Abstract
The alteration of soil chemical composition and soil-to-plants micronutrients transfer are crucial for agricultural sustainability and food safety. This study aims to investigate Cu, Zn, and Mo contents in agricultural soils and assess their accumulation in edible parts of plants within the main agricultural areas of the Ararat region, Armenia. The contents of the studied elements were determined using a combination of atomic absorption and X-ray fluorescence spectrometry. To investigate the relationship between soil and plant contents of Cu, Zn, and Mo, and to assess the influence of major and minor elements (Fe, Mn, Ca, K, Sr, Rb, Ti, Zr) on their distribution, hierarchical clustering was applied. The contents of studied micronutrients in soils followed the order Zn(55.6-219 mg/kg) > Cu(42.1-189 mg/kg) > Mo(2.5-5 mg/kg), and in plant samples the same pattern was observed: Zn(0.39-10.1 mg/kg) > Cu(0.015-1.89 mg/kg) > Mo(0.001-0.039 mg/kg). The results revealed species-dependent differences in micronutrient uptake, with vegetables excluding watermelon and melon (i.e., tomato, eggplant and cucumber) showing higher Cu accumulation. In contrast, watermelon and melon showed higher Mo accumulation. Hierarchical clustering indicated that Cu and Zn were associated with Fe/Mn oxides, whereas Mo was linked to Ca-rich soils. Soil-to-plant transfer factors indicated no excessive bioaccumulation, and dietary exposure assessments confirmed that Cu, Zn, and Mo daily intakes of through fruit and vegetable consumption remained below the EFSA’s upper and adequate intake levels. These findings contribute to a better understanding of soil-plant micronutrient interactions and provide essential data for optimizing agricultural soil fertility management.
1. Introduction
Achieving the United Nations Sustainable Development Goals (SDGs), particularly those related to food security, environmental sustainability and human health, requires development of resilient agricultural systems that ensure both productive soils and safe food chains. 1 Within this framework, the European Green Deal and the associated EU Mission “A Soil Deal for Europe” recognize soil health as a fundamental factor for ecosystem stability and the sustainable production of safe and nutritious food.2,3 These initiatives highlight the necessity of improving scientific understanding of soil processes related to nutrient availability and soil-plant interactions, thereby supporting evidence-based strategies for sustainable land management. In this context, the investigation of soil-to-plant transfer of micronutrients represents a critical area of research for enhancing agricultural productivity and environmental sustainability. 4 Essential trace elements such as copper (Cu), zinc (Zn), and molybdenum (Mo) play important roles in plant physiology, contributing to enzyme activation, photosynthesis, and nitrogen fixation. 5 Although required in small quantities, these mictonutrients are essential for plant growth, metabolic function and overall development.
The balance of these micronutrients is important as deficiencies can impair plant growth and development processes, whereas excessive levels may induce toxicity. The availability and accumulation of Cu, Zn, and Mo in plant tissues are influenced by complex factors, including major soil elements (Fe, Ca, K etc.), pH, soil organic matter (SOM) content, and anthropogenic inputs such as fertilizers. In addition, climatic conditions and irrigation practices can impact these processes as well.6,7 In particular, soil pH influences the solubility and mobility of Cu, Zn, and Mo, with acidic soils often increasing the solubility of Cu and Zn while reducing Mo availability. Mobile anions of Mo can coprecipitate with CaCO3, and Pb2+, Cu2+, Zn2+, Mn2+, and Ca2+ cations. 8 SOM also plays a crucial role by forming complexes with micronutrients, thereby affecting their uptake by plants. 9
The average Cu, Zn, and Mo contents in the upper continental crust are 39 mg/kg, 78, and 1.5 mg/kg, respectively. 10 In the case of soil, Cu ranges from 14-109 mg/kg and Zn: 60-89 mg/kg. The mean Mo content in soil is 1.1 mg/kg. 8 In regions with significant importance for agricultural production, investigations of soil Cu, Zn, and Mo contents and their uptake by plants is particularly significant. In addition, among these elements Mo is an emergin pollutant 11 and compared to Cu and Zn little information is available about its distribution, availability, soil-plant transfer and accumulation characteristics.
