Purification and characterization of barley-aleurone xylanase

Xylanase (-1,4-D-xylan xylanohydrolase; EC 3.2.1.8) from aleurone layers of barley (Hordeum vulgare L. cv. Himalaya) was purified and characterized. Purification was by preparative isoelectric focusing and a Sephadex G-200 column. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the enzyme showed a single protein band with an apparent molecular weight (Mr)=34000 daltons. The isoelectric point of the enzyme was 4.6. The enzyme had maximum activity on xylan at pH 5.5 and at 35° C. It was most stable between pH 5 and 6 and at temperatures between 0 and 4° C. The K_m was 0.86 mg xylan·ml^-1.Abbreviations GA₃ gibberellic acid - kDa kilodalton - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis     

Selective expression of a probable amylase/protease inhibitor in barley aleurone cells: Comparison to the barley amylase/subtilisin inhibitor

We have cloned and sequenced a 650-nucleotide cDNA from barley (Hordeum vulgare L.) aleurone layers encoding a protein that is closely related to a known -amylase inhibitor from Indian finger millet (Eleusine coracana Gaertn.), and that has homologies to certain plant trypsin inhibitors. mRNA for this probable amylase/protease inhibitor (PAPI) is expressed primarily in aleurone tissue during late development of the grain, as compared to that for the amylase/subtilisin inhibitor, which is expressed in endosperm during the peak of storage-protein synthesis. PAPI mRNA is present at high levels in aleurone tissue of desiccated, mature grain, and in incubated aleurone layers prepared from rehydrated mature seeds. Its expression in those layers is not affected by either abscisic acid or gibberellic acid, hormones that, respectively, increase and decrease the abundance of mRNA for the amylase/subtilisin inhibitor. PAPI mRNA is almost as abundant in gibberellic acid-treated aleurone layers as that for -amylase, and PAPI protein is synthesized in that tissue at levels that are comparable to -amylase. PAPI protein is secreted from aleurone layers into the incubation medium.Abbreviations ABA abscisic acid - ASI barley amylase/subtilisin inhibitor - bp nucleotide base pairs - Da dalton - dpa days post anthesis - GA₃ gibberellic acid - PAPI probable amylase/protease inhibitor - poly(A)RNA polyadenylated RNA - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis     

Chemotactic factor causes rapid decreases in phosphatidylinositol,4,5-bisphosphate and phosphatidylinositol 4-monophosphate in rabbit neutrophils

Stimulation of rabbit neutrophils prelabeled with 32P by the synthetic chemotactic peptide f-Met-Leu-Phe induces a rapid decrease in the radioactivity in both phosphatidylinositol, 4,5 bis phosphate and phosphatidylinositol 4-monophosphate. The mean +/- standard error of the mean values of the maximum decrease in phosphatidylinositol, 4,5 bis phosphate occurred at 10 seconds following stimulation and is equal to 19 +/- 3% of the control value. The corresponding value for phosphatidylinositol 4-monophosphate occurred at 60 seconds following stimulation and is equal to 37 +/- 7% of the control value. On the other hand, the radioactivity in phosphatidic acid and lysophospholipids increased continuously with time following stimulation. The relationship of these changes to calcium release and neutrophil activation is discussed.     

Analysis of brain and myelin lipids by high-performance thin layer chromatography and densitometry

We have developed a simple method involving high-performance thin layer chromatographic separation of total brain and myelin lipids. Only two solvent systems consisting of chloroform: methanol: acetic acid and water at different concentrations were needed. The plate was then stained with three sequential procedures to visualize phospholipids, cholesterol and galactolipids. Densitometric procedure at each step of staining was utilized to obtain quantitative analysis of brain and myelin samples.     

