RESUMO
Quinoa is a facultative halophyte with excellent tolerance to salinity. In this study, the epidermal bladder cell complex (EBCc) of quinoa leaves was studied to determine their cellular characteristics and involvement in salt tolerance. We used light microscopy, confocal RAMAN microscopy, confocal fluorescence microscopy, transmission electron microscopy, and environmental scanning electron microscopy complemented by energy dispersive X-ray analysis. Ionic content was quantified with flame atomic absorption spectroscopy and with flame emission photometry. Results show that: (i) the number of EBCcs remains constant but their density and area vary with leaf age; (ii) stalk cells store lipids and exhibit thick walls, bladder cells present carotenes in small vesicles, oxalate crystals in vacuoles and lignin in their walls and both stalk and bladder cells have cuticles that differ in wax and cutin content; (iii) chloroplasts containing starch can be found on both stalk and bladder cells, and the latter also presents grana; (iv) plasmodesmata are observed between the stalk cell and the bladder cell, and between the epidermal cell and the stalk cell, and ectodesmata-like structures are observed on the bladder cell. Under high salinity conditions, (v) there is a clear tendency to accumulate greater amounts of K+ with respect to Na+ in the bladder cell; (vi) stalk cells accumulate similar amounts of K+ and Na+; (vii) Na+ accumulates mainly in the medullary parenchyma of the stem. These results add knowledge about the structure, content, and role of EBCc under salt stress, and surprisingly present the parenchyma of the stem as the main area of Na+ accumulation.
Assuntos
Chenopodium quinoa , Epiderme Vegetal , Chenopodium quinoa/metabolismo , Chenopodium quinoa/química , Epiderme Vegetal/ultraestrutura , Epiderme Vegetal/citologia , Epiderme Vegetal/metabolismo , Estresse Salino , Cátions , Folhas de Planta/ultraestrutura , Folhas de Planta/metabolismo , SalinidadeRESUMO
In willow seeds, photooxidative damage is mainly restricted to the outer cotyledonary tissues, significantly reducing normal germination. Here we analyzed the damage generated in cotyledonary tissues and investigated whether the increase in reactive oxygen species (ROS) generation in seedlings from photooxidized seeds can affect the morphogenetic capacity of the shoot apical meristem. Seeds were photooxidized under different light intensities and the evolution of the damage during seedling growth was studied by light and transmission electron microscopies. The level of lipid peroxidation and changes in antioxidant capacity were measured following the time course of superoxide dismutase, catalase, ascorbate peroxidase and guaiacol peroxidase enzyme activities, and the effect of photooxidative stress on the genesis of new leaf primordia and lateral roots was examined. Early and active endocytosis and autophagy, changes in chloroplast morphology, as well as the accumulation and diffusion of ROS all play important roles in the early cell death observed in cotyledonary tissues. Following germination, seedlings from photooxidized seeds anticipated the emergence of first leaves, which complemented the altered functionality of the damaged cotyledons.
Assuntos
Cotilédone/crescimento & desenvolvimento , Folhas de Planta/crescimento & desenvolvimento , Salix/crescimento & desenvolvimento , Plântula/crescimento & desenvolvimento , Sementes/crescimento & desenvolvimento , Análise de Variância , Antioxidantes/metabolismo , Ascorbato Peroxidases/metabolismo , Catalase/metabolismo , Cloroplastos/metabolismo , Cotilédone/metabolismo , Cotilédone/ultraestrutura , Germinação , Luz , Peroxidação de Lipídeos , Malondialdeído/metabolismo , Microscopia Eletrônica de Transmissão , Oxirredução/efeitos da radiação , Peroxidase/metabolismo , Folhas de Planta/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Salix/metabolismo , Plântula/metabolismo , Plântula/ultraestrutura , Sementes/metabolismo , Sementes/efeitos da radiação , Estresse Fisiológico/efeitos da radiação , Superóxido Dismutase/metabolismo , Fatores de TempoRESUMO
Salix nigra seeds subjected to increased humidification show a decrease in normal germination (NG) during early imbibition followed by a recovery in that parameter at increasing imbibition times. Since photo-oxidized seeds contain high levels of reactive oxygen species (ROS), it is possible to infer that the atypical decrease in NG is a consequence of a higher ROS mobilization at early imbibition and the subsequent recovery from an increase in antioxidant activity. In this study, several oxidative stress indicators were evaluated in photo-oxidized seeds subjected to priming. ROS production was studied using electronic spin resonance spectroscopy, spontaneous chemiluminescence (SCL), spectrophotometry (with XTT), and histochemical (with DAB and NBT) and cytochemical (with CeCl(3)) techniques. Four indicators of molecular damage were monitored: lipid peroxidation, pigment destruction, protein oxidation, and membrane integrity. Antioxidant activity was evaluated by changes in the enzymes SOD, CAT, APX, and POX. The results revealed that the decrease in NG at the beginning of priming occurs by an oxidative burst, as determined by increases in both SCL and superoxide anion radical (O2(·-)) Such oxidative burst generates lipid peroxidation, protein oxidation, and a decrease in both pigment content and enzyme activities. With increasing hydration, damages are progressively reversed and NG restored, which coincides with the increased activity of antioxidant defences. It is proposed that these novel observations regarding the occurrence of an oxidative burst are related to the high basal ROS levels and the high membrane content retained in the mature embryo tissues.
