NBR1-mediated selective autophagy of ARF7 modulates root branching.

Auxin dictates root architecture via the Auxin Response Factor (ARF) family of transcription factors, which control lateral root (LR) formation. In Arabidopsis, ARF7 regulates the specification of prebranch sites (PBS) generating LRs through gene expression oscillations and plays a pivotal role during LR initiation. Despite the importance of ARF7 in this process, there is a surprising lack of knowledge about how ARF7 turnover is regulated and how this impacts root architecture. Here, we show that ARF7 accumulates in autophagy mutants and is degraded through NBR1-dependent selective autophagy. We demonstrate that the previously reported rhythmic changes to ARF7 abundance in roots are modulated via autophagy and might occur in other tissues. In addition, we show that the level of co-localization between ARF7 and autophagy markers oscillates and can be modulated by auxin to trigger ARF7 turnover. Furthermore, we observe that autophagy impairment prevents ARF7 oscillation and reduces both PBS establishment and LR formation. In conclusion, we report a novel role for autophagy during development, namely by enacting auxin-induced selective degradation of ARF7 to optimize periodic root branching.

PAT mRNA decapping factors are required for proper development in Arabidopsis.

Evolutionarily conserved protein associated with topoisomerase II (PAT1) proteins activate mRNA decay through binding mRNA and recruiting decapping factors to optimize posttranscriptional reprogramming. Here, we generated multiple mutants of pat1, pat1 homolog 1 (path1), and pat1 homolog 2 (path2) and discovered that pat triple mutants exhibit extremely stunted growth and all mutants with pat1 exhibit leaf serration while mutants with pat1 and path1 display short petioles. All three PATs can be found localized to processing bodies and all PATs can target ASYMMETRIC LEAVES 2-LIKE 9 transcripts for decay to finely regulate apical hook and lateral root development. In conclusion, PATs exhibit both specific and redundant functions during different plant growth stages and our observations underpin the selective regulation of the mRNA decay machinery for proper development.

Going through changes - the role of autophagy during reprogramming and differentiation.

Somatic cell reprogramming is a complex feature that allows differentiated cells to undergo fate changes into different cell types. This process, which is conserved between plants and animals, is often achieved via dedifferentiation into pluripotent stem cells, which have the ability to generate all other types of cells and tissues of a given organism. Cellular reprogramming is thus a complex process that requires extensive modification at the epigenetic and transcriptional level, unlocking cellular programs that allow cells to acquire pluripotency. In addition to alterations in the gene expression profile, cellular reprogramming requires rearrangement of the proteome, organelles and metabolism, but these changes are comparatively less studied. In this context, autophagy, a cellular catabolic process that participates in the recycling of intracellular constituents, has the capacity to affect different aspects of cellular reprogramming, including the removal of protein signatures that might hamper reprogramming, mitophagy associated with metabolic reprogramming, and the supply of energy and metabolic building blocks to cells that undergo fate changes. In this Review, we discuss advances in our understanding of the role of autophagy during cellular reprogramming by drawing comparisons between plant and animal studies, as well as highlighting aspects of the topic that warrant further research.

The mRNA decapping machinery targets LBD3/ASL9 to mediate apical hook and lateral root development.

Multicellular organisms perceive and transduce multiple cues to optimize development. Key transcription factors drive developmental changes, but RNA processing also contributes to tissue development. Here, we report that multiple decapping deficient mutants share developmental defects in apical hook, primary and lateral root growth. More specifically, LATERAL ORGAN BOUNDARIES DOMAIN 3 (LBD3)/ASYMMETRIC LEAVES 2-LIKE 9 (ASL9) transcripts accumulate in decapping deficient plants and can be found in complexes with decapping components. Accumulation of ASL9 inhibits apical hook and lateral root formation. Interestingly, exogenous auxin application restores lateral roots formation in both ASL9 over-expressors and mRNA decay-deficient mutants. Likewise, mutations in the cytokinin transcription factors type-B ARABIDOPSIS RESPONSE REGULATORS (B-ARRs) ARR10 and ARR12 restore the developmental defects caused by over-accumulation of capped ASL9 transcript upon ASL9 overexpression. Most importantly, loss-of-function of asl9 partially restores apical hook and lateral root formation in both dcp5-1 and pat triple decapping deficient mutants. Thus, the mRNA decay machinery directly targets ASL9 transcripts for decay, possibly to interfere with cytokinin/auxin responses, during development.

