Abundance,contribution, and possible driver of ammonia-oxidizing archaea (AOA) in various types of aquatic ecosystems

Journal of Soils and Sediments - In some habitats, ammonia oxidation highly depends on the activity of ammonia-oxidizing archaea (AOA), which are therefore important for studying biogeochemical...     

Contributions of ammonia-oxidizing archaea and bacteria to nitrification in Oregon forest soils

Ammonia oxidation, the first step of nitrification, is mediated by both ammonia-oxidizing archaea (AOA) and bacteria (AOB); however, the relative contributions of AOA and AOB to soil nitrification are not well understood. In this study we used 1-octyne to discriminate between AOA- and AOB-supported nitrification determined both in soil-water slurries and in unsaturated whole soil at field moisture. Soils were collected from stands of red alder (Alnus rubra Bong.) and Douglas-fir (Pseudotsuga menziesii Mirb. Franco) at three sites (Cascade Head, the H.J. Andrews, and McDonald Forest) on acidic soils (pH 3.9–5.7) in Oregon, USA. The abundances of AOA and AOB were measured using quantitative PCR by targeting the amoA gene, which encodes subunit A of ammonia monooxygenase. Total and AOA-specific (octyne-resistant) nitrification activities in soil slurries were significantly higher at Cascade Head (the most acidic soils, pH < 5) than at either the H.J. Andrews or McDonald Forest, and greater in red alder compared with Douglas-fir soils. The fraction of octyne-resistant nitrification varied among sites (21–74%) and was highest at Cascade Head than at the other two locations. Net nitrification rates of whole soil without NH₄⁺ amendment ranged from 0.4 to 3.3 mg N kg⁻¹ soil d⁻¹. Overall, net nitrification rates of whole soil were stimulated 2- to 8-fold by addition of 140 mg NH₄⁺-N kg⁻¹ soil; this was significant for red alder at Cascade Head and the H.J. Andrews. Red alder at Cascade Head was unique in that the majority of NH₄⁺-stimulated nitrifying activity was octyne-resistant (73%). At all other sites, NH₄⁺-stimulated nitrification was octyne-sensitive (68–90%). The octyne-sensitive activity—presumably AOB—was affected more by soil pH whereas the octyne-resistant (AOA) activity was more strongly related to N availability.     

Prevalence of ammonia-oxidizing bacteria over ammonia-oxidizing archaea in sediments as related to nutrient loading in Chinese aquaculture ponds

Purpose

Nitrogen (N) application in excess of assimilatory capacity for aquaculture ponds can lead to water-quality deterioration through ammonia accumulation with toxicity to fish. Ammonia-oxidizing archaea (AOA) and bacteria (AOB) potentially process extra ammonium, so their abundance and diversity are of great ecological significance. This study aimed to reveal variations in communities of AOA and AOB as affected by aquaculture activities.

Materials and methods

From June to September 2012, water and sediments were sampled monthly in three ponds feeding Mandarin fish in a suburb of Wuhan City, China. Molecular methods based on ammonia monooxygenase (amoA) gene were used to determine abundance and diversity of AOA and AOB in the sediments.

Results and discussion

The pond with the highest fish stock had the highest nutrient loadings in terms of different forms of N and carbon (C) in both sediment and water. The abundance and diversity of AOB were significantly higher than those of AOA in the sediment. The AOB abundance showed a significantly positive relationship to concentration of soluble reactive phosphorus (SRP) in interstitial water, and both abundance and diversity of AOA were significantly negative to concentration of ammonium in interstitial water. Furthermore, AOA species affiliated to Nitrososphaera-like and Nitrosophaera Cluster was distinguishable from those observed in other aquaculture environments.

Conclusions

Nutrients in sediment were enriched by intensive aquaculture activity, among which organic N and C, together with ammonium and SRP, shaped the communities of ammonia oxidizers, with AOB dominating over AOA in terms of abundance and diversity.

