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  • 1
    ISSN: 1432-2048
    Schlagwort(e): Amyloid (seed) ; Endo-β-glucanase ; β-Galactosidase ; Germination (seed) ; β-Glucosidase ; Tropaeolum (amyloid mobilisation) ; Xyloglucan ; α-Xylosidase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The levels of cell-wall xyloglucan (amyloid) in nasturtium (Tropaeolum majus L.) cotyledons were monitored during a 28-d period covering seed imbibition, germination and early seedling development. The activities of the following enzymes capable of hydrolysing the glycosidic linkages in the xyloglucan were assayed in cotyledon extracts over the same period: endo-(1→4)-β-glucanase (EC 3.2.1.4), β-glucosidase (EC 3.2.1.21), α-xylosidase and β-galactosidase (EC 3.2.1.23). The endo-β-glucanase was assayed viscometrically using xyloglucan as substrate, and the three glycosidases using appropriate p-nitrophenylglycosides. Alpha xylosidase and β-galactosidase, the enzymes which would be expected to hydrolyse the side-chains from the xyloglucan molecule, were also assyed using xyloglucan as substrate. Under our culture conditions, xyloglucan levels remained constant at 30 mg per cotyledon pair for 7 d, that is until 3 d after germination: thereafter, the amount of xyloglucan diminished to zero in a 12-d period. The most rapid period of depletion was between days 9 and 13. The mobilisation of all reserve substances from the cotyledons resulted in a weight-loss of 92 mg: xyloglucan, therefore, is an important storage substance, representing 33% by weight of the seed's substrate reserves. It is a cell-wall storage polysaccharide. Xyloglucan mobilisation was accompanied by a 17-fold increase in endo-β-glucanase activity, a 7-fold increase in β-galactosidase and an 8-fold increase in α-xylosidase activities, all determined using xyloglucan as substrate. All three activities began to increase at day 5, peaked at days 12–14 when the most rapid phase of xyloglucan breakdown was over, and had declined to zero by days 22–25. The levels of theses enzymes have been shown to be consistent with their being responsible for xyloglucan hydrolysis in vivo. Nitrophenyl-β-galactosidase activity increased up to day 3, remained constant and then increased again 2.5-fold from day 5, peaking at day 11. Nitrophenyl-β-glucosidase remained relatively constant up to day 16 and then decreased to zero by day 25. Nitrophenyl-α-xylosidase activity was not detected.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 2
    ISSN: 1432-2048
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Summary The activities of α-galactosidase, β-mannosidase and α-mannosidase were determined in extracts from the endosperm and from the embryo of fenugreek seeds at different stages of germination. Endosperm homogenates contained little or no activity of the above enzymes in the early stages of germination, before the reserve galactomannan began to be mobilised. The onset of galactomannan breakdown coincided with the appearance of α-galactosidase and β-mannosidase activities, which increased throughout the period of galactomannan degradation and then remained constant. A similar rise in α-galactosidase and β-mannosidase activities occurred during galactomannan breakdown in dry-isolated endosperms incubated under germination conditions. The increase could be suppressed by metabolic inhibitors which also inhibit galactomannan breakdown. Embryo homogenates contained high α-galactosidase, high α-mannosidase and some β-mannosidase activity at all stages of germination. No “oligomannosyl β-1,4 phosphorylase” activity could be detected either in the endosperm or in the embryo. It is concluded that the galactomannan of fenugreek is broken down by a series of hydrolases secreted by the aleurone layer of the endosperm. They include α-galactosidase, β-mannosidase and probably also endo-β-mannanase.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 3
    ISSN: 1432-2048
    Schlagwort(e): Endosperm (galactomannan breakdown) ; Galactomannan breakdown ; Seed (water stress) ; Trigonella ; Water stress (seed)
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Imposition of water stress on germinated fenugreek (Trigonella foenum-graecum L.) seeds and isolated fenugreek endosperms after the beginning of galactomannan mobilisation caused a reduction in the rate of breakdown of the polysaccharide relative to unstressed controls. The activities, measured in vitro, of the three hydrolytic enzymes involved in the breakdown process (α-d-galactosidase, EC 3.2.1.22;endo-β-d-mannanase, EC 3.2.1.78;exo-β-d-mannanase, EC 3.2.1.25) were not decreased. Although there was some accumulation of galactomannan-hydrolysis products in endosperms under stress, there was no clear correlation between sugar levels and the inhibition of galactomannan breakdown. When water stress was applied to fenugreek seeds after germination but before the beginning of