Connecting genes to whole plants in dilution effect of target-site ALS inhibitor resistance of Schoenoplectiella juncoides (Roxb.) Lye (Cyperaceae).

This study focuses on dilution effect of target-site resistance (TSR) to acetolactate synthase (ALS) inhibitors in Schoenoplectiella juncoides, which harbors two ALS genes, ALS1 and ALS2. We assessed gene expression, enzyme activity, and whole-plant resistance profiles across four S. juncoides lines: the susceptible line, the parental resistant lines with a homozygous mutation in either ALS1 or ALS2, and the bred progeny line with homozygous mutations in both ALS1 and ALS2. Gene expression and enzyme function showed a proportional relationship that the expression ratios of ALS1 to ALS2, approximately 70:30, were consistent with the functional ratio predicted by the double-sigmoidal plateau positions observed in enzyme assays. However, at the whole-plant level, resistance did not correlate to the putative abundance of susceptible enzyme, but the parental lines showed similar resistance to each other despite different enzyme-level resistances. This suggests a non-proportional mechanism in the reflection of physiological enzymatic profiles to whole-plant resistance profiles. These findings highlight the complexity of herbicide resistance and the need for further research to understand the mechanisms that influence resistance outcomes. Understanding these relationships is essential for developing strategies to manage herbicide resistance effectively.

Effect of plant-growth-promoting rhizobacteria on phytoremediation efficiency of Scirpus triqueter in pyrene-Ni co-contaminated soils.

The aim of this study was to investigate whether the plant-growth-promoting rhizobacteria (PGPR) could enhance phytoremediation efficiency of Scirpus triqueter (S.triqueter) in the pyrene-Ni co-contaminated soil. We also expected to reveal the possible mechanism for the affected phytoremediation efficiency induced by PGPR. We used three kinds of contaminated soils (Ni-contaminated soil, pyrene-contaminated soil and pyrene-Ni co-contaminated soil) to conduct this pot study. After harvest, plants growth indicators, polyphenol oxidase (PPO) activity and soil microbial community structure of each treatment were investigated to explain the different dissipation rates of pyrene and removal rates of Ni between treatments with and without PGPR. The results showed that PGPR-inoculated S. triqueter increased dissipation rates of pyrene and removal rates of Ni in all three contaminated soils, among which Ni removal rates in Ni single contaminated soil was elevated most significantly, from 0.895‰ to 8.8‰, increasing nearly 9 folds. However, Ni removal rate efficiency in co-contaminated soil was weakened because more toxic and complicated co-contaminated soil restrained plant growth and Ni absorption. We also observed that co-contamination harmed the soil microbial community more severely than that in single pyrene or Ni contaminated soil through phospholipid fatty acids analysis. Furthermore, dissipation rates of pyrene and removal rates of Ni were found positively correlated to the PPO activity and the abundance of branched and saturated fatty acids reflected by Pearson correlation analysis.

A high-throughput, modified ALS activity assay for Cyperus difformis and Schoenoplectus mucronatus seedlings.

Cyperus difformis L. (CYPDI) and Schoenoplectus mucronatus (L.) Palla (SCHMU) are major weeds of California (CA) rice, where resistance to acetolactate synthase (ALS)-inhibitors was identified in several CYPDI and SCHMU populations that have also evolved resistance to photosystem II (PSII)-inhibiting herbicides. The mechanism of ALS resistance in these populations remains to be clarified but this information is crucial in a weed management program, especially in a scenario where resistance to multiple herbicides has been identified. ALS activity assays are commonly used to diagnose resistance to ALS-inhibitors, but protocols currently available are burdensome for the study of CYPDI and SCHMU, as they require large amounts of plant material from young seedlings and have low yields. Our objective was to investigate the ALS resistance mechanism in suspected ALS-resistant (R) CYPDI and SCHMU biotypes using a modified ALS activity assay that requires less plant material. ALS enzymes from suspected R biotypes were at least 10,000-fold less sensitive to bensulfuron-methyl than susceptible (S) cohorts, indicating ALS resistance that is likely due to an altered target-site. Protein concentration (mgg-1 tissue) did not differ between R and S biotypes within each species, suggesting that R biotypes do not over produce ALS enzymes. CYPDI biotypes had up to 4-fold more protein per mg of tissue than SCHMU biotypes, but up to 7-fold more acetoin per mg-1 protein was quantified in SCHMU, suggesting greater ALS catalytic ability in SCHMU biotypes, regardless of their herbicide resistance status. Our optimized protocol to measure ALS activity allowed for up to a 3-fold increase in the number of assays performed per g of leaf tissue. The modified assay may be useful for measuring ALS activity in other weed species that also produce small amount of foliage in early growth stages when protein in tissue is most abundant.

