A Solvent for the Paper Chromatographic of Separation of Glucose and Sorbitol

Rees, W. R.; Reynolds, TM

doi:10.1038/181767a0

Cited by 107 publications

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“…Basic compounds were separated using 1-butanol-glacial acetic acid-water, 60:15:25, (v/v) and phenol-water-NH40H plus 0.1% KCN (26). Neutral compounds were separated using 2-butanone-glacial acetic acidsaturated aqueous H3BO3, 9:1:1 (v/v) (18) …”

Section: Methodsmentioning

confidence: 99%

Uptake and Metabolism of d-Glucose by Neocosmospora vasinfecta E. F. Smith

Budd

1976

Plant Physiol.

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Freshly harvested, nongrowing mycelium of Neocosmospora vasinfecta E. F. Smith rapidly absorbed exogenous glucose but converted a greater proportion to trehalose and glucan than to respiratory CO2. This effect was accentuated in mycelium preincubated for 3.5 hours in water before exposure to glucose. Glucose was absorbed via two uptake systems, both apparently constitutive, with apparent Km values for glucose of 0.02 mM (high affinity) and 2 mM (low affinity). The glucose derivative 3-0-methylgloucose (3-0-MG) was also absorbed by two apparently constitutive systems with apparent Km values for 3-0-MG of 0.065 mM and 1.9 mM. Absorption of 3-0-MG by both freshly harvested and preincubated mycelium led to its accumulation. Freshly harvested mycelium lost accumulated 3-0-MG rapidly to water, whereas preincubated mycelium showed reduced or no leakage. The reduction in leakage due to preincubation was prevented by 5 ag/nd cycloheximide in the preincubation medium. Glucose competitively inhibited 3-0-MG uptake via the high affinity system and induced loss of previously accumulated 3-0-MG from preincubated mycelium. The uptake of both glucose and 3-0-MG was associated with a transient alkalinization of the uptake medium. It is concladed that uptake of both glucose and 3-0-MG by at least the high affinity system is energy-linked and probably mediated by proton cotransport.Soluble sugars are among the most widely available carbon sources for the growth of microorganisms, and indeed for all living cells. The utilization of externally supplied sugars is dependent upon the presence of specialized sugar-absorption mechanisms in the cell-surface membrane, as well as on the presence of the appropriate intracellular enzymes. Such absorption mechanisms, intensively studied in bacterial cells and membrane preparations (e.g. refs. 2 and 9), have been described in a variety of plant cells and also in fungi (3, 14, 16, 20-23, 25, 28).Metabolic influences on sugar transport are evident from many of these studies. Conversely, sugar transport should influence, directly or indirectly, many other cellular processes. A novel interaction between absorbed sugars and fluxes of the chloride ion was recently observed in the mycelial ascomycete Neocosmospora vasinfecta (10,11,13). This, together with an almost total lack of information on glucose uptake and metabolism in this organism, prompted the present studies. MATERIALS AND METHODSThe organism and the methods of growth and harvesting were as previously described (6). The sole carbon source in the growth medium was i-glucose, provided at an initial level of 3% (167 ' Supported by Operating Grant NRC A2813 from the National Research Council of Canada. mM). The washed mycelium was resuspended in demineralized H20 at a density of 0.3 to 0.7 mg dry matter/ml and aerated with moist air. From this mycelial suspension, samples of about 30 to 70 mg dry weight were withdrawn either immediately ("fresh" mycelium) or after a period of aeration ("preincubated" mycelium). Normally, preincubation ...

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Section: Methodsmentioning

confidence: 99%

Uptake and Metabolism of d-Glucose by Neocosmospora vasinfecta E. F. Smith

Budd

1976

Plant Physiol.

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“…Neutral sugar fractions were analyzed by descending paper chromatography on Whatman No. 1 paper with the following solvent systems: solvent (1), M-propanol:ethyl acetate:water (7:1:2) (Smith, 1960); solvent (2), ethylmethyl ketone:acetic acid:saturated boric acid solution (9:1:1) (Rees and Reynolds, 1958); solvent (3), ethyl acetate:acetic acid:water (14:3:3) (Smith, 1960); and solvent (4), ethyl acetate:acetic acid:pyridine:water (50:12:18:10) (Jarvis and Duncan, 1974). Radioactive compounds on chromatograms were detected by X-ray film (Kodak, Ltd).…”

Section: Analysis Of Lipids Neutral Sugars and Cell Wall Fractionsmentioning

confidence: 99%

TRANSPORT, TRANSLOCATION AND METABOLISM OF ¹⁴C‐PHOTOSYNTHATES AT THE HOST‐PARASITE INTERFACE OF PISUMSATIVUM AND ERYSIPHE PISI