galactomannan hydrolysis, both galactomannan breakdown and the development of the hydrolytic enzyme activities were inhibited. Washing of newly germinated seeds for 2 h in water prior to the imposition of stress gave partial relief of the inhibition of galactomannan mobilisation, partial recovery ofendo-β-d-mannanase levels, and full recovery of α-d-galactosidase levels. It is argued: 1) that water stress after germination but before the beginning of galactomannan hydrolysis inhibits the production of hydrolytic enzymes in the endosperm, probably via decreased removal at lowered water content of diffusible inhibitory substances; and 2) that water stress after the beginning of galactomannan hydrolysis decreases the rate of galactomannan breakdown in vivo principally via decreased diffusion at lowered water content of enzymes from the aleurone layer through the storage tissue of the endosperm.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 4
    Digitale Medien
    Digitale Medien
    Springer
    Planta 178 (1989), S. 41-51 
    ISSN: 1432-2048
    Schlagwort(e): Cyamopsis ; Endosperm ; Galacto-mannan biosynthesis ; Galactosyltransferase ; Mannosyltransferase ; Polysaccharide biosynthesis ; Seed development ; Trigonella
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Particulate enzyme preparations were isolated from developing fenugreek (Trigonella foenum-graecum L.) and guar (Cyamopsis tetragonoloba [L.] Taub.) seed endosperms during the period of galactomannan deposition in vivo. These preparations catalysed the formation of polysacharide products from guanosine 5′-diphosphate (GDP)-mannose, from uridine 5′-diphosphate (UDP)-galactose and from mixtures of the two nucleotides. The products were analysed by solubility, by complete acid hydrolysis, and by selective enzymatic cleavage using pure enzymes of known specificity. With GDP-[U-14C]-d-mannose as substrate and a divalent metal cation (Mg+2, Mn+2, or Ca+2) a highly efficient transfer of labelled d-mannosyl residues was obtained to give a product identified as linear (1→4)-β-linked d-mannan. No transfer of galactosyl residues was obtained when GDP-[U-14C]-d-galactose was the only substrate, although very low and variable amounts of an unidentified product which released labelled glucose on acid hydrolysis were formed. In the presence of UDP-galactose, GDP-mannose and Mn+2 ions, products were formed which have been characterised as galactomanans — a linear (1→4)-β-d-mannan backbone carrying d-galactopyranosyl substituents linked (1→6)-α to mannose. The degree of galactose substitution of the d-mannan backbone was manipulated in vitro by varying GDP-mannose concentrations at constant (saturating) UDP-galactose levels. The transfer of d-galactosyl residues from UDP-galactose to galactomannan was absolutely dependent upon the simultaneous transfer of D-mannosyl residues from GDP-mannose. d-Mannan sequences pre-formed in situ using the mannosyltransferase in the absence of UDP-galactose could not become galactose-substituted in a subsequent incubation either with UDP-galactose alone or with UDP-galactose plus GDP-mannose A model for the interaction of GDP-mannose mannosyltransferase and UDP-galactose galactosyltransferase in galactomannan biosynthesis is proposed.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 5
    ISSN: 1432-2048
    Schlagwort(e): Cell wall ; Endo-(1→4)-β-d-glucanase (xyloglucan-specific) ; Storage polysaccharide (seed) ; Tropaeolum (xyloglucan mobilisation) ; Xyloglucan — α-Xylosidase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The α-xylosidase which is involved in the postgerminative mobilisation of xyloglucan in nasturtium seed cotyledons has now been purified to apparent homogeneity by a facile procedure involving lectin affinity chromatography. The purified enzyme, a glycoprotein, moved as a single band (apparent molecular weight 85000) on sodium dodecyl sulphate-gel electrophoresis, whilst isoelectric focusing gave a number of enzymatically active protein bands spanning the range pI = 5.0 to 7.1 (maximum activity at pI = 6.1). The enzyme did not hydrolyse the simple α-xylosides p-nitrophenyl-α-d-xylopyranoside and woprimeverose (α-d-Xyl(1→6)-d-Glc), or polymeric tamarind-seed xyloglucan. It released xylose from a complex mixture of oligosaccharides produced by exhaustive hydrolysis of tamarind seed xyloglucan using the xyloglucan-specific endo-(1→4)-β-d-glucanase from germinated nasturtium seeds (M. Edwards et al. 1986, J. Biol. Chem., 261. 9489–9494). The three xyloglucan oligosaccharides of lowest molecular size were purified from this mixture and were shown by 1H-nuclear magnetic resonance (1H-NMR) and enzymatic analysis to have the structures:
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 6
    ISSN: 1432-2048
    Schlagwort(e): Alpha galactosidase ; Cell wall storage polysaccharide ; Cyamopsis ; Galactomannan (biosynthesis) ; Senna ; Trigonella
