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  • 1
    ISSN: 0931-1890
    Keywords: Key words Chalcone synthase ; Flavonoids ; Phenylalanine ammonia-lyase ; Scots pine ; UV-B radiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract  Cultivation of Scots pine (Pinus sylvestris L.) seedlings under simulated global radiation including the UV-B band (280 – 320 nm; 220 mW m–2 UV-BBE) led to increased formation of the diacylated flavonol glucosides 3″,6″-di-p-coumaroyl-astragalin and 3″,6″-di-p-coumaroyl-isoquercitrin in primary and cotyledonary needles, respectively. 3″,6″-Di-p-coumaroyl-astragalin was also the main constitutive diacylated flavonol glucoside in both needle types. This compound predominantly accumulated in primary needles upon UV-B irradiation, and reached concentrations of 2.4 μmol g–1 fresh weight (fw). Its concentration was only weakly affected in cotyledonary needles. 3″,6″-Di-p-coumaroyl-isoquercitrin was mainly induced in cotyledonary needles with maximum concentrations of 0.8 to 0.9 μmol g–1 fw, but was virtually unaffected in primary needles under the same irradiation conditions. Pulse labelling with L-(U-14C)phenylalanine revealed that these metabolites were formed de novo. Phenylalanine ammonia-lyase (EC 4.3.1.5) and chalcone synthase (EC 2.3.1.74) were only slightly induced by the UV-B treatment. The results described here represent the first report on UV-B-induced flavonoid biosynthesis in a conifer species.
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  • 2
    ISSN: 0931-1890
    Keywords: Key words Beech ; Ozone ; Photosynthesis ; Closed chambers ; Critical Levels
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract  An exposure - response study with proportional-to-ambient ozone levels was conducted in closed chambers on 3-year-old European beech (Fagus sylvatica L.) of montane origin. The fumigation started in April 1990 and lasted for a single growing season. Climate data and ozone concentrations monitored at an experimental station of the Institute for Applied Plant Biology, Schönenbuch, Switzerland were simulated in the exposure chambers 12 days later (1*O3). To test exposure-response relations three additional treatments were applied, subambient (0.2*O3) and two proportionally increased ozone treatments (1.5*O3 and 2*O3). The photosynthetic behaviour of the trees in August revealed the light reactions to be less affected than parameters which are related to the dark reactions of photosynthesis. Assimilation (A350), apparent carboxylation efficiency (CE), and maximum photosynthetic capacity (A2500) were reduced with increasing ozone concentration. For the ozone response of CE and A2500′Critical Levels′ were calculated.
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  • 3
    ISSN: 0931-1890
    Keywords: Sirococcus shoot blight Controlled inoculation PCR detection PCR quantification
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract. Controlled inoculation of spruce seedling needle crowns and of shoots of 4-year-old spruce trees by Sirococcus conigenus led to disease symptoms (discoloration and necrosis) and to the induction of phenolic metabolites. Even upon complete infection, as proved by re-isolation of the pathogen from inoculated seedlings, only 40% of the plants developed visible disease symptoms after 38 days. A Sirococcus-specific polymerase chain reaction (PCR) primer pair, SIRO1 and SIRO6, was designed based on sequences of a RAPD fragment. The primer pair permitted the detection of 1 pg fungal DNA (10–40 genomes) in 1 mg fresh weight spruce tissues (needles, bark, wood), regardless of visible disease symptoms. The amounts of the major phenolic compound of spruce needles, catechin, increased significantly in all of the five spruce provenances as a response to inoculation with Sirococcus. The second major phenolic compound, picein, increased in three of the provenances, whereas the remaining two had high concentrations to begin with and showed no reaction. Minor phenolic compounds increased in response to infection regardless of provenance. In a preliminary field study, Sirococcus infection of spruce was detectable by PCR even in the presence of massive infection by other fungi, such as Rhizospaera spp. and Lophodermium spp.
