Global protein security is increasingly challenged by the growing demand for sustainable alternatives to animal-derived proteins. Although plant proteins are central to this transition, they remain limited by imbalanced amino acid profiles, reduced digestibility, and inferior techno-functional properties, restricting their nutritional equivalence. Recent studies have explored processing strategies to address these limitations; however, these approaches are often evaluated independently, with limited integration of structural mechanisms and phytochemical-protein interactions. This review presents a comparative and mechanistic synthesis based on cross-study evaluation, integrating protein structure, processing-induced modifications, and phytochemical-assisted interactions. Processing strategies including extrusion, fermentation, enzymatic hydrolysis, and pH shifting primarily enhance protein accessibility and reduce antinutritional constraints. In contrast, phytochemicals modulate protein conformation and interfacial behavior through both non-covalent and covalent interactions. Evidence across studies indicates that these effects are strongly concentration-dependent: moderate phytochemical interactions promote partial unfolding and improved functionality, whereas excessive interactions induce aggregation and reduce digestibility. Notably, the combined application of bioprocessing and phytochemical strategies yields greater improvements in solubility, emulsification, and bioavailability than individual approaches, although variability in protein source and processing conditions remains a key limitation. Overall, this review establishes a mechanistic framework linking protein structure, processing dynamics, and phytochemical interactions, highlighting concentration-dependent effects and synergistic strategies for improving plant protein functionality, while identifying variability and optimization challenges for future applications.
Globally, the pathogen-driven enteric diseases remain a leading cause of morbidity and mortality, disproportionately affecting populations in low-income regions. While significant strides have been made in vaccine research, challenges such as high production costs, complex manufacturing processes, and inefficient distribution continue to hinder global vaccine accessibility. To overcome these challenges, plant-based edible vaccines have emerged as a promising and innovative alternative, offering advantages such as affordability, ease of administration, and reduced reliance on cold-chain infrastructure. This study presents a comprehensive overview of recent advances in the development of edible plant-derived vaccines, highlighting the strategic evolution from conventional genetic engineering to more refined and scalable biotechnological methods. In the work, various applications targeting diseases such as measles, hepatitis B, rabies, dengue, and norovirus, are discussed as well as veterinary vaccines for livestock and aquaculture. The significant progress in creating vaccines targeting prevalent human pathogens is examined and the discussion is extended to include edible vaccines developed for livestock, underscoring their role in both public and veterinary health. Furthermore, the review emphasizes the transformative potential of 'omics' technologies, such as genomics, proteomics, and metabolomics, as well as artificial intelligence in streamlining vaccine design, improving antigen expression, and accelerating development timelines. This assessment review was conducted by performing a comprehensive literature survey of peer-reviewed articles. By integrating multidisciplinary insights, the work underscores the feasibility and future prospects of edible plant vaccines as a sustainable solution to global immunization challenges, with practical applications in improving vaccine accessibility, enhancing outbreak preparedness, and supporting immunization efforts in resource-limited settings. It aims to inform and inspire continued research and collaborative innovation in this emerging field with significant implications for global public health.
Modern military performance increasingly depends on cognitive resilience and neurological health, making nutrition a strategic determinant of combat readiness. This review integrates nutritional neuroscience and military medicine to explore duckweed as a next-generation neuro-nutritional resource. Duckweed species possess protein content (25-45% dry weight) and a complete essential almost amino acid (EAA) profile that may contribute to pathways involved in neurotransmitter synthesis, neuroplasticity, and stress adaptation. Beyond amino acids, duckweed delivers neuroprotective compounds such as lutein, β-carotene, polyphenols, and vitamin B₁₂ (pseudo-cobalamins) that mitigate oxidative stress, inflammation, and cognitive decline induced by fatigue or trauma. Its growth rate, minimal land and water requirements make it a sustainable, field-deployable protein source suitable for remote bases or disaster zones. This review synthesizes evidence on duckweed's biochemical composition, digestibility, and safety, highlighting its potential as a sustainable substitute for soy or Spirulina in military rations. The convergence of plant biotechnology, cognitive nutrition, and defense health positions duckweed as a strategic component of future military and space nutrition systems, supporting nutritional strategies relevant to brain health in demanding operational contexts.