Different responses of alpine plants to nitrogen addition: effects on plant-plant interactions.

The different responses of plant species to resource stress are keys to understand the dynamics of plant community in a changing environment. To test the hypothesis that nitrogen (N) increase would benefit N competitive species, rather than N stress-tolerant species, to compete with neighbours, we conducted an experiment with neighbour removal, N addition and soil moisture as treatments in an alpine grassland on the southeastern Tibetan Plateau. Both growths and competitive-response abilities (CRA, the ability to tolerate the inhibitory effects of neighbors) of Kobresia macrantha, Polygonum viviparum and Potentilla anserine in wet site were facilitated by N addition, conversely, both growths and CRA of Taraxacum mongolicum and Ligularia virgaurea were suppressed by N addition, indicating that the responses of CRA of target species under N addition were consistent with the N utilization strategies of them. Moreover, the facilitative effects of N addition on competitive-response abilities of Kobresia macrantha and Polygonum viviparum were not found at the dry site, illustrating that soil moisture can alter the changes of neighbour effects caused by N addition. Life strategy of dominant species in plant community on the undisturbed southeastern Tibetan Plateau may shift from N stress-tolerant to N competitive, if the N increases continuously.

The inhibition and adaptability of four wetland plant species to high concentration of ammonia wastewater and nitrogen removal efficiency in constructed wetlands.

Four plant species, Typha orientalis, Scirpus validus, Canna indica and Iris tectorum were selected to assess their physiological response and effects on nitrogen and COD removal to high total ammoniacal nitrogen (TAN) in constructed wetlands. Results showed that high TAN caused decreased relative growth rate, net photosynthetic rate, and leaf transpiration. C. indica and T. orientalis showed higher TAN adaptability than S. validus and I. tectorum. Below TAN of 200 mg L(-1), growth of C. indica and T. orientalis was less affected or even stimulated at TAN range 100-200 mg L(-1). However, S. validus and I. tectorum was obviously suppressed when TAN was above 100 mg L(-1). High TAN generated obvious oxidative stress showing increased proline and malondialdehyde contents, and superoxide dismutase was inhibited. It indicated that the threshold for plant self-bioremediation against high TAN was 200 mg L(-1). What's more, planted CWs showed higher nitrogen and COD removal. Removal rate of C. indica and T. orientalis was higher than S. validus and I. tectorum.

Plant Functional Diversity Can Be Independent of Species Diversity: Observations Based on the Impact of 4-Yrs of Nitrogen and Phosphorus Additions in an Alpine Meadow.

Past studies have widely documented the decrease in species diversity in response to addition of nutrients, however functional diversity is often independent from species diversity. In this study, we conducted a field experiment to examine the effect of nitrogen and phosphorus fertilization ((NH4)2 HPO4) at 0, 15, 30 and 60 g m-2 yr-1 (F0, F15, F30 and F60) after 4 years of continuous fertilization on functional diversity and species diversity, and its relationship with productivity in an alpine meadow community on the Tibetan Plateau. To this purpose, three community-weighted mean trait values (specific leaf area, SLA; mature plant height, MPH; and seed size, SS) for 30 common species in each fertilization level were determined; three components of functional diversity (functional richness, FRic; functional evenness, FEve; and Rao's index of quadratic entropy, FRao) were quantified. Our results showed that: (i) species diversity sharply decreased, but functional diversity remained stable with fertilization; (ii) community-weighted mean traits (SLA and MPH) had a significant increase along the fertilization level; (iii) aboveground biomass was not correlated with functional diversity, but it was significantly correlated with species diversity and MPH. Our results suggest that decreases in species diversity due to fertilization do not result in corresponding changes in functional diversity. Functional identity of species may be more important than functional diversity in influencing aboveground productivity in this alpine meadow community, and our results also support the mass ratio hypothesis; that is, the traits of the dominant species influenced the community biomass production.