These aforementioned issues are particularly relevant in intensively cultivated agricultural lands, such as the area of the Ararat Valley, a significant agricultural area partially situated in the Ararat region (Armenia). Previous regional-scale soil survey of the Ararat region revealed distinct geochemical patterns, identifying both natural and anthropogenic sources of elements such as Cr, V, Ti, As, Zn, Cu, and Pb. 12 Moreover, the first assessment of dietary exposure to lead (Pb) and arsenic (As) revealed that, although local fruits and vegetables do not bio-accumulate these toxic elements, the total dietary intake from multiple sources contributes significantly to tolerable daily doses. 13 Considering the growing concerns over micronutrient imbalances, the limited understanding of Cu, Zn, and Mo uptake by plants, and the influence of local geochemical factors, a deeper investigation is warranted. Therefore, this study aimed to investigate the contents and soil-to-plant transfer of Cu, Zn, and Mo in agricultural soils of the Ararat region (Armenia).
2. Materials and Methods
2.1. Study Area
The study area was part of the Ararat Valley, which is located in the Ararat region of Armenia. This area is characterized by favorable climatic conditions, with hot (24-26 °C) and dry summers and low annual precipitation (200-250 mm), which support the production of high-quality vegetables, fruits, and wine. 14 The geological base of the area consists of alluvial, diluvial, and lacustrine sediments of the Upper Plieocene-Pleistocene on which Anthrosols and Solonetzes-Solomchakes soil types have developed. 15 In this area, there are several towns, including Masis (23.1 thousand inhabitants as of early 2025), Artashat (22.8 thousand), Ararat (17.6 thousand) and Vedi (12.1 thousand). Moreover, some industrial units (Ararat Cement Factory and Ararat Gold Factory) close to the study site have been identified as potential sources of some elements. 12
2.2. Sampling and Sample Preparation
The spatial distribution of the soil and adjacent fruit and vegetable sampling points are shown in Figure 1. Soil sampling was done from the comparatively large agricultural lands of the study area, that exportsits production to the main major markets of the country’s capital city of Yerevan, which has more than 1 milion population. Sampling was carried out following the SOPs (SOPs EGD-01, EDG-10) developed by the Environmental Geochemistry Department of the CENS using international ISO standards,16,17 the US EPA
18
and methodological guidelines.19-22 In total, 23 agricultural soil samples were collected (Figure 1) from the top 20 cm (after removing grass cover if such was presented) using a stainless steel spade. Five subsamples were mixed and the composite sample was placed into plastic bags and transported to the laboratory where the soil samples were air-dried at 20°C, crushed with a ceramic mortar and sieved through a 2 mm mesh. The obtained fraction (<2 mm) was pulverized using a Vibrating grinder VG 6. Soil and plant sampling sites - the spatial distribution map
At each agricultural plot, fresh fruits and vegetables were randomly sampled from the same location as the soil samples according to the procedures described in the ISO 874-1980 standard 23 and Codex Alimentarius Commission guidelines. 24 From each agricultural plot, to 3-5 subsamples were collected and combined to create composite samples for each plant species. The analysis included 8 plant species (apple, cherry, grape, tomato, eggplant, cucumber, watermelon and melon) with 3 composite samples for each species, resulting in 24 plant samples in total. The plant species classification was performed according to fruit and vegetable groups, while taking into consideration their botanical families. Specifically, fruits included apple and cherry (Rosaceae), and grape (Vitaceae), whereas vegetables included tomato and eggplant (Solanaceae), and cucumber, melon and watermelon (Cucurbitaceae).
2.3. Chemical Analysis, Quality Assurance and Control
The contents of Fe, Mn, Ti, Zr, Ca, K, Sr, Rb,
2.4. Data Treatment and Geospatial Mapping
Descriptive Statistics of the Studied Elements (mg/kg) in Soil and Plants
Note. W/M - watermelon and melon.
2.5. The Soil-Plant Transfer Factor (TF) and Dietary Exposure of Trace Elements
The transfer of trace elements (Zn, Cu and Mo) from the soil to the edible parts of each plant species was expressed by the transfer factor (TF), which was calculated using the following equation:
To assess dietary exposure to Zn, Cu and Mo through the consumption of the studied plant species (i.e. fruits and vegetables) by the adult population residing in the study area, the estimated daily intake (EDI) was calculated using the following equation:
In the case of combined consumption of all studied plant species, the total estimated daily intake (∑EDI) was calculated by summing the mean EDI values of each investigated plant species (Equation (3)):
The calculated EDIs were compared with dietary reference values (DRVs) set by the European Food Safety Authority (EFSA). 28
3. Results and Discussions
3.1. Content and Spatial Distribution of the Studied Elements in Soil and Plants
Descriptive statistics of the studied elements in both media are presented in Table 1. Table 1 demonstrates that the skewness values differ from 0 indicating a deviation from the normal distribution. However, the Shapiro-Wilk test showed that Cu and Mo contents in soil display a lognormal distribution whereas in the case of plants, only Mo showed a lognormal distribution. Despite these cases, an abnormal distribution was observed.