Synthesis, Characterization and Decomposition of Potassium Jarosite for Adsorptive As(V) Removal in Contaminated Water: Preliminary Study.

Jarosite-type compounds precipitated in the zinc industry for iron control can also incorporate arsenic and can be used for wastewater treatment for As elimination. According with the last, this work is related to arsenic incorporation at room temperature in decomposed potassium jarosite. The work began with the synthesis of the compound at 75 °C for 9 h using Fe2(SO4)3 and K2SO4 at a pH of 1.1. Once jarosite was obtained, solids were subjected to an alkaline decomposition using NaOH at pH 10 for 30 min, and then As was added to the solution as HAsNaO4 and the pH modified by adding HNO3 until it reached a value of 1.1. The initial, intermediate, and final products were wholly characterized by scanning electron microscopy (SEM) in conjunction with energy dispersive spectrometry (EDS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy (RS), and X-ray photoelectron spectrometry (XPS). The obtained results show that As(V) can be adsorbed by ionic exchange in the amorphous FeOH structure of decomposed jarosite and when pH decreased to 1.1, the compound recrystallized, incorporating up to 6% As on average, which is indicative that this process can be used to reduce As in contaminated waters.

Leaching of Copper Contained in Waste Printed Circuit Boards, Using the Thiosulfate-Oxygen System: A Kinetic Approach.

The present work is related to the treatment of crushed waste of printed circuit boards (WPCBs) from electrical and electronic devices (WEEE), carrying out the recovery of copper in solution. In the first stage, the studied material was characterized by AAS, SEM-EDS, and XRD. The results revealed significantly high amounts of copper (744.42 mg/g), compared with the rest of the metals present in the sample, mainly iron and zinc. In the second stage of the work, alkali dynamic leaching experiments were carried out in the S2O3−2− O2 medium, evaluating important kinetic variables in order to verify the controlling step of the system and adjust the data to a kinetic model. According to the results obtained from the various experimental tests executed, it was found that in the studied system of S2O3−2− O2, the leaching of copper was preferably adjusted to the model of spherical particles with a shrinking core finding a mixed chemical−diffusive control, with values of Ea = 25.78 kJ/mol and n = 0.22 (for the leaching reagent), indicating that the reaction was controlled by the oxygen transport to the solid−liquid interface and also by the chemical reaction in the surface of particles, obtaining up to 99.82% copper in solution.

mRNA Decapping Factors LSM1 and PAT Paralogs Are Involved in Turnip Mosaic Virus Viral Infection.

Turnip mosaic virus is a devastating potyvirus infecting many economically important brassica crops. In response to this, the plant host engages its RNA silencing machinery, involving AGO proteins, as a prominent strategy to restrain turnip mosaic virus (TuMV) infection. It has also been shown that the mRNA decay components DCP2 and VCS partake in viral infection suppression. Here, we report that the mRNA decapping components LSM1, PAT1, PATH1, and PATH2 are essential for TuMV infection. More specifically, lsm1a/lsm1b double mutants and pat1/path1/path2 triple mutants in summ2 background exhibit resistance to TuMV. Concurrently, we observed that TuMV interferes with the decapping function of LSM1 and PAT proteins as the mRNA-decay target genes UGT87A2 and ASL9 accumulate during TuMV infection. Moreover, as TuMV coat protein can be specifically found in complexes with PAT proteins but not LSM1, this suggests that TuMV "hijacks" decapping components via PAT proteins to support viral infection.[Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Overexpression of ATG8/LC3 enhances wound-induced somatic reprogramming in Physcomitrium patens.

Animal and plant somatic cells have the capacity to switch states or reprogram into stem cells to adapt during stress and injury. This ability to deal with stochastic changes or reprogramming of somatic cells also needs macroautophagy/autophagy. Here, we expand on this notion and provide a primary example of how overexpression of ATG8/LC3 in the moss Physcomitrium patens enhances the ability to reprogram somatic cells into stem cells when subjected to severe wounding. This observation suggests that autophagy is not only required for cells to dedifferentiate but also makes cells more competent to do so.Abbreviation: ATG: autophagy related; atg5: AUTOPHAGY 5; ATG8/LC3: AUTOPHAGY 8/microtubule associated protein 1 light chain 3; GFP: green fluorescent protein.