Phytotoxicity and bioaccumulation of ZnO nanoparticles in Schoenoplectus tabernaemontani.

The rapid development of nanotechnology will inevitably result in an increasing release of engineered nanoparticles (NPs) to wastewaters. In this study we investigated the fate and toxicity of ZnO NPs in aquatic plant mesocosms, as well as the potential for root accumulation and root-to-shoot translocation of these Zn NPs in the wetland plant Schoenoplectus tabernaemontani exposed to ZnO NPs. The growth of S. tabernaemontani in these hydroponic mesocosms was significantly inhibited by ZnO NPs (1000 mg L(-1)) compared to a control. Levels of Zn in the plant roots for the ZnO NP treatment ranged from 402 to 36513 µg g(-1), while values ranged from 256 to 9429 µg g(-)(1) (dry weight) for Zn(2+) treatment, implying that the uptake of Zn from ZnO NPs was substantially greater than that for Zn(2+). The root uptake (of the initial mass of Zn in the solution) for ZnO NP treatment ranged from 8.6% to 43.5%, while for Zn(2+) treatment they were 1.66% to 17.44%. The low values of the translocation factor for both ZnO NP (0.001-0.05) and Zn(2+) (0.05-0.27) treatments implied that the potential for translocation of Zn NPs from roots to shoots was limited. ZnO NP distribution in the root tissues of S. tabernaemontani was confirmed by scanning electron microscopy (SEM). Transmission electron microscopy (TEM) demonstrated that ZnO NPs could pass through plant cell walls, and were present within the plant cells of S. tabernaemontani.

Ammonium tolerance and toxicity of Actinoscirpus grossus--a candidate species for use in tropical constructed wetland systems.

Actinoscirpus grossus, a native species in tropical wetlands of South-East Asia, North Australia and the Pacific islands, has been reported to perform well in experimental scale constructed wetland (CW) systems. However, little is known about how high NH4(+) concentrations prevailing in wastewater affect growth and performance of this species. We examined growth, morphological and physiological responses of A. grossus to NH4(+) concentrations of 0.5, 2.5, 5, 10 and 15mM under hydroponic growth conditions. The relative growth rates (RGR) of the plants were highest at 2.5mM NH4(+) but significantly reduced at 10 and 15mM NH4(+). The roots of the plants were stunted and produced subepidermal lignified-cell layers at exposure to 10 and 15mM NH4(+). The photosynthetic rates did not differ between treatments (average An=21.3±0.4µmolCO2m(-2)s(-1)) but the photosynthetic nitrogen and carbon use efficiency (PNUE and PCUE) were significantly depressed at 10 and 15mM NH4(+) treatments. The concentration of NH4(+) in the roots, but not in the leaves, reflected the NH4(+) concentration in the growth medium suggesting that the species is unable to regulate the NH4(+) uptake. The high root respiration rates in concert with high tissue NH4(+) and declined C/N ratio at 10 and 15mM NH4(+) suggest that the NH4(+) assimilation occurs primarily in the roots and the plant has inadequate C-skeletons for NH4(+) assimilation and exudation at high NH4(+) concentration in the external solution. The concentrations of mineral cations were generally reduced and the root membrane permeability increased at high external NH4(+) concentrations. Our study shows that A. grossus tolerates NH4(+) concentrations up to 5mM which is characteristic of most types of wastewater. Hence, A. grossus is a good native candidate species for use in CW systems in tropical and subtropical climates in South-East Asia, North Australia and the Pacific islands.

Remotely-sensed indicators of N-related biomass allocation in Schoenoplectus acutus.

