Isolation and characterization of free glycans of the oligomannoside type from the extracellular medium of a plant cell suspension

The oligosaccharides Man₅GlcNAc and Man₃(Xyl)GlcNAc(Fuc)GlcNAc presumed to originate fromN-glycosyl proteins have been purified from an extracellular medium (concentration: 2–5 mg/l of 14 day cultures) of white campion (Silene alba) suspension culture. Their primary structures have been determined by¹H-400-MHz NMR spectroscopy and FAB-MS spectrometry. They are probably the result of an autophagic process including protein catabolism due to sucrose starvation. Additional identification of digalactosylglycerol (galactolipid breakdown) argues for this hypothesis.Abbreviations Fuc l-fucose - Man d-mannose - Xyl d-xylose - GlcNAc N-acetyl-d-glucosamine - Gal d-galactose - Glc d-glucose - FAB-MS fast atom bombardment mass spectrometry - NMR nuclear magnetic resonance     

Desmarestia antarctica (Desmarestiales,Phaeophyceae), a new ligulate Antarctic species with an endophytic gametophyte

Desmarestia antarctica Moe & Silva, a new species from the Antarctic Peninsula and the Scotia Arc, is distinguished from all other ligulate members of the genus by the arrangement of zoosporangia, which are produced together with paraphyses in a raised sorus. Among all species in the genus, onlyD. anceps Montagne shares this feature.Desmarestia antarctica is the first species of the genus for which an endophytic gametophyte is demonstrated, the usual host beingCurdiea racovitzae De Wildeman, a red alga of the familyGracilariaceae. The sporophyte contains only a moderate concentration of acid compared to Northern Hemisphere ligulate species, while physodes that probably contain polyphenolic compounds form noticeable speckles and dark margins in preserved plants. Often more than one axis arises from a single holdfast, probably as the result of compounding rather than proliferation.Dedicated to Prof. DrLothar Geitler on the occasion of the 90th anniversary of his birthday.     

Prehistoric Pinus woodland dynamics in an upland landscape in northern Scotland: the roles of climate change and human impact

Pollen, microscopic charcoal, palaeohydrological and dendrochronological analyses are applied to a radiocarbon and tephrochronologically dated mid Holocene (ca. 8500–3000 cal b.p.) peat sequence with abundant fossil Pinus (pine) wood. The Pinus populations on peat fluctuated considerably over the period in question. Colonisation by Pinus from ca. 7900–7600 cal b.p. appears to have had no specific environmental trigger; it was probably determined by the rate of migration from particular populations. The second phase, at ca. 5000–4400 cal b.p., was facilitated by anthropogenic interference that reduced competition from other trees. The pollen record shows two Pinus declines. The first at ca. 6200–5500 cal b.p. was caused by a series of rapid and frequent climatic shifts. The second, the so-called pine decline, was very gradual (ca. 4200–3300 cal b.p.) at Loch Farlary and may not have been related to climate change as is often supposed. Low intensity but sustained grazing pressures were more important. Throughout the mid Holocene, the frequency and intensity of burning in these open Pinus–Calluna woods were probably highly sensitive to hydrological (climatic) change. Axe marks on several trees are related to the mid to late Bronze Age, i.e., long after the trees had died.     

Mechanism of l-methionine overproduction by Escherichia coli: the replacement of Ser-54 by Asn in the MetJ protein causes the derepression of l-methionine biosynthetic enzymes

We derived l-methionine-analogue-resistant mutants from Escherichia coli JM109 strain by mutagenesis with N-methyl-N′-nitro-N-nitrosoguanidine and selected the potent l-methionine-overproducing strains by microbioassay using lactic acid bacteria. One of the mutants, strain TN1, produced approximately 910 mg l-methionine/l following the addition of 0.1% yeast extract to fundamental medium containing glucose and ammonium sulfate. The l-methionine biosynthetic enzymes, cystathionine γ-synthase and cystathionine β-lyase, of the l-methionine-overproducing mutants were little repressed by l-methionine. To analyse the mechanism of l-methionine overproduction in the mutant strains, the metJ gene coding for the E. colimet repressor, MetJ protein, was cloned and sequenced by the polymerase chain reaction. The same single-amino-acid subsitution (wild-type Ser → Asn) at position 54 was observed in four independent l-methionine-producing mutants. When the wild-type metJ gene was then introduced into strain TN1 having the mutant metJ gene, the level of enzyme synthesis and the l-methionine productivity in the transformants were found to revert to those of the wild-type. It was therefore considered that only one point mutation in the metJ gene occurred in the l-methionine-producing mutants. These results demonstrate the important role of residue 54 of the MetJ protein in l-methionine overproduction, probably because of the derepression of l-methionine biosynthetic enzymes. Received: 6 January 1999 / Received last revision: 19 February 1999 / Accepted: 26 February 1999     