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Galactomannan deposition was investigated in developing endosperms of three leguminous species representative of taxonomic groups which have galactomannans with high, medium and low galactose content. These were fenugreek (Trigonella foenum-graecum L.; mannose/galactose (Man/Gal) = 1.1), guar (Cyamopsis tetragonoloba (L.) Taub.; Man/Gal = 1.6) and Senna occidentalis (L.) Link. (Man/Gal = 3.3), respectively. Endosperms were analysed at different stages of seed development for galactomannan content and the levels, in cell-free extracts, of a mannosyltransferase and a galactosyltransferase which have been shown to catalyse galactomannan biosynthesis in vitro (M. Edwards et al., 1989, Planta 178, 41–51). There was a close correlation in each case between the levels of the biosynthetic mannosyl- and galactosyltransferases and the deposition of galactomannan. The relative in vitro activities of the mannosyl- and galactosyltransferases in fenugreek and guar were similar, and almost constant throughout the period of galactomannan deposition. In Senna the ratio mannosyltransferase/galactosyltransferase was always higher than in the other two species, and it increased substantially throughout the period of galactomannan deposition. In fenugreek and guar the galactomannans present in the endosperms of seeds at different stages of development had the Man/Gal ratios characteristic of the mature seeds. By contrast the galactomannan present in Senna endosperms at the earliest stages of deposition had a Man/Gal ratio of about 2.3. During late deposition this ratio increased rapidly, stabilising at about 3.3, the ratio characteristic of the mature seed. The levels of α-galactosidase in the developing endosperms of fenugreek and guar were low and remained fairly constant throughout the deposition of the galactomannan. In Senna, α-galactosidase activity in the endosperm was low during early galactomannan deposition, but increased subsequently, peaking during late galactomannan deposition. The developmental patterns of the α-galactosidase activity and of the increase in Man/Gal ratio of the Senna galactomannan were closely similar, indicating a cause-and-effect relationship. The endosperm α-galactosidase activity in Senna was capable, in vitro, of removing galactose from guar galactomannan without prior depolymerisation of the molecule. In fenugreek and in guar the genetic control of the Man/Gal ratio in galactomannan is not the result of a post-depositional modification, and must reside in the biosynthetic process. In Senna, the Man/Gal ratio of the primary biosynthetic galactomannan product is controlled by the biosynthetic process. Yet the final Man/Gal ratio of the galactomannan in the mature seed is, to an appreciable extent, the result of galactose removal from the primary biosynthetic product by an α-galactosidase activity which is present in the endosperm during late galactomannan deposition.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 7
    Digitale Medien
    Digitale Medien
    Springer
    Planta 195 (1995), S. 489-495 
    ISSN: 1432-2048
    Schlagwort(e): Biosynthesis (computer simulation) ; Cell wall (plant) ; Cyamopsis ; Galactomannan ; Senna ; Trigonella
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Membrane-bound enzymes from developing legume-seed endosperms catalyse galactomannan biosynthesis in vitro from GDP-mannose and UDP-galactose. A mannosyltransferase [mannan synthase] catalyses the extension of the linear (1→4)-β-linked d-mannan backbone towards the non-reducing end. A specific α-galactosyltransferase brings about the galactosyl-substitution of the backbone by catalysing the transfer of a (1→6)-α-d-galactosyl residue to an acceptor mannosyl residue at or close to the non-reducing terminus of the growing backbone. Labelled galactomannans with a range of mannose/galactose (Man/Gal) ratios were formed in vitro from GDP-[14C]mannose and UDP-[14C]galactose using membrane-bound enzyme preparations from fenugreek (Trigonella foenum-graecum L.), guar (Cyamopsis tetragonoloba (L.) Taub.) and senna (Senna occidentalis (L.) Link.), species which in vivo, form galactomannans with Man/Gal ratios of 1.1, 1.6 and 3.3 respectively. The labelled galactomannans were fragmented using a structure-sensitive endo-(1→4)-β-d-mannanase and the quantitative fragmentation data were processed using a computer algorithm which simulated the above model for galactomannan biosynthesis on the basis of a second-order Markov chain process, and also the subsequent action of the endo-mannanase. For each galactomannan data-set processed, the algorithm generated a set of four conditional probabilities required by the Markov model. The need for a second-order Markov chain description indicated that the galactomannan subsite recognition sequence of the galactosyltransferase must encompass at least three backbone mannose residues, i.e. the site of substitution and the two preceding ones towards the reducing end of the growing galactomannan chain. Data-sets from the three plant species generated three distinctly different sets of probabilities, and hence galactose-substitution rules. For each species, the maximum degree of galactose-substitution consistent with these rules was closely similar to that observed for the primary product of galactomannan biosynthesis in vivo. The data provide insight into the mechanism of action and the spatial organisation of membrane-bound polysaccharide synthases.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 8