Use of the O2-Thiosemicarbazide System, for the Leaching of: Gold and Copper from WEEE & Silver Contained in Mining Wastes.

Environmental pollution today is a latent risk for humanity, here the need to recycle waste of all kinds. This work is related to the kinetic study of the leaching of gold and copper contained in waste electrical and electronic equipment (WEEE) and silver contained in mining wastes (MW), using the O2-thiosemicarbazide system. The results obtained show that this non-toxic leaching system is adequate for the leaching of said metals. Reaction orders were found ranging from 0 (Cu), 0.93 (Ag), and 2.01 (Au) for the effect of the reagent concentration and maximum recoveries of 77.7% (Cu), 95.8% (Au), and 60% (Ag) were obtained. Likewise, the activation energies found show that the leaching of WEEE is controlled by diffusion (Cu Ea = 9.06 and Au Ea = 18.25 kJ/Kmol), while the leaching of MW (Ea = 45.55 kJ/Kmol) is controlled by the chemical reaction. For the case of stirring rate, it was found a low effect and only particles from WEEE and MW must be suspended in solution to proceed with the leaching. The pH has effect only at values above 8, and finally, for the case of MW, the O2 partial pressure has a market effect, going the Ag leaching from 33% at 0.2 atm up to 60% at a 1 atm.

Autophagy mediates temporary reprogramming and dedifferentiation in plant somatic cells.

Somatic cells acclimate to changes in the environment by temporary reprogramming. Much has been learned about transcription factors that induce these cell-state switches in both plants and animals, but how cells rapidly modulate their proteome remains elusive. Here, we show rapid induction of autophagy during temporary reprogramming in plants triggered by phytohormones, immune, and danger signals. Quantitative proteomics following sequential reprogramming revealed that autophagy is required for timely decay of previous cellular states and for tweaking the proteome to acclimate to the new conditions. Signatures of previous cellular programs thus persist in autophagy-deficient cells, affecting cellular decision-making. Concordantly, autophagy-deficient cells fail to acclimatize to dynamic climate changes. Similarly, they have defects in dedifferentiating into pluripotent stem cells, and redifferentiation during organogenesis. These observations indicate that autophagy mediates cell-state switches that underlie somatic cell reprogramming in plants and possibly other organisms, and thereby promotes phenotypic plasticity.

Genotoxic endpoints in a Pb-accumulating pea cultivar: insights into Pb2+ contamination limits.

Lead (Pb) persists among the most hazardous contaminant metals. Pb-induced genotoxic effects remain a matter of debate as they are a major cause of plant growth impairment, but assessing Pb genotoxicity requires the selection of Pb-sensitive genotoxic biomarkers. Seedlings of the ecotoxicological model species Pisum sativum L. were exposed to Pb2+ (≤ 2000 mg L-1). Flow cytometry (FCM) revealed that 28 days after, Pb2+ arrested root cell cycle at G2 but no eu/aneuploidies were found. Comet assay and FCM-clastogenicity assays showed that Pb2+ increased DNA breaks in roots at concentrations as low as 20 mg L-1. Leaves showed no variation in DNA-ploidy or cell cycle progression but had increased DNA breaks at the highest Pb2+ dose. We conclude that both Comet assay and the full-peak coefficient of variation (FPCV) were the most relevant endpoints of Pb-phytogenotoxicity. Also, the Pb-induced DNA breaks may be related with the arrest at the G2-checkpoint. Data will be relevant to better define Pb2+ ecogenotoxicological effects and their measuring tools and may contribute to a regulatory debate of this pollutant limits.

Individual components of paired typical NLR immune receptors are regulated by distinct E3 ligases.