Tansley Review No. 59 Leaf boundary layers

Studies of heat and mass exchange between leaves and their local environment are central to our understanding of plant-atmosphere interactions. The transfer across aerodynamic leaf boundary layers is generally described by non-dimensional expressions which reflect largely empirical adaptations of engineering models derived for flat plates. This paper reviews studies on leaves, and leaf models with varying degrees of abstraction, in free and forced convection. It discusses implecations of finding for leaf morphology as it affects – and is affected by – the local microclimate. Predictions of transfer from many leaves in plant communities are complicated by physical and physiological feedback mechanisms between leaves and their environment. Some common approaches, and the current challenge of integrating leaf-atmosphere interactions into models of global relevance, are also briefly addressed.     

The effect of water flow on photosynthetic processes of the alga Ulva lactuca L.

Photosynthetic rates of aquatic macrophytes are affected by the diffusion of carbon through the boundary layer which decreases with increasing flow velocities. This is shown by a significant increase in photosynthetic rates of Ulva lactuca fronds exposed to friction velocities (u _*) between 0.0 and 0.3 cm s^–1. No further increase in photosynthetic rates is observed at u _* values between 0.3 and 1.0 cm s^–1. Stagnating flow conditions at saturating light levels caused a 29% reduction in photosynthetic rates. Experiments in a closed system indicate that Ulva may utilize internal carbon sources during periods of stagnation. On a subtidal rock directly exposed to wave action, Ulva lactuca is not exposed to such conditions while in a wave flushed intertidal pool stagnation occurs during very short time intervals based on in situ measurements of u _* levels on fronds of this species using a microprocessor-controlled, autonomous recorder and hot-film sensors.     

Physico-chemical characteristics of a permanent Spanish hypersaline lake: La Salada de Chiprana (NE Spain)

La Salada de Chiprana Lake, located in the Ebro River basin, northeastern Spain, is the only permanent and deep water hypersaline ecosystem in all of western Europe. With a total surface of 31 ha and a maximum depth of 5.6 m, it has several basins bounded by elongated sandstone-bodies or ribbons which are paleochannels of Miocene age. Its salinity varied from 30 to 73 g 1^–1 during the 1989 hydrological cycle and the most abundant ions were magnesium and sulphate. Depth-time distributions of major physico-chemical variables demonstrated that the lake was stratified in two distinctive layers during most of the year. The chemocline disappeared only in October, with the complete overturn of the water column. In the deep water, three conditions occurred which allowed development of green sulphur bacteria populations: (1) oxygen depletion, (2) presence of hydrogen sulphide and (3) presence of light. Benthic microbial mats covered the sediments of shallow shores of moderate slope.     

The ability of U.K. salmonid alevins to emerge through a sand layer

Artificial redds were used to examine the effect of layers of sand at the gravel surface upon the emergence of trout and salmon. Alevins of both species can negotiate sand layers up to 8 cm thick.     

The Cyanelles (Organelles of a Low Evolutionary Scale) Possess a Phosphate-Translocator and a Glucose-Carrier in Cyanophora paradoxa

Cyanelles from Cyanophora paradoxa can easily be isolated and assayed for their carrier composition by the silicone oil filtering technique. The present investigation demonstrates a P_i-translocator transferring phosphate, dihydroxyacetone phosphate and 3-phosphoglycerate in a counter exchange mode in cyanelles as in chloroplasts of higher plants. The uptake of P_i is inhibited by dihydroxyacetone phosphate, phosphoglycerate and glucose-6-P, only poorly by phosphoenolpyruvate and not by 2-phosphoglycerate. The inhibitors pyridoxalphosphate and 4,4′diisothiocyanostilbene-2,2K'disulfonic acid at low concentration also affect P_i-uptake. Cyanelles probably transport photosynthate (reductant and ATP) by triosephosphates. This is the first demonstration of a phosphate translocator in an organism of a low evolutionary scale. Cyanelles also transport glucose which proceeds in two phases. In the lower concentration range (≤ 2.5 mM), glucose penetrates by facilitated diffusion, whereas transport follows first-order kinetics at higher amounts (> 2.5 mM). In the low concentration range, glucose-transport is affected by high concentrations of 3-O-methylglucose and fructose. The physiological role of the glucose-transport carrier in Cyanophora is doubtful. It may function in transporting glucose into cyanelles if the carbon level inside them becomes limiting, e.g. in dark periods.