     

Microbial contribution to SOM quantity and quality in density fractions of temperate arable soils

The formation of soil organic matter (SOM) very much depends on microbial activity. Even more, latest studies identified microbial necromass itself being a significant source of SOM and found microbial products to initiate and enhance the formation of long-term stabilized SOM. The objectives of this study were to investigate the microbial contribution to SOM in pools of different stability and its impact on SOM quality. Hence, four arable soils of widely differing properties were density-fractionated into free and occluded particulate organic matter (fPOM, oPOM < 1.6 g cm⁻³ and oPOM < 2.0 g cm⁻³) and mineral associated organic matter (MOM > 2.0 g cm⁻³) by using sodium polytungstate. These fractions were characterized by in-source pyrolysis-field ionization mass spectrometry (Py-FIMS). Main SOM compound classes of the fractions were determined and further SOM properties were derived (polydispersity, thermostability). The contribution of microbial derived input to arable soil OM was estimated from the hexose to pentose ratio of the carbohydrates and the ratio of C₄–C₂₆ to C₂₆–C₃₆ fatty acids. Additionally, selected samples were investigated by scanning electron microscopy (SEM) for visualizing structures as indicators for the origin of OM. Results showed that, although the samples differed significantly regarding soil properties, SOM composition was comparable and almost 50% of identifiable SOM compounds of all soils types and all density fractions were assigned to phenols, lignin monomers and alkylaromatics. Most distinguishing were the high contents of carbohydrates for the MOM and of lipids for the POM fractions. Qualitative features such as polydispersity or thermostability were not in general assignable to specific compounds, density fractions or different mean residence times. Only the microbial derived part of the soil carbohydrates could be shown to be correlated with high SOM thermostability (r² = 0.63**, n = 39). Microbial derived carbohydrates and fatty acids were both enriched in the MOM, showing that the relative contribution of microbial versus plant-derived input to arable SOM increased with density and therefore especially increased MOM thermostability. Nevertheless, the general microbial contribution to arable SOM is suggested to be high for all density fractions; a mean proportion of about 1:1 was estimated for carbohydrates. Despite biomolecules released from living microorganisms, SEM revealed that microbial mass (biomass and necromass) is a considerable source for stable SOM which is also increasing with density.     

Preliminary study on the distribution of ammonia oxidizers and their contribution to potential ammonia oxidation in the plant-bed/ditch system of a constructed wetland

Purpose

Ammonia oxidation—as the rate-limiting step of nitrification—has been found to be performed by both ammonia-oxidizing archaea (AOA) and bacteria (AOB). However, how ammonium content and oxidation–reduction status regulate the distribution of ammonia oxidizers in constructed wetlands and their contribution to potential ammonia oxidation rate are still in dispute. This study aimed to explore the effects of ammonium content and oxidation–reduction status on the abundances of AOA/AOB and examine the contributions of AOA and AOB populations to ammonia oxidation rates in the plant-bed/ditch system of a constructed wetland.

Materials and methods

Sampling was carried out in the plant-bed/ditch system of the Shijiuyang Constructed Wetland, China. Three plant-bed soil cores were collected using a soil auger and sampled at depths of 0, 20, and 50 cm in 5-cm increments. Five ditch surface sediments (0–5 cm) were collected along the water flow direction. The abundances of AOA and AOB were investigated by quantitative polymerase chain reaction based on amoA genes. The potential ammonia oxidation rate was determined using the chlorate inhibition method.

Results and discussion

The results showed that AOA outnumbered AOB in the plant-bed surface soil which had lower ammonium content (4.67–7.63 mg kg^?1), but that AOB outnumbered AOA in the ditch surface sediment which had higher ammonium content (14.0–22.9 mg kg^?1). Ammonium content was found to be the crucial factor influencing the relative abundances of AOA and AOB in the surface samples of the plant-bed/ditch system. In the deep layers of the plant bed, AOA abundance outnumbered AOB, though much lower oxidation–reduction potential occurred along the water flow direction. Thus, the oxidation–reduction potential may be another factor influencing the distributions of AOA and AOB in the deep layers of the plant bed without significant difference in ammonium content (p?<?0.05). Moreover, the potential ammonia oxidation rate was significantly dominated by AOB rather than AOA in the plant-bed/ditch system.

Conclusions

The high ammonium content in the ditch sediment likely favored AOB. AOA seemed to persist more readily even under low oxidation–reduction potential in the deep layers of the plant bed. Ammonium content and the oxidation–reduction potential were important parameters influencing the distribution of AOA and AOB in the plant-bed/ditch system of Shijiuyang Constructed Wetland. AOB contributed more to ammonia oxidation than AOA, both in the plant-bed soils (r?=?0.592, p?=?0.0096) and in the ditch sediments (r?=?0.873, p?=?0.0002).     

