Cross-resistance mechanisms to ACCase-inhibiting herbicides in short-spike canarygrass (Phalaris brachystachys).

Herbicides that inhibit acetyl-coenzyme A carboxylase (ACCase) are commonly used to control weedy grasses such as short-spike canarygrass (Phalaris brachystachys). Two resistant biotypes of P. brachystachys (R1 and R2) were found in different winter wheat fields in Iran. This study was done to confirm the suspected resistance observed in the field and to elucidate the resistance mechanisms involved. The results indicated that the both resistant biotypes showed cross-resistance to diclofop-methyl (DM), pinoxaden (PN) and cycloxydim (CD) herbicides. Based on the herbicide dose that inhibited 50% of the ACCase activity (I50), the ACCase activity of the resistant biotypes was less sensitive than the S biotype to DM, CD, and PN. No differences in translocation were detected between biotypes; most of the herbicide remained in the treated leaves. The 14C-DM metabolites were identified using thin-layer chromatography. Pre-treatment with the cytochrome P450 inhibitor ABT inhibited 14C-DM metabolism in the R1 biotype, indicating that metabolism is involved in the DM resistance in the R1 biotype. DNA sequencing studies found an Ile-1781-Thr change in both resistant biotypes, conferring cross-resistance to ACCase inhibitors. In general, in the R1 biotype which showed a higher level of resistance than that of the R2 biotype, cross-resistance was observed because of mutation and DM metabolism, while in the R2 biotype, the mutation confers resistance to ACCase-inhibiting herbicides. This is the first reported evidence of the mechanisms responsible for the resistance to ACCase herbicides in P. brachystachys. These results could be useful for improved management of resistant biotypes carrying similar mutations.

Phytochemical screening, total phenolic content and phytotoxic activity of corn (Zea mays) extracts against some indicator species.

Allelopathic effects of corn (Zea mays) extracts was studied, against seed germination and seedling growth of Phalaris minor, Helianthus annuus, Triticumaestivum, Sorghum halepense, Z. mays. Bioassay results showed that aqueous extracts of corn root and shoot, markedly affected seed germination, and other parameters compared with related controls. Preliminary phytochemical screening revealed the presence of various phytochemicals such as tannins, phlobatannins, flavonoids, terpenoids and alkaloids in both roots and shoot aqueous extracts. However, saponins were only present in the shoot aqueous extract, while in shoot ethanol extracts, only terpenoids and alkaloids were detected. Additionally, total polyphenolic (TPC) content in aqueous extracts of corn root and shoot, plus ethanol extracts of corn shoot were determined using an Ultraviolet-visible spectroscopy. Results revealed TPC content of the corn shoot aqueous extract showed the highest yield, compared to other extracts. These findings suggest that phytochemicals present in Z. mays extracts may contribute to allelopathy effect.

Impacts of silicon-based grass defences across trophic levels under both current and future atmospheric CO2 scenarios.

Silicon (Si) has important functional roles in plants, including resistance against herbivores. Environmental change, such as increasing atmospheric concentrations of CO2, may alter allocation to Si defences in grasses, potentially changing the feeding behaviour and performance of herbivores, which may in turn impact on higher trophic groups. Using Si-treated and untreated grasses (Phalaris aquatica) maintained under ambient (400 ppm) and elevated (640 and 800 ppm) CO2 concentrations, we show that Si reduced feeding by crickets (Acheta domesticus), resulting in smaller body mass. This, in turn, reduced predatory behaviour by praying mantids (Tenodera sinensis), which consequently performed worse. Despite elevated CO2 decreasing Si concentrations in P. aquatica, this reduction was not large enough to affect the feeding behaviour of crickets or their predator. Our results suggest that Si-based defences in plants have adverse impacts on both primary and secondary trophic taxa, and these are not likely to decline under future climate change scenarios.

Antifungal and phytotoxic activity of essential oil from root of Senecio amplexicaulis Kunth. (Asteraceae) growing wild in high altitude-Himalayan region.

