Asialo-α1-acid glycoprotein resialylated with 9-amino-5-N-acetyl-d-neuraminic acid is resistant towards bacterial,viral and mammalian sialidases

We demonstrate that 9-amino-NeuAc transferred to asialo-₁-acid glycoprotein resists cleavage by bacterial, viral and mammalian sialidases. This is the first synthetic sialic acid analogue, which can be activated and transferred to glycoprotein, but is not a sialidase (EC 3.2.1.18) substrate.Abbreviations HPLC high performance liquid chromatography - BSA bovine serum albumin - NeuAc N-acetyl-d-neuraminic acid, 5-acetamido-3,5-dideoxy-d-glycero-d-galacto-non-2-ulosonic acid - 9-Amino-NeuAc 9-amino-5-N-acetyl-d-neuraminic acid, 5-acetamido-9-trideoxy-d-glycero-d-galacto-non-2-ulosonic acid - CMP-NeuAc cytidine-5-monophospho-N-acetyl-d-neuraminic acid - CMP-9-amino-NeuAc cytidine-5-monophospho-9-amino-5-N-acetyl-d-neuraminic acid - 9-azido-NeuAc 5-acetamido-9-azido-3,5,9-trideoxy-d-glycero-d-galacto-non-2-ulosonic acid. Enzymes EC 3.2.1.18 sialidase, acylneuraminylhydrolase - EC 2.4.99.1 Galß1-4GlcNAc a(2-6)-sialytransferase     

In vitro propagation of Rauwolfia micrantha, a rare medicinal plant

Shoot tip and single node explants from young shoots of 1-year old flowering plants of Rauwolfia micrantha Hook. f. were cultured on Murashige & Skoog (MS) medium variously supplemented with 6-benzyladenine (BA) and -naphthaleneacetic acid (NAA). A combination of 13.2 M BA and 2.68 M NAA induced high frequency (77%) formation of up to 3 shoots from each node in 8 weeks. The regeneration of shoot tips from the field-grown plants and in vitro shoots placed horizontally differed. Repeated subculturing of the shoot tips and single nodes at 6-week intervals for over a year in combination of 4.4 M BA and 0.27 M NAA enabled mass multiplication of shoots without any evidence of decline. Rooting of the excised shoots on medium containing 2.6 M NAA was preceded by callus formation. The rooted plants were removed off the callus, hardened off and 80% established in pots. Micropropagated plants displayed uniform morphological, growth, flowering, fruiting and seed germination characteristics.Abbreviations BA 6-benzyladenie - IAA indole-3-acetic acid - IBA indole-3-butyrie acid - 2-ip 2-isopentenyladenine - MS Murashige & Skoog (1962) - NAA -naphthaleneacetic acid     

Specificity ofAmaranthus leucocarpus lectin

We have demonstrated thatAmaranthus leucocarpus lectin hemagglutinating activity was powerfully inhibited by the T-antigen, containing Gal(1–3)GalNAc(1–3)Ser/Thr, and the T_n-antigen, which contains GalNAc(1–3)Ser/Thr. This suggests that the acetamido group at C-2 and the axial -OH at C-4 of theN-acetyl-D-galactopyranosylamine ring are important for lectin binding. The hemagglutination assays also established that desialylated and Pronase-treated human typeO erythrocytes with an M phenotype were better recognized than erythrocytes from all other blood groups. The recognition was dependent on pH and ionic strength.     

Transport of basic amino acids in Riccia fluitans: Evidence for a second binding site

The transport of several amino acids with different side-chain characteristics has been investigated in the aquatic liverwort Riccia fluitans. i) The saturation of system I (neutral amino acids) by addition of excess -aminoisobutyric acid to the external medium completely eliminated the electrical effects which are usually set off by neutral amino acids. Under these conditions arginine and lysine significantly depolarized the plasmalemma. ii) L- and D-lysine/arginine were discriminated against in favour of the L-isomers. iii) Increasing the external proton concentration in the interval pH 9 to 4.5 stimulated plasmalemma depolarization, electrical net current, and uptake of [¹⁴C]-basic amino acids. iv) Uptake of [¹⁴C]-glutamic acid took place only at acidic pHs. v) [¹⁴C]-histidine uptake had an optimum between pH 6 and 5.5. vi) Overlapping of the transport of basic, neutral, and acidic amino acids was common. It is suggested that besides system I, a second system (II), specific for basic amino acids, exists in the plasmalemma of Riccia fluitans. It is concluded that the amino-acid molecule with an uncharged side chain is the substrate for system I, which also binds and transports the neutral species of acidic amino acids, whereas system II is specific for amino acids with a positively charged side chain. The possibility of system II being a proton cotransport is discussed.Abbreviation AiB -aminoisobutyric acid     