两种农田土壤中的氨氧化细菌和氨氧化古菌对硝化抑制剂DCD和DMPP的响应-盆栽试验

Taking two important agricultural soils with different pH, brown soil （Hap-Udic Luvisol） and cinnamon soil （Hap-Ustic Luvisol）, from Northeast China, a pot culture experiment with spring maize （Zea mays L.） was conducted to study the dynamic changes in the abundance and diversity of soil ammonia-oxidizing bacteria （AOB） and ammonia-oxidizing archaea （AOA） populations during maize growth period in response to the additions of nitrification inhibitors dicyandiamide （DCD） and 3,4-dimethylpyrazole phosphate （DMPP） by the methods of real-time polymerase chain reaction （PCR） assay, PCR-denaturing gradient gel electrophoresis （DGGE）, and construction of clone library targeting the amoA gene. Four treatments were established, i.e., no urea （control）, urea, urea plus DCD, and urea plus DMPP. Both DCD and DMPP inhibited growth of AOB significantly, compared to applying urea alone. Soil bacterial amoA gene copies had a significant positive linear correlation with soil nitrate content, but soil archaeal amoA gene copies did not. In both soils, all AOB sequences fell within Nitrosospira or Nitrosospira-like groups, and all AOA sequences belonged to group 1.1b crenaxchaea. With the application of DCD or DMPP, community composition of AOB and AOA in the two soils had less change except that the AOB community composition in Hap-Udic Luvisol changed at the last two growth stages of maize under the application of DCD. AOB rather than AOA likely dominated soil ammonia oxidation in these two agricultural soils.     

Seasonal aspects of sciarid emergence in arable land (Diptera: Sciaridae)

In April-October 1990-95 soil sciarids emerging in two Danish barley fields were recorded by means of emergence traps. In 1990-92 the fields were conventionally tilled, in the autumn of 1992 one of the fields was abandoned as annual set-aside (1993) and in 1994-95 most of the field was recropped. In the cropped and uncropped fields 480-527 ind. m⁻² and 369-433 ind. m⁻² emerged, respectively. Ten sciarid species were recorded, eight of which are well-known farmland inhabitants. Apparently, abandonment affected Scatopsciara atomaria negatively, whereas Corynoptera perpusilla was encouraged. The predominant species were probably bivoltine or trivoltine. In Lycoriella castanescens, Scatopsciara atomaria, Corynoptera perpusilla and Bradysia trivittata the female proportion increased significantly during the season. It is assumed that the primary sex ratio of the species is spanandrous. The seasonal change in sex ratio might arise from the unusual modes of reproduction demonstrated in Sciaridae. In Lycoriella castanescens, Bradysia trivittata and Corynoptera perpusilla a significant seasonal increase in wing length was recorded, whereas the wing length of Scatopsciara atomaria and Bradysia rufescens decreased significantly during the season. This suggests a seasonality in the production of large, potentially migrant females. In arable fields sciarid life history traits such as sex ratio, adult biometry and fecundity might be facultative.     

Distinct responses of ammonia-oxidizing bacteria and archaea to green manure combined with reduced chemical fertilizer in a paddy soil

Journal of Soils and Sediments - Ammonia oxidation is the rate-limiting step in nitrification. Ammonia-oxidizing bacteria (AOB) and archaea (AOA) are the key drivers of the nitrogen cycle. Chinese...     

Responses of ammonia-oxidizing bacteria and archaea to nitrogen fertilization and precipitation increment in a typical temperate steppe in Inner Mongolia

As the first and rate-limiting step of nitrification, ammonia oxidation can be realized either by ammonia-oxidizing bacteria (AOB) or archaea (AOA). However, the key factors driving the abundance, community structure and activity of ammonia oxidizers are still unclear, and the relative importance of AOA and AOB in ammonia oxidation is unresolved. In the present study, we examined the effects of long-term (6 years) nitrogen (N) addition and simulated precipitation increment on the abundance and community composition of AOA and AOB based on a field trial in a typical temperate steppe of northern China. We used combined approaches of quantitative PCR, terminal-restriction fragment length polymorphism (T-RFLP) and clone library analyses of amoA genes. The study objective was to determine (1) AOA and AOB diversity and activity in response to N addition and increased precipitation and (2) the relative contributions of AOA and AOB to soil ammonia oxidation in the typical temperate steppe. The results showed that the potential nitrification rate (PNR) increased with N addition, but decreased with increased precipitation. Both N addition and increased precipitation significantly increased AOB but not AOA abundance, and a significant correlation was only observed between PNR and AOB amoA gene copies. The T-RFLP analysis showed that both N and precipitation were key factors in shaping the composition of AOB, while AOA were only marginally influenced. Phylogenetic analysis indicated that all AOA clones fell within the soil and sediment lineage while all AOB clones fell within the Nitrosospira. The study suggested that AOA and AOB had distinct physiological characteristics and ecological niches. AOB were shown to be more sensitive to N and precipitation than AOA, and the ammonia oxidation process was therefore supposed to be mainly driven by AOB in this temperate steppe.     

