Structural Evidence of Programmed Cell Death Induction by Tungsten in Root Tip Cells of Pisum sativum

Previous studies have shown that excess tungsten (W), a rare heavy metal, is toxic to plant cells and may induce a kind of programmed cell death (PCD). In the present study we used transmission electron microscopy (TEM) and confocal laser scanning microscopy (CLSM) to investigate the subcellular malformations caused by W, supplied as 200 mg/L sodium tungstate (Na2WO4) for 12 or 24 h, in root tip cells of Pisum sativum (pea), The objective was to provide additional evidence in support of the notion of PCD induction and the presumed involvement of reactive oxygen species (ROS). It is shown ultrastructurally that W inhibited seedling growth, deranged root tip morphology, induced the collapse and deformation of vacuoles, degraded Golgi bodies, increased the incidence of multivesicular and multilamellar bodies, and caused the detachment of the plasma membrane from the cell walls. Plastids and mitochondria were also affected. By TEM, the endoplasmic reticulum appeared in aggregations of straight, curved or concentric cisternae, frequently enclosing cytoplasmic organelles, while by CLSM it appeared in bright ring-like aggregations and was severely disrupted in mitotic cells. However, no evidence of ROS increase was obtained. Overall, these findings support the view of a W-induced vacuolar destructive PCD without ROS enhancement

Chromium-Induced Ultrastructural Changes and Oxidative Stress in Roots of Arabidopsis thaliana

Chromium (Cr) is an abundant heavy metal in nature, toxic to living organisms. As it is widely used in industry and leather tanning, it may accumulate locally at high concentrations, raising concerns for human health hazards. Though Cr effects have extensively been investigated in animals and mammals, in plants they are poorly understood. The present study was then undertaken to determine the ultrastructural malformations induced by hexavalent chromium [Cr(VI)], the most toxic form provided as 100 μM potassium dichromate (K2Cr2O7), in the root tip cells of the model plant Arabidopsis thaliana. A concentration-dependent decrease of root growth and a time-dependent increase of dead cells, callose deposition, hydrogen peroxide (H2O2) production and peroxidase activity were found in Cr(VI)-treated seedlings, mostly at the transition root zone. In the same zone, nuclei remained ultrastructurally unaffected, but in the meristematic zone some nuclei displayed bulbous outgrowths or contained tubular structures. Endoplasmic reticulum (ER) was less affected under Cr(VI) stress, but Golgi bodies appeared severely disintegrated. Moreover, mitochondria and plastids became spherical and displayed translucent stroma with diminished internal membranes, but noteworthy is that their double-membrane envelopes remained structurally intact. Starch grains and electron dense deposits occurred in the plastids. Amorphous material was also deposited in the cell walls, the middle lamella and the vacuoles. Some vacuoles were collapsed, but the tonoplast appeared integral. The plasma membrane was structurally unaffected and the cytoplasm contained opaque lipid droplets and dense electron deposits. All electron dense deposits presumably consisted of Cr that is sequestered from sensitive sites, thus contributing to metal tolerance. It is concluded that the ultrastructural changes are reactive oxygen species (ROS)-correlated and the malformations observed are organelle specific

Hydrogen Peroxide Production by the Spot-Like Mode Action of Bisphenol A

Bisphenol A (BPA), an intermediate chemical used for synthesizing polycarbonate plastics, has now become a wide spread organic pollutant. It percolates from a variety of sources, and plants are among the first organisms to encounter, absorb, and metabolize it, while its toxic effects are not yet fully known. Therefore, we experimentally studied the effects of aqueous BPA solutions (50 and 100 mg L-1, for 6, 12, and 24 h) on photosystem II (PSII) functionality and evaluated the role of reactive oxygen species (ROS) on detached leaves of the model plantArabidopsis thaliana. Chlorophyll fluorescence imaging analysis revealed a spatiotemporal heterogeneity in the quantum yields of light energy partitioning at PSII inArabidopsisleaves exposed to BPA. Under low light PSII function was negatively influenced only at the spot-affected BPA zone in a dose- and time-dependent manner, while at the whole leaf only the maximum photochemical efficiency (Fv/Fm) was negatively affected. However, under high light all PSII photosynthetic parameters measured were negatively affected by BPA application, in a time-dependent manner. The affected leaf areas by the spot-like mode of BPA action showed reduced chlorophyll autofluorescence and increased accumulation of hydrogen peroxide (H2O2). When H(2)O(2)was scavengedviaN-acetylcysteine under BPA exposure, PSII functionality was suspended, while H(2)O(2)scavenging under non-stress had more detrimental effects on PSII function than BPA alone. It can be concluded that the necrotic death-like spots under BPA exposure could be due to ROS accumulation, but also H(2)O(2)generation seems to play a role in the leaf response against BPA-related stress conditions