Coastal marshes depend on belowground biomass of roots and rhizomes to contribute to peat and soil organic carbon, accrete soil and alleviate flooding as sea level rises. For nutrient-limited plants, eutrophication has either reduced or stimulated belowground biomass depending on plant biomass allocation response to fertilization. Within a freshwater wetland impoundment receiving minimal sediments, we used experimental plots to explore growth models for a common freshwater macrophyte, Schoenoplectus acutus. We used N-addition and control plots (4 each) to test whether remotely sensed vegetation indices could predict leaf N concentration, root:shoot ratios and belowground biomass of S. acutus. Following 5 months of summer growth, we harvested whole plants, measured leaf N and total plant biomass of all above and belowground vegetation. Prior to harvest, we simulated measurement of plant spectral reflectance over 164 hyperspectral Hyperion satellite bands (350-2500 nm) with a portable spectroradiometer. N-addition did not alter whole plant, but reduced belowground biomass 36% and increased aboveground biomass 71%. We correlated leaf N concentration with known N-related spectral regions using all possible normalized difference (ND), simple band ratio (SR) and first order derivative ND (FDN) and SR (FDS) vegetation indices. FDN(1235, 549) was most strongly correlated with leaf N concentration and also was related to belowground biomass, the first demonstration of spectral indices and belowground biomass relationships. While S. acutus exhibited balanced growth (reduced root:shoot ratio with respect to nutrient addition), our methods also might relate N-enrichment to biomass point estimates for plants with isometric root growth. For isometric growth, foliar N indices will scale equivalently with above and belowground biomass. Leaf N vegetation indices should aid in scaling-up field estimates of biomass and assist regional monitoring.

Effects of lead on anatomy, ultrastructure and concentration of nutrients in plants Oxycaryum cubense (Poep. & Kunth) Palla: a species with phytoremediator potential in contaminated watersheds.

Lead (Pb) has been highlighted as a major pollutant of both terrestrial and aquatic ecosystems, causing negative impacts to these environments. The concentration of Pb in plants has increased in recent decades, mainly due to anthropogenic activities. This study has as a hypothesis that the species Oxycaryum cubense (Poep. & Kunth) Palla, abundant in aquatic environments, has the potential to be used a phytoremediator. The plants were grown in a hydroponic system with Pb in increasing concentrations (0, 4, 8, 16 and 32 mg l(-1)) for 15 days. Inductively coupled mass spectrometer (ICP OES) was used to determine the concentration of mineral nutrients and lead. Optical and transmission electron microscopy were used for the analysis of cellular damage induced by lead in roots and leaves. Ultrastructural alterations were observed as disorganization of thylakoids in the chloroplast and disruption of mitochondrial membranes in cells of leaf tissues of plants subjected to increasing Pb concentrations. There was accumulation of Pb, especially in the root system, affecting the absorption and translocation of some mineral nutrients analysed. In roots, there was reduction in the thickness of the epidermis in plants treated with Pb. This species was shown to be tolerant to the Pb concentrations evaluated, compartmentalizing and accumulating Pb mainly in roots. Due to these results, it may be considered a species with phytoremediation capacity for Pb, with potential rizofiltration of this metallic element in contaminated watersheds.

Effects of sub-lethal glyphosate concentrations on growth and photosynthetic performance of non-target species Bolboschoenus maritimus.

Glyphosate use has increased over the last decades for the control of invasive plant species in wetland ecosystems. Although glyphosate has been considered 'environmentally' safe, its repeated use could increase the toxicological risk derived from diffuse pollution of surface and groundwater on non-target vegetation. A glasshouse study was designed to determine the effect produced by the addition of different sub-lethal doses of glyphosate herbicides (5-30 mg L(-1)) to the nutrient solution on the growth and photosynthetic apparatus of Bolboschoenus maritimus. Although B. maritimus plants were able to grow and survive after 20 d of exposure to glyphosate, the presence of this herbicide affected their growth, through a direct interaction with the root system. Particularly, at 30 mg L(-1) glyphosate, B. maritimus showed ca. 30% of biomass decrease. The reduction in B. maritimus growth was due to a decrease in net photosynthetic rate (A), which ranged between values ca. 11.5 and 5.5 µmol m(-2)s(-1) CO2 for the control and the highest glyphosate treatment, respectively. The response of A to glyphosate could be largely accounted for by non-stomatal limitations, since stomatal conductance was similar in all glyphosate treatments. Thus, A decrease was prompted by the negative impact of herbicide on photochemical (PSII) apparatus, the reduction in the absorption of essential nutrients, the reduction of photosynthetic pigments and possibly the reduction in Rubisco carboxilation capacity. Moreover, glyphosate excess caused photoinhibitory damage. In conclusion, in this study we have shown that herbicide water pollution could be a source of indirect phytotoxicity for B. maritimus.