This work was aimed to evaluate the essential oil from root of medicinally important plant Senecio amplexicaulis for chemical composition, antifungal and phytotoxic activity. The chemical composition analysed by GC/GC-MS showed the presence of monoterpene hydrocarbons in high percentage with marker compounds as α-phellandrene (48.57%), o-cymene (16.80%) and ß-ocimene (7.61%). The essential oil exhibited significant antifungal activity against five phytopathogenic fungi, Sclerotium rolfsii, Macrophomina phaseolina, Rhizoctonia solani, Pythium debaryanum and Fusarium oxysporum. The oil demonstrated remarkable phytotoxic activity in tested concentration and significant reduction in seed germination percentage of Phalaris minor and Triticum aestivum at higher concentrations. The roots essential oil showed high yield for one of its marker compound (α-phellandrene) which makes it important natural source of this compound.

Ile-1781-Leu and Asp-2078-Gly Mutations in ACCase Gene, Endow Cross-resistance to APP, CHD, and PPZ in Phalaris minor from Mexico.

Herbicides that inhibit acetyl coenzyme A carboxylase (ACCase) are commonly used in Mexico to control weedy grasses such as little seed canarygrass (Phalaris minor). These herbicides are classified into three major families (ariloxyphenoxypropionates (APP), cyclohexanodiones (CHD), and, recently, phenylpyrazolines (PPZ)). In this work, the resistance to ACCase (APP, CHD, and PPZ) inhibiting herbicides was studied in a biotype of Phalaris minor (P. minor) from Mexico, by carrying out bioassays at the whole-plant level and investigating the mechanism behind this resistance. Dose-response and ACCase in vitro activity assays showed cross-resistance to all ACCase herbicides used. There was no difference in the absorption, translocation, and metabolism of the (14)C-diclofop-methyl between the R and S biotypes. The PCR generated CT domain fragments of ACCase from the R biotype and an S reference were sequenced and compared. The Ile-1781-Leu and Asp-2078-Gly point mutations were identified. These mutations could explain the loss of affinity for ACCase by the ACCase-inhibing herbicides. This is the first report showing that this substitution confers resistance to APP, CHD, and PPZ herbicides in P. minor from Mexico. The mutations have been described previously only in a few cases; however, this is the first study reporting on a pattern of cross-resistance with these mutations in P. minor. The findings could be useful for better management of resistant biotypes carrying similar mutations.

Interference and economic threshold level of little seed canary grass in wheat under different sowing times.

Little seed canary grass (LCG) is a pernicious weed of wheat crop causing enormous yield losses. Information on the interference and economic threshold (ET) level of LCG is of prime significance to rationalize the use of herbicide for its effective management in wheat fields. The present study was conducted to quantify interference and ET density of LCG in mid-sown (20 November) and late-sown (10 December) wheat. Experiment was triplicated in randomized split-plot design with sowing dates as the main plots and LCG densities (10, 20, 30, and 40 plants m(-2)) as the subplots. Plots with two natural infestations of weeds including and excluding LCG were maintained for comparing its interference in pure stands with designated densities. A season-long weed-free treatment was also run. Results indicated that composite stand of weeds, including LCG, and density of 40 LCG plants m(-2) were more competitive with wheat, especially when crop was sown late in season. Maximum weed dry biomass was attained by composite stand of weeds including LCG followed by 40 LCG plants m(-2) under both sowing dates. Significant variations in wheat growth and yield were observed under the influence of different LCG densities as well as sowing dates. Presence of 40 LCG plants m(-2) reduced wheat yield by 28 and 34% in mid- and late-sown wheat crop, respectively. These losses were much greater than those for infestation of all weeds, excluding LCG. Linear regression model was effective in simulating wheat yield losses over a wide range of LCG densities, and the regression equations showed good fit to observed data. The ET levels of LCG were 6-7 and 2.2-3.3 plants m(-2) in mid- and late-sown wheat crop, respectively. Herbicide should be applied in cases when LCG density exceeds these levels under respective sowing dates.

Essential oil composition, phytotoxic and antifungal activities of Ruta chalepensis L. leaves from High Atlas Mountains (Morocco).

This study aimed at the determination of chemical composition of essential oil obtained by hydrodistillation, and to evaluate their phytotoxic and antifungal activities. Leaves of Ruta chalepensis L. were collected from the region of Tensift Al Haouz (High Atlas Mountains) Marrakech, Morocco. The essential oil (oil yield is 0.56%) was analysed by GC-FID and GC/MS. Twenty-two compounds were identified and accounted for 92.4% of the total oil composition. The major components were undecan-2-one (49.08%), nonan-2-one (33.15%), limonene (4.19%) and decanone (2.71%). Antifungal ability of essential oils was tested by disc agar diffusion against five plant pathogenic fungi: Fusarium proliferatum, Fusarium pseudograminearum, Fusarium culmorum, Fusarium graminearum and Fusarium polyphialidicum. The oils were also tested in vitro for herbicidal activity by determining their influence on the germination and the shoot and root growth of two weed species, Triticum durum and Phalaris canariensis L.

