The DroughtBox: A new tool for phenotyping residual branch conductance and its temperature dependence during drought.

Xylem hydraulic failure is a major driver of tree death during drought. However, to better understand mortality risk in trees, especially during hot-drought events, more information is required on both rates of residual water-loss from small branches (gres ) after stomatal closure, as well as the phase transition temperature (Tp ), beyond which gres significantly increases. Here, we describe and test a novel low-cost tool, the DroughtBox, for phenotyping gres and Tp across species. The system consists of a programmable climatically controlled chamber in which branches dehydrate and changes in the mass recorded. Test measurements show that the DroughtBox maintains stable temperature and relative humidity across a range of set points, a prerequisite for getting accurate gres and Tp values. Among a study group of four conifer and one angiosperm species, we observed a range of gres (0.44-1.64 mmol H2 O m-2 s-1 ) and Tp (39.4-43.8°C) values. Furthermore, the measured time to hydraulic failure varied between two conifers species and was shortened in both species following a heatwave event. The DroughtBox is a reliable and customizable tool for phenotyping gres and Tp , as well as for testing models of time to hydraulic failure that will improve our ability to assess climate change impacts on plants.

Abundance and diversity of ammonia-oxidizing prokaryotes in the root-rhizosphere complex of Miscanthus × giganteus grown in heavy metal-contaminated soils.

Mine wastes have been considered as a source of heavy metal (HM) contamination in the environment and negatively impact many important ecosystem services provided by soils. Plants like Miscanthus, which tolerate high HM concentrations in soil, are often used for phytoremediation and provide the possibility to use these soils at least for the production of energy crops. However, it is not clear if plant growth at these sites is limited by the availability of nutrients, mainly nitrogen, as microbes in soil might be affected by the contaminant. Therefore, in this study, we investigated in a greenhouse experiment the response of ammonia-oxidizing microbes in the root-rhizosphere complex of Miscanthus × giganteus grown in soils with different levels of long-term arsenic (As) and lead (Pb) contamination. Quantitative PCR of the ammonia monooxigenease gene (amoA) was performed to assess the abundance of ammonia-oxidizing bacteria (AOB) and archaea (AOA) at two different points of plant growth. Furthermore, bulk soil samples before planting were analyzed. In addition, terminal restriction fragment length polymorphism (T-RFLP) analysis was used to investigate the diversity of archaeal amoA amplicons. Whereas high concentrations of As and Pb in soil (83 and 15 g/kg, respectively) resulted independent from plant growth in a clear reduction of AOA and AOB compared to the control soils with lower HM contents, in soils with contamination levels of 10 g/kg As and 0.2 g/kg Pb, only AOB were negatively affected in bulk soil samples. Diversity analysis of archaeal amoA genes revealed clear differences in T-RFLP patterns in response to the degree of HM contamination. Therefore, our results could clearly prove the different response patterns of AOA and AOB in HM-contaminated soils and the development of archaeal amoA phylotypes which are more tolerant towards HMs in soil samples from the areas that were impacted the most by mining waste, which could contribute to functional redundancy of ammonia-oxidizing microbes in soils and stability of nitrification pattern.

Expression of stress-related genes in tomato plants exposed to arsenic and chromium in nutrient solution.

The molecular responses of hydroponically cultivated tomato plants to As(V) or Cr(VI) were assessed by transcript accumulation analysis of genes coding for products potentially involved in heavy metal tolerance. A quantitative real-time PCR experiment was performed with Hsp90-1, MT2- and GR1-like protein genes using RNA isolated from tomato roots or shoots treated for 24h with As(V) or Cr(VI) at concentrations ranging from 80 to 640 microM. Both transient metallic treatments induced Hsp90-1 transcript accumulation in tomato plants. MT2- and GR1-like transcripts accumulated in tomato roots treated with As(V) but were only slightly affected by Cr(VI) treatment. Tomatoes showed phenotypic symptoms to heavy metal toxicity when plants were exposed to Cr(VI) but not As(V). Plant lethality was observed at 1280 microM Cr(VI), indicating that tomatoes were more tolerant to As than Cr stress under the experimental conditions used here.

Photodegradation product of sulcotrione and the physiological response of maize (Zea mays) and white mustard (Sinapis alba).