Plant-based proteins are novel nutrient sources that are widely available in nature and cost-effective to produce compared to animal proteins, while contributing to lower greenhouse gas emissions and reducing risks to both environmental and human health. Thus, the present work comprehensively reviews plant-based proteins as an alternative to conventional / animal proteins. The review article highlights why plant proteins should be considered a novel nutrient source and an alternative to animal proteins. Various plant protein-derived sources and their corresponding extraction methods, along with their associated limitations, are reviewed. In addition, the review article highlights plant protein modification methods, interfacial behavior, protein structure, interactions, and functional properties. The effects of different physical and environmental conditions and factors on the functional and nutritional properties of plant proteins are also discussed. In addition, the challenges associated with plant proteins, such as allergenicity, consumer acceptance and perceptions, food safety, regulatory issues, and technological challenges, are also explored.
The increasing global demand for functional foods rich in nutrients has spurred the exploration of agricultural practices aimed at enhancing the nutritional quality of crops. This study explored the effects of potassium biofortification on quinoa microgreens (QMG). The results showed that treatment with 8 mmol·L- 1 KCl increased fresh weight by 60.25-fold, reduced nitrate and free amino acids, and increased the contents of soluble sugars, proteins, vitamin C, fats, and cellulose. The application of potassium also increased the contents of polyphenols, flavonoids, and saponins, as well as the activity of antioxidant enzymes. A comprehensive evaluation indicated that QMG harvested 25 days after germination had optimal nutritional value and lower nitrate content. This study demonstrates that potassium biofortification is an effective strategy to improve the yield, nutritional quality, and health-promoting value of QMG, providing a theoretical basis for its development as a functional food.
Plant-based cheese alternatives (PBCAs) are considered sustainable substitutes for dairy-based cheese (DB); however, their consumer acceptance remains limited, particularly in dairy-dominant cultural contexts such as Uruguay, where specific consumer insights are scarce. This study investigated the influence of sensory experience, product information, and conceptual expectations on consumer perceptions of commercially available mozzarella-style PBCAs in Uruguay. Three sessions were conducted: blind and informed tastings (n = 110) and an expectation survey with a separate participant group (n = 236). Liking, purchase intention, and conceptual associations were evaluated using hedonic scales, CATA questions, and cluster analyses. Dairy-based cheese consistently achieved the highest liking and purchase intention scores. Positive sensory drivers for acceptance included a smooth mouthfeel, creaminess, and shiny appearance, whereas artificial flavors and strong aftertastes reduced acceptance. One PBCA showed an acceptable texture but lacked flavor. Crucially, providing product information did not enhance PBCA acceptance, underscoring the importance of sensory experiences. Cluster analysis revealed two consumer segments: "high purchase intention group" (n = 176), who are younger, and associate PBCAs with sustainability and health; and "low purchase intention group" (n = 60), who were older and perceived PBCAs as artificial and unappealing. PBCAs hold potential among sustainability-oriented consumers, but improving flavor authenticity and sensory quality is crucial. Addressing these sensory barriers and strategically positioning products around health and environmental benefits are crucial for broadening overall acceptance and facilitating sustainable dietary shifts in the population.
Pregelatinized flours are gaining prominence in the gluten-free food industry due to their ability to tailor functional properties to specific technological requirements and end-product needs. This study aimed to characterize the physicochemical and techno-functional properties of pregelatinized rice (PRF), buckwheat (PBF), green (PGPF), and yellow pea (PYPF) flours to evaluate their potential as functional ingredients for gluten-free premixes. Comprehensive insights into nutritional and technological behavior of these materials are pivotal for developing novel foods. Results indicated that pregelatinization significantly altered the techno-functional profile of the flours. Specifically, water holding capacity increased by 408% for PRF, 281% for PBF, 222% for PYPF, and 171% for PGPF, while water absorption capacity increased by 549, 296, 243, and 257%, respectively. These enhancements are critical for improving texture and moisture retention in gluten-free matrices. Furthermore, this pretreatment generally increased the capacity to absorb organic molecules (hydrophobic compounds), whereas emulsification properties decreased. Microstructural analysis confirmed disruption of starch granules, explaining the enhanced water interactions. Additionally, the flours exhibited a high mineral content (Ca, Fe, P, and K), which is essential for enriching the nutritional profile of gluten-free diets, with a notable increase in Ca concentration after the pregelatinization. Principal component analysis demonstrated that pregelatinization primarily enhanced water and oil holding capacities, underscoring its efficacy in optimizing functional attributes within gluten-free systems. These findings confirm the significant potential of pregelatinized rice, buckwheat, and pea flours as versatile ingredients for the industry, offering a solution to texture and elasticity challenges in gluten-free bakery products.