Changes in vitamin A added to a fermented total mixed ration prepared with reed canarygrass (Phalaris arundinacea†L.).

A fermented total mixed ration (TMR) was prepared by adding a vitamin premix containing vitamin A and enzyme to reed canarygrass roughage. The vitamin A levels were then determined after 30 days of fermentation at 20 or 30°C. The vitamin A contents had decreased in roughage fermented at both storage temperatures, and decreased further when an enzyme supplement was included. Because the majority of the added vitamin A was destroyed during fermentation and storage, the addition of vitamin A at the beginning of preparation of fermented TMR is not recommended.

ß-Pinene inhibited germination and early growth involves membrane peroxidation.

ß-Pinene, an oxygenated monoterpene, is abundantly found in the environment and widely occurring in plants as a constituent of essential oils. We investigated the phytotoxicity of ß-pinene against two grassy (Phalaris minor, Echinochloa crus-galli) and one broad-leaved (Cassia occidentalis) weeds in terms of germination and root and shoot growth. ß-Pinene (0.02-0.80 mg/ml) inhibited the germination, root length, and shoot length of test weeds in a dose-response manner. The inhibitory effect of ß-pinene was greater in grassy weeds and on root growth than on shoot growth. ß-Pinene (0.04-0.80 mg/ml) reduced the root length in P. minor, E. crus-galli, and C. occidentalis over that in the control by 58-60, 44-92, and 26-85 %, respectively. In contrast, shoot length was reduced over the control by 45-97 % in P. minor, 48-78 % in E. crus-galli, and 11-75 % in C. occidentalis at similar concentrations. Further, we examined the impact of ß-pinene on membrane integrity in P. minor as one of the possible mechanisms of action. Membrane integrity was evaluated in terms of lipid peroxidation, conjugated diene content, electrolyte leakage, and the activity of lipoxygenases (LOX). ß-Pinene (≥0.04 mg/ml) enhanced electrolyte leakage by 23-80 %, malondialdehyde content by 15-67 %, hydrogen peroxide content by 9-39 %, and lipoxygenases activity by 38-383 % over that in the control. It indicated membrane peroxidation and loss of membrane integrity that could be the primary target of ß-pinene. Even the enhanced (9-62 %) activity of protecting enzymes, peroxidases (POX), was not able to protect the membranes from ß-pinene (0.04-0.20 mg/ml)-induced toxicity. In conclusion, our results show that ß-pinene inhibits root growth of the tested weed species through disruption of membrane integrity as indicated by enhanced peroxidation, electrolyte leakage, and LOX activity despite the upregulation of POX activity.

Mechanism of isoproturon resistance in Phalaris minor: in silico design, synthesis and testing of some novel herbicides for regaining sensitivity.

Isoproturon, 3-p-cumenyl-1 dimethylurea was the only herbicide controlling Phalaris minor, a major weed growing in wheat fields till the early 1980s. Since it has acquired resistance against isoproturon, like other substituted urea herbicides, where the identified target site for isoproturon is in the photosynthetic apparatus at D1 protein of Photosystem-II (PS-II). Nucleotide sequence of susceptible and resistant psbA gene of P. minor has been reported to have four point mutations. During the present work D1 protein of both susceptible and resistant biotypes of P Minor has been modeled. Transmembrane segments of amino acids were predicted by comparing with the nearest homolog of bacterial D1 protein. Volume and area of active site of both susceptible and resistant biotypes has been simulated. Isoproturon was docked at the active site of both, susceptible and resistant D1 proteins. Modeling and simulation of resistance D1 protein indicates that the resistance is due to alteration in secondary structure near the binding site, resulting in loss in cavity area, volume and change in binding position, loss of hydrogen bonds, hydrophobic interaction and complete loss of hydrophobic sites. To regain sensitivity in resistant biotype new derivatives of isoproturon molecules have been proposed, synthesized and tested. Among the 17 derivatives we found that the N-methyl triazole substituted isoproturon is a potential substitute for isoproturon.