Arbuscular Mycorrhizal Symbiosis Enhances Photosynthesis in the Medicinal Herb Salvia fruticosa by Improving Photosystem II Photochemistry

We investigated the influence of Salvia fruticosa colonization by the arbuscular mycorrhizal fungi (AMF) Rhizophagus irregularis on photosynthetic function by using chlorophyll fluorescence imaging analysis to evaluate the light energy use in photosystem II (PSII) of inoculated and non-inoculated plants. We observed that inoculated plants used significantly higher absorbed energy in photochemistry (ΦPSII) than non-inoculated and exhibited significant lower excess excitation energy (EXC). However, the increased ΦPSII in inoculated plants did not result in a reduced non-regulated energy loss in PSII (ΦNO), suggesting the same singlet oxygen (1O2) formation between inoculated and non-inoculated plants. The increased ΦPSII in inoculated plants was due to an increased efficiency of open PSII centers to utilize the absorbed light (Fv’/Fm’) due to a decreased non-photochemical quenching (NPQ) since there was no difference in the fraction of open reaction centers (qp). The decreased NPQ in inoculated plants resulted in an increased electron-transport rate (ETR) compared to non-inoculated. Yet, inoculated plants exhibited a higher efficiency of the water-splitting complex on the donor side of PSII as revealed by the increased Fv/Fo ratio. A spatial heterogeneity between the leaf tip and the leaf base for the parameters ΦPSII and ΦNPQ was observed in both inoculated and non-inoculated plants, reflecting different developmental zones. Overall, our findings suggest that the increased ETR of inoculated S. fruticosa contributes to increased photosynthetic performance, providing growth advantages to inoculated plants by increasing their aboveground biomass, mainly by increasing leaf biomass

Leaf Age-Dependent Photoprotective and Antioxidative Response Mechanisms to Paraquat-Induced Oxidative Stress in Arabidopsis thaliana

Exposure of Arabidopsis thaliana young and mature leaves to the herbicide paraquat (Pq) resulted in a localized increase of hydrogen peroxide (H2O2) in the leaf veins and the neighboring mesophyll cells, but this increase was not similar in the two leaf types. Increased H2O2 production was concomitant with closed reaction centers (qP). Thirty min after Pq exposure despite the induction of the photoprotective mechanism of non-photochemical quenching (NPQ) in mature leaves, H2O2 production was lower in young leaves mainly due to the higher increase activity of ascorbate peroxidase (APX). Later, 60 min after Pq exposure, the total antioxidant capacity of young leaves was not sufficient to scavenge the excess reactive oxygen species (ROS) that were formed, and thus, a higher H2O2 accumulation in young leaves occurred. The energy allocation of absorbed light in photosystem II (PSII) suggests the existence of a differential photoprotective regulatory mechanism in the two leaf types to the time-course Pq exposure accompanied by differential antioxidant protection mechanisms. It is concluded that tolerance to Pq-induced oxidative stress is related to the redox state of quinone A (QA)

High anthocyanin accumulation in poinsettia leaves is accompanied by thylakoid membrane unstacking, acting as a photoprotective mechanism, to prevent ROS formation

In poinsettia (Euphorbia pulcherrima Willd. ex Klotzsch.), green leaves turn reddish and then red, due to vacuolar anthocyanin accumulation. Reddish leaves accumulate anthocyanins mainly in the adaxial (upper) epidermis, and less in the mesophyll cells, while red, in both adaxial and abaxial (lower) epidermides, and the adjacent mesophyll cells. In green leaves, the photoprotective mechanism of non-photochemical quenching (NPQ) is sufficient under low light (LL), but not under high light (HL), which results in a more reduced redox state of the plastoquinone (PQ) pool compared to reddish. In red leaves, higher anthocyanin accumulation is accompanied by unstacking of thylakoid membranes that results in loss of photosystem II (PSII) complexes and undetectable hydrogen peroxide (H2O2). Superoxide dismutase (SOD) activity is enhanced in the reddish compared to green leaves, while it decreases significantly in the red ones. Anthocyanin accumulation was significantly correlated to the redox state of the PQ pool and the higher accumulation in reddish compared to green leaves was responsible for the diminished H2O2 production, since ascorbate peroxidase (APX) activity remained unchanged in all leaves. We suggest that H2O2 production regulated by the redox state of the PQ pool induces anthocyanin biosynthesis that is accompanied by thylakoid membrane unstacking and loss of PSII complexes, serving as a photoprotective mechanism to HL, preventing the formation of reactive oxygen species (ROS)