Transformation of Atriplex gmelini plants from callus lines using Agrobacterium tumefaciens

Atriplex gmelini plants were regenerated via organogensis from hypocotyl explants. Callus lines were induced from the hypocotyl explants on Linsmaier and Skoog (LS) medium supplemented with 1 M benzyladenine and 5 M -naphthaleneacetic acid in the dark. Shoots were regenerated from the callus lines on LS medium supplemented with 20 M thidiazuron and 0.1 M -naphthaleneacetic acid under a high-intensity light condition (450 mol m^–2 s^–1). The regenerated shoots were rooted on LS medium without growth regulators to obtain fully developed plants. We succeeded in transforming Atriplex gmelini from callus lines using Agrobacterium tumefaciens.     

Notes on the genusLinum in Madagascar

Linum emirnense Bojer subsp.marojejyense Humb. is elevated to a species and the heterostylous condition thought to exist inL. emirnense is determined to be an artifact resulting from a fungal infection of the fruit.     

Heteromerikarpie und Fruchtpolymorphismus beiMicroparacaryum,gen. nov., (Boraginaceae)

Microparacaryum (M. Pop. exH. Riedl)Hilger & Podlech is described as a new genus of theBoraginaceae-Cynoglosseae. It comprises the annual species hitherto included inParacaryum (DC.)Boiss. andMattiastrum (Boiss.)Brand. Distribution maps are given for all 3 genera.Microparacaryum consists of two species,M. salsum (Boiss.)Hilger & Podlech (M. s.) andM. intermedium (Fresen.)Hilger & Podlech (M. i.). ParticularlyM. i. is a very variable species, and most of the species formerly recognized belong here. Scattered all over the range of the genus, plants occur with nutlets exhibiting flat or incurved marginal wings, often in mixed populations. This fruit polymorphism is taxonomically treated by recognizing formae. In addition, the following new infraspecific taxa and combinations are described:M. i. var.intermedium formaparacaryoides Hilger & Podlech,M. i. var.stellatum (H. Riedl)Hilger & Podlech,M. i. var.stellatum formamattiastroides Hilger & Podlech,M. s. formamattiastroides Hilger & Podlech.

     

Chemical variation in two species of theLecanora subfusca group (Lecanoraceae,lichenizedAscomycotina)

The chemical variation inLecanora epibryon andL. subimmersa, two species of theL. subfusca group, has been examined. In both species the chemical differences are correlated with geographical distribution, but not with morphological differences. As a consequence the chemotypes are recognized at subspecific level. InL. epibryon three chemical races are segregated according to the various chemosyndromes present. Subspeciesepibryon, containing atranorin and triterpenoids, occurs in the northern hemisphere and South America, while the other two subspecies occur only in the southern hemisphere. The subsp.broccha (Nyl.)Lumbsch, comb. nova, contains atranorin, the stictic acid and the 2,5,7-trichloro-3-O-methylnorlichexanthone chemosyndromes, while subsp.xanthophora Lumbsch, subsp. nova, is similar but lacks the stictic acid chemosyndrome. Two chemical races occur in the pantropical speciesL. subimmersa. While subsp.subimmersa contains atranorin and zeorin, subsp.ramboldii Lumbsch & Elix, subsp. nova, contains an additional ten chlorinated xanthones.L. impressa is reduced to synonymy toL. subimmersa.     

Agaroids from New Zealand members of the Gracilariaceae (Gracilariales,Rhodophyta) — a novel dimethylated agar

Polysaccharide extracts from four New Zealand members of the Gracilariaceae have been characterized by ¹³C-NMR spectroscopy and GLC analysis of alditol acetate derivatives prepared using a new double hydrolysis-reduction procedure. All were based on variously substituted repeating disaccharide units of agarobiose and 20% of its precursor containing l-galactose-6-sulfate. Gracilaria truncata yielded a firm gelling agar with 67% methylation on the 6-position of the d-galactose residues. The other extracts belong to a new class of agar molecules having methylation on both the 6-position of the d-galactose units and the 2-position of the l-sugar units. The Curdiea coriacea polysaccharide displayed this double methylation almost completely ( 96 %); the alkali-modified polymer thus had only two free hydroxy-groups per disaccharide repeat unit, yet still gave a firm gel. The Curdiea flabellata and Melanthalia abscissa extracts had this double methylation pattern but to a lesser extent, and additional xylosyl branch units on up to 18% of the repeating disaccharide units.     