In plants and animals, nucleotide-binding leucine-rich repeat (NLR) proteins serve as intracellular immune receptors. Defence signalling by NLRs often requires the formation of NLR heteropairs. Our knowledge of the molecular mechanism regulating this process is limited. In a reverse genetic screen to identify the partner of the Arabidopsis typical NLR, SUPRESSOR OF NPR1, CONSTITUTIVE 1 (SNC1), we discovered three NLRs that are redundantly required for SNC1-mediated defence, which were named SIDEKICK SNC1 1 (SIKIC1), SIKIC2 and SIKIC3. Immunoprecipitation-mass spectrometry analyses revealed that SIKIC2 physically associates with SNC1. We also uncovered that the protein level of SIKIC2 is under the control of two previously uncharacterized redundant E3 ubiquitin ligases MUSE1 and MUSE2. As SNC1 accumulation has previously been shown to be regulated by the E3 ubiquitin ligase SCFCPR1, this report provides evidence that the homeostasis of individual components of partnered typical NLRs is subjected to differential regulation via ubiquitin-mediated protein degradation.

Inorganic Hg toxicity in plants: A comparison of different genotoxic parameters.

Inorganic Mercury (Hg) contamination persists an environmental problem, but its cyto- and genotoxicity in plants remains yet unquantified. To determine the extent of Hg-induced cyto- and genotoxicity, and assess most sensitive endpoints in plants, Pisum sativum L. seedlings were exposed for 14 days to different HgCl2 concentrations up to 100 µM. Shoots and roots from hydroponic exposure presented growth impairment and/or morphological disorders for doses >1 µM, being the roots more sensitive. Plant growth, ploidy changes, clastogenicity (HPCV), cell cycle dynamics (G1-S-G2), Comet-tail moment (TM), Comet-TD, Mitotic-index (MI) and cell proliferation index (CPI) were used to evaluate Hg-induced cyto/genotoxicity. Both leaf and root DNA-ploidy levels, assessed by flow cytometry (FCM), remained unaltered after exposure. Root cell cycle impairment occurred at lower doses (≥1 µM) than structural DNA damages (≥10 µM). Cytostatic effects depended on the Hg concentration, with delays during S-phase at lower doses, and arrests at G1 at higher ones. This arrest was paralleled with decreases of both mitotic index (MI) and cell proliferation index (CPI). DNA fragmentation, assessed by the Comet assay parameters of TD and TM, could be visualized for conditions ≥10 µM, while FCM-clastogenic parameter (FPCV) and micronuclei (MNC) were only altered in roots exposed to 100 µM. We demonstrate that inorganic-Hg induced cytostaticity is detectable even at 1 µM (a value found in contaminated sites), while structural DNA breaks/damage are only visualized in plants at concentrations ≥10 µM. We also demonstrate that among the different techniques tested for cyto- and genotoxicity, TD and TM Comet endpoints were more sensitive than FPCV or MNC. Regarding cytostatic effects, cell cycle analysis by FCM, including the difference in % cell cycle phases and CPI were more sensitive than MI or MNC frequency. Our data contribute to better understand Hg cyto- and genotoxicity in plants and to understand the information and sensitivity provided by each of the genotoxic techniques used.

DNA damage as a consequence of NLR activation.

DNA damage observed during plant immune responses is reported to be an intrinsic component of plant immunity. However, other immune responses may suppress DNA damage to maintain host genome integrity. Here, we show that immunity-related DNA damage can be abrogated by preventing cell death triggered by Nucleotide-binding, Leucine-rich-repeat immune Receptors (NLRs). SNI1 (suppressor of npr1-1, inducible 1), a subunit of the structural maintenance of chromosome (SMC) 5/6 complex, was reported to be a negative regulator of systemic acquired resistance (SAR) and to be necessary for controlling DNA damage. We find that cell death and DNA damage in sni1 loss-of-function mutants are prevented by mutations in the NLR signaling component EDS1. Similar to sni1, elevated DNA damage is seen in other autoimmune mutants with cell death lesions, including camta3, pub13 and vad1, but not in dnd1, an autoimmune mutant with no visible cell death. We find that as in sni1, DNA damage in camta3 is EDS1-dependent, but that it is also NLR-dependent. Using the NLR RPM1 as a model, we also show that extensive DNA damage is observed when an NLR is directly triggered by effectors. We also find that the expression of DNA damage repair (DDR) genes in mutants with cell death lesions is down regulated, suggesting that degraded DNA that accumulates during cell death is a result of cellular dismantling and is not sensed as damaged DNA that calls for repair. Our observations also indicate that SNI1 is not directly involved in SAR or DNA damage accumulation.

Matching NLR Immune Receptors to Autoimmunity in camta3 Mutants Using Antimorphic NLR Alleles.

