biotic nature. The abiotic space environment is characterized by ionizing radiation and altered gravity, geomagnetic field (GMF), pressure, and light conditions. Biotic interactions include both pathogenic and beneficial interactions. Here, we provide an overall picture of the effects of abiotic and biotic space-related factors on plant physiology. The knowledge required for the success of future space missions will lead to a better understanding of fundamental aspects of plant physiological responses, thus providing useful tools for plant breeding and agricultural practices on Earth.

Phytopathogens are well known for their devastating activity that causes worldwide significant crop losses. However, their exploitation for crop welfare is relatively unknown. Here, we show that the microbial volatile organic compound (mVOC) profile of the bacterial phytopathogen, Erwinia amylovora, enhances Arabidopsis thaliana shoot and root growth. GC-MS head-space analyses revealed the presence of typical microbial volatiles, including 1-nonanol and 1-dodecanol. E. amylovora mVOCs triggered early signaling events including plasma transmembrane potential Vm depolarization, cytosolic Ca2+ fluctuation, K+-gated channel activity, and reactive oxygen species (ROS) and nitric oxide (NO) burst from few minutes to 16 h upon exposure. These early events were followed by the modulation of the expression of genes involved in plant growth and defense responses and responsive to phytohormones, including abscisic acid, gibberellin, and auxin (including the efflux carriers PIN1 and PIN3). When tested, synthetic 1-nonanol and 1-dodecanol induced root growth and modulated genes coding for ROS. Our results show that E. amylovora mVOCs affect A. thaliana growth through a cascade of early and late signaling events that involve phytohormones and ROS.

The geomagnetic field (GMF) is a natural component of the biosphere, and, during evolution, all organisms experienced its presence while some evolved the ability to perceive magnetic fields (MF). We studied the response of 14-3-3 proteins and the plasma membrane (PM) proton pump H+-ATPase to reduced GMF values by lowering the GMF intensity to a near-null magnetic field (NNMF). Seedling morphology, H+-ATPase activity and content, 14-3-3 protein content, binding to PM and phosphorylation, gene expression, and ROS quantification were assessed in maize (Zea mays) dark-grown seedlings. Phytohormone and melatonin quantification were also assessed by LG-MS/MS. Our results suggest that the GMF regulates the PM H+-ATPase, and that NNMF conditions alter the proton pump activity by reducing the binding of 14-3-3 proteins. This effect was associated with both a reduction in H2O2 and downregulation of genes coding for enzymes involved in ROS production and scavenging, as well as calcium homeostasis. These early events were followed by the downregulation of IAA synthesis and gene expression and the increase in both cytokinin and ABA, which were associated with a reduction in root growth. The expression of the homolog of the MagR gene, ZmISCA2, paralleled that of CRY1, suggesting a possible role of ISCA in maize magnetic induction. Interestingly, melatonin, a widespread molecule present in many kingdoms, was increased by the GMF reduction, suggesting a still unknown role of this molecule in magnetoreception.

The genus Coffea is known for the two species C. arabica (CA) and C. canephora (CC), which are used to prepare the beverage coffee. Proper identification of green beans of coffee varieties is based on phenotypic and phytochemical/molecular characteristics. In this work, a combination of chemical (UV/Vis, HPLC-DAD-MS/MS, GC-MS, and GC-FID) and molecular (PCR-RFLP) fingerprinting was used to discriminate commercial green coffee accessions from different geographical origin. The highest content of polyphenols and flavonoids was always found in CC accessions, whereas CA showed lower values. ABTS and FRAP assays showed a significant correlation between phenolic content and antioxidant activity in most CC accessions. We identified 32 different compounds, including 28 flavonoids and four N-containing compounds. The highest contents of caffeine and melatonin were detected in CC accessions, whereas the highest levels of quercetin and kaempferol derivatives were found in CA accessions. Fatty acids of CC accessions were characterized by low levels of linoleic and cis octadecenoic acid and high amounts of elaidic acid and myristic acid. Discrimination of species according to their geographical origin was achieved using high-throughput data analysis, combining all measured parameters. Lastly, PCR-RFLP analysis was instrumental for the identification of recognition markers for the majority of accessions. Using the restriction enzyme AluI on the trnL-trnF region, we clearly discriminated C. canephora from C. arabica, whereas the cleavage performed by the restriction enzymes MseI and XholI on the 5S-rRNA-NTS region produced specific discrimination patterns useful for the correct identification of the different coffee accessions. This work extends our previous studies and provides new information on the complete flavonoid profile, combining high-throughput data with DNA fingerprinting to assess the geographical discrimination of green coffee.