Phenolic Compounds and Peroxidases in Sunflower Near-Isogenic Lines After Downy Mildew Infection

Two near-isogenic lines of sunflower, resistant (+Pl6 gene) and susceptible (-P16 gene) to downy mildew were used in this paper. Secondary infection with a suspension of Plasmopara halstedii spores was done on the plants in the phase of first pair of leaves. In the samples taken 12 h after infection, content of phenolic compounds in methanolic extracts from frozen leaves was analysed by HPLC. POD activity was determined spectrophotmetrically and POD isoforms by isoelectrophoresis. Constitutive level of phenolic compounds and their accumulation after infection were higher in the susceptible than in the resistant line. Increased POD amount in leaf, constitutively present in NS-H-26R, was in correlation with increased guaiacol-dependent POD activity and low total phenolics contents. After infection, guaiacol- and chlorogenic acid-dependent POD activity significantly increased in both lines. Scopoletin-dependent POD activity was induced upon infection only in NS-H-26R. IEF electrophoresis revealed existance of four anionic isoforms of peroxidase in leaves of both lines. The main isoform with pI 5 was particularly intesified in the resistant line. In conclusion, scopoletin-dependent POD activity that was induced upon infection only in resistant NIL indicates a specific role of POD in coumarin metabolism that is possibly connected with the presence of Pl6 gene

WHEAT STRAW CONVERSION BY ENZYMATIC SYSTEM OF GANODERMA LUCIDUM

The purpose of this study was to resolve the question of whether various nitrogen sources and concentrations affect characteristics of selected G. lucidum ligninolytic enzymes participating in wheat straw fermentation. This is the first study reporting the presence of versatile peroxidase activity in crude extract of G. lucidum culture, as well as isoforms profile of Mn-oxidizing peroxidases. NH4NO3 was the optimum nitrogen source for laccase and Mn-dependent peroxidase activity, while peptone was the optimum one for versatile peroxidase activity. Four bands with laccase activity were obtained by native PAGE and IEF separations from medium enriched with inorganic nitrogen source, and only two bands from medium containing organic source. Medium composition was not shown to affect isoenzyme patterns of Mn-oxidizing peroxidases. Four isoforms of Mn-dependent peroxidase and three of versatile peroxidase were obtained on native PAGE. By IEF separation, five isoforms of Mn-dependent peroxidase and only two of versatile peroxidase were observed. The results demonstrated that G. lucidum has potential for mineralization and transformation of various agricultural residues and should take more significant participation in large-scale biotechnological processes

Comparison of Photoacoustic Signals in Photosynthetic and Nonphotosynthetic Leaf Tissues of Variegated Pelargonium zonale

Green-white variegated leaves of Pelargonium zonale were studied using the photoacoustic method. Our aim was to characterize photosynthetically active green tissue and nonphotosynthetically active white tissue by the photoacoustic amplitude signals. We observed lower stomatal conductance and higher leaf temperature in white tissue than in green tissue. Besides these thermal differences, significantly higher absorbance in green tissue was based on chlorophyll and carotenoids which were absent in white tissue. However, optical properties of epidermal layers of both tissues were equal. The photoacoustic amplitude of white tissue was over four times higher compared to green tissue, which was correlated with lower stomatal conductance. In addition, at frequencies gt 700 Hz, the significant differences between the photoacoustic signals of green and white tissue were obtained. We identified the photoacoustic signal deriving from photosynthetic oxygen evolution in green tissue, using high intensity of red light modulated at 10 Hz. Moreover, the photoacoustic amplitude of green tissue increased progressively with time which corresponded to the period of induction of photosynthetic oxygen evolution. For the first time, very high frequencies (1 kHz to 5 kHz) were applied on leaf material

Antioxidative response in variegated Pelargonium zonale leaves and generation of extracellular H2O2 in (peri)vascular tissue induced by sunlight and paraquat

In this study we exposed variegated leaves of Pelargonium zonale to strong sunlight (>1100 mu mol m(-2) s(-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

