Evaluation of trace metal accumulation in six vegetable crops intercropped with phytostabilizing plant species, in a French urban wasteland.

The extensive development of agriculture in urban and peri-urban wastelands polluted with several trace elements (TE) poses risks to human health through contaminated food products. The objective was to explore the accumulation of TE in the various parts of vegetable crop plants (tomato, French bean, radish, potato, spinach, and leek) intercropped with phytostabilizing plant species (ryegrass and white clover, respectively). Field studies were conducted in a multicontaminated French urban wasteland with Cd, Cu, Pb and Zn, and an alkaline soil pH. Analyses of the respective non-edible parts of monocultured vegetable crops showed accumulation of all TE, mostly Zn, then Pb and Cu, and finally Cd. The corresponding TE accumulation factors (soil to plant) were all below 0.25. In the edible parts, average concentrations for TE were above the limit values, according to European and Chinese standards. TE contents in the phytostabilizing species chosen were in the same orders of magnitude and the same ranking as described for vegetable crops and most accumulation was in the roots. Unexpectedly, the presence of the phytostabilizing plants had a very strong positive impact on the soil to plant accumulation factor. Moreover, the edible plant parts were poorly impacted by the co-cropping with phytostabilizing plants.

Impacts of metallic trace elements on an earthworm community in an urban wasteland: Emphasis on the bioaccumulation and genetic characteristics in Lumbricus castaneus.

Metallic trace elements (MTEs) soil pollution has become a worldwide concern, particularly regarding its impact on earthworms. Earthworms, which constitute the dominant taxon of soil macrofauna in temperate regions and are crucial ecosystem engineers, are in direct contact with MTEs. The impacts of MTE exposure on earthworms, however, vary by species, with some able to cope with high levels of contamination. We combined different approaches to study the effects of MTEs at different levels of biological organisation of an earthworm community, in a contaminated urban wasteland. Our work is based on field collection of soil and earthworm samples, with a total of 891 adult earthworms from 8 species collected, over 87 quadrats across the study plot. We found that MTE concentrations are highly structured at the plot scale and that some elements, such as Pb, Zn, and Cu, are highly correlated. Comparing species assemblage to MTE concentrations, we found that the juvenile and adult abundances, and community composition, were significantly affected by pollution. Along the pollution gradient, as species richness decreased, Lumbricus castaneus became more dominant. We thus investigated the physiological response of this species to a set of specific elements (Pb, Zn, Cu, and Cd) and studied the impacts of MTE concentrations at the plot scale on its population genetic. These analyses revealed that L. castaneus is able to bioaccumulate high quantities of Cd and Zn, but not of Cu and Pb. The population genetic analysis, based on the genotyping of 175 individuals using 8 microsatellite markers, provided no evidence of the role of the heterogeneity in MTE concentrations as a barrier to gene flow. The multidisciplinary approach we used enabled us to reveal the comparatively high tolerance of L. castaneus to MTE concentrations, suggesting that this is a promising model to study the molecular bases of MTE tolerance.

Potential use of the PRI and active fluorescence for the diagnosis of the physiological state of plants under ozone exposure and high atmospheric vapor pressure deficit.

Assessing photosynthesis rates with remote sensing is important for tracking the physiological state of plants. The photochemical reflectance index (PRI) is a good estimator of short-term light-use efficiency (LUE) at the leaf scale but its responses to environmental factors are poorly understood. In this study, we assessed changes in the responses of the PRI to ozone exposure and to an increase in atmospheric drought (separately and combined) in oak (Quercus robur) and holm oak (Quercus ilex) that were planted in climatic cells under controlled conditions. The aim was to evaluate the ability of the PRI as a relevant indicator to assess the impact of abiotic factors on photosynthesis. Leaf-scale measurements of biochemical, physiological and spectral properties, including the PRI in dim light on dark-adapted leaves (PRI0), kinetics of PRI responses to PAR variations (photosynthetically active radiation), and leaf chlorophyll fluorescence parameters were performed. The results show that PRI0 is a good proxy of the leaf chlorophyll content, and is correlated to chlorophyll fluorescence parameters on dark adapted leaves (Fo, Fm). The correction of the PRI from the leaf chlorophyll content variations (PRIc) significantly improves correlations between the PRI and NPQ (non-photochemical quenching). The variability of PARsat (estimated PAR value at PRI saturation using PRI vs. PAR relationships) depends on ozone exposure and on the increase in atmospheric vapor pressure deficit. For Quercus robur, results highlight that PARsat is linked to abiotic stress indicating that the PRI may be used as a relevant indicator of abiotic factors limiting the photosynthesis. Quercus ilex did not show significant variability in PRI0 and PARsat, which suggest that it is a more drought resistant species than Q. robur.