To establish infection, pathogens deploy effectors to modify or remove host proteins. Plant immune receptors with nucleotide-binding, leucine-rich repeat domains (NLRs) detect these modifications and trigger immunity. Plant NLRs thus guard host "guardees." A corollary is that autoimmunity may result from inappropriate NLR activation because mutations in plant guardees could trigger corresponding NLR guards. To explore these hypotheses, we expressed 108 dominant-negative (DN) Arabidopsis NLRs in various lesion mimic mutants, including camta3, which exhibits autoimmunity. CAMTA3 was previously described as a negative regulator of immunity, and we find that autoimmunity in camta3 is fully suppressed by expressing DNs of two NLRs, DSC1 and DSC2. Additionally, expression of either NLR triggers cell death that can be suppressed by CAMTA3 expression. These findings support a model in which DSC1 and DSC2 guard CAMTA3, and they suggest that other negative regulators of immunity may similarly represent guardees.

Making sense of plant autoimmunity and 'negative regulators'.

Genetics studies the structure/function of genes via the characterization of their mutant phenotypes. In plants, a readily scorable mutant phenotype comprises macroscopic lesions symptomatic of disease in the absence of pathogens. Such mutants therefore exhibit autoimmune phenotypes. Many of these mutants are considered to be associated with immunity and the corresponding genes have been described as 'negative regulators' of immunity and/or cell death. Pathogens deliver effectors into host cells to increase infectivity by modifying or removing host proteins. Plants detect effectors via nucleotide-binding, leucine-rich repeat (NLR) immune receptors, which monitor host effector targets. In response to effector-mediated target tampering, NLR proteins potentiate immunity. The guard hypothesis proposes that NLRs 'guard' host 'guardees' targeted by pathogen effectors. An obvious corollary to this guard model is that forms of plant autoimmunity are a result of inappropriate NLR protein activation. In this review, we discuss what is known about some of the 'negative regulators' of immunity, and propose simple strategies that may help to characterize autoimmune mutants.

Photosynthesis light-independent reactions are sensitive biomarkers to monitor lead phytotoxicity in a Pb-tolerant Pisum sativum cultivar.

Lead (Pb) environmental contamination remains prevalent. Pisum sativum L. plants have been used in ecotoxicological studies, but some cultivars showed to tolerate and accumulate some levels of Pb, opening new perspectives to their use in phytoremediation approaches. However, the putative use of pea plants in phytoremediation requires reliable toxicity endpoints. Here, we evaluated the sensitivity of a large number of photosynthesis-related biomarkers in Pb-exposed pea plants. Plants (cv. "Corne de Bélier") were exposed to Pb concentrations up to 1,000 mg kg(-1) soil during 28 days. The photosynthetic potential biomarkers that were analyzed included pigments, chlorophyll (Chl) a fluorescence, gas exchange, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) activity, and carbohydrates. Flow cytometry (FCM) was also used to assess the morpho-functional status of chloroplasts. Finally, Pb-induced nutrient disorders were also evaluated. Net CO2 assimilation rate (A) and RuBisCO activity decreased strongly in Pb-exposed plants. Plant dry mass (DM) accumulation, however, was only reduced in the higher Pb concentrations tested (500 and 1,000 mg kg(-1) soil). Pigment contents increased solely in plants exposed to the largest Pb concentration, and in addition, the parameters related to the light-dependent reactions of photosynthesis, Fv/Fm and ΦPSII, were not affected by Pb exposure. In contrast to this, carbohydrates showed an overall tendency to increase in Pb-exposed plants. The morphological status of chloroplasts was affected by Pb exposure, with a general trend of volume decrease and granularity increase. These results point the endpoints related to the light-independent reactions of photosynthesis as more sensitive predictors of Pb-toxicity than the light-dependent reactions ones. Among the endpoints related to the light-independent photosynthesis reactions, RuBisCO activity and A were found to be the most sensitive. We discuss here the advantages of using these parameters as biomarkers for Pb toxicity in plants. Finally, we report that, despite showing physiological disorders, these cultivar plants survived and accumulated high doses of Pb, and their use in environmental/decontamination studies is open to debate.

Autophagy deficiency leads to accumulation of ubiquitinated proteins, ER stress, and cell death in Arabidopsis.

