ABSTRACT
In this study we exposed variegated leaves of Pelargonium zonale to strong sunlight (>1100µmolm-2s-1 of photosynthetically active radiation) with and without paraquat (Pq), with the aim to elucidate the mechanisms of H2O2 regulation in green and white tissues with respect to the photosynthetically-dependent generation of reactive oxygen species (ROS). Sunlight induced marked accumulation of H2O2 in the apoplast of vascular and (peri)vascular tissues only in green sectors. This effect was enhanced by the addition of Pq. In the presence of diphenyl iodide, an NADPH oxidase inhibitor, H2O2 accumulation was abolished. Distinct light-induced responses were observed: in photosynthetic cells, sunlight rapidly provoked ascorbate (Asc) biosynthesis and an increase of glutathione reductase (GR) and catalase activities, while in non-photosynthetic cells, early up-regulation of soluble ascorbate peroxidase, dehydroascorbate reductase (DHAR) and GR activities was observed. Paraquat addition stimulated DHAR and GR activities in green sectors, while in white sectors activities of monodehydroascorbate reductase, DHAR and class III peroxidases, as well as Asc content rapidly increased. Differential antioxidative responses in the two tissues in the frame of their contrasting metabolisms, and the possible role of (peri)vascular H2O2 in signaling were discussed.
Subject(s)
Antioxidants/metabolism , Hydrogen Peroxide/metabolism , Paraquat/toxicity , Pelargonium/drug effects , Pelargonium/metabolism , Plant Leaves/metabolism , Plant Vascular Bundle/metabolism , Sunlight , Ascorbic Acid/metabolism , Catalase/metabolism , Extracellular Space/metabolism , Glutathione/metabolism , Pelargonium/radiation effects , Peroxidases/metabolism , Photosynthesis/drug effects , Photosynthesis/radiation effects , Plant Leaves/drug effects , Plant Leaves/enzymology , Plant Leaves/radiation effects , Plant Stomata/drug effects , Plant Stomata/physiology , Plant Stomata/radiation effects , Plant Vascular Bundle/drug effects , Plant Vascular Bundle/radiation effectsABSTRACT
Changes in cell walls have been previously observed in the mature infection organ, or haustorium, of the parasitic angiosperm Cuscuta, but are not equally well charted in young haustoria. In this study, we focused on the molecular processes in the early stages of developing haustoria; that is, before the parasite engages in a physiological contact with its host. We describe first the identification of differentially expressed genes in young haustoria whose development was induced by far-red light and tactile stimuli in the absence of a host plant by suppression subtractive hybridization. To improve sequence information and to aid in the identification of the obtained candidates, reference transcriptomes derived from two species of Cuscuta, C. gronovii and C. reflexa, were generated. Subsequent quantitative gene expression analysis with different tissues of C. reflexa revealed that among the genes that were up-regulated in young haustoria, two xyloglucan endotransglucosylase/hydrolase (XTH) genes were highly expressed almost exclusively at the onset of haustorium development. The same expression pattern was also found for the closest XTH homologues from C. gronovii. In situ assays for XTH-specific action suggested that xyloglucan endotransglucosylation was most pronounced in the cell walls of the swelling area of the haustorium facing the host plant, but was also detectable in later stages of haustoriogenesis. We propose that xyloglucan remodelling by Cuscuta XTHs prepares the parasite for host infection and possibly aids the invasive growth of the haustorium.
Subject(s)
Cuscuta/anatomy & histology , Cuscuta/enzymology , Glycosyltransferases/metabolism , Host-Parasite Interactions , Pelargonium/parasitology , Cell Wall/genetics , Cell Wall/radiation effects , Cuscuta/genetics , Gene Expression Profiling , Gene Expression Regulation, Plant/radiation effects , Genes, Plant , Host-Parasite Interactions/radiation effects , Light , Molecular Sequence Annotation , Pelargonium/radiation effects , Phylogeny , RNA, Messenger/genetics , RNA, Messenger/metabolism , Real-Time Polymerase Chain Reaction , Reproducibility of Results , Sequence Analysis, RNA , Species Specificity , Transcriptome/genetics , Transcriptome/radiation effectsABSTRACT
Light absorption was quantified in the following systems: isolated chloroplasts and leaves of spinach (Spinacea oleracea L.), a mutant of geranium (Pelargonium zonale L.) widely differing in pigment content, and coleus (Coleus blumei Benth.) at different stages of leaf ontogenesis. For these species and pea (Pisum sativum L.), scattering-compensated absorption spectra of chloroplast suspensions are presented. Comparison of leaf and chloroplast spectra showed considerable changes in the extent of the 'package' effect and the lengthening of the effective optical path in a leaf. The difference between leaf and isolated chloroplast absorption could be quantitatively described by adapting Duysens's treatment of flattening. It was found that the accumulation of chlorophyll in leaves is accompanied by a monotonous enhancement of the package effect. The results are discussed with special reference to the role of light scattering in leaf optics, light utilization in photosynthesis and wavelength-dependent light gradients in a leaf.