Late glacial and Holocene vegetation,climate and fire history inferred from Lagoa Nova in the southeastern Brazilian lowland

The pollen and charcoal record of a 1,500-cm-long lake sediment core from Lagoa Nova allows a reconstruction of vegetation, climate and fire history in the Atlantic lowland of southeastern Brazil. Today the potential vegetation of the study region would be dense and tall semi-deciduous forest, related to a dry season of 4 months and 1,250 mm precipitation per year. The lowermost core section of Lagoa Nova is probably of late Glacial age and contains only few, poorly preserved pollen grains, indicating dry environmental conditions. Pollen preservation was good above 1,005-cm core depth. Extrapolating from four radiocarbon dates, this interval represents about the last 10,000 years. During the early Holocene (until about 8,500 b.p.), the landscape was dominated by savanna of the campo cerrado type (open shrub woodland), with frequent trees of Curatella americana and small areas of gallery forests along the drainage system. Savanna fires, probably natural, were frequent at that time. The palaeovegetation pattern is consistent with a long dry season of about 6 months and annual precipitation lower than today. Then, up to about 7,560 b.p., gallery forests expanded in the valleys, reflecting a shorter dry season of about 5 months and somewhat higher annual rainfall. Fire was less frequent during this wetter period. Between about 7,560 and 6,060 b.p., savanna expanded and gallery forests retreated, indicating a return to drier climatic conditions of probably between 5 and 6 months dry season and lower precipitation. Fire was again frequent, but not as frequent as during the early Holocene. During the mid to late Holocene period between about 6,060 and 2,180 b.p., the valleys were covered by semi-deciduous forest, but on the hills cerrado vegetation continued to grow. The dry season was probably around 5 months and rainfall was higher than in the preceding period. During the following period between about 2,810 and 600 b.p., the open cerrado on the hills changed to closed cerrado, reflecting wetter conditions with a shorter dry season than in the previous period. It was only after about 600 b.p. that dense semi-deciduous forests expanded throughout the study region, indicating the start of modern, wet climatic conditions with an annual dry season of about 4 months. Fire was nearly absent during this period. The reconstructed palaeoenvironment from the Lagoa Nova record is similar to that of Lago do Pires, indicating that changes in past vegetation, climate and fire frequency are regionally consistent in southeastern Brazil.     

Veronica allahuekberensis (Scrophulariaceae), eine neue Art aus der Türkei

Veronica allahuekberensis A. Öztürk, spec. nova, probably endemic to the Allahuekber mountain in N.E. Anatolia, belongs toV. orientalis group of sectionVeronica and resemblesV. cuneifolia D. Don subsp.cuneifolia, endemic to S.W. Anatolia.

     

Taxonomic status of the hominine cranium KNM-ER 1813 (primates: Homininae) from the Plio/Pleistocene of Koobi Fora

The hominine cranium KNM-ER 1813, from the late Plio/Pleistocene of Koobi Fora, has been regarded recently by some authors as a female ofHomo habilis Leakey, Tobias, andNapier, 1964 and by others as an enigma. Reassessment of its cranial morphology, dental metrics, proportions, and a new detailed determination of its sex indicates that it does not conform with the diagnosis forH. habilis, and is probably a male. It is sympatric withH. habilis yet shows more primitive features and rather a closer affinity to the smaller, more primitive chronospeciesH. antiquus Ferguson, 1984, and is thus the first, nearly complete skull of our oldest known human ancestor.     

Die europäischen Arten der GattungCallitula spinola (Hymenoptera: Pteromalidae)

Summary The author reviews the European species of the Pteromalid genusCallitula Spinola and keys them out. All four species develop as parasites, probably all on small Diptera: two species [pyrrhogaster (Walk.) andbicolor spin.] on Cecidomyids and Chloropids in grass stems, two species are associated withPhragmites communis [C. ferrierei sp. n. andelongatus (Thoms)]. From the latter two one is described as new and in the other (elongatus) the so-far-unknown male is described. *** DIRECT SUPPORT *** A5180147 00002      

Differences in the substrate specificity of glycosyltransferases involved in landomycins A and E biosynthesis