One of the strategies for decreasing the consumption of herbicides consists in improving their uptake and efficiency. It was suggested that the photodegradation of herbicides due to sunlight results in a greater demand of herbicides to be introduced into the environment in order to ensure the plant protection activity. Moreover, an ecotoxicological effect of the photoproducts needs to be clarified. The physiological response of Zea mays and Sinapis alba (weed) to sulcotrione and its main photoproduct, called chromone (xanthene-1,9-dione-3,4-dihydro-6-methylsulfonyl), was evaluated under controlled conditions in a growth chamber. The dose-response effects were determined on Z. mays and S. alba. Using the sulcotrione (doses ranging from 1 to 9mg per plant), the physiological parameters indicated a decrease of photosynthesis for the S. alba species while the Z. mays species were only slightly affected. On the contrary, the chromone had no herbicide activity on both species. The sulcotrione is known to block 4-hydroxyphenyl pyruvate dioxygenase (HPPD) enzyme. The differences between the parent herbicide and the photoproduct could be ascribed to drastic structural modifications. We have shown that the chromone probably do not block the HPPD active site.

Photolysis of the herbicide sulcotrione: formation of a major photoproduct and its toxicity evaluation.

BACKGROUND: Sulcotrione is a selective herbicide marketed for use in maize since 1993, but its environmental fate is not yet fully elucidated. A major metabolite resulting from cleavage between the two ring moieties, leading to 2-chloro-4-mesylbenzoic acid (CMBA), has been identified; it presents a rather low toxicity. In photochemical studies this compound has also been claimed to be formed in high proportions. The present authors recently found that, under irradiation, sulcotrione mainly yields a cyclization product (CP). Thus, Sulcotrione photochemistry is still a matter of debate. The aim of the present work was to give an unequivocal answer to this issue. The potential toxicity of CP, CMBA and sulcotrione towards three organisms considered as representative of aquatic ecosystems was also evaluated. RESULTS: The main transformation product of sulcotrione is the cyclization product (CP), and CMBA is formed in smaller amounts. For the toxicological approach, the tested organisms were a bacterium, Vibrio fischeri (Bejerinck) Lehmann & Neumann, an alga, Pseudokirchneriella subcapitata (Korshikov) Hindak, and a protozoan, Tetrahymena pyriformis (Ehrenberg) Lwoff. Sulcotrione is more harmful towards the alga, but CP is more toxic to the bacterium and the protozoan. It must be noted that the measured toxicities are nonetheless rather low. CONCLUSION: On irradiation, sulcotrione mainly gives the photocyclization product, which presents a higher toxicity than sulcotrione and CMBA. This cyclization product should thus be considered in sulcotrione environmental risk assessment.

Effect of chromium species on phytochemical and physiological parameters in Datura innoxia.

In soil, chromium can be found in two main valence states: hexavalent Cr(VI) and trivalent Cr(III). In this study, we investigated the impact of Cr on photosynthetic gas exchange, photosystem II (PSII) activity, Cr translocation and accumulation, proline content and alkaloids production, i.e. scopolamine and hyoscyamine, in Datura innoxia. Cr uptake was influenced by its oxidation state and its concentration in growth medium. The plant roots were determined as being the main organ of Cr accumulation. Cr(VI) was more toxic than Cr(III) as indicated by reduction in plant biomass and net photosynthesis. The stomatal conductance showed a similar trend to that of photosynthetic capacity. Cr(III) and Cr(VI) had a different impact on substomatal CO(2) concentration then Cr toxicity was related to its oxidation states. In plants stressed with a Cr(VI) excess, a down regulation of PSII activity was observed with an impairment of photochemical activity. Indeed, the maximum quantum yield of PSII (F(v)/F(m)), the quantum yield of PSII (PhiPSII) and the efficiency of excitation capture by open centers (F'(v)/F'(m)) decreased. Cr(III) had little effects on PSII primary photochemistry, whatever its form induces an increase of scopolamine content without changes in hyoscyamine content in leaves of D. innoxia. These results provide that chromium contamination can change the secondary metabolites composition of leaves, thereby, impacting the quality, safety and efficacy of natural plant products synthesized by D. innoxia plants.

Effect of citric acid and EDTA on chromium and nickel uptake and translocation by Datura innoxia.