Phytotoxicity of wastewater containing lead (Pb) effects Scirpus grossus.

Phytoremediation is an environment-friendly and cost-effective method to clean the environment of heavy metal contamination. A prolonged phytotoxicity test was conducted in a single exposure. Scirpus grossus plants were grown in sand to which the diluted Pb (NO3)2 was added, with the variation of concentration were 0, 100, 200, 400, 600, and 800 mg/L. It was found that Scirpus grossus plants can tolerate Pb at concentrations of up to 400 mg/L. The withering was observed on day-7 for Pb concentrations of 400 mg/L and above. 100% of the plants withered with a Pb concentration of 600 mg/L on day 65. The Pb concentration in water medium decreased while in plant tissues increased. Adsorption of Pb solution ranged between 2 to 6% for concentrations of 100 to 800 mg/L. The Bioaccumulation Coefficient and Translocation Factor of Scirpus grossus were found greater than 1, indicating that this species is a hyperaccumulator plant.

Tidal marsh plant responses to elevated CO2 , nitrogen fertilization, and sea level rise.

Elevated CO2 and nitrogen (N) addition directly affect plant productivity and the mechanisms that allow tidal marshes to maintain a constant elevation relative to sea level, but it remains unknown how these global change drivers modify marsh plant response to sea level rise. Here we manipulated factorial combinations of CO2 concentration (two levels), N availability (two levels) and relative sea level (six levels) using in situ mesocosms containing a tidal marsh community composed of a sedge, Schoenoplectus americanus, and a grass, Spartina patens. Our objective is to determine, if elevated CO2 and N alter the growth and persistence of these plants in coastal ecosystems facing rising sea levels. After two growing seasons, we found that N addition enhanced plant growth particularly at sea levels where plants were most stressed by flooding (114% stimulation in the + 10 cm treatment), and N effects were generally larger in combination with elevated CO2 (288% stimulation). N fertilization shifted the optimal productivity of S. patens to a higher sea level, but did not confer S. patens an enhanced ability to tolerate sea level rise. S. americanus responded strongly to N only in the higher sea level treatments that excluded S. patens. Interestingly, addition of N, which has been suggested to accelerate marsh loss, may afford some marsh plants, such as the widespread sedge, S. americanus, the enhanced ability to tolerate inundation. However, if chronic N pollution reduces the availability of propagules of S. americanus or other flood-tolerant species on the landscape scale, this shift in species dominance could render tidal marshes more susceptible to marsh collapse.

A phytotoxicity test of bulrush (Scirpus grossus) grown with diesel contamination in a free-flow reed bed system.

In this study, bulrush (Scirpus grossus) was subjected to a 72 day phytotoxicity test to assess its ability to phytoremediate diesel contamination in simulated wastewater at different concentrations (0, 8700, 17,400 and 26,100mg/L). Diesel degradation by S. grossus was measured in terms of total petroleum hydrocarbon (TPH-D). The TPH-D concentration in the synthetic wastewater was determined with the liquid-liquid extraction method and gas chromatography. S. grossus was found to reduce TPH-D by 70.0 and 80.2% for concentrations of 8700 mg/L and 17,400mg/L, respectively. At a diesel concentration of 26,100mg/L, S. grossus died after 14 days. Additionally, the biomass of S. grossus plants was found to increase throughout the phytotoxicity test, confirming the ability of the plant to survive in water contaminated with diesel at rates of less than 17,400mg/L.

Changes in pH and organic acids in mucilage of Eriophorum angustifolium roots after exposure to elevated concentrations of toxic elements.