Combined Effects of Ozone and Drought on the Physiology and Membrane Lipids of Two Cowpea (Vigna unguiculata (L.) Walp) Cultivars.

The interactive effects of drought and ozone on the physiology and leaf membrane lipid content, composition and metabolism of cowpea (Vigna unguiculata (L.) Walp.) were investigated in two cultivars (EPACE-1 and IT83-D) grown under controlled conditions. The drought treatment (three-week water deprivation) did not cause leaf injury but restricted growth through stomatal closure. In contrast, the short-term ozone treatment (130 ppb 12 h daily during 14 day) had a limited impact at the whole-plant level but caused leaf injury, hydrogen peroxide accumulation and galactolipid degradation. These effects were stronger in the IT83-D cultivar, which also showed specific ozone responses such as a higher digalactosyl-diacylglycerol (DGDG):monogalactosyldiacylglycerol (MGDG) ratio and the coordinated up-regulation of DGDG synthase (VuDGD2) and ω-3 fatty acid desaturase 8 (VuFAD8) genes, suggesting that membrane remodeling occurred under ozone stress in the sensitive cultivar. When stresses were combined, ozone did not modify the stomatal response to drought and the observed effects on whole-plant physiology were essentially the same as when drought was applied alone. Conversely, the drought-induced stomatal closure appeared to alleviate ozone effects through the reduction of ozone uptake.

Ozone exposure and flux-based response functions for photosynthetic traits in wheat, maize and poplar.

Ozone exposure- and dose-response relationships based on photosynthetic leaf traits (CO2 assimilation, chlorophyll content, Rubisco and PEPc activities) were established for wheat, maize and poplar plants grown in identical controlled conditions, providing a comparison between crop and tree species, as well as between C3 and C4 plants. Intra-specific variability was addressed by comparing two wheat cultivars with contrasting ozone tolerance. Depending on plant models and ozone levels, first-order, second-order and segmented linear regression models were used to derive ozone response functions. Overall, flux-based functions appeared superior to exposure-based functions in describing the data, but the improvement remained modest. The best fit was obtained using the POD0.5 for maize and POD3 for poplar. The POD6 appeared relevant for wheat, although intervarietal differences were found. Our results suggest that taking into account the dynamics of leaf antioxidant capacity could improve current methods for ozone risk assessment for plants.

The impact of light intensity on shade-induced leaf senescence.

Plants often have to cope with altered light conditions, which in leaves induce various physiological responses ranging from photosynthetic acclimation to leaf senescence. However, our knowledge of the regulatory pathways by which shade and darkness induce leaf senescence remains incomplete. To determine to what extent reduced light intensities regulate the induction of leaf senescence, we performed a functional comparison between Arabidopsis leaves subjected to a range of shading treatments. Individually covered leaves, which remained attached to the plant, were compared with respect to chlorophyll, protein, histology, expression of senescence-associated genes, capacity for photosynthesis and respiration, and light compensation point (LCP). Mild shading induced photosynthetic acclimation and resource partitioning, which, together with a decreased respiration, lowered the LCP. Leaf senescence was induced only under strong shade, coinciding with a negative carbon balance and independent of the red/far-red ratio. Interestingly, while senescence was significantly delayed at very low light compared with darkness, phytochrome A mutant plants showed enhanced chlorophyll degradation under all shading treatments except complete darkness. Taken together, our results suggest that the induction of leaf senescence during shading depends on the efficiency of carbon fixation, which in turn appears to be modulated via light receptors such as phytochrome A.

Mitochondrial malate dehydrogenase lowers leaf respiration and alters photorespiration and plant growth in Arabidopsis.