Use of the RothC model to estimate the carbon sequestration potential of organic matter application in Japanese arable soils

We estimated the carbon (C) sequestration potential of organic matter application in Japanese arable soils at a country scale by applying the Rothamsted carbon (RothC) model at a 1-km resolution. After establishing the baseline soil organic carbon (SOC) content for 1990, a 25-year simulation was run for four management scenarios: A (minimum organic matter application), B (farmyard manure application), C (double cropping for paddy fields) and D (both B and C). The total SOC decreased during the simulation in all four scenarios because the C input in all four scenarios was lower than that required to maintain the baseline 1990 SOC level. Scenario A resulted in the greatest depletion, reflecting the effects of increased organic matter application in the other scenarios. The 25-year difference in SOC accumulation between scenario A and scenarios B, C and D was 32.3, 11.1 and 43.4 Mt C, respectively. The annual SOC accumulation per unit area was similar to a previous estimate, and the 25-year averages were 0.30, 0.10 and 0.41 t C ha⁻¹ year⁻¹ for scenarios B, C and D, respectively. The system we developed in the present study, that is, linking the RothC model and soil spatial data, can be useful for estimating the potential C sequestration resulting from an increase in organic matter input to Japanese arable soils, although more feasible scenarios need to be developed to enable more realistic estimation.     

Use of the RothC model to estimate the carbon sequestration potential of organic matter application in Japanese arable soils

Abstract

We estimated the carbon (C) sequestration potential of organic matter application in Japanese arable soils at a country scale by applying the Rothamsted carbon (RothC) model at a 1-km resolution. After establishing the baseline soil organic carbon (SOC) content for 1990, a 25-year simulation was run for four management scenarios: A (minimum organic matter application), B (farmyard manure application), C (double cropping for paddy fields) and D (both B and C). The total SOC decreased during the simulation in all four scenarios because the C input in all four scenarios was lower than that required to maintain the baseline 1990 SOC level. Scenario A resulted in the greatest depletion, reflecting the effects of increased organic matter application in the other scenarios. The 25-year difference in SOC accumulation between scenario A and scenarios B, C and D was 32.3, 11.1 and 43.4?Mt?C, respectively. The annual SOC accumulation per unit area was similar to a previous estimate, and the 25-year averages were 0.30, 0.10 and 0.41?t?C?ha^?1?year^?1 for scenarios B, C and D, respectively. The system we developed in the present study, that is, linking the RothC model and soil spatial data, can be useful for estimating the potential C sequestration resulting from an increase in organic matter input to Japanese arable soils, although more feasible scenarios need to be developed to enable more realistic estimation.     

Conversion of forest to agricultural land affects the relative contribution of bacteria and fungi to nitrification in humid subtropical soils

Land-use and management practices can affect soil nitrification. However, nitrifying microorganisms responsible for specific nitrification process under different land-use soils remains unknown. Thus, we investigated the relative contribution of bacteria and fungi to specific soil nitrification in different land-use soils (coniferous forest, upland fields planted with corn and rice paddy) in humid subtropical region in China. ¹⁵N dilution technique in combination with selective biomass inhibitors and C₂H₂ inhibition method were used to estimate the relative contribution of bacteria and fungi to heterotrophic nitrification and autotrophic nitrification in the different land-use soils in humid subtropical region. The results showed that autotrophic nitrification was the predominant nitrification process in the two agricultural soils (upland and paddy), while the nitrate production was mainly from heterotrophic nitrification in the acid forest soil. In the upland soils, streptomycin reduced autotrophic nitrification by 94%, whereas cycloheximide had no effect on autotrophic nitrification, indicating that autotrophic nitrification was mainly driven by bacteria. However, the opposite was true in another agricultural soil (paddy), indicating that fungi contributed to the oxidation of NH₄⁺ to NO₃^?. In the acid forest soil, cycloheximide, but not streptomycin, inhibited heterotrophic nitrification, demonstrating that fungi controlled the heterotrophic nitrification. The conversion of forest to agricultural soils resulted in a shift from fungi-dominated heterotrophic nitrification to bacteria- or fungi-dominated autotrophic nitrification. Our results suggest that land-use and management practices, such as the application of N fertilizer and lime, the long-term waterflooding during rice growth, straw return after harvest, and cultivation could markedly influence the relative contribution of bacteria and fungi to specific soil nitrification processes.     