Herbicidal activity of culture filtrates of Trichoderma spp. against two problematic weeds of wheat.

The herbicidal potential of culture filtrates of four Trichoderma spp., namely Trichoderma harzianum Rifai, Trichoderma pseudokoningii Rifai, Trichoderma reesei Simmons and Trichoderma viride Pers., was evaluated against two problematic weeds of wheat, Phalaris minor L. and Rumex dentatus L. In laboratory bioassays, generally, metabolites of all four Trichoderma species significantly reduced various root and shoot growth parameters of the two target weed species. The original concentrations of the culture filtrates of all Trichoderma spp., except T. harzianum, significantly reduced various parameters of root and shoot growth of wheat seedlings. In a foliar spray bioassay, the culture filtrates of all four Trichoderma spp. significantly diminished root and shoot biomass of R. dentatus. The effect of these filtrates on the shoot growth of P. minor and wheat was not significant. Culture filtrates of the four Trichoderma species were successively extracted with butanol, n-hexane, chloroform and ethyl acetate. In detached leaf injection bioassays, n-hexane fractions (3 mg mL(-1)) of T. pseudokoningii, T. reesei and T. viride, and ethyl acetate fractions of T. horzianum and T. pseudokoningii were found to be toxic against R. dentatus. This study concludes that the culture filtrates of Trichoderma species have herbicidal potential in the control of R. dentatus.

Herbicidal activity of Withania somnifera against Phalaris minor.

Herbicidal activity of Withania somnifera (L.) Dunal. was studied against Phalaris minor Retz., one of the most problematic weeds of wheat in Pakistan. In laboratory bioassays the aqueous, methanol and n-hexane extracts of 5, 10 and 15% w/v (fresh weight basis) of the roots and shoots of W. somnifera were applied. Extracts in the different solvents exhibited markedly variable herbicidal activities against germination and seedling growth of the target weed species. The methanol extracts showed the highest toxicity. Different concentrations of methanol shoot and root extracts declined the germination of P. minor by 21-71%, its shoot length by 40-72%, its root length by 50-99% and the plant biomass by 32-83%. The aqueous extracts proved to be comparatively less toxic than the methanol extracts, where generally the highest concentration of 15% exhibited pronounced toxicity against the target weed species. There was up to 48, 51, 99 and 55% suppression of the weed's germination, shoot length, root length and plant biomass, respectively, due to the 15% aqueous root and shoot extracts. Generally, the n-hexane extracts of both roots and shoots exhibited insignificant or stimulatory effects against weed shoot length and plant biomass. In a foliar spray bioassay, aqueous and methanol shoot extracts of 10% w/v (dry weight basis) concentration were sprayed on one- and two-week old pot grown P. minor seedlings. Two subsequent sprays were carried out at five day intervals each. The aqueous extract significantly reduced the shoot and root dry biomass of one-week old P. minor plants. In a residue incorporation bioassay, crushed shoots of W. somnifera were incorporated in the soil at 1, 2, ... 5% w/w. Phalaris minor seeds were sown one week after residue incorporation and plants were harvested 45 days after sowing. The lower concentrations of 2 and 3% significantly reduced, while higher concentrations of 4 and 5% of residue incorporation completely arrested, the germination of P. minor. The present study concludes that both roots and shoots of W. somnifera contain herbicidal constituents against P. minor.

Interaction of 8-hydroxyquinoline with soil environment mediates its ecological function.

BACKGROUND: Allelopathic functions of plant-released chemicals are often studied through growth bioassays assuming that these chemicals will directly impact plant growth. This overlooks the role of soil factors in mediating allelopathic activities of chemicals, particularly non-volatiles. Here we examined the allelopathic potential of 8-hydroxyquinoline (HQ), a chemical reported to be exuded from the roots of Centaurea diffusa. METHODOLOGY/PRINCIPAL FINDINGS: Growth bioassays and HQ recovery experiments were performed in HQ-treated soils (non-sterile, sterile, organic matter-enriched and glucose-amended) and untreated control soil. Root growth of either Brassica campestris or Phalaris minor was not affected in HQ-treated non-sterile soil. Soil modifications (organic matter and glucose amendments) could not enhance the recovery of HQ in soil, which further supports the observation that HQ is not likely to be an allelopathic compound. Hydroxyquinoline-treated soil had lower values for the CO(2) release compared to untreated non-sterile soil. Soil sterilization significantly influenced the organic matter content, PO(4)-P and total organic nitrogen levels. CONCLUSION/SIGNIFICANCE: Here, we concluded that evaluation of the effect of a chemical on plant growth is not enough in evaluating the ecological role of a chemical in plant-plant interactions. Interaction of the chemical with soil factors largely determines the impact of HQ on plant growth.