EDTA and citric acid were tested to solubilize metals and enhance their uptake by Datura innoxia, chosen because of its ability to accumulate and tolerate metals. Two application modes were used on an industrial soil contaminated mainly by Cr and Ni. The results showed that citric acid was the most effective at increasing the uptake of Cr and EDTA for Ni. These results are consistent with the effectiveness of both chelants in solubilizing metals from the soil. The translocation factor (TF) of Ni was 1.6- and 6.7-fold higher than the control, respectively, for one and two applications of 1mmolkg(-1) EDTA. After two applications of 5 and 10mmolkg(-1) citric acid, the TF of Cr increased 2- and 3.5-fold relative to the control. Whatever the concentration, the application of EDTA modified the plant physiology significantly. For citric acid this was only observed with the highest dose (10mmolkg(-1)).

Interaction of bioaccumulation of heavy metal chromium with water relation, mineral nutrition and photosynthesis in developed leaves of Lolium perenne L.

Contamination by chromium (Cr) is widespread in agricultural soils and industrial sites. This heavy metal represents a risk to human health. In order to gain fundamental insights into the nature of the adaptation to Cr excess, the characterisation of physiological indices, including responses of photosynthetic gas exchange and chlorophyll a fluorescence along with changes in mineral nutrient contents and water status were studied in ray grass (Lolium perenne L.). Increased concentrations of Cr(VI) (0-500 microM Cr) in the Coïc and Lessaint nutrient solution were applied. The growth of Lolium perenne is decreased by chromium and the leaves have lost their pigments. Chromium accumulation was greater in roots than in leaves and reached 2450 and 210 microg g(-1) DW, respectively with 500 microM Cr(VI) in nutrient medium. The physiological parameters were severely reduced by this heavy metal. Cr induced toxicity arising from 100 microM Cr(VI) and resulted in a modification of mineral content in roots and leaves, especially for Ca, Mg and Fe. The chromium stress decreased CO2 assimilation rates mainly due to stomatal closure, which reduced water loss by transpiration without decreasing the cellular available CO2. The fluorescence parameters associated with photosystem II (PSII) activity and the photochemical activity are modified by chromium. Non-radiative energy dissipation mechanisms were triggered during stress since non-photochemical quenching was increased and efficiency of excitation capture by open centers was reduced.

Photosynthetic activity of Lolium perenne as a function of endophyte status and zinc nutrition.

Grass infection by endophyte fungi can confer host resistance to different types of stress, but limited evidence is available on the related effects on the photosynthetic mechanism of the grasses. Zinc has direct and indirect effects on this mechanism and is one of the more important environmental pollutants. To measure whether photosynthesis of the host plant is affected by fungal infection when endophyte-free and endophyte-infected grasses contain similar excess zinc concentrations in their leaves, two batches of Lolium perenne L. cv. Apollo plants were established, one batch was infected with Neotyphodium lolii, the other was not. Both batches were then treated for 8 d with a nutrient solution containing 0, 1, 5, 10 or 20 mm ZnSO4. The increase in zinc concentration induced some reduction in photosystem II (PSII) activity but not enough to account for the total drop in the net photosynthetic rate. Endophyte fungus favoured maintenance of the PSII activity, but did not significantly modify the net photosynthesis and similar zinc concentration levels were observed in leaves of both types of plant. Interactive effects of zinc and light induced less photodamage to the PSII of the host, which is able to react to an increase in photon flux density (PFD). In endophyte-free plants, the reaction centre contributed more than antenna complexes to energy dissipation. In endophyte-infected plants, the quenching of the reaction centre and antenna complexes rose simultaneously and at a constant rate, as zinc concentrations increased.

Use of commercial plant species in a hydroponic system to treat domestic wastewaters.

The objectives in this work were to investigate a conceptual layout for an inexpensive and simple system that would treat primary municipal wastewater to discharge standards. A commercial hydroponic system was adapted for this study and the wastewater was used to irrigate wooly digitalis (Digitalis lanata Ehrh.) and foxglove (Digitalis purpurea L.). These plants are medicinal and produce cardenolide compounds. Influent and effluent samples were collected once a month for six months and analyzed to determine the various parameters relating to water quality. The legal discharge levels for total suspended solids (SS), biochemical oxygen demand (BOD5), and chemical oxygen demand (COD) were reached for the two tested plants after 48 h of wastewater treatment; the removal was 82, 93, and 79%, respectively, for wooly digitalis and 92, 92, and 84%, respectively, for foxglove. Similar results were obtained during a 6-mo period although the sewage composition varied widely. The system tended to be unable to remove N and P to concentrations below regulated levels. Compared with the nutrient solution composition, the wastewater was more concentrated in Na+ and Cl- and less in N, K+, and Ca2+. These variations can lead to the decline of wooly digitalis plants. Foxglove developed a significant root system to increase mineral absorption wastewater being used as the unique nutritive source. After 10 wk all the wooly digitalis seedlings were dead. Despite this fact, however, the root system remained in place for a significant time (< 4 mo), thus continuing to filter wastewater and to be used as a bacterial support thus making it possible to have a security period to replace the dead plants.