Use of long-term monitoring data to derive a relationship between nitrogen surplus and nitrate leaching for grassland and arable land on well-drained sandy soils in the Netherlands

The decrease in nitrogen (N) use in agriculture led to improvement of upper groundwater quality in the Sand region of the Netherlands in the 1991–2009 period. However, still half of the farms exceeded the European nitrate standard for groundwater of 50 mg/l in the 2008–2011 period. To assure that farms will comply with the quality standard, an empirical model is used to derive environmentally sound N use standards for sandy soils for different crops and soil drainage conditions. Key parameters in this model are the nitrate-N leaching fractions (NLFs) for arable land and grassland on deep, well-drained sandy soils. NLFs quantify the fraction of the N surplus on the soil balance that leaches from the root zone to groundwater and this fraction represents N available for leaching and denitrification. The aim of this study was to develop a method for calculating these NLFs by using data from a random sample of commercial arable farms and dairy farms that were monitored in the 1991–2009 period. Only mean data per farm were available, which blocked a direct derivation of NLFs for unique combinations of crop type, soil type and natural soil drainage conditions. Results showed that N surplus leached almost completely from the root zone of arable land on the most vulnerable soils, that is, deep, well-drained sandy soils (95% confidence interval of NLF 0.80–0.99), while for grassland only half of the N surplus leached from the root zone of grassland (0.39–0.49). The NLF for grassland decreased with 0.015 units/year, which is postulated to be due to a decreased grazing and increased year-round housing of dairy cows. NLFs are positively correlated with precipitation surplus (0.05 units/100 mm for dairy farms and 0.10 units/100 mm for arable farms). Therefore, an increase in precipitation due to climate change may lead to an increase in leaching of nitrate.     

Dispersal power,home range and habitat preference of cantharid larvae (Coleoptera: Cantharidae) in arable land

The dispersal ability, home range size and habitat preference of sixth instar larvae of the widespread cantharid species Cantharis fusca (L.) and C. livida (L.) were studied in a mark–recapture experiment in a meadow–field (winter grain) area between autumn and spring in 1999/2000. The main results and conclusions were: (i) The mean dispersal velocity of C. fusca/C. livida larvae was 1.4/1.6 m d^–1 with a maximum of 3.2/2.3 m d^–1. The larvae were able to disperse more than 100 m during their larval development, demonstrating that larvae and not only adults contribute to spreading. (ii) The average home range area of seven C. fusca individuals was 12.9 m² (minimum 8 m²/maximum 19 m²). The low number of multiple recaptures and the large distances larvae can cover indicate that the real home range size was underestimated. (iii) C. fusca larvae significantly preferred the meadow area compared to the bare ground of the field. This can be explained by the meadow's higher plant cover and humidity C. livida specimens that were released one month later and recaptured only in low numbers showed no such preference. (iv) Due to the high dispersal ability of soldier beetle larvae, immigration from meadows and grass bulks of boundary strips into the crop margins and inner field areas is possible; it can be augmented by creating constant plant cover, e.g. through winter grain or cover crops.     

三种农田土壤中氨氧化细菌amoA基因多样性比较分析

比较分析中国东部不同季风气候区中海伦黑土(HL)、封丘潮土(FQ)和鹰潭红壤(YT)3种土壤氨氧化细菌amoA基因多样性。采用非培养方法直接从土壤中提取微生物总DNA,用氨氧化细菌amoA基因特异引物扩增总DNA,构建了3种土壤amoA基因文库,并对文库进行限制性长度多态性(RFLP)分析。HL、FQ和YT的amoA基因文库克隆数量分别为49、50和48个,相应的RFLP类型数为10、10和14个OTUs,其中有4个OTUs为三种土壤共有;YT中氨氧化细菌amoA基因多样性指数最高,FQ最低;HL和FQ群落的相似为70%,HL与YT的相似度为50%,而FQ和YT之间仅为42%,说明氨氧化细菌具有地理分布的规律:17个amoA基因序列可以被聚成6个cluster,分属Nitrosospira和Nitrosomonas两个属。三种农田土壤中均存在丰富的氨氧化细菌,表明氨氧化细菌在农田土壤氮素循环中具有重要作用。     