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  • 4
    ISSN: 1365-2486
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Geography
    Notes: Elevated levels of both ozone and UV-B radiation are typical for high-altitude sites. Few studies have investigated their possible interaction on plants. This study reports interactive effects of O3 and UV-B radiation in four-year-old Norway spruce and Scots pine trees. The trees were cultivated in controlled environmental facilities under simulated climatic conditions recorded on Mt Wank, an Alpine mountain in Bavaria, and were exposed for one growing season to simulated ambient or twice-ambient ozone regimes at either near ambient or near zero UV-B radiation levels. Chlorotic mottling and yellowing of current year needles became obvious under twice-ambient O3 in both species at the onset of a high ozone episode in July. Development of chlorotic mottling in relation to accumulated ozone concentrations over a threshold of 40 nL L–1 was more pronounced with near zero rather than ambient UV-B radiation levels. In Norway spruce, photosynthetic parameters at ambient CO2 concentration, measured at the end of the experiment, were reduced in trees cultivated under twice-ambient O3, irrespective of the UV-B treatment. Effects on photosynthetic capacity and carboxylation efficiency were restricted to trees exposed to near zero levels of UV-B radiation, and twice-ambient O3. The data indicate that UV-B radiation, applied together with O3, ameliorates the detrimental effects of O3. The data also demonstrate that foliar symptoms develop more rapidly in Scots pine than in Norway spruce at higher accumulated ozone concentrations. Symbols and abbreviations: LSD, least significant difference; PAS300, UV-B irradiance weighted according to the plant action spectrum of Green et al. (1974) normalized at 300 (nm); AOT40, (AOT = accumulated over threshold) reflects the sum of hourly ozone concentrations above 40 nL L–1 during daylight hours (〉 50 Wm–2) ( Kärenlampi & Skärby 1996); A350, net photosynthesis at ambient CO2; G350, stomatal conductance for water vapour at ambient CO2; A2500, net photosynthesis at saturating CO2 (maximal potential photosynthetic activity); CE, carboxylation efficiency; ROS, reactive oxygen species; RuBP, ribulose 1,5-bisphosphate; Rubisco, ribulose 1,5-bisphosphate carboxylase/oxygenase; GLM, general linear model.
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  • 5
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Physiologia plantarum 100 (1997), S. 0 
    ISSN: 1399-3054
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Tropospheric ozone will increase during the next decades and will reach the critical threshold concentration of 40 nl l−1 more often in rural regions than in urban areas. Damage inflicted on plants by ozone can be divided into chronic and acute damage, reflecting different defence strategies of the plants. Chronic damage, somewhat similar to a premature senescence, follows disturbances in sugar metabolism, inhibition of photosynthesis, disbalances in the redox-status and ROS-production in the chloroplast stroma. The acute damage resembles the hypersensitive response which occurs after a pathogen attack. Either the accumulation of reactive oxygen species (ROS) or the efficiency of scavenging systems in the apoplast may govern the spread of cell death. Probably, different signalling pathways mediate the plant responses, such as ethylene, salicylate and ROS. ROS production and lipid peroxidation take place either on the plasma membrane or in the chloroplastic membrane and lipid hydroperoxides and derivatives, such as jasmonate, can act as signals for subsequent plant reactions. Both kinds of plant responses, i.e., acute as well as chronic damage, can affect the plants' abilities to respond to further stress attacks.
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  • 6
    ISSN: 1365-3040
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: To test the involvement of ethylene in mediating ozone-induced cell death and leaf loss in European beech (Fagus sylvatica L.), tree seedlings were exposed to proportionally increased or decreased field ozone levels for up to 6 months. Ozone treatment caused cell death and accelerated leaf loss at higher than ambient levels, but had only minor effects at ambient and no effects at subambient ozone levels. The emission of ethylene, the levels of its precursor, 1-aminocyclopropane-1-carboxylate (ACC), and mRNA levels of specific ACC synthase (FS-ACS2) and ACC oxidase (FS-ACO1) isoforms showed a persistent increase and preceded cell death by approximately 2 weeks. Inhibition of ethylene biosynthesis led to reduced lesion formation whereas application of ACC accelerated ozone-induced cell death and leaf loss. Similar results were obtained when adult beech trees were exposed to 2 × ozone by a whole tree free-air canopy exposure system. The results suggest a role of ethylene in amplifying ozone effects under field conditions in this major European broad-leaved tree species.
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  • 7
    ISSN: 1432-2048
    Keywords: Elicitor ; Enzyme induction ; Lophodermium ; Pinus ; Stilbene metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Treatment of Pinus sylvestris L. cell-suspension cultures with an elicitor preparation from the pine needle pathogen Lophodermium seditiosum, resulted in a severalhundredto thousandfold accumulation of the stilbenes pinosylvin and pinosylvin 3-O-methyl ether in methanolic cell extracts. There was a simultaneous induction of the biosynthetic enzymes phenylalanine ammonia-lyase (E.C. 4.3.1.5.) and stilbene synthase (pinosylvin-forming, E.C. 2.3.1.146). For the first time, an incorporation of stilbenes into the cell wall fraction as well as stilbene excretion into the extracellular space was demonstrated in addition to intracellular accumulation.