Residue and bio-efficacy evaluation of controlled release formulations of metribuzin against weeds in wheat.

Controlled release formulations of metribuzin in polyvinyl chloride, (emulsion); carboxy methyl cellulose, CMC and carboxy methyl cellulose- kaolinite composite, CMC-KAO are reported. The MET-CMC-KAO-3 (T(9)) formulation provided a superior control (76.1%) of weeds in field grown wheat in comparison to metribuzin 75 DF (57.14%) at the dose (350 g a.i. ha(-1)) after 90 days of sowing. The treatment (T(9)) reduced the dry weight of the weed flora after 30 days of sowing (4.0 g m(-2)) and significantly superior over metribuzin 75 DF (6.0 g m(-2)) and control (17.72 g m(-2)). There were nil to negligible metribuzin residue in soil at harvest of wheat crop and were within prescribed limit of 10 mg L(-1) in drinking water (EPA).

Taking ecological function seriously: soil microbial communities can obviate allelopathic effects of released metabolites.

BACKGROUND: Allelopathy (negative, plant-plant chemical interactions) has been largely studied as an autecological process, often assuming simplistic associations between pairs of isolated species. The growth inhibition of a species in filter paper bioassay enriched with a single chemical is commonly interpreted as evidence of an allelopathic interaction, but for some of these putative examples of allelopathy, the results have not been verifiable in more natural settings with plants growing in soil. METHODOLOGY/PRINCIPAL FINDINGS: On the basis of filter paper bioassay, a recent study established allelopathic effects of m-tyrosine, a component of root exudates of Festuca rubra ssp. commutata. We re-examined the allelopathic effects of m-tyrosine to understand its dynamics in soil environment. Allelopathic potential of m-tyrosine with filter paper and soil (non-sterile or sterile) bioassays was studied using Lactuca sativa, Phalaris minor and Bambusa arundinacea as assay species. Experimental application of m-tyrosine to non-sterile and sterile soil revealed the impact of soil microbial communities in determining the soil concentration of m-tyrosine and growth responses. CONCLUSIONS/SIGNIFICANCE: Here, we show that the allelopathic effects of m-tyrosine, which could be seen in sterilized soil with particular plant species were significantly diminished when non-sterile soil was used, which points to an important role for rhizosphere-specific and bulk soil microbial activity in determining the outcome of this allelopathic interaction. Our data show that the amounts of m-tyrosine required for root growth inhibition were higher than what would normally be found in F. rubra ssp. commutata rhizosphere. We hope that our study will motivate researchers to integrate the role of soil microbial communities in bioassays in allelopathic research so that its importance in plant-plant competitive interactions can be thoroughly evaluated.

How nitrogen and sulphur addition, and a single drought event affect root phosphatase activity in Phalaris arundinacea.

Conservation and restoration of fens and fen meadows often aim to reduce soil nutrients, mainly nitrogen (N) and phosphorus (P). The biogeochemistry of P has received much attention as P-enrichment is expected to negatively impact on species diversity in wetlands. It is known that N, sulphur (S) and hydrological conditions affect the biogeochemistry of P, yet their interactive effects on P-dynamics are largely unknown. Additionally, in Europe, climate change has been predicted to lead to increases in summer drought. We performed a greenhouse experiment to elucidate the interactive effects of N, S and a single drought event on the P-availability for Phalaris arundinacea. Additionally, the response of plant phosphatase activity to these factors was measured over the two year experimental period. In contrast to results from earlier experiments, our treatments hardly affected soil P-availability. This may be explained by the higher pH in our soils, hampering the formation of Fe-P or Fe-Al complexes. Addition of S, however, decreased the plants N:P ratio, indicating an effect of S on the N:P stoichiometry and an effect on the plant's P-demand. Phosphatase activity increased significantly after addition of S, but was not affected by the addition of N or a single drought event. Root phosphatase activity was also positively related to plant tissue N and P concentrations, plant N and P uptake, and plant aboveground biomass, suggesting that the phosphatase enzyme influences P-biogeochemistry. Our results demonstrated that it is difficult to predict the effects of wetland restoration, since the involved mechanisms are not fully understood. Short-term and long-term effects on root phosphatase activity may differ considerably. Additionally, the addition of S can lead to unexpected effects on the biogeochemistry of P. Our results showed that natural resource managers should be careful when restoring degraded fens or preventing desiccation of fen ecosystems.