Autophagy is a homeostatic degradation and recycling process that is also involved in defense against microbial pathogens and in certain forms of cellular suicide. Autophagy has been proposed to negatively regulate plant immunity-associated cell death related to the hypersensitive response (HR), as older autophagy-deficient mutants are unable to contain this type of cell death 5 to 10 d after infection. Such spreading cell death was found to require NPR1 (nonexpressor of PR genes 1), but surprisingly did not occur in younger atg mutants. In contrast, we find that npr1 mutants are not impaired in rapid programmed cell death activation upon pathogen recognition. Furthermore, our molecular evidence suggests that the NPR1-dependent spreading cell death in older atg mutants may originate from an inability to cope with excessive accumulation of ubiquitinated proteins and ER stress which derive from salicylic acid (SA)-dependent signaling (e.g., systemic acquired resistance). We also demonstrate that both senescence and immunity-related cell death seen in older atg mutants can be recapitulated in younger atg mutants primed with ER stress. We therefore propose that the reduction in SA signaling caused by npr1 loss-of-function is sufficient to alleviate the stress levels accumulated during aging in autophagy deficient cells which would otherwise become insurmountable and lead to uncontrolled cell death.

Zonal responses of sensitive vs. tolerant wheat roots during Al exposure and recovery.

Aluminium (Al) irreversibly inhibits root growth in sensitive, but not in some tolerant genotypes. To better understand tolerance mechanisms, seedlings from tolerant ('Barbela 7/72' line) and sensitive ('Anahuac') Triticum aestivum L. genotypes were exposed to AlCl(3) 185 µM for: (a) 24 h followed by 48 h without Al (recovery); (b) 72 h of continuous exposure. Three root zones were analyzed (meristematic (MZ), elongation (EZ) and hairy (HZ)) for callose deposition, reserves (starch and lipids) accumulation, endodermis differentiation and tissue architecture. Putative Al-induced genotoxic or cytostatic/mytogenic effects were assessed by flow cytometry in root apices. Tolerant plants accumulated less Al, presented less root damage and a less generalized callose distribution than sensitive ones. Starch and lipid reserves remained constant in tolerant roots but drastically decreased in sensitive ones. Al induced different profiles of endodermis differentiation: differentiation was promoted in EZ and HZ, respectively, in sensitive and tolerant genotypes. No ploidy changes or clastogenicity were observed. However, differences in cell cycle blockage profiles were detected, being less severe in tolerant roots. After Al removal, only the 'Barbela 7/72' line reversed Al-induced effects to values closer to the control, mostly with respect to callose deposition and cell cycle progression. We demonstrate for the first time that: (a) cell cycle progression is differently regulated by Al-tolerant and Al-sensitive genotypes; (b) Al induces callose deposition >3 cm above root apex (in HZ); (c) callose deposition is a transient Al-induced effect in tolerant plants; and (d) in HZ, endodermis differentiation is also stimulated only in tolerant plants, probably functioning in tolerant genotypes as a protective mechanism in addition to callose.

Chromium (VI) induces toxicity at different photosynthetic levels in pea.

In order to comprehensively characterize the effects of Cr (VI) on the photosynthetic performance of Pisum sativum, plants irrigated with Cr solutions (ranging from 20 to 2000 mg l(-1)) were evaluated using the following classical endpoints: gas exchange parameters, chlorophyll a (Chl a) fluorescence, leaf pigments, Rubisco activity, soluble sugars and starch content. Flow cytometry (FCM) was applied in an innovative approach to evaluate the morphological and fluorescence emission status of chloroplasts from plants exposed to Cr (VI). The parameters related to gas exchange, net CO(2) assimilation rate (A) and Rubisco activity were severally affected by Cr exposure, in some cases even at the lowest dosage used. While all biomarkers used to measure Chl a fluorescence indicated a decrease in fluorescence at the maximum dosage, pigment contents significantly increased in response to Cr (VI). The morphology of chloroplasts also was altered by Cr (VI) exposure, as a volume decrease was observed. Soluble sugars and starch showed an overall tendency to increase in Cr (VI) exposed plants, but sucrose and glucose decreased highly when exposed to 2000 mg l(-1). In conclusion, our results indicate that Cr (VI) affects photosynthesis at several levels, but the most Cr (VI)-sensitive endpoints were chloroplast morphology and biochemical processes; only at higher dosages the photochemical efficiency is compromised.