An integral use approach of chia seeds was applied through microencapsulation, combining chia oil as the core with seed-derived proteins and mucilage as wall components for the development of potential functional food ingredients. Oil-in-water emulsions were formulated and dried by spray-drying and freeze-drying to evaluate how their distinct drying mechanisms influence the physicochemical, structural, and functional properties of chia oil microparticles. Both drying methods successfully produced chia oil microparticles, yielding powders with low moisture content, high encapsulation efficiency (approximately 91%, w/w), and high protein content. However, the drying method influenced the structural and functional properties of the microparticles. Scanning electron micrographs revealed lamellar structures in freeze-dried samples and spherical particles in spray-dried ones. After reconstitution, emulsion particle size increased during storage, particularly in freeze-dried systems, whereas spray-dried microparticles showed superior dispersibility and instant properties. Regarding oxidative stability, freeze-dried microparticles provided greater protection against lipid oxidation, reaching a peroxide value of 2.90 meq/kg oil after 45 days of storage. Besides, the α-linolenic acid content remained stable in freeze-dried microparticles, whereas a significant decrease was observed in spray-dried samples over the same period. A preliminary sensory evaluation of a beverage fortified with freeze-dried microparticles indicated that 94% of panelists were able to identify the fortified sample, which showed significantly lower sensory acceptance for taste, color, and appearance than the non-fortified product. Overall, these findings demonstrate the feasibility of using seed-derived proteins and mucilage as wall materials for chia oil encapsulation and highlight the potential of freeze-dried microparticles as ingredients for functional beverages.
The Traditional Mexican Diet (TMexD) is a culturally rooted, plant-forward dietary pattern derived from Mesoamerican agriculture and culinary practice, built on minimally processed staples-maize-based preparations, legumes, vegetables, fruits, and herbs. Characteristic techniques such as nixtamalization and fermentation alter starch structure, mineral availability, and gut microbial activity. Through these effects, the pattern shapes metabolic responses relevant to carcinogenesis. This narrative review clarifies defining features of the TMexD across Mexico's regional diversity and synthesizes epidemiologic evidence from Mexican and Mexican-heritage populations linking closer alignment with the TMexD to favorable lipid and insulin profiles and lower markers of inflammation. Site-specific observations suggest inverse associations for multiple cancers, with the most direct evidence for breast and colorectal cancer, supportive component-level evidence for gastric cancer, and indirect evidence for lung and prostate cancer. Convergent mechanistic pathways include improved insulin/IGF signaling, modulation of oxidative stress and bile-acid metabolism, short-chain fatty acid production that reinforces mucosal integrity, and enhanced immune surveillance; traditional dishes contribute additional phytochemicals with antioxidant and anti-inflammatory activity. We also examine how globalization, migration, acculturation, and urban food environments influence access to heritage staples and day-to-day practice of the pattern, noting that some regional staples remain robust while others are declining. Although definitions of "traditional" vary and measurement of preparation methods is inconsistent across studies, the convergent epidemiologic and mechanistic evidence positions the TMexD as a credible, culturally concordant framework for cancer prevention in contemporary Mexican and diaspora settings.
Traditional wheat-based pasta is widely consumed; however, its reformulation with functional plant-based ingredients remains challenging because technological quality, bioactive properties, and consumer acceptance must be balanced. This study aimed to optimize and validate a functional pasta formulation by partially replacing wheat flour (WF) with lupin flour (LF) and flaxseed flour (FF). A 2² central composite rotatable design was applied, using LF (2-12%) and FF (5-15%) as independent variables. Instrumental color, cooking properties, texture, total soluble phenolic compounds (TSPC), antioxidant capacity (ORAC), and sensory acceptance were evaluated. Flour substitution significantly affected pasta quality, particularly color, cooking behavior, and texture. Increasing FF reduced lightness and increased redness, whereas higher LF contributed to yellowness. Although cooking and texture properties were modified, several formulations maintained acceptable technological performance, with low cooking loss and suitable texture-related attributes. Enriched pasta showed significantly higher TSPC and ORAC values than the control, indicating enhanced bioactive potential. Sensory evaluation revealed moderate to high acceptance, although excessive substitution negatively affected color and texture perception. Sensory acceptance was mainly associated with texture, stickiness, and cooking time. Numerical optimization identified an optimal formulation with 12.0% LF and 12.13% FF, achieving an overall desirability of 77.3%. Validation assays showed good agreement between predicted and experimental values, confirming model adequacy. Overall, lupin and flaxseed flours are promising ingredients for developing functional pasta with improved bioactive potential and acceptable technological and sensory quality.