A lanGT4 mutant of the landomycin A producer Streptomyces cyanogenus S136 was constructed, leading to the production of landomycin D with two deoxy sugars in the side chain and proving that LanGT4 is responsible for attaching the third deoxy sugar of the hexasaccharide side chain. Heterologous expression of lndGT4 of the landomycin E producer Streptomyces globisporus 1912 in the lanGT4 mutant restored landomycin A production, indicating that LndGT4, like LanGT4, also has the ability to work iteratively. A S. cyanogenus S136 mutant with a mutation in lanGT1, encoding a d-olivosyltransferase, was shown to produce landomycin I with one deoxy sugar and, surprisingly, a new landomycin derivative (landomycin L) containing a d-olivose followed by an l-rhodinose. Heterologous expression of lndGT1 of S. globisporus 1912 in the lanGT1 mutant did not restore landomycin A production but led to the formation of a second new landomycin derivative (landomycin K) containing an unusual pentasaccharide chain (d-olivose–d-olivose–l-rhodinose–d-olivose–l-rhodinose). The formation of landomycin L and landomycin K is most probably attributed to the high substrate flexibility of the rhodinosyltransferase LanGT4. A. Erb and C. Krauth contributed equally to this work.     

Two new species ofPhyllagathis related toP. tuberculata (Melastomataceae) from Peninsular Malaysia

Phyllagathis tuberculata King and two closely allied new species,P. magnifica A. Weber andP. stonei A. Weber, are described and illustrated. Within the genus, these species form a distinct and isolated group which is restricted to Peninsular Malaysia.P. tuberculata occurs in Perak (probably confined to G. Bujang Melaka),P. magnifica andP. stonei are found in the mountains on the Pahang/Selangor border (Genting Highlands, Gombak valley). The distinctive characters of the three species are listed and some general information relating to inflorescence morphology, tubercle anatomy, fruit structure and seed dispersal is provided.     

The outer membrane TolC is involved in cysteine tolerance and overproduction in <Emphasis Type="Italic">Escherichia coli</Emphasis>

l-Cysteine is an important amino acid in terms of its industrial applications. We previously found marked production of l-cysteine directly from glucose in recombinant Escherichia coli cells by the combination of enhancing biosynthetic activity and weakening the degradation pathway. Further improvements in l-cysteine production are expected to use the amino acid efflux system. Here, we identified a novel gene involved in l-cysteine export using a systematic and comprehensive collection of gene-disrupted E. coli K-12 mutants (the Keio collection). Among the 3,985 nonessential gene mutants, tolC-disrupted cells showed hypersensitivity to l-cysteine relative to wild-type cells. Gene expression analysis revealed that the tolC gene encoding the outer membrane channel is essential for l-cysteine tolerance in E. coli cells. However, l-cysteine tolerance is not mediated by TolC-dependent drug efflux systems such as AcrA and AcrB. It also appears that other outer membrane porins including OmpA and OmpF do not participate in TolC-dependent l-cysteine tolerance. When a low-copy-number plasmid carrying the tolC gene was introduced into E. coli cells with enhanced biosynthesis, weakened degradation, and improved export of l-cysteine, the transformants exhibited more l-cysteine tolerance and production than cells carrying the vector only. We concluded that TolC plays an important role in l-cysteine tolerance probably due to its export ability and that TolC overexpression is effective for l-cysteine production in E. coli. Natthawut Wiriyathanawudhiwong and Iwao Ohtsu contributed equally to this work.     

An 11th century a.d. burnt granary at La Gravette, south-western France: preliminary archaeobotanical results

A thick layer of carbonised seeds was encountered in an 11th century a.d. room situated in the seigneurial part of the village of La Gravette. This paper presents the first results of charcoal and seed analyses which give information on the food products stored in the granary and on their arrangement there. Triticum aestivum/durum/turgidum was by far the most important stored crop, while Avena sp., then Hordeum vulgare, Secale cereale, Triticum monococcum and Vitis vinifera were secondary. Weeds were poorly represented. Charcoals were dominated by deciduous Quercus sp., and 11 additional wood taxa were recorded, including especially Fagus sylvatica, Fraxinus sp., Rosaceae, Corylus avellana, Acer campestre and Ulmus sp. According to the charcoal distribution, Quercus and Fagus were probably building materials while most of other taxa would have been used for basketry, wattling or joinery work. In the western part of the granary, naked wheat was stored in bulk. In the eastern part, various crops (at least naked wheat, barley, rye, oat and grape) were stored in small amounts, most of which were probably separated by light wooden structures. The cereal crops had largely been processed and cleaned. The stored products probably represent taxes paid to the lord who owned the granary.     

