Identification of pterostilbene as a phytoalexin from Vitis vinifera leaves

An inducible antifungal compound in grapevine leaves (Vitis vinifera L., cv Cabernet-Sauvignon) has been identified as trans-pterostilbene (3,5-dimethoxy-4′-hydroxy stilbene). It is only a minor component of the phytoalexin response of V. vinifera but its antifungal activity is relatively high by comparison with resveratrol and the viniferins, stress metabolites which have been identified previously in grapevine. Methods for the quantitative analysis of pterostilbene, resveratrol, ε- and α-viniferins by HPLC are described.     

Starch synthetases from Vitis vinifera and zea mays

ADPglucose: α-1,4-glucan α-4-glucosyltransferases (starch synthetases) from leaves of Vitis vinifera and leaves and kernels of Zea mays were chromatographed on DEAE-cellulose columns. One form of the enzyme was present in grape leaves having activity both in the presence and absence of primer. Two forms were present in both leaves and kernels of maize. The second peak of activity in maize leaves and the first peak in maize kernels synthesized a polyglucan in the absence of primer. A peak of branching enzyme (Q-enzyme) occurred between the two starch synthetase peaks with both tissues. When fractions containing starch synthetase and branching enzyme were added to the first leaf starch synthetase peak, up to 100-fold activation of the unprimed reaction occurred. Branching enzyme did not stimulate the unprimed activity of the first kernel peak and no branching enzyme could be detected in this peak. The unprimed product was a branched polyglucan with mainly α-1,4-links.     

Temperature and enzymic control of malate metabolism in berries of Vitis vinifera

The specific activities of grape enzymes concerned with malic acid metabolism were determined at various stages of berry development under two temperature regimes. It was found that the lower acidity levels occurring in high temperature grapes at maturity are due to a changed pattern in acid breakdown rather than to reduced malic acid accumulation. The differences in acid disappearance cannot be explained by a shift in relative enzyme activities towards malate consumption with rising temperature.     

Control of glycolysis in ripening berries of Vitis vinifera

The concentrations of glycolytic intermediates, acid components and adenosine nucleotides were determined at half-weekly intervals during development and ripening of grape berries. Based on distinctive non-equilibrium conditions and enzymic activities which are not controlled by substrate availability at the levels of phosphoenolpyruvate/pyruvate and fructose-6-phosphate/fructose diphosphate it is concluded that these two sites represent the major control points in the reaction sequences between sugar and acid pools in this fruit.     

Enzymatic activities of starch synthesis in potato tubers of different sizes

The objective of this study was to determine the relationship between tuber weight and enzymatic activities involved in tuber starch synthesis. As tuber weight increased, the activities of sucrose synthetase, UDPG pyrophosphorylase, and granular starch synthetase escalated, whereas the activities of soluble starch synthetase and ADPG pyrophosphorylase stayed constant and that of phosphorylase declined. This suggests that when samples are taken to determine specific enzymatic activities, the sampling procedure should ensure that results do not vary because of differences in the tuber weight or size distribution.     

Enzymes of starch and sucrose metabolism in Zea mays leaves

Mesophyll and bundle sheath cells of maize leaves were separated and enzymes of starch and sucrose metabolism assayed. The starch content and activities of ADPglucose (ADPG) starch synthetase and phosphorylase expressed both on a chlorophyll and a protein basis were much lower in mesophyll cells compared to bundle sheath preparations. Exposure of the leaves to continuous illumination for 2·5 days caused the starch content of mesophyll cells to rise greatly and led to considerable increases in ADPG starch synthetase and phosphorylase activity. In glasshouse grown leaves the bulk of invertase, sucrose phosphate synthetase, sucrose phosphatase, UDPglucose pyrophosphorylase and amylase was situated in the mesophyll layer. Sucrose synthetase, ADPG starch synthetase and phosphorylase were largely confined to the bundle sheath. No enzyme could be completely assigned to one particular cell layer. Upon continuous illumination both ADPG starch synthetase and phosphorylase increased in the mesophyll bythe same relative amount. The mesophyll is likely to be a major site for sucrose synthesis in maize leaves.     

Enzymes of starch metabolism in developing grains of high lysine barley mutant

The absolute activities of ADPG(UDPG)-pyrophosphorylase, starch phosphorylase, ADPG(UDPG)-starch synthetase, NDP-kinase and inorganic pyrophosphatase have been studied in high lysine mutant barley Notch-2 and its parent NP 113 grains during development. In general, mutant Notch-2 grains had higher average activities of UDPG-pyrophosphorylase and starch phosphorylase and lower activity of ADPG(UDPG)-starch synthetase per grain than the parent NP 113 during grain development. Activities of NDP-kinase, ADPG-pyrophosphorylase and inorganic pyrophosphatase differed only to a small extent between the mutant Notch-2 and NP 113. It is suggested that the lower activity of ADPG(UDPG)-starch synthetase might be responsible for the reduced accumulation of starch in the mutant Notch-2 grain as compared with parent NP 113 during development.     