This study investigated germination as a pre-isolation modification method to enhance the techno-functional and nutritional properties of common buckwheat (Fagopyrum esculentum Moench) protein. Germination was conducted at two different temperatures (20 and 30 °C) for 24, 48, and 72 h, and isolated proteins were analyzed for their chemical, structural, and functional properties. Results showed that the highest protein yield was achieved at 30 °C for 24 h, though durations exceeding 48 h reduced purity. SDS-PAGE and FTIR analyses confirmed that endogenous protease activity degraded high-molecular-weight proteins and increasing random coil proportions, indicating partial unfolding. SEM imaging revealed that germination transformed the dense protein surface into a porous morphology. Increasing germination time and temperature led to decreased lightness (L*) and hue angle, along with moderate increases in redness (a*), indicating notable changes in the visual appearance of the protein isolates. DSC results showed a decrease in thermal stability (Td) due to structural changes. In addition, germination significantly improved solubility, emulsifying activity, foaming capacity, and water/oil absorption, particularly at the 24 h of germination. Most notably, germination at 30 °C for 24 h increased in vitro protein digestibility from 75.67 to 82.19%. The results show that the germination is a promising approach to enhance the functionality and digestibility of buckwheat protein for potential application in food industry.
Polygonum cognatum is an underutilized edible plant rich in dietary fiber, minerals, and bioactive compounds with potential applications in functional foods. This study evaluated the effects of Polygonum cognatum powder (PCP) incorporation (0-15%) on the nutritional, bioactive, technological, sensory, and glycemic properties of gluten-free and wheat-based crackers. PCP addition significantly (p < 0.05) increased protein, total dietary fiber, phenolic content, antioxidant capacity, and mineral composition (Ca, Fe, Mg, K, and P), while reducing phytic acid levels in both formulations. Antioxidant activity (DPPH, FRAP, and CUPRAC) and total phenolic content increased markedly in a dose-dependent manner, indicating the high bioactive potential of PCP. In vitro starch digestion analysis showed that PCP incorporation effectively reduced the estimated glycemic index (eGI), with values decreasing from 92.45 to 57.98 in wheat-based crackers and from 96.57 to 60.12 in gluten-free crackers. Gluten-free samples consistently exhibited slightly higher eGI values, likely due to the higher digestibility of starch-based matrices. PCP addition increased spread ratio and hardness, Spread ratio and hardness increased above 3% PCP substitution level. Sensory evaluation indicated that 3-6% substitution provided optimal acceptability, whereas higher levels (≥ 12%) negatively affected color, taste, and crispness. Overall, PCP is a promising functional ingredient for developing nutritionally enhanced and lower-glycemic bakery products.
Muicle (Justicia spicigera) is a traditional Mexican medicinal plant rich in phenolic compounds and pigments, yet its extraction conditions have not been systematically optimized. Using a 3² factorial response surface methodology (RSM) design, solvent type (water, ethanol, hexane) and extraction technique (stirring, decoction, sonication) were evaluated in leaves and flowers. Total phenolic content ranged from 2.77 to 97.39 mg GAE/100 g in leaves and reached up to 139.82 mg GAE/100 g in flowers, while ABTS antioxidant capacity was remarkably high in both tissues. RSM models (R² ≥ 0.70) identified water decoction as optimal for leaves and water decoction sonication for flowers, maximizing phenolic recovery and natural coloration. Metabolomic profiling putatively identified 27 metabolites including chalcones, flavonoids, stilbenes, secoiridoids, and omega phospholipids supporting the observed bioactivity. These results provide validated extraction protocols and a metabolomic baseline for muicle based functional foods, natural colorants, and nutraceuticals.