Phytotoxic withanolides from Jaborosa rotacea.

Twelve new withanolides were isolated from the aerial part of Jaborosa rotacea: five had a spiranoid delta-lactone (1-5); one contained a 26,12-delta-lactone and a C-12-C-23 bond (6); five corresponded to trechonolide-type withanolides with configuration at C-23 opposite of those previously isolated (7, 8, 10-12); two of these have an additional oxido-bridge between C-21 and C-24; finally a withanolide with a hemiketal ring formed between a 21-hydroxyl and a 12-ketone (13) and the closely related jaborosalactone R were also isolated. New compounds were fully characterized by a combination of spectroscopic methods (1D and 2D NMR and MS). The structures of the spiranoid withanolide and of the epimer of trechonolide A were confirmed by X-ray diffraction studies. Compounds 4, 5, 6, and 8 showed selective phytotoxicity toward monocotyledoneous and dicotyledoneous species.

Induction of glutathione S-transferase and glutathione by toxic compounds and elicitors in reed canary grass.

Treatment of read canary grass leaves with phenol, 4-chlorophenol, naphthalic anhydride and phenylethylisothiocyanate increased glutathione S-transferase activity by 1.4-2.4-fold (control 17 U g(-1) DW). Benzothiadiazole, beta-aminobutyric acid and salicylic acid increased activity by 1.3-1.8-fold. Total glutathione pool was increased by the toxic compounds by 1.2-2-fold and by the elicitors 1.4-1.6-fold (control 593 nmol g(-1) DW). Unlike the other compounds, benzothiadiazole and salicylic acid did not decrease the redox state. Benzothiadiazole acted synergistically with chlorophenol on glutathione S-transferase and glutathione levels and counteracted the decrease in redox state caused by the xenobiotic. Reed canary grass thus has a strong potential to neutralize toxic compounds, which may be further enhanced by elicitors.

Melatonin acts as a growth-stimulating compound in some monocot species.

In a recent study melatonin (N-acetyl-5-methoxytryptamine), a well-investigated animal molecule but minimally studied in plants, was seen to have a physiological role as growth-promoting molecule in lupin hypocotyls. In the present study, the role of melatonin as a growth promoter is extended to coleoptiles of canary grass, wheat, barley and oat, in which it shows a relative auxinic activity [with respect to indole-3-acetic acid (IAA), the main auxin in plants] of between 10 and 55%. In addition, melatonin is seen to have an important inhibitory growth effect on roots similar to that played by auxin. The quantitation by liquid chromatography with electrochemical detection and identification by tandem mass spectrometry of melatonin and IAA in etiolated coleoptiles of the monocots assayed showed that both compounds are present in similar levels in these tissues. These results point to the co-existence of auxin and melatonin in tissues and raises the possibility of their co-participation in some physiological actions as auxinic hormones in plants.

Screening the wetland plant species Alisma plantago-aquatica, Carex rostrata and Phalaris arundinacea for innate tolerance to zinc and comparison with Eriophorum angustifolium and Festuca rubra Merlin.

Several wetland plant species appear to have constitutive metal tolerance. In previous studies, populations from contaminated and non-contaminated sites of the wetland plants Typha latifolia, Phragmites australis, Glyceria fluitans and Eriophorum angustifolium were found to be tolerant to high concentrations of metals. This study screened three other species of wetland plants: Alisma plantago-aquatica, Carex rostrata and Phalaris arundinacea for innate tolerance to zinc. The degree of tolerance was compared to known zinc-tolerant E. angustifolium and Festuca rubra Merlin. It was found that A. plantago-aquatica and P. arundinacea did not posses innate tolerance to zinc, but that C. rostrata was able to tolerate elevated levels of zinc, at levels comparable to those tolerated by E. angustifolium and F. rubra Merlin. The findings support the theory that some wetland angiosperm species tend to be tolerant to exposure to high levels of metals, regardless of their origin.