Manners

1982

New Phytologist

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SUMMARYHaustorial complex, mycelial and host (leaf without mycelia) samples were obtained from leaves of Pisum sativum infected with Erysiphe pisi either immediately following a 20 min exposure to i^COa ('pulse') or after a further 20 h 'chase' in i^COg. After the 'pulse', labelled ethanolsoluble sugars (glucose, fructose and sucrose) and mannitol predominated in the host and mycelial fractions respectively and lipids (mainly triglycerides) and an ethanol-insoluble glucan (not a-1,4 linked) contained most of the ^*C in the haustorial complexes. Ethanol-soluble carbohydrates contained only 3 % of the total ^*C in the haustorial complexes with sucrose and glycerol as the major components. After the ' chase', the total ^*C in host and mycelium declined but the proportions in starch or glycogen, protein, cell wall and nucleic acid fractions increased. Total ^^C in haustorial complexes increased but that contained in sucrose and glycerol decreased and that in mannitol increased. During both 'pulse' and 'chase', the quantity of ^*C in the haustorial complexes was small compared with that in the mycelial and leaf fractions.These results indicate that transport of host sugars to the fungus is mediated by haustoria. Sucrose and glycerol are either translocates or metabolic intermediates in this process and mannitol is the major primary sink of the fungus. Glycogen and other macromolecules are produced at a later stage in the fungus. Polysaccharides and lipids are important sinks at the host-haustorial interface. These results are discussed in relation to the regulation of transport at the host-parasite interface.

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“…For chromatography, aliquots of the soluble fraction were dried under a cold-air streatn. Descending paper chromatography was on Whatman 4 paper in butan-2-one: acetic acid:water saturated with boric acid (9:1:1) (Rees and Reynolds, 1958). Radioactivity was detected hy the scanning apparatus of Loughman and Martin (1957).…”

Section: Analytical Proceduresmentioning

confidence: 99%

ECOLOGICAL PHYSIOLOGY OF THE LICHEN HYPOGYMNIA PHYSODES

Farrar

1976

New Phytologist

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SUMMARY The effect of constant saturation with water upon Hypogymnia physodes for periods of up to 7 days was investigated in either light or dark, at either 20°C (representative of most laboratory experiments on lichen physiology) or 5°C (more representative of field conditions). In all cases the capacity for photosynthesis, phosphate uptake, and total polyol content declined over 7 days; the decline was greater at 20°C than 5°C, and usually greater in the dark than in the light. In general, both alga and fungus reacted similarly to the treatments. Previously fixed 14C mostly remained in the polyol pool during the treatments, with relatively little entering the insoluble fraction. The substantial loss of previously fixed 14C during 7 days saturation was almost entirely from the methanol soluble fraction. Within the insoluble fraction, there was also no loss of 14C during the 7‐day period if protein had been previously labelled with 14C‐leucine. Two other lichens, Xanthoria aureola and Peltigera horizontalis, also showed broadly similar declines to Hypogymnia physodes in various physiological characteristics.

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A Solvent for the Paper Chromatographic of Separation of Glucose and Sorbitol

Cited by 107 publications

References 0 publications

Uptake and Metabolism of d-Glucose by Neocosmospora vasinfecta E. F. Smith

Uptake and Metabolism of d-Glucose by Neocosmospora vasinfecta E. F. Smith

TRANSPORT, TRANSLOCATION AND METABOLISM OF ¹⁴C‐PHOTOSYNTHATES AT THE HOST‐PARASITE INTERFACE OF PISUMSATIVUM AND ERYSIPHE PISI

ECOLOGICAL PHYSIOLOGY OF THE LICHEN HYPOGYMNIA PHYSODES

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A Solvent for the Paper Chromatographic of Separation of Glucose and Sorbitol

Cited by 107 publications

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Uptake and Metabolism of d-Glucose by Neocosmospora vasinfecta E. F. Smith

Uptake and Metabolism of d-Glucose by Neocosmospora vasinfecta E. F. Smith

TRANSPORT, TRANSLOCATION AND METABOLISM OF 14C‐PHOTOSYNTHATES AT THE HOST‐PARASITE INTERFACE OF PISUMSATIVUM AND ERYSIPHE PISI

ECOLOGICAL PHYSIOLOGY OF THE LICHEN HYPOGYMNIA PHYSODES

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TRANSPORT, TRANSLOCATION AND METABOLISM OF ¹⁴C‐PHOTOSYNTHATES AT THE HOST‐PARASITE INTERFACE OF PISUMSATIVUM AND ERYSIPHE PISI