The replacement of wheat flour (WF) by legume flours allows to obtain functional baked goods, but generally leads to technological and sensory defects. Applying thermal treatments to legumes can lead to increased nutrient digestibility and water absorption, reduced unpleasant flavors, and various textural modifications. This work evaluated the technological aspects of dough and breads made with thermally treated lentil flour (LF). In this study, LF was produced from raw lentils that were soaked, cooked (30 min - boiling water), and dried (12 h - 60 °C). WF was replaced by different percentages of LF: 0 (Control), 10 (LF10), 20 (LF20) and 30% (LF30). Hydration properties, color, specific volume, alveolar parameters, rheological characteristics, nutritional composition, and sensory acceptability were evaluated. The LF dough reached lower volumes during fermentation, leading to inferior bread specific volume, with this effect being more pronounced when LF substitution level increased. LF20 and LF30 were labelled as good sources of fiber (with values of 10.2% and 13.1% respectively), and LF30 can also be considered as good source of protein (with a value of 17.2%). Results shows that even the smallest amounts of LF significantly improved the nutritional profile of the bread. LF breads were perceived as nutritive, artisanal, and with the ability of satiate by the consumers, and the flavor lentil was only perceived with LF > 10%. Lentil addition was shown to be a viable alternative for improving the nutritional profile of traditional breads, leading to products with high acceptance. LF20 bread could be considered as products with a good balance between different technological, nutritional and sensory attributes.
Broccoli sprouts are abundant in bioactive antioxidants and are recognized as a novel class of functional food. This study explored the mechanisms by which selenium-sulfur coupled with heat treatment (HSS) affects the antioxidant capacity and accumulation of bioactive compounds in broccoli microgreens. The results showed that, compared to the control group, 4-day-old broccoli sprouts treated with HSS exhibited a 27.16% increase in total phenolic content and a 26.63% increase in anthocyanin content. Activities of catalase, superoxide dismutase, and peroxidase increased by 27.97, 30.54, and 46.48%, respectively. These changes enhance the antioxidant capacity of broccoli sprouts. HSS induced the biosynthesis of glucosinolates (GLs) and isothiocyanates (ITCs) by increasing myrosinase activity in the sprouts and upregulating the expression levels of genes associated with the GLs and ITCs biosynthetic pathways. iTRAQ proteomics analysis further revealed that the GLs biosynthetic pathway is the key pathway for ITCs accumulation induced by combined selenium-thiocyanate and heat stress. In addition, MAM1, APS3, SULTR1;2, and CYP79B3 were identified as potential key proteins. These proteins contribute to ITCs formation in sprouts under the combined treatment. These findings support the HSS as a novel regulatory strategy to enhance the nutritional value of broccoli sprouts and lay the foundation for the development of functional foods based on broccoli sprouts.
The transition toward sustainable protein sources has driven growing interest in plant, insects, and microalgae proteins as alternatives to animal-derived ingredients. However, their variable amino acid composition, functional and sensory characteristics limit their application in high-quality food formulations. Computational approaches such as linear programming, mathematical optimization, and artificial intelligence offer powerful tools to design nutritionally balanced and functionally feasible protein blends. By integrating data on protein quality, techno-functional properties, and sensory thresholds, these methods enable the rational selection and combination of ingredients. This review summarizes current advances in computational food design, highlighting how data-driven formulation can accelerate innovation in sustainable protein products and bridge the gap between empirical experimentation and predictive modeling.
Food-derived biopeptides have attracted increasing attention due to their potential health benefits and favorable safety profiles. In this study, rice protein hydrolysates (< 3 kDa) and their derived peptides were investigated for their protective effects against tumor necrosis factor (TNF)-α/interferon (IFN)-γ-induced injury in human keratinocyte cells (HaCaT). The hydrolysates significantly enhanced cell viability and migration, and five peptides were identified by LC-MS/MS and in-silico analysis. These peptides improved the viability of damaged cells, with PG9 (PSWVAFTGG) showing the greatest activity. PG9 significantly downregulated the mRNA expression of pro-inflammatory cytokines, including Regulated on Activation, Normal T-cell Expressed and Secreted (RANTES), Interleukin (IL)-1β, IL-6, and IL-23, while also markedly promoting keratinocyte migration. Molecular dynamics simulations revealed that PG9 formed stable hydrogen-bonds with key residues (Lys745 and Asp855) within the epidermal growth factor receptor (EGFR) binding site, suggesting its involvement in EGFR-mediated wound healing signaling. In addition, PG9 regulated the PI3K/AKT/mTOR pathway by reducing the expression of PI3K, AKT, mTOR, and inhibiting AKT phosphorylation. These results identify PG9 as a stable EGFR-binding peptide capable of modulating inflammatory signaling and supporting its potential as a natural agent for skin repair.