Effect of temperature on lipid,starch and enzymes of starch metabolism in grape,tomato and broad bean leaves

Grapevine (Vitis vinifera cv Cabernet Sauvignon) leaves have previously been shown to accumulate starch at temperatures of ca 18/13° (day/nig     

Isoelectric point changes in Vitis vinifera catechol oxidase

A comparison between electrophoretic and isoelectric focussing patterns of grape catechol oxidase is reported. A relationship exists between changes in     

Multiple forms of Vitis vinifera catechol oxidase

Grape catechol oxidase shows multiple forms upon ion exchange chromatography, acrylamide gel electrophoresis and gel filtration. Conversion of some bands into others, which occurs during isolation and storage, is enhanced by dilution and by treatment with urea or acid pH. Estimation of MWs suggested that the conversions might be due to dissociation of the enzyme into subunits, but attempts to induce reassociation were unsuccessful. The effects of urea and acid pH could be imitated by partial enzymic proteolysis of the enzyme. Analysis of the various bands observed in gel electrophoresis suggested that some of the enzyme forms have the same MWs but differ in charge distribution.     

Enzymes of starch and sugar phosphate metabolism in achlorophyllous ribosome-deficient plastids from high-temperature-grown rye leaves

Several enzymes of non–photosynthetic sugar phosphate and starch metabolism were measured in gradient–purified chloroplasts from normal rye leaves ( Secale cereale L. cv. Halo) grown at 22°C and in the non-photosynthetic plastids isolated from 70S ribosome-deficient rye leaves grown at a non–permissive elevated temperature of 32°C. Activities of the enzymes phosphoglycerate kinase (EC 2.7.2.3), hexokinase (EC 2.7.1.1), phosphoglucose isomerase (EC 5.3.1.9), phosphoglucomutase (EC 2.7.5.1), glucose-6-phosphate dehydrogenase (EC 1.1.1.49), 6-phosphogluconate de-hydrogenase (EC 1.1.1.46), ADPglucose pyrophosphorylase (EC 2.7.7.27), starch synthase (EC 2.4.1.21), and phosphorylase (EC 2.4.1.1) were present in ribosome-deficient plastids from 32°C-grown leaves indicating a cytoplasmic origin of the plastid-specific forms of these enzymes. While the photosynthetic marker enzyme NADP⁺-dependent glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.13) was considerably diminished, both the specific activities and the total activities per leaf of the plastid-specific forms of hexokinase, phosphoglucose isomerase and phosphoglucomutase were markedly increased in the ribosome–deficient plastids, relative to normal chloroplasts. The results demonstrate that after elimination of functional protein synthesis in the chloroplasts the supply of chloroplast–specific enzymes by the cytoplasm is not generally suppressed as observed for many enzymes and proteins involved in photosynthesis, but may even be increased in accord with changed metabolic demands.     

Phosphorylase activity in relation to starch synthesis in developing Hordeum distichum grain

Activity, control and primer requirements of starch phosphorylase in developing barley endosperm were investigated. Phosphorylase was detected in endosperm extracts from 3 days after anthesis. Unprimed activity was predominant between 2 and 10 days after anthesis, when it constituted 70–80% of total activity, but this proportion declined rapidly as the grain developed. The existence of at least 2 isoenzymes was indicated by studies of pH dependence and phosphate inhibition, and was further supported by acrylamide gel electrophoresis and column chromatography using DEAE-cellulose. The two isoenzymes which ere possibly both glyco proteins, appear in barley endosperm soon after anthesis. One appears capable of unprimed activity, and may be associated with the initiation of a-1,2 glucans, which then serve as primers for starch synthetase. This disappears by 13–15 days after anthesis. The other isoenzyme is capable of some unprimed activity but undergoes modification between 15 and 20 days after anthesis, resulting in the loss of unprimed activity. The relevance of the results to initiation of starch synthesis and to starch synthetase in amyloplasts is discussed.     

Enzymes involved in starch synthesis in the developing mung bean seed

The levels of free sugars, starch and enzymes involved in starch metabolism—sucrose synthetase, UDP and ADP glucose pyrophosphorylase, phosphorylase and starch synthetase—were assayed during seed development of three cultivars of mung bean (Vigna radiata). Free sugars and starch increased with increasing seed weight. Changes in levels of sucrose synthetase, UDP- and ADP-glucose pyrophosphorylases, and phosphorylase were paralleled by changes in starch accumulation. After the maximum activity levels of these enzymes had been reached, maximum activities of soluble starch synthetase and starch granule-bound starch synthetase occurred. There were high activities of sucrose synthetase and phosphorylase at maximum rates of starch accumulation. Thus, starch could be synthesized via the ADP glucose pathway in mung bean seeds. However, phosphorylase may account for the starch accumulation in the early stages of mung bean seed development.     