Eine blütenmorphologische Analyse derLagoecieae (Apiaceae)

An analysis of the morphology, anatomy and ontogeny of the flowers, particularly of the gynoecium ofLagoecieae is presented. 1. The gynoecial model of angiosperms can be applied to all three generaArctopus, Lagoecia andPetagnia. 2. In the case ofArctopus an additional Apikalseptum is developed. 3. In the synascidiate region of the gynoecium the adaxial carpel is reduced inArctopus andPetagnia, the abaxial inLagoecia. 4. The reduced carpel produces either one mature ovule inArctopus, a rudimentary ovule inPetagnia, or none inLagoecia. 5.Petagnia andLagoecia have a completely pseudomonomerous gynoecium. 6.Arctopus displays many flower characteristics which lack in theSaniculoideae but occur in theHydrocotyloideae. 7. ForPetagnia andLagoecia an independent phylogenetic development within theSaniculoideae is assumed.

Herrn Univ.-Prof. Dr.Walter Leinfellner zum 70. Geburtstag gewidmet.     

Stachys beckeana (Labiatae) in Albanien und Jugoslawien

Morphological and cytological investigations were carried out inStachys beckeana Dörfler & Hayek andS. recta L. subsp.sarajevensis sensuHayek. These two taxa have the same morphological characteristics (shape and indumentum of leaves; morphology of sepals, petals and fruits) and both have 2n = 2x = 34 chromosomes. They cannot be considered as different taxa;S. beckeana Dörfler & Hayek is the valid name (synonyms:S. recta L. subsp.sarajevensis sensuHayek,S. hayekii Malý,S. recta L. subsp.hayekii Malý).

Erster Beitrag zur Neubearbeitung der Artengruppe derStachys recta L.     

Identification of aryl-phospho-β-<Emphasis Type="SmallCaps">d</Emphasis>-glucosidases in<Emphasis Type="Italic"> Bacillus subtilis</Emphasis>

Four aryl-phospho--d-glucosidases were identified in Bacillus subtilis by using 4-methylumbelliferyl-phospho--d-glucopyranoside as a substrate. Two of these enzymes are the products of the bglA and bglH genes, previously suggested to encode aryl-phospho--d-glucosidases, while the other enzymes are encoded by the yckE and ydhP genes. Together, these four genes account for >99.9% of the glucosidase activity in B. subtilis on aryl-phospho--d-glucosides. yckE was expressed at a low and constant level during growth, sporulation, and spore germination, and was not induced by aryl--d-glucosides. ydhP was also not induced by aryl--d-glucosides. However, while ydhP was expressed at only a very low level in exponential-phase cells and germinating spores, this gene was expressed at a higher levels upon entry into the stationary phase of growth. Strains lacking yckE or ydhP exhibited no defects in growth, sporulation, or spore germination or in growth on aryl--d-glucosides. However, a strain lacking bglA, bglH and yckE grew poorly if at all on aryl--d-glucosides as the sole carbon source.Abbreviations MU 4-Methylumbelliferone - MUG 4-Methylumbelliferyl--d-glucopyranoside - MUGal 4-Methylumbelliferyl--d-galactopyranoside - MUG-P 4-Methylumbelliferyl--d-glucopyranoside-6-phosphate     

Classical morphological features of centrospermous families

The orderCentrospermae (Caryophyllales, Chenopodiales) as treated inA. Englers Syllabus, 12th edition (1964), is compared with several other modern and older systems with the result that no less than 11–13 (and more) families are considered to be centrospermous in the strict sense; to these may be added thePolygonales and, doubtfully, thePlumbaginales andBatidales. As indicated by their name Centrospermae their main character is the central or basal placentation in combination with campylotropy (or amphitropy) of the ovules, seeds with perisperm, and coiled or curved embryos in peripheral position. Other outstanding features are found in the embryology; the ovules are bitegmic-crassinucellate, a nucellar cap is present, as well as an endostome and air spaces; the pollen is trinucleate. Anomalous secondary thickening in stems and roots often occurs. The pollen morphology, specific P-type sieve-element plastids, and the presence or absence of betalains are also important characters. Other floral features, especially the structure of the gynoecium, the androecium, the perianth and the receptacle, as well as the morphology of the inflorescences are of taxonomic importance. The putative relationships of theCaryophyllidae can perhaps best be resolved on the basis of more detailed morphological investigations (e.g. the so-called apocarpy, the development of the androecium, the pollen morphology, chromosome numbers, etc.).Presented in the Symposium Evolution of Centrospermous Families, during the XIIth International Botanical Congress, Leningrad, July 8, 1975.     