Cacti by-products are rich in bioactive compounds, yet they remain largely underexplored in foods. This study investigated the nutritional and functional potential of red pitaya (Selenicereus monacanthus) peel flour in an innovative chocolate truffle composition. The phenolic composition, antioxidant activity, in vitro bioaccessibility, and intestinal permeability of flour were evaluated. It showed a significant content of bioactive flavonoids, with isorhamnetin, rutin, myricetin, quercetin, and luteolin as major identified compounds. In vitro passive permeability of polyphenol from flour was moderate (Papp = 5.24 ± 0.85 × 10- 6 cm/s). Chocolate truffles made with pitaya peel flour (BPF) showed an 86% increase in fiber, a 23% increase in ash content, a 27% reduction in lipids, and a 30% decrease in carbohydrates compared to those without addition. Adding peel flour increased the food's antioxidant activity (3.5-fold) and phenolic composition (2.6-fold) compared to the control. Under simulated digestion, polyphenols' bioaccessibility in BPF was higher in the ileum, and antioxidant activity was higher in the duodenal stage, compared to the gastric stage. Data indicates that pitaya peel flour is a promising input for developing foods.
Sprouts, due to their high nutritional value and suitable environmental conditions during the germination process, are an ideal medium for the growth of pathogenic microorganisms. Primary sprout contamination usually occurs through seeds and post-harvest washing. Therefore, effective seed and sprout disinfection can be an essential requirement for the food safety of edible sprouts. In this systematic review, 146 studies published between 2010 and 2024 were evaluated, and the effectiveness of different chemical and non-chemical disinfection methods was investigated. In traditional chemical methods, which usually use sodium hypochlorite (NaOCl)/calcium hypochlorite (Ca (OCl)₂) and hydrogen peroxide, a reduction in microbial load was observed, but there are disadvantages, such as possible negative effects on seed viability and quality. There are concerns about chemical residues and their effectiveness against pathogens internalized or protected in biofilms is limited. However, chlorine dioxide has been introduced as a promising alternative, which has higher oxidation power, lower dependence on pH, and produces fewer carcinogenic byproducts. Non-chemical methods have also received much attention due to better preservation of seed quality and the absence of chemical residues. Physical and biological non-chemical methods are also effective in disinfection. However, the most effective strategy is the combination of several methods, called "Hurdle" technology. This multiple-stress approach overwhelms the repair mechanisms of the microorganism and leads to further microbial reduction, reduced side effects on the seed and sprout, and increased overall efficiency. On the other hand, the use of hurdle technology is the most promising approach of procuring the best possible safety at the same time as keeping the quality and germination of the end product.
Basil (Ocimum basilicum L.) is a globally consumed herb whose hydroxycinnamic acid esters and flavonoids contribute to dietary antioxidant intake. This study aimed to characterise the phenolic composition, antioxidant capacity, and developmental-stage variability of twelve Turkish basil genotypes grown across three ecologically contrasting sites. Twelve genotypes were evaluated in a two-year field experiment (Year 1 and Year 2) at three ecologically contrasting locations in Türkiye: Bursa, Eskişehir, and Tokat. Agronomic performance, nine-compound HPLC-TOF phenolic profiles (Year 1), total phenolic content (TPC), and antioxidant capacity (ABTS, DPPH, and FRAP) were measured. The effects of developmental stage on the phenolic composition of plants from Bursa (Year 1) were characterised. Location was the dominant source of variation in phenolic quality. Plants from Tokat ranked highest for individual phenolic concentrations (Year 1 HPLC-TOF data) and for all three antioxidant assays (two-year means). Rosmarinic acid varied 4.4-fold (59.0-261.8 mg 100 g⁻¹ DW; Year 1), with the highest values recorded for genotypes R-3k, R-19, R-15, and R-4 from Tokat. ABTS antioxidant capacity was 113% higher in plants from Tokat than in those from Bursa (two-year means: 483 vs. 227 µmol TE g⁻¹ DW). Analysis of rosmarinic acid in plants from Bursa showed an irregular, stage-dependent pattern, whereas TPC remained broadly stable across developmental stages (vegetative to full flowering: +3.9%). The genotype × stage interaction accounted for 44-50% of the total variance. PCA explained 89.3% of the variance across two components, with Tokat combinations separating clearly from those of the other sites. Under the tested field conditions, genotypes R-23 and R-4 emerged as strong candidates for phenolic-acid-targeted cultivation, while Y-15 recorded the highest two-year mean TPC at Tokat.