Assuntos
Estresse Oxidativo , Salix/metabolismo , Sementes/crescimento & desenvolvimento , Germinação , Proteínas de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Salix/crescimento & desenvolvimento , Sementes/metabolismo , Água/metabolismoRESUMO
The production of reactive oxygen species (ROS) in specific regions of Salix seedlings roots seems essential for the normal growth of this organ. We examined the role of different ROS in the control of root development in Salix nigra seedlings, and explored possible mechanisms involved in the regulation of ROS generation and action. Root growth was not significantly affected by OH quenchers, while it was either partially or completely inhibited in the presence of H2O2 or O2·â» scavengers, respectively. O2·â» production was elevated in the root apex, particularly in the subapical meristem and protodermal zones. Apical O2·â» generation activity was correlated to a high level of either Cu/Zn superoxide dismutase protein as well as carbonylated proteins. While NADPH-oxidase (NOX) was probably the main source of O2·â» generation, the existence of other sources should not be discarded. O2·â» production was also high in root hairs during budding, but it markedly decreased when the hair began to actively elongate. Root hair formation increased in the presence of H2O2 scavengers, and was suppressed when H2O2 or peroxidase inhibitors were supplied. The negative effect of H2O2 was partially counteracted by a MAPKK inhibitor. Possible mechanisms of action of the different ROS in comparison with other plant model systems are discussed.
Assuntos
Raízes de Plantas/crescimento & desenvolvimento , Espécies Reativas de Oxigênio/metabolismo , Salix/crescimento & desenvolvimento , Plântula/crescimento & desenvolvimento , Western Blotting , Eletroforese em Gel de Poliacrilamida , Peróxido de Hidrogênio/metabolismo , NADPH Oxidases/metabolismo , Peroxidase/metabolismo , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Salix/metabolismo , Plântula/metabolismo , Superóxidos/metabolismoRESUMO
BACKGROUND AND AIMS: Salix nigra seeds are desiccation-tolerant, as are orthodox seeds, although in contrast to other orthodox seeds they lose viability in a few weeks at room temperature. They also differ in that the chloroplasts of the embryo tissues conserve their chlorophyll and endomembranes. The aim of this paper was to investigate the role of chlorophyll in seed deterioration. METHODS: Seeds were aged at different light intensities and atmospheric conditions. Mean germination time and normal and total germination were evaluated. The formation of free radicals was assessed using electronic spin resonance spectroscopy, and changes in the fatty acid composition from phospholipids, galactolipids and triglycerides using gas-liquid chromatography. Membrane integrity was studied with electronic spin resonance spin probe techniques, electrolyte leakage and transmission electron microscopy. KEY RESULTS: Light and oxygen played an important role in free-radical generation, causing a decrease in normal germination and an increase in mean germination time. Both indices were associated with a decrease in polyunsaturated fatty acids derived from membrane lipids as phospholipids and galactolipids. The detection of damage in thylakoid membranes and an increase in plasmalemma permeability were consistent with the decrease in both types of lipids. Triglycerides remained unchanged. Light-induced damage began in outermost tissues and spread inwards, decreasing normal germination. CONCLUSIONS: Salix nigra seeds were very susceptible to photooxidation. The thylakoid membranes appeared to be the first target of the photooxidative process since there were large decreases in galactolipids and both these lipids and the activated chlorophyll are contiguous in the structure of that membrane. Changes in normal germination and mean germination time could be explained by the deteriorative effects of oxidation.
Assuntos
Luz , Membranas/efeitos da radiação , Salix/efeitos da radiação , Sementes/efeitos da radiação , Adaptação Fisiológica , Clorofila/efeitos da radiação , Espectroscopia de Ressonância Magnética/métodos , Membranas/fisiologia , Oxirredução , Oxigênio/metabolismo , Fotoquímica , Salix/fisiologia , Sementes/fisiologiaRESUMO
The Zfhx1a gene expresses two different isoforms; the full length Zfhx1a-1 and a truncated isoform termed Zfhx1a-2 lacking the first exon. Deletion analysis of the Zfhx1a-1 promoter localized cell-specific repressors, and a proximal G-string that is critically required for transactivation. Transfection of Zfhx1a-1 cDNA, but not Zfhx1a-2, downregulates Zfhx1a-1 promoter activity. Mutation of an E2-box disrupted the binding of both Zfhx1a isoforms. Consistent with this, transfected Zfhx1a-1 does not regulate the transcriptional activity of the E-box mutated Zfhx1a-1 promoter. Competitive EMSAs and transfection assays show that Zfhx1a-2 can function as a dominant negative isoform since it is able to compete and displace Zfhx1a-1 from its binding site and overcome Zfhx1a-1 induced repression of the Zfhx1a-1 promoter in cells. Hence, the Zfhx1a-1 gene is autoregulated in part by negative feedback on its own promoter which is, in turn, modified by the availability of the negative dominant isoform Zfhx1a-2.