Giemsa karyotypes and their evolutionary significance inScilla bifolia,S. drunensis,andS. vindobonensis (Liliaceae)

Quantitatively evaluated C-banded and conventional karyograms are presented for the related speciesScilla bifolia subsp.danubialis Speta,S. drunensis (Speta)Speta, andS. vindobonensis Speta. On the basis of banding patterns and karyotype structureS. bifolia subsp.danubialis (2n = 18, 2×) andS. drunensis (2n = 36, 4×) are quite similar, whileS. vindobonensis (2n = 18, 2×) is entirely different. There is a moderate degree of karyotypic variation withinS. bifolia subsp.danubialis andS. drunensis. However, inS. vindobonensis karyotypes and banding patterns are almost completely stable over a geographical range of about 500 km. The present results confirm the recent taxonomic separation ofS. vindobonensis fromS. bifolia, and suggest a considerable phylogenetic distance between these two diploid species. The results are discussed with reference to the morphological characters of the species and their geographical distribution.Evolution ofScilla and Related Genera, II.Dedicated to Univ.-Prof. Dr.Elisabeth Tschermak-Woess on the occasion of her 60th birthday.     

Insect Induced Self-Pollination

Significant increase of pod production occurs inLupinus palaestinus Boiss. andL. pilosus Murr. following insect visits. The cause of this increase is investigated through (1) examination of the biology of pollination, (2) examination of pod production under various pollination conditions, (3) examination of cross pollination by genetical markers. All data strongly suggest that Insect Induced Self Pollination is the main factor in the increase of pod production of these species in nature.     

Isolation and chemical characterization of lipopolysaccharides from four Mycoplana species (M. bullata,M. segnis,M. ramosa and M. dimorpha)

Lipopolysaccharides (LPS), isolated from four Mycoplana species, i.e. the type strains of M. bullata, M. segnis, M. ramosa and M. dimorpha, were characterized onto their chemical composition and their respective lipid A-types. Those of M. bullata and M. segnis showed on DOC-PAGE an R-type character and had lipid A's of the Lipid A_DAG-type which exclusively contained 2,3-diamino-2,3-dideoxy-d-glucose as lipid A sugar. LPS's of M. ramosa and M. dimorpha showed, although only weakly expressed, ladder-like patterns on DOC-PAGE indicating some S-type LPS's and lipid A of the d-glucosamine type (Lipid A_GlcN). M. bullata LPS contained mannose and glucose in major amounts and additionally l-glycero-d-mannoheptose, whereas M. segnis LPS was composed of rhamnose, mannose and glucose together with both, d-glycero-d-manno- and l-glycero-d-manno-heptoses in a molar ratio of 1:2. All LPS's contained 2-keto-3-deoxy-octonic acid (Kdo), phosphate and an unidentified acidic component X. In addition to X, M. segnis LPS contained glucuronic and galacturonic acids, whereas M. ramosa LPS contained only galacturonic acid. Acetic acid hydrolysis of the LPS resulted in splitting off lipid A moieties, very rich in 3-hydroxy fatty acids, in particular in 3-OH-12:0 (in Lipid A_DAG), or in 3-OH-14:0 (in Lipid A_GlcN). Analysis of the 3-acyloxyacyl residues revealed major amounts of amide-linked 3-OH(3-OH-13:0)12:0 in lipid A of M. bullata and 3-OH(12:0)12:0 in lipid A of M. segnis. The rare 4-oxo-myristic acid (4-oxo-14:0) was observed only in M. bullata LPS, where it is ester-linked. Amide linked diesters could not be traced in M. ramosa and M. dimorpha. All four lipid A's lacked erster-bound acyloxyacyl residues.Non-standard abbreviations DAG 2,3-diamino-2,3-dideoxy-d-glucose - Kdo 2-keto-3-deoxy-octonate - LPS lipopolysaccharide - PITC phenyl isothiocyanate - NANA N-acetyl neuraminic acid     

Aurantiosporium,a new genus forUstilago subnitens (Ustilaginales)

A new genus,Aurantiosporium Piepenbring, Vánky & Oberwinkler (Ustilaginales), is proposed for the smut speciesUstilago subnitens Schröter & Hennings onScleria melaleuca Reichb. The soral morphology, teliospore development, the ultrastructure of the teliospore wall and teliospore germination ofAurantiosporium subnitens, studied on collections from Costa Rica, are described for the first time. The character set ofA. subnitens including intercellular teliospore development, spores in irregular groups and light coloured spore walls with numerous layers in TEM is neither known fromUstilago norCintractia nor any other smut species.Part 113