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  • 8
    ISSN: 1432-2048
    Keywords: 1-Aminocyclopropane-1-carboxylic acid ; Chitinase ; β-1,3-Glucanase ; Ethylene ; Helianthus cell-suspension cultures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Auxin-dependent, heterotrophic suspension cells of sunflower (Helianthus annuus L. C.K. Spanners All-zweck) showed, on a cell-protein basis, a seven-fold increase in chitinase activity, which began 5 d after treatment with 10−5 mol·L−1 of the triazole-type growth retardant BAS 111.W. In proportion to this increase, chitinase activity appeared to be excreted into the culture medium. The intracellular activity of β-1,3-glucanase, assayed fluorimetrically with laminarin as the substrate, was only slightly enhanced. Dose-response experiments with BAS 111.W showed that the onset of the induction of chitinase activity coincided with an inhibition of ethylene formation and an accumulation of endogenous 1-aminocyclopropane-1-carboxylic acid (ACC) as a result of blocking the conversion of ACC to ethylene. Other nitrogen-heterocyclic growth retardants (e.g. tetcyclacis, ancymidol), the triazole-type fungicide BAS 480.F, salicylic acid, CoCl2 and 2,4-dichlorophenoxy-acetic acid, which also increased the ACC/ethylene ratio, similarly induced chitinase activity. In contrast, aminoethoxy vinylglycine, which simultaneously lowered endogenous ACC and ethylene formation, did not stimulate chitinase activity. However, after addition of BAS 111.W and ACC, an accumulation of endogenous ACC was accompanied by a strong induction of the enzymatic activity. This effect did not correlate with changes in the cell culture growth nor in the cellular contents of immunoreactive abscisic acid, 3-indoleacetic acid, gibberellins or cytokinins. Furthermore, ethephon, which chemically generates ethylene, led to a slight reduction in ACC levels and tended to decrease chitinase activity relative to the control. In conclusion, it is hypothesized that the induction of chitinase activity in sunflower cell suspensions is antagonistically regulated by ethylene and ACC. At least at higher production rates, ethylene appears to function as an inhibiting factor whereas ACC may be a promoting one. The stimulation of chitinase and β-1,3-glucanase activity, caused by the retardant BAS 111.W and the fungicide BAS 480.F, is discussed as an additional effect of both compounds which possibly leads to an increased resistance of plants to fungal infections.
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  • 9
    ISSN: 1432-2048
    Keywords: Key words: Detoxification of formaldehyde ; Formal- dehyde dehydrogenase (glutathione-dependent) ; Protein sequence ; Zea (formaldehyde dehydrogenase)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Glutathione-dependent formaldehyde dehydrogenase (FDH; EC 1.2.1.1) has been purified 3900-fold from maize cell-suspension cultures to a specific activity of 4.68 μmol (mg protein)−1 min−1. The homogeneous enzyme consisted of two identical subunits with a molecular mass of 42 kDa, and an isoelectric point of 5.8. Eight tryptic peptides were sequenced and gave a perfect fit to the protein sequence derived from maize Fdh cDNA (J. Fliegmann and H. Sandermann, 1997, Plant Mol Biol 34: 843–854). There was 62% identity with the eucaryotic FDH consensus sequence. Michaelis constants of approx. 20 μm (formaldehyde), approx. 50 μm (glutathione) and approx. 31 μm (NAD+) were determined for the maize enzyme as well as for FDH partially purified from dog lung. Besides S-hydroxymethylglutathione, pentanol-1, octanol-1, and ω-hydroxyfatty acids served as substrates for both FDH preparations. The unusual substrate specificity indicates that FDH may be involved in the detoxification of long-chain lipid peroxidation products.
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  • 10
    ISSN: 1432-2048
    Keywords: Anthocyanin ; L-α-Aminooxy-β-phenylpropionic acid ; t-Cinnamic acid ; Daucus ; Phenylalanine ammonia lyase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cells of Daucus carota grown in a liquid medium produced large amounts of cyanidin as the only flavonoid aglycon. After inoculation in fresh medium a maximum activity of phenylalanine ammonia lyase (PAL; EC 4.3.1.5) was observed within 24 h. L-α-aminooxy-β-phenylpropionic acid (L-AOPP), thought to be a competitive inhibitor of PAL, inhibited cyanidin accumulation up to 80%. In order to study the regulatory role of PAL, the effects of L-AOPP and t-cinnamic acid, the product of the deamination of phenylalanine, were investigated. Cinnamic acid, applied in vivo (10-4 M), was not able to compensate for the inhibition of cyanidin production caused by L-AOPP (10-4 M) in the same sample. Carrot cells treated with L-AOPP exhibited a “super-induction” of PAL already described for gherkin hypocotyls (Amrhein and Gerhardt 1979). This effect was not influenced by t-cinnamic acid. L-AOPP seems to be a very specific inhibitor since it affected neither growth nor soluble protein content, whereas t-cinnamic acid inhibited both. Investigations on the content of soluble amino acids in L-AOPP-treated cells revealed a specific accumulation of soluble phenylalanine, whereas treatment with t-cinnamic acid led to an increase of amino acids in general, thus indicating that the latter compound has a rather unspecific effect on cellular metabolism. In vitro studies with PAL isolated from Daucus carota revealed that L-AOPP inhibited the enzyme at very low doses (K I=2.4·10-9), whereas t-cinnamic acid, by comparison, affected the enzyme at high concentrations (K I=1.8·10-4).
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