Enzymes of starch metabolism in Nicotiana tabacum callus

Activities of enzymes of starch metabolism were determined in tobacco callus grown in light or darkness and in the presence or absence of gibberellic acid. There was a higher rate of starch turnover in light-grown cultures, as judged by the activities of synthetic and degradative enzymes. Gibberellic acid-treated tissues contained a lower level of starch than the corresponding control tissue. This decrease could be correlated with the activity of phosphorylase. Cultured tissue and seedling material were found to have comparable levels of activity for the starch metabolizing enzymes.     

The relationship of starch metabolism to grain size in wheat

The present investigation was aimed at determining the levels of important enzymes of starch metabolism at different stages of grain development in wheats differing in final grain size and starch content per grain, to ascertain whether these enzymes have some relationship with grain size and/or starch content. Active starch synthesis in these varieties started from 14 days onward and continued till 35 days after anthesis. Invertase was active only during initial stages of grain development. Sucrose-UDP-glucosyltransferase had maximum activity at the 14 and/or 21 day stage and was present throughout the period of grain development. UDP- and ADP-glucose pyrophosphorylases and amylase were most active during the period of active starch synthesis and at the same time tended to parallel grain size and starch content at different stages of grain development.     

Modulation of activities of steryl glucoside hydrolase and UDPG: Sterol glucosyltransferase from Sinapis alba by detergents and lipids

Interactions of detergents and lipid compounds on the activity of delipidated preparations of UDPG: sterol glucosyltransferase and steryl β-d-glucoside hydrolase (SG hydrolase) isolated from white mustard seedlings were studied. It has been found that various lipids exert diverse effects on the activity of SG hydrolase. This activity was distinctly stimulated by several neutral, relatively unpolar compounds such as phytol, tripalmitoylglycerol, methyl stearate or cholesteryl acetate and, to a lesser extent, by free fatty acids. On the other hand a number of phospho- and glycolipids were inhibitory. A particularly strong inhibition was observed with charged, zwitterionic phospholipids such as PC, PE or their 2-lyso derivatives. These results point to the possibility of in vivo regulation of the membrane-bound SG hydrolase by its lipid microenvironment. In contrast to SG hydrolase no evidence was found for a clear-cut effect of lipids on the activity of UDPG: sterol glucosyltransferase even after a pretreatment of the enzyme preparation with phospholipase C.     

Antioxidative responses in Vitis vinifera infected by grapevine fanleaf virus

The antioxidative response of grapevine leaves (Vitis vinifera cv. Trebbiano) affected by the presence of grapevine fanleaf virus was studied during the summer of 2010 at three different harvest times (July 1st and 26th, and August 30th). At the first and second harvest, infected leaves showed increases in the concentration of superoxide radical and hydrogen peroxide, the latter increasing for enhanced activity of superoxide dismutase. In contrast, at the last harvest time, increases in the ascorbate pool and ascorbate peroxidase activity maintained hydrogen peroxide to control levels. The glutathione pool was negatively affected as summer progressed, showing a decrease in its total and reduced form amounts. At the same time, increases in the ascorbate pool were observed, making antioxidant defenses of grapevine effective also at the last harvest time. Increases in phenolic acids, and in particular in p-hydroxybenzoic acid, at the first and second harvest might have enhanced the efficiency of the antioxidant system through an interrelation between a peroxidase/phenol/ascorbate system and the NADPH/glutathione/ascorbate cycle. The lack of increase in p-hydroxybenzoic acid at the third harvest could be due instead to the enhanced utilization of this acid for hydrogen peroxide detoxification. With time, grapevine plants lost their capacity to contrast the spread of grapevine fanleaf virus, but acquired a greater ability to counteract pathogen-induced oxidative stress, being endowed with more reduced antioxidant pools.     

Novel monoterpene disaccharide glycosides of Vitis vinifera grapes and wines

β-Rutinosides (6-O-α-l-rhamnopyranosyl-β-d-glucopyranosides) and 6-O-α-l-arabinofuranosyl-β-d-glucopyranosides of geraniol, nerol and linalol have been isolated and characterized in Muscat of Alexandria grapes and wine. These monoterpene disaccharide glycosides, which are precursors of linalol oxidation state monoterpenes of the grape, are also present in another non-muscat Vitis vinifera var. Rhine Riesling.     

Cold-induced fructosan synthesis in leaves of Dactylis glomerata

Dactylis glomerata accumulated fructosan more rapidly at 5° than at 15—20°. The pattern of incorporation of ¹⁴CO₂ into fructosan was determined in plants grown at 5°. During the major period of fructosan synthesis there was initial incorporation of label into mono- and disaccharides, and progressive synthesis of polymeric material occurred subsequently. Rates and levels of synthesis were much lower in leaf blades than in leaf bases. The MW distribution of the polymeric material in leaf bases differed from that in the blades and from that observed in plants which synthesize inulin.