Cytochemical localization and function of the 3-linked glucan callose in the developing cotton fibre cell wall

Summary Several recent biochemical studies concerning the hemicellulosic content of the developing cotton fibre wall have pointed to an important increase of 1,3-linked glucans at the onset of the secondary wall formation and their slow decrease until the end of fibre development (Meinert andDelmer 1977,Huwyler et al. 1978, 1979,Maltby et al. 1979). These almost insoluble glucans are extra-cellular and possibly associated with the S₁ or winding layer, but no other data on their exact localization were given.By means of a specific fluorescence method, using a 0.05% decolourized aniline blue solution, we show that one of these 3-linked glucans,callose, is always localized, independently of the fibre age, in the innermost wall layer bordering the cell lumen, from the onset of the secondary thickening up to the end of fibre development. Some possible roles assumed by these callose deposits are suggested and discussed. They may be involved in the normal mechanism of cellulose biosynthesis, as being effectively consumed by turnover or, more probably, as forming a permanently restored interface or matrix where cellulose microfibrils undergo a sort of maturation and are oriented before their definitive incorporation in the organized cell wall. They are not to be confused with the wound callose deposits which characterize damaged or immature fibres.     

Uptake of3H-adrenaline and14C-noradrenaline into neuronal and extraneuronal tissue compartments in the perfused gas gland of the swimbladder of the Atlantic cod,Gadus morhua

Summary The neuronal and extraneuronal accumulation of radiolabelledl-adrenaline andl-noradrenaline was studied in the gas gland of the swimbladder of the Atlantic cod,Gadus morhua. Both amines are taken up into the tissue compartments, and a preference for noradrenaline for both uptake processes was demonstrated. A relatively high neuronal accumulation compared to earlier results (cod spleen; Ungell and Nilsson 1984) was seen and this is probably due to the more dense innervation of the swimbladder gas gland. A higher extraneuronal accumulation may be due to the presence of arterial smooth muscle.It is concluded that the adrenergic nerves of the swimbladder gas gland of the cod preferentially accumulate noradrenaline, a situation similar to that in mammals.Abbreviations A/NA adrenaline/noradrenaline ratio - PBA phenoxybenzamine - PNMT phenylethanolamine-N-methyl tranferase     

Observations on the micro-anatomy of crystallolithus hyalinus gaarder and markali

Summary The fine structure of Crystallolithus hyalinus (Gaarder and Markali), known to be the motile stage of Coccolithus pelagicus (Wallich) Schiller (Parke and Adams 1960), has been investigated for purposes of comparison with Chrysochromulina and Prymnesium, recently recognised as probably related to it. Anoptral contrast light microscopy of living cells and electron microscopy of thin sections and shadow-cast whole mounts show that there is a close resemblance to species of Chrysochromulina not only in cell structure but also in the details of haptonema structure and scale structure. The origin of the scales from internal vesicles has been traced as in other genera but the holococcoliths (crystalloliths) do not appear to arise in this way. The mode of origin of the crystalloliths is thus unknown, but it is suggested that they may be developed outside the cell from calcite secreted in solution into the appropriate cavity. Their constant position between a layer of scales and an outer enveloping skin is consistent with this and one unfamiliar cytoplasmic organelle, composed of peculiar thick-walled tubes within some large superficial vesicles, is described as perhaps involved in this process. Apart from these special features the resemblance to both Chrysochromulina and Prymnesium is sufficient to add considerable weight to Christensen's suggestion (Christensen 1962) that a new group should be defined to accommodate these genera.     

Effect of mercury vapour on the non-protein thiol content and respiration rate of eggs of the cattle tickBoophilus microplus (Acari: Ixodiae)

With continuous and ultimately lethal exposure of eggs ofboophilus microplus to mercury vapour, respiration and non-protein thiol decreased to low levels. With sub-lethal exposure, respiration decreased initially and then increased, whereas non-protein thiol (largely glutathione) increased initially and then decreased. This initial increase in glutathione (GSH) which probably resulted from an attempt by the organism to control or reverse the biochemical lesion, did not arise from the oxidized form (gssg) through a shift in the equilibrium from right to left thus: 2gshgssg+2H⁺+2e^–.The significance of the results are discussed in relation to theories ofgsh function and of Hg toxicity. The possible application of mercurials in tick control is discussed.     