Comparison of Photoacoustic Signals in Photosynthetic and Nonphotosynthetic Leaf Tissues of Variegated Pelargonium zonale

Green-white variegated leaves of Pelargonium zonale were studied using the photoacoustic method. Our aim was to characterize photosynthetically active green tissue and nonphotosynthetically active white tissue by the photoacoustic amplitude signals. We observed lower stomatal conductance and higher leaf temperature in white tissue than in green tissue. Besides these thermal differences, significantly higher absorbance in green tissue was based on chlorophyll and carotenoids which were absent in white tissue. However, optical properties of epidermal layers of both tissues were equal. The photoacoustic amplitude of white tissue was over four times higher compared to green tissue, which was correlated with lower stomatal conductance. In addition, at frequencies >700 Hz, the significant differences between the photoacoustic signals of green and white tissue were obtained. We identified the photoacoustic signal deriving from photosynthetic oxygen evolution in green tissue, using high intensity of red light modulated at 10 Hz. Moreover, the photoacoustic amplitude of green tissue increased progressively with time which corresponded to the period of induction of photosynthetic oxygen evolution. For the first time, very high frequencies (1 kHz to 5 kHz) were applied on leaf material

Antifungal activity of quinhydrone against saccharomyces cerevisiae

Quinhydrone (QH) is a redox active charge transfer complex commonly used as a redox standard. Information on quinhydrone generation in plants is scarce and its physiological role is still unclear. Recently we have showed that excess zinc may induce oxidative stress through QH accumulation in the cell wall and stabilization of phenoxyl radicals [1]. The aim of our research was to investigate the antifungal activity of quinhydrone against yeast Saccharomyces cerevisiae (112, Hefebank Weihenstephan). Saccharomyces cerevisiae was grown on the Sabouraud maltose broth (HiMedia, Mumbai, India) in the presence of different concentrations of QH ranging from 75 mu M to 500 mu M. Concentrations of QH greater than 300 mu M had complete inhibitory effect on yeast growth, while lower concentrations (up to 200 mu M) did not affect the growth. QH had a significant impact on antioxidative defense enzymatic systems, indicated by the changes in the activity in catalase (CAT) and superoxide dismutase (SOD). CAT activities increased by 43% (150 mu M QH) and SOD activities by 122% and 60% at 150 mu M and 220 mu M QH respectively. However, abrupt inhibition of both enzymes was observed at concentrations higher than 220 mu M QH (>= 70%), to be almost completely diminished at 280 mu M QH. High molecular weight genomic DNA without any laddering or smearing was detected in both control and QH treated yeast cells, indicating the absence of apoptosis

Comparative analysis of phenolic profiles of ovipositional fluid of Rhinusa pilosa (Mecinini, Curculionidae) and its host plant Linaria vulgaris (Plantaginaceae)

Rhinusa pilosa (Gyllenhal) is a highly specific weevil that induces stem galls on the common toadflax Linaria vulgaris Mill. females oviposit the eggs near the apex of a growing shoot. The act of oviposition is accompanied by secretion of an ovipositional fluid, which is considered to be cecidogen, directly involved in gall induction. The remains of cecidogenic fluid were collected from the surface of the oviposition point on the stem. We performed a comparative analysis of the phenolics extracted from cecidogen, the stem and galls of L. vulgaris and adult and larva of R. pilosa by HPLC-DAD. One compound with A (max) at 273, 332 nm (R (t) 30.65 min) was exclusively found in the methanol extract of cecidogen. To further characterize the cecidogen and stem phenolic profiles, we used UHPLC coupled with an OrbiTrap mass analyzer. Among 49 phenolic compounds extracted from both the ovipositional fluid and the plant, protocatechuic acid and two phenolic glycosides were exclusively found in cecidogen: diosmetin-O-acetylrutinoside and an unidentified compound. The unknown compound produced an MS2 base peak at 387 and 327 and 267 m/z base peaks at MS3 and MS4 fragmentation, respectively, and had the molecular formula C32H31O18. The plausible role of phenolic compounds in the induction of gall formation on L. vulgaris is discussed