The presence of Eriophorum angustifolium in mine tailings of pyrite maintains a neutral pH, despite weathering, thus lowering the release of toxic elements into acid mine drainage water. We investigated if the presence of slightly elevated levels of free toxic elements triggers the plant rhizosphere to change the pH towards neutral by increasing organic acid contents. Plants were treated with a combination of As, Pb, Cu, Cd, and Zn at different concentrations in nutrient medium and in soil in a rhizobox-like system for 48-120 h. The pH and organic acids were detected in the mucilage dissolved from root surface, reflecting the rhizospheric solution. Also the pH of root-cell apoplasm was investigated. Both apoplasmic and mucilage pH increased and the concentrations of organic acids enhanced in the mucilage with slightly elevated levels of toxic elements. When organic acids concentration was high, also the pH was high. Thus, efflux of organic acids from the roots of E. angustifolium may induce rhizosphere basification.

Characterization of sulfonylurea-resistant Schoenoplectus juncoides having a target-site Asp(376)Glu mutation in the acetolactate synthase.

Schoenoplectus juncoides, a noxious weed for paddy rice, is known to become resistant to sulfonylurea (SU) herbicides by a target-site mutation in either of the two acetolactate synthase (ALS) genes (ALS1 and ALS2). SU-resistant S. juncoides plants having an Asp376Glu mutation in ALS2 were found from a paddy rice field in Japan, but their resistance profile has not been quantitatively investigated. In this study, dose-response of the SU-resistant accession was compared with that of a SU-susceptible accession at in vivo whole-plant level as well as at in vitro enzymatic level. In whole-plant tests, resistance factors (RFs) based on 50% growth reduction (GR50) for imazosulfuron (ISF), bensulfuron-methyl (BSM), metsulfuron-methyl (MSM), bispyribac-sodium (BPS), and imazaquin (IMQ) were 176, 40, 14, 5.2 and 1.5, respectively. Thus, the accession having an Asp376Glu mutation in ALS2 was highly resistant to the three SU herbicides and moderately resistant to BPS, but was not substantially resistant to IMQ. This is slightly different from the earlier results reported from other weeds with an Asp376Glu mutation, in which the mutation confers resistance to broadly all the chemical classes of ALS-inhibiting herbicides. In enzymatic tests, ALS2 of S. juncoides was expressed in E. coli; the resultant ALS2 was subjected to an in vitro assay. RFs of the mutated ALS2 based on 50% enzymatic inhibition (I50) for ISF, BSM, MSM, BPS, and IMQ were 3699, 2438, 322, 80, and 4.8, respectively. The RFs of ALS2 were highly correlated with those of the whole-plant; this suggests that the Asp376Glu mutation in ALS2 is a molecular basis for the whole-plant resistance. The presence of two ALS genes in S. juncoides can at least partially explain why the whole-plant RFs were less than those of the expressed ALS2 enzymes.

Response characteristics of Scirpus trioueter and its rhizosphere to pyrene contaminated soils at different growth stages.

Scirpus triqueter (Triangular club-rush), a typical wetland species, is used to study the response characteristics to pyrene. A pot experiment was conducted to investigate the growth parameters (height, diameter, shoot number, total volume, underground biomass, above-ground biomass and total biomass), and enzymes (catalase and superoxide dismutase) of S. triqueter. The characteristics of soil enzymes (catalase and polyphenol oxidase) and microorganisms (bacteria and fungi) were also assessed after pyrene treatment. Elevated pyrene concentration (80 mgkg(-1)) in the soil reduced the shoot number and biomass significantly, especially at the early growth stage. In root tissue, the enzyme catalase was activated at 80 mgkg(-1) of pyrene. Compared to roots, shoots had higher enzyme activities. Catalase activities in the rhizosphere increased throughout the growth period of S. triqueter. Polyphenol oxidase activities in the rhizosphere were higher than those in the bulk soil and unplanted soil. The populations of bacteria (total bacteria, pyrene-tolerant bacteria, and actinomyces) and fungi decreased under the stress of high pyrene concentration, while that of pyrene-tolerant bacteria increased with the increasing pyrene concentration. The presence of pyrene did not benefit the growth of S. triqueter. S. triqueter and soil enzymes varied within the growth stages. The presence of S. triqueter could improve the activity of soil enzymes and facilitate the propagation of microorganisms which could help eliminate pyrene contamination.

Direct and indirect effects of ammonia, ammonium and nitrate on phosphatase activity and carbon fluxes from decomposing litter in peatland.