Malate dehydrogenase (MDH) catalyzes a reversible NAD(+)-dependent-dehydrogenase reaction involved in central metabolism and redox homeostasis between organelle compartments. To explore the role of mitochondrial MDH (mMDH) in Arabidopsis (Arabidopsis thaliana), knockout single and double mutants for the highly expressed mMDH1 and lower expressed mMDH2 isoforms were constructed and analyzed. A mmdh1mmdh2 mutant has no detectable mMDH activity but is viable, albeit small and slow growing. Quantitative proteome analysis of mitochondria shows changes in other mitochondrial NAD-linked dehydrogenases, indicating a reorganization of such enzymes in the mitochondrial matrix. The slow-growing mmdh1mmdh2 mutant has elevated leaf respiration rate in the dark and light, without loss of photosynthetic capacity, suggesting that mMDH normally uses NADH to reduce oxaloacetate to malate, which is then exported to the cytosol, rather than to drive mitochondrial respiration. Increased respiratory rate in leaves can account in part for the low net CO(2) assimilation and slow growth rate of mmdh1mmdh2. Loss of mMDH also affects photorespiration, as evidenced by a lower postillumination burst, alterations in CO(2) assimilation/intercellular CO(2) curves at low CO(2), and the light-dependent elevated concentration of photorespiratory metabolites. Complementation of mmdh1mmdh2 with an mMDH cDNA recovered mMDH activity, suppressed respiratory rate, ameliorated changes to photorespiration, and increased plant growth. A previously established inverse correlation between mMDH and ascorbate content in tomato (Solanum lycopersicum) has been consolidated in Arabidopsis and may potentially be linked to decreased galactonolactone dehydrogenase content in mitochondria in the mutant. Overall, a central yet complex role for mMDH emerges in the partitioning of carbon and energy in leaves, providing new directions for bioengineering of plant growth rate and a new insight into the molecular mechanisms linking respiration and photosynthesis in plants.

Ozone-induced changes in photosynthesis and photorespiration of hybrid poplar in relation to the developmental stage of the leaves.

Young poplar trees (Populus tremula Michx. x Populus alba L. clone INRA 717-1B4) were subjected to 120 ppb of ozone for 35 days in phytotronic chambers. Treated trees displayed precocious leaf senescence and visible symptoms of injury (dark brown/black upper surface stippling) exclusively observed on fully expanded leaves. In these leaves, ozone reduced parameters related to photochemistry (Chl content and maximum rate of photosynthetic electron transport) and photosynthetic CO(2) fixation [net CO(2) assimilation, Rubisco (ribulose-1,5-bisphosphate carboxylase oxygenase) activity and maximum velocity of Rubisco for carboxylation]. In fully expanded leaves, the rate of photorespiration as estimated from Chl fluorescence was markedly impaired by the ozone treatment together with the activity of photorespiratory enzymes (Rubisco and glycolate oxidase). Immunoblot analysis revealed a decrease in the content of serine hydroxymethyltransferase in treated mature leaves, while the content of the H subunit of the glycine decarboxylase complex was not modified. Leaves in the early period of expansion were exempt from visible symptoms of injury and remained unaffected as regards all measured parameters. Leaves reaching full expansion under ozone exposure showed potential responses of protection (stimulation of mitochondrial respiration and transitory stomatal closure). Our data underline the major role of leaf phenology in ozone sensitivity of photosynthetic processes and reveal a marked ozone-induced inhibition of photorespiration.

Ozone risk assessment for plants: central role of metabolism-dependent changes in reducing power.

The combination of stomatal-dependent ozone flux and total ascorbate level is currently presented as a correct indicator for determining the degree of sensitivity of plants to ozone. However, the large changes in carbon metabolism could play a central role in the strategy of the foliar cells in response to chronic ozone exposure, participating in the supply of reducing power and carbon skeletons for repair and detoxification, and modifying the stomatal mode of functioning. To reinforce the accuracy of the definition of the threshold for ozone risk assessment, it is proposed to also consider the redox pool (NAD(P)H), the ratio between carboxylases and the water use efficiency as indicators of the differential ozone tolerance of plants.

A DIGE analysis of developing poplar leaves subjected to ozone reveals major changes in carbon metabolism.

Tropospheric ozone pollution is described as having major negative effects on plants, compromising plant survival. Carbon metabolism is especially affected. In the present work, the effects of chronic ozone exposure were evaluated at the proteomic level in developing leaves of young poplar plants exposed to 120 ppb of ozone for 35 days. Soluble proteins (excluding intrinsic membrane proteins) were extracted from leaves after 3, 14 and 35 days of ozone exposure, as well as 10 days after a recovery period. Proteins (pI 4 to 7) were analyzed by 2-D DIGE experiments, followed by MALDI-TOF-TOF identification. Additional observations were obtained on growth, lesion formation, and leaf pigments analysis. Although treated plants showed large necrotic spots and chlorosis in mature leaves, growth decreased only slightly and plant height was not affected. The number of abscised leaves was higher in treated plants, but new leaf formation was not affected. A decrease in chlorophylls and lutein contents was recorded. A large number of proteins involved in carbon metabolism were identified. In particular, proteins associated with the Calvin cycle and electron transport in the chloroplast were down-regulated. In contrast, proteins associated with glucose catabolism increased in response to ozone exposure. Other identified enzymes are associated with protein folding, nitrogen metabolism and oxidoreductase activity.