The comparison of the average Cu content detected in the agricultural soils of the study areas with the highest mean Cu content (49.26 mg/kg in vineyards), 29 the highest among all land use categories across the European Union (EU), revealed a 1.7-fold excess relative to the EU average. For Zn, a 2.1-fold excess was observed compared with the average Zn content in EU soils. 30 Regarding Mo, literature reports a typical range of 0.8-3.3 mg/kg 31 for agricultural soils, and the average content observed in our study approaches the upper limit of this range (3.3 mg/kg).
A comparatively low coefficient of variation (CV) for Cu, Zn, and Mo was observed in the soil, indicating homogeneous distribution of these elements across the studied agricultural plots. Compared to the soil, the CV of Cu, Zn, and Mo were higher in plants, which can be attributed to species-specific differences in the accumulation of these elements. Variability in the uptake and translocation mechanisms among plant species likely contributes to the observed fluctuations in trace metal concentrations. Moreover, the CV of Zn was 131% suggesting a high level (>100%) of variation in this element in the plants of the study site and their potential anthropogenic origin in soil. 32 A comparison of the mean value of different plant groups showed that the mean value of Cu in vegetables (excluding watermelon/melon) exceeded that in fruit and watermelon/melon (W/M) by 2.5 and 2.2 times, respectively. The mean value of Zn was the highest in W/M exceeding those in fruit and vegetables by 10.3 and 4.8 times, respectively. For Mo, the highest mean values observed in W/M exceeded those in fruits and vegetables by 3.8 and 1.6 times, respectively.
The box-plots (Figure 2) show that all the studied elements displayed outliers and extreme values in both soil and plants. Particularly, a single positive outlier of Mo was observed in cucumber. Song et al (2019) stated that cucumber plants absorb Mo in its ionic form, making it more bioavailable for physiological processes.
33
Research has further indicated that Mo interacts with other micronutrients, particularly iron (Fe), which significantly affects its uptake and distribution within the plant.
34
Boxplots of Cu, Zn, and Mo contents in soil and plants
In the case of Zn, all three extreme values (Figure 2) were detected for watermelon, whereas two outliers and one extreme value of Cu were observed in the tomato samples. Cheng 35 stated that the bioavailability of Zn in watermelons is significantly influenced by its chemical form. Studies have indicated that the red flesh of watermelon contains high levels of bioavailable Zn, with organic acids in watermelon extracts enhancing its solubility and absorption. Furthermore, owing the presence of hydroxyl (cellulose) and carboxyl (pectin) groups, watermelon rind can serve as an effective binding agent for metals. 36 Similarly, Othman et al 37 demonstrated the potential of watermelon rind as a biosorbent for Zn(II) removal from wastewater.
The higher uptake of Cu by tomato plants can be attributed to various physiological and biochemical mechanisms that facilitate its absorption and translocation. This characteristic is particularly relevant in phytoremediation, in which tomato plants are used to mitigate Cu contamination in soils. Tomatoes have demonstrated hyperaccumulator potential for Cu, with the ability to absorb and accumulate the metals in their aerial parts, achieving a removal rate of 87.7%, and demonstrating their effectiveness in Cu phytoremediation. 38
The outliers and extreme values of the studied elements observed in the plants form a spatial cluster situated in the areas close to the sites where cement production and the gold plant are located (Figure 3). These are also areas where comparatively high contents (Figure 3) of the studied elements were observed in soils.