Here we investigate the response of soils and litter to 5 years of experimental additions of ammonium (NH4), nitrate (NO3), and ammonia (NH3) to an ombrotrophic peatland. We test the importance of direct (via soil) and indirect (via litter) effects on phosphatase activity and efflux of CO2. We also determined how species representing different functional types responded to the nitrogen treatments. Our results demonstrate that additions of NO3, NH4 and NH3 all stimulated phosphatase activity but the effects were dependent on species of litter and mechanism (direct or indirect). Deposition of NH3 had no effect on efflux of CO2 from Calluna vulgaris litter, despite it showing signs of stress in the field, whereas both NO3 and NH4 reduced CO2 fluxes. Our results show that the collective impacts on peatlands of the three principal forms of nitrogen in atmospheric deposition are a result of differential effects and mechanisms on individual components.

Characterisation of ALS genes in the polyploid species Schoenoplectus mucronatus and implications for resistance management.

BACKGROUND: The polyploid weed Schoenoplectus mucronatus (L.) Palla has evolved target-site resistance to ALS-inhibiting herbicides in Italian rice crops. Molecular and genetic characterisation of the resistance mechanism is relevant to the evolution and management of herbicide resistance. The authors aimed (a) to study the organisation of the target-site loci in two field-selected S. mucronatus populations with different cross-resistance patterns, (b) to identify the mutations endowing resistance to ALS inhibitors and determine the role of these mutations by using transgenesis and (c) to analyse the implications for the management of the S. mucronatus populations. RESULTS: Two complete ALS genes (ALS1 and ALS2) having an intron and a third partial intronless ALS gene (ALS3) were identified. The presence of multiple ALS genes was confirmed by Southern blot analyses, and ALS loci were characterised by examining cytosine methylation. In S. mucronatus leaves, the transcripts of ALS1, ALS2 and ALS3 were detected. Two mutations endowing resistance (Pro(197) to His and Trp(574) to Leu) were found in both resistant populations, but at different frequencies. Tobacco plants transformed with the two resistant alleles indicated that the Pro(197)-to-His substitution conferred resistance to SU and TP herbicides, while the allele with the Trp(574)-to-Leu substitution conferred cross-resistance to SU, TP, IMI and PTB herbicides. CONCLUSION: Schoenoplectus mucronatus has multiple ALS genes characterised by methylated sites that can influence the expression profile. The two mutated alleles proved to be responsible for ALS resistance. At population level, the resistance pattern depends on the frequency of various resistant genotypes, and this influences the efficacy of various ALS-inhibiting herbicides.

Differences in phytotoxicity and dissipation between ionized and nonionized oil sands naphthenic acids in wetland plants.

Naphthenic acids (NAs) are composed of alkyl-substituted acyclic and cycloaliphatic carboxylic acids and, because they are acutely toxic to fish, are of toxicological concern. During the caustic hot-water extraction of oil from the bitumen in oil sands deposits, NAs become concentrated in the resulting tailings pond water. The present study investigated if dissipation of NAs occurs in the presence of hydroponically grown emergent macrophytes (Typha latifolia, Phragmites australis, and Scirpus acutus) to determine the potential for phytoremediation of these compounds. Plants were grown with oil sands NAs (pKa approximately 5-6) in medium at pH 7.8 (predominantly ionized NAs) and pH 5.0 (predominantly nonionized NAs) to determine if, by altering their chemical form, NAs may be more accessible to plants and, thus, undergo increased dissipation. Whereas the oil sands NA mixture in its nonionized form was more toxic to wetland plants than its ionized form, neither form appeared to be sequestered by wetland plants. The present study demonstrated that plants may selectively enhance the dissipation of individual nonionized NA compounds, which contributes to toxicity reduction but does not translate into detectable total NA dissipation within experimental error and natural variation. Plants were able to reduce the toxicity of a NA system over 30 d, increasing the median lethal concentration (LC50; % of hydroponic solution) of the medium for Daphnia magna by 23.3% +/- 8.1% (mean +/- standard error; nonionized NAs) and 37.0% +/- 2.7% (ionized NAs) as determined by acute toxicity bioassays. This reduction in toxicity was 7.3% +/- 2.6% (nonionized NAs) and 45.0% +/- 6.8% (ionized NAs) greater than that in unplanted systems.