    ISSN: 1432-2048
    Schlagwort(e): Endosperm ; Galactomannan ; Secretion ; Ultrastructure ; Trigonella
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The mode of deposition (secretion) of galactomannan in the cells of the seed endosperm ofTrigonella foenum-graecum has been studied by electron microscopy. In cells which are just beginning to secrete galactomannan there are stacks of rough endoplasmic reticulum (ER). The intracisternal space (containing the enchylema) of the rough ER then swells, becomes vacuolated and forms a voluminous network, with “pockets” of cytoplasm entrapped within poculiform rough ER. The enchylema contains material which reacts with periodate-thiocarbohydrazidesilver proteinate in a very similar manner to the galactomannan already deposited in the cell wall. It appears that the galactomannan is formed in the intracisternal space of the rough endoplasmic reticulum and then expelled outside the plasmalemma. This mode of deposition contrasts with that of other plant cell wall polysaccharides whose secretion is mediated by Golgi vesicles.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 9
    ISSN: 1432-2048
    Schlagwort(e): Endosperm (galactomannan) ; Germination (seeds) ; Lipid ; Phytate ; Storage proteins ; Trigonella
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract Changes in total nitrogen, soluble amino nitrogen, lipid and phytate contents, and in the activities of proteinase (pH 7.0), isocitrate lyase and phytase were followed in the endosperm, cotyledons, and axis during germination of fenugreek seeds and subsequent growth of the seedlings. The endosperm is comprised largely of cell-wall galactomannans: the majority of the seed total nitrogen, lipid and phytate (5%, 8%, 0.44% of seed dry weight respectively) is localised within the cotyledons as stored reserves. Germination is completed after 10–14 h from the start of imbibition, but the major reserves are not mobilised during the first 24 h. Then the total nitrogen content of the cotyledons starts to decrease and that of the axis increases; there is a concomitant accumulation of soluble amino nitrogen in both cotyledons and axis. An increase in proteinase activity in the cotyledons correlates well with the depletion of total nitrogen therein. Depletion of lipid and phytate reserves in the different seed tissues constitutes a late event, occurring after 50 h from the start of imbibition, and is coincident with the final disintegration of the endosperm tissue. The depletion of phytate and stored lipids is accompanied by an increase in phytase and isocitrate lyase activity. It appears that the products of lipid hydrolysis are converted by gluconeogenesis to serve as the major source of sugars for the growing axis after the endosperm galactomannan has been completely mobilised.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 10
    ISSN: 1432-2048
    Schlagwort(e): Galactomannan ; Endosperm ; Polysaccharide (biosynthesis, storage) ; Mannosyltransferase ; Seed (development) ; Trigonella
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: Abstract The time-course of galactomannan and stachyose (digalactosyl-sucrose) deposition in the fenugreek seed endosperm has been determined, and correlated with standard parameters of seed development. During, and only during, the period of galactomannan deposition, endosperm homogenates are capable of catalysing the transfer of labelled d-mannosyl residues from guanosine 5′-diphosphate d-[U-14C]mannose to a soluble polysaccharide product indistinguishable from galactomannan. The mannosyltransferase activity peaks twice, once at the beginning of galactomannan deposition, and again in the middle of the most rapid phase of galactomannan deposition. The enzyme in the later peak sediments with grossly particulate material (1,000 g pellet), whereas the earlier peak contains a considerable proportion of a particulate enzyme sedimenting at 100,000 g. These observations are discussed in the light of existing information on the ultrastructural aspects of galactomannan deposition. The mannosyltransferase is clearly involved in galactomannan formation in vivo, but the status of an accompanying galactosyltransferase is less clear.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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