Treatment of domestic wastewater by an hydroponic NFT system.

The objectives in this work were to investigate a conceptual layout for an inexpensive and simple system that would treat primary municipal wastewater to discharge standards. A commercial hydroponic system was adapted for this study and the wastewater was used to irrigate Datura innoxia plants. Influent and effluent samples were collected once a month for six months and analysed to determine the various parameters relating to the water quality. The legal discharge levels for total suspended, biochemical oxygen demand and chemical oxygen demand were reached with the plant system after 24 h of wastewater treatment. Total nitrogen and total phosphorus reduction were also obtained. NH4(+)-N was reduced by 93% with nitrification proving to be the predominant removal process. Significant nitrification occurred when the BOD5 level dropped 45 mg/l. Similar results were obtained for six months although the sewage composition varied widely. D. innoxia develops and uses the wastewater as the unique nutritive source.

Treatment of domestic wastewater using the nutrient film technique (NFT) to produce horticultural roses.

Removal of organic matter, nitrogen and phosphorus was investigated in a pilot based on the nutrient film technique system used for horticultural production. Rosebushes producing commercial roses were set on an inclined impermeable surface over which a thin film of domestic wastewater flowed directly through the root matrix. The roses produced with wastewater as nutrient solution were qualitatively and quantitatively similar to those produced traditionally and can be marketable. In the presence of the rosebushes, the lowering of chemical oxygen demand (COD), biological oxygen demand (BOD5) and suspended solids (SS) increased with their influent strengths in a linear fashion correlation. Whatever the pollutant load of the influent, within the range studied, the level of organic pollution required to allow the discharge of the effluent into water courses was reached after 24 h of treatment. At this time, the COD reached 39 +/- 13 mg L(-1), the BOD5 7 +/- 4 mg L(-1) and the SS 8 +/- 6 mg L(-1). The removal percentages were 89, 95 and 94, respectively, whereas without plants they were 55, 33 and 53, respectively. The rosebushes had a beneficial effect on the removal of phosphorus from 20% to 23% greater in their presence than without. In presence of plants, the nitrate was the principal form of nitrogen from 48 h of treatment whereas without rosebushes, nitrite represent more than 20% of initial nitrogen whatever the treatment duration. The root matrix served not only as a filter for the organic matter, but also provided a microhabitat suitable for nitrifying bacteria and heterotrophic micro-organisms, which responded rapidly to the pollutant load.

Urban wastewater treatment by a nutrient film technique system with a valuable commercial plant species (Chrysanthemum cinerariaefolium Trev.).

Urban wastewater causes rapid eutrophication of natural waters and requires treatment before discharge. This is expensive and produces huge quantities of sludge. In the European Community, it will no longer be lawful to dispose of this sludge as landfill after 2005 (European Directive 91/271/CEE of May 21, 1991). Wastewater treatment by the Chrysanthemum cinerariaefolium plants in horizontal flow was investigated using the nutrient film technique (NFT), a widely used hydroponic system in the commercial greenhouse industry. After a 48 h plant treatment, the purification efficiency was 95%, 91%, and 99% with respect to suspended solids (SS), biochemical oxygen demand (BOD5), and chemical oxygen demand (COD), and the elimination of nutrients (total nitrogen and total phosphorus) varied between 40% and 80%. SS and thus indirectly BOD5 and COD were removed by filtration and adsorption; the solids trapped in the root systems were then decomposed and mineralized. The system with 25 plants purified 30 L of wastewater in 48 h. One-hundred people communities wastewater could be treated with a 6 m2 area of production. Pyrethrin contents and chlorophyll a fluorescence of plants grown on raw urban waters were not significantly different from those grown on a standard nutrient solution.