Assessing the potential of ammonia oxidizing bacteria to produce nitrous oxide in soils of a high arctic lowland ecosystem on Devon Island, Canada

The contribution of nitrifiers (ammonia-oxidizing bacteria (AOB)) and denitrifiers to nitrous oxide (N₂O) emission from arctic soils remains inconclusive. Based on preliminary experiments, we hypothesized that AOB are the primary producers of N₂O in a high arctic lowland ecosystem on Devon Island, Nunavut, Canada. In part 1 of the study, flux chambers were installed in a catena to determine in situ fluxes of gases (N₂O and carbon dioxide (CO₂)) from 16 June to 13 July 2004. Although fluxes were low, N₂O production occurred in the wettest area of the landscape when ammonium levels were high. As ammonium, but not nitrate, levels declined in the wet sedge meadow, N₂O emissions correspondingly decreased. In part 2, the contribution of nitrification and denitrification to N₂O production was assessed by Acetylene Inhibition Assay and ¹⁵N isotopically enriched incubations. Ammonium fertilization stimulated N₂O emissions to a greater extent than nitrate, and acetylene had a greater impact on N₂O emissions in ammonium-fertilized soils than in nitrate-amended soils. Stable isotope analysis indicated that at 50-55% water filled pore space, nitrification was the dominant (>80%) N₂O emitting process. In part 3, molecular analyses of the two N₂O producing groups indicated the both nitrifiers and denitrifiers did not differ between landforms. Our results suggest nitrifier denitrification is the dominant process occurring in these arctic soils and that the role of denitrifiers in N₂O release from arctic soils needs to be re-evaluated.     

31P-NMR-spectroscopic investigations of NaOH-extracts from soils with different land use in Yucatan (Mexico)

In aqueous NaOH-extracts of top-soils (0-10 cm) from typic chromuderts, phosphorus was determined by NMR-spectroscopy. In addition to inorganic orthophosphate, orthophosphate monoesters and diesters were detected. Pyrophosphate was only found in traces after clear-cutting and burning of the forests. The result of clear-cutting and agricultural land use is an increase of inorganic P from 19% (forest) to 33% (maize field). Compost gardens and mature tree gardens contain only 9-13% inorganic P in the NaOH-extract. The organic P-fraction consists mainly of orthophosphate monoesters (57-72%). The orthophosphate diester content is only 10–21%. It could be shown, that the increase of inorganic P in the course of clear-cutting and ploughing is correlated with a decrease of orthophosphate mono- and diesters.     

Chemical analysis of Teucrium species (Lamiaceae) growing on serpentine soils in Bulgaria

Trace elemental concentrations (Fe, Ni, Mn, Cr, Co, Cu, Zn, Pb, Cd, and As) in aerial biomass (stems, leaves, and flowers) of the medicinal plants Teucrium chamaedrys, T. montanum, and T. polium growing on serpentine soils in Bulgaria were examined in relation to the total element contents in their respective rhizospheric soils. The objectives of the study were to evaluate: (1) elemental concentrations in plant tissues and associated soil samples together with the plants ability to tolerate/accumulate metal concentrations; (2) correlations between total metal concentrations in plants and their rhizospheric soils. Metal concentrations varied across species and sites. Their levels in plant tissues from nonserpentine sites were always lower compared to those from serpentine ones. The highest concentrations for Fe, Ni, Mn, Cr, As, Cu, and Pb were found in T. polium. Positive correlation coefficients were established between Ni, Cr, Fe, Co, and Cu in plants and Ca in the soil. None of the species tested hyperaccumulated metals although the metal concentration in some of them exceeded the range naturally found in plants. The Teucrium species can be considered as “excluders”, containing relatively low metal concentrations in their aerial parts (stems, leaves, and flowers) even in cases of high elemental concentrations in the rhizospheric soil. In all three medicinal plants, metal concentrations for toxic elements were above the permissible limits for pharmaceutical purposes. Therefore, plants found on serpentine bedrock are not recommended to be collected for pharmaceutical purposes.