Syntaxonomy and ecology of plant communities of the Carici rupestris-Kobresietea bellardii in the western carpathians

We present a syntaxonomic account of the communities of the alliances of Oxytropido-Elynion Br.-Bl. 1949 and Festucion versicoloris Krajina 1933 from Western Carpathians. Both alliances comprise naked-rush, cushion form and dwarf-shrub heath communities typical of wind-exposed habitats occurring at the highest altitudes of the Tatra Mts. They represent a relic vegetation of the cold stages of the Pleistocene (probably Late Glacial Maximum) and they can be classified within the class of Carici rupestris-Kobresietea bellardii Ohba 1974. A set of relevés was subject to numerical-classification analysis. Floristics and ecology of the communities were characterised and the relationships to similar syntaxa were discussed. The Oxytropido-Elynion is restricted to the extreme ridge positions in the highest altitudes of the Belianske Tatry Mts. Five associations were distinguished, such as the Pyrolo carpaticae-Salicetum reticulatae, the Festuco versicoloris-Oreochloetum distichae, the Festucetum versicoloris, the Oxytropido carpaticae-Elynetum myosuroides and the Drabo siliquosae-Festucetum versicoloris. The Festucion versicoloris is limited to the mylonite zone of the alpine and subnival belt of the Vysoké Tatry and Západné Tatry Mts (and found as rare in the Nízke Tatry Mts). The stands of these communities prefer terraces of steep rocky faces and cliffs and stabilised small-grained screes below the cliffs. Within this alliance, three associations were described, including the Agrostio alpinae-Festucetum versicoloris, the Silenetum acaulis and the Salicetum kitaibelianae.     

Floral mechanisms in the tribeSalpiglossidae (Solanaceae)

SinceDelpino (1869),Juel (1894, 1911), andMüller (inMöller 1921: 164), the flowers ofSolanaceae have received little attention with regard to function and pollination syndromes. The present paper deals with representatives of 6 of the 9 known salpiglossidean genera. Previous observations are updated and discussed at the tribal level. Most species studied are butterfly- or moth-pollinated. With the exception ofSalpiglossis, the fertile floral parts are concealed in the corolla tube, and their arrangement is specially suited for deposit of the pollen on the lepidopteran tongue. Particularly notable are (a) abundant stigmatic secretion that makes the pollen sticky, and (b) versatile anthers that optimize contact between the tongue and the thecae.Brunfelsia andBrowallia exhibit a mechanism analogous to that ofApocynaceae, however, with two entrances instead of five. When the tongue is inserted, it is forced to contact the stigma and becomes glued with its secretion. When the tongue is pulled out, it touches the anthers and causes slight balancing movement. InStreptosolen, very probably an ornithophilous descendant of theBrowallia stock, the mechanism is much simplified.Leptoglossis andHunzikeria bear a novel device for pollen deposition: there are two fertile wheel-like anthers that are capable of full rotation up to eight turns.     

Mid- and late-Holocene vegetation and fire history at Biviere di Gela, a coastal lake in southern Sicily, Italy

The vegetation and fire history of few coastal sites has been investigated in the Mediterranean region so far. We present the first paleoecological reconstruction from coastal Sicily, the largest island in the Mediterranean Sea. We analysed pollen and charcoal in the sediments of Biviere di Gela, a lake (lagoon) on the south coast of Sicily. Our data suggest that the area became afforested after a marine transgression at ca. 7200 cal b.p. (5250 b.c.). Build-up of forest and shrublands took ca. 200–300 years, mainly with the deciduous trees Quercus, Ostrya and Fraxinus. Juniperus expanded ca. 6900 cal b.p. (4950 b.c.), but declined again 6600 cal b.p. (4650 b.c.). Afterwards, evergreen trees (Q. ilex-type and Olea) became dominant in the forest and Pistacia shrublands were established. Forest and shrubland reached a maximum ca. 7000–5000 cal b.p. (5050–3050 b.c.); subsequently forest declined in response to human impact, which was probably exacerbated by a general trend towards a more arid climate. During the Neolithic, fire was used to open the landscape, significantly reducing several arboreal taxa (Q. ilex, Fraxinus, Juniperus) and promoting herbs and shrubs (Achillea, Cichorioideae, Brassicaceae, Ephedra). Final forest disruption occurred around 2600 cal b.p. (650 b.c.) with the onset of the historically documented Greek colonization. We conclude that the open maquis and garrigue vegetation of today is primarily the consequence of intensive land-use over millennia. Under natural or near-natural conditions arboreal taxa such as Q. ilex, Olea and Pistacia would be far more important than they are today, even under the hot and rather dry coastal conditions of southern Sicily.