12
Spatial distribution maps of Cu, Zn, and Mo contents in soil and plants
3.2. Soil and Plant Cu, Zn, and Mo Relationship With Other Elements in Soils
Considering the importance of major and minor elements in the fixation and precipitation of Cu, Zn, and Mo in soil environments, Fe, Mn, Ca, K, Ti, Zr, Rb, and Sr were studied to provide a better understanding of the complex geochemical processes that may influence the soil-to-plant transfer of Cu, Zn, and Mo. The relationship between Cu, Zn, and Mo contents in plants with the soil contents of Fe, Mn, Ti, Zr, Ca, K, Sr, Rb, Cu, and Zn were studied by Spearman rank correlation (SM Table 2). In addition, considering the presence of outliers and extreme values (Figure 2) scatterplot matrix was also created to visualize and better understand the peculiarities of the linear relationship (SM Figure 1). The correlation analysis showed that a negative significant correlation was observed only between plants Mo and soil Zn contents (-0.473, p<0.05) and plants Zn and soil K contents (-0.452, p<0.05). The scatterplots showed that there is an absence of distinct relationshipbetween Cu, Zn, and Mo contents in soil and plant. However, subdatasets (subsamples) were identified (Figure 4) indicating about possible linear relationship between these elements in the studied media (i.e., soil and plants). Particlularly, after elimination of some of the samples a linear relationship was observed between soil and plant Mo and Zn contents. Moreover, a significant level of positive correlation was detected (Zn: 0.710 and Mo: 0.738, p<0.05) (Figure 4). This can be explained by peculiarities of local geological bases which is characterized by the volcanic and sedimentary rocks conditioning a site specific influence of Fe and Ca on micronutrients geochemical behavior having a negative significant correlation (-0.653, p<0.01). For the Cu a significant correlation was not observed. Scatterplots of Mo and Zn relationship in plants and soil with full dataset (above) and after elimination (below) of some samples (in red: Subsample selected from the full dataset)
To unveil the link between soil chemical composition and Cu, Zn, and Mo contents in studied plants, hierarchical clustering (Figure 5) of these elements with some major and minor elements in soil (Fe, Mn, Ca, K, Sr, Rb, Ti, and Zr) was performed. Dendrogram of hierarchical cluster analysis
Hierarchical clustering identified two distinct groups at rescaled distance of 15. The first cluster includes Ti, Fe, Zr, Rb, Zn, Cu, Mn, and K. In this group,the element pairs of Ti-Zr, and K-Rb have similar physicochemical properties and are mainly inherited from the parent materials.39,40 In addition, the presence of Fe and Mn in this group highlights the dominance of Fe/Mn oxides/hydroxides in the absorption of Cu and Zn in soils. 8 The second group included soil Ca, Sr and Zn, Mo, and Cu contents in the studied plants. In this group, the pair of elements with similar geochemical behavior is Ca-Sr, because Sr is naturally associated with Ca and has analogous geochemical and biogeochemical properties. 8 Therefore, it can be suggested that the transfer of Zn, Mo, and Cu from soil to plants can be conditioned by the dominant role of carbonates, which can fix and control the availability of these elements to plants. This finding is in line with the local geological basis composed of alluvial, deluvial, and proluvial sediments and other sedimentary rocks (travertines, limestone, clays, etc.) dominating in the study sites. 15 Moreover, some anthropogenic enrichment of carbonatic rock can be observed due to the exploitation of travertines, limestone, clays quarries in this study area.
3.3. Soil to Plant TF and Dietary Intake of Cu, Zn, and Mo
The TF of the studied elements ranges from 0.0001 to 0.026 with a mean of 0.008 for Cu, from 0.002 to 0.117 with a mean of 0.024 for Zn, and from 0.001 to 0.08 with a mean of 0.04 for Mo. Comparatively higher mean TF values for Cu, Zn, and Mo were observed in vegetables (0.011 for eggplant, tomato, and cucumber), while the highest values were recorded for watermelon and melon (0.082 and 0.08 mg/kg, respectively). No bioaccumulation of Cu, Zn or Mo was observed in any of the cases studied. However, a direct relationship between soil and Mo content was identified in watermelon and melon (W/M)the, indicating a potential capacity of these plants to accumulate Mo (Figure 6). Scatterplot of Mo contents in soil and W/M (watermelon and melon)
This pattern is an important consideration for future studies, as in the biogeochemical provinces of Armenia, where naturally high Mo levels occur, watermelon and melon (W/M) may accumulate significant amounts of Mo and therefore represent potential dietary sources of this element. Based on this context, dietary intake of Mo, together with Cu and Zn was estimated and compared with established dietary reference values (DRVs) in order to assess their contribution to diet through the consumption of fruits and vegetables from the study area. The main purpose of this estimation was to highlight that the investigated elements may have both beneficial effects as essential nutrients and adverse effects at excessive exposure levels. Therefore, two types of DRVs were considered for comparison, including adequate intake (AI) from a nutritional perspective and tolerable upper intake level (UL) from a toxicological perspective.
According to the dietary reference values (DRVs) set by the EFSA, the adequate intake (AI) of Mo for adult males and females is 0.065 mg/day. Additionally, EFSA has established a tolerable upper intake level (UL) of 0.6 mg/day for the adult population. The estimated daily intake (EDI) values of Mo from the consumption of the studied fruits and vegetables were significantly below the AI and, consequently, did not exceed the UL. In the case of combined consumption of all the studied plant species, the total estimated daily intake (∑EDI) of Mo was 0.0189 mg/day (SM Table 2), accounting for 29% of the AI. Similar results were obtained for dietary exposure to Cu in comparison with DRVs, including a UL of 5 mg/day for the adult population. When considering the combined consumption of all the studied plant species, the ∑EDI of Cu was 0.8019 mg/day (SM Table 2), accounting for approximately 50.1% of the AI of 1.6 mg/day for males and 61.7% of the AI of 1.3 mg/day for females. Both individual and total estimated daily intake (EDI) values of zinc (Zn) for each plant species and for combined consumption were well below the tolerable upper intake level (UL) of 25 mg/day. Unlike molybdenum (Mo) and copper (Cu), the adequate intake (AI) for Zn, as set by EFSA, varies not only by sex but also based on levels of phytate intake (LPI). Although no data on levels of phytate intake (LPI) are available for the population in this study to precisely determine the appropriate adequate intake (AI) for comparison, all calculated EDI values, including the total estimated daily intake (∑EDI) of 3.3392 mg/day (SM Table 2), remained within the reference values and were relatively higher than those obtained for Mo and Cu.
Overall, the estimated Cu, Zn and Mo intake levels through the consumption of studied agricultural products from the Ararat Valley primarily reflect their contribution to human nutrition, rather than indicating a potential health concern. These findings highlight the role of the studied plant species as important dietary sources of essential micronutrients.
4. Conclusion
This study provides the first insight into Cu, Zn, and Mo transfer from soil-to-plants in an important agricultural area in Armenia. The results showed that the average Cu and Zn contents in the soils of the study area exceeded those found in EU soils by factors of 1.7 (highest mean in vineyard soils) and 2.1 (EU average), respectively. Outliers and extreme values were observed in both soil and plants. Moreover, the outliers and extreme values of Cu, Zn, and Mo observed in plants form a spatial cluster located close to urban areas and industrial units (Cement production, the Ararat gold plant etc.). It was found that while soil-to-plants transfer of the studied elements varies across species, no excessive bioaccumulation was observed. At the same time, a linear relationship between soil and plant Mo and Zn contents were identified. Hierarchical clustering showed that the soil mineral composition (Fe/Mn oxides and Ca minerals) plays an important role in the uptake of the studied elements by plants.
The dietary exposure assessment confirmed that the intake of Cu, Zn, and Mo through locally grown fruits and vegetables by the adult population residing in the study area remained well below the tolerable upper intake levels, while simuntaneously contributing to dietary intake as important sources of essential micronutrients.
Overall, this research contributes to the understanding of the processes and complex interactions between soil chemical composition and the transfer of essential micronutrients to plants and provides a valuable scientific basis for developing sustainable soil management practices in agricultural regions.
Supplemental Material
Supplemental material - Micronutrient (Cu, Zn, Mo) Contents and Soil-To-Plant Transfer in Agricultural Lands of the Ararat Region, Armenia
Supplemental material for Micronutrient (Cu, Zn, Mo) Contents and Soil-To-Plant Transfer in Agricultural Lands of the Ararat Region, Armenia by Pipoyan D., Beglaryan M., Gevorgyan A., Sahakyan L. and , Tepanosyan G. in Environmental Health Insights.
Supplemental Material
Supplemental material - Micronutrient (Cu, Zn, Mo) Contents and Soil-To-Plant Transfer in Agricultural Lands of the Ararat Region, Armenia
Supplemental material for Micronutrient (Cu, Zn, Mo) Contents and Soil-To-Plant Transfer in Agricultural Lands of the Ararat Region, Armenia by Pipoyan D., Beglaryan M., Gevorgyan A., Sahakyan L. and , Tepanosyan G. in Environmental Health Insights.
Footnotes
Acknowledgments
The research was supported by the Higher Education and Science Committee of the MESCS RA (Research project №24FP-4C006).
Author Contributions
Funding
The authors disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This work was supported by the Higher Education and Science Committee of MESCS RA (Research project №24FP-4C006).
Declaration of Conflicting Interests
The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
Data Availability Statement
The datasets used and/or analyzed during the current study are available from the corresponding author at reasonable request.
Supplemental Material
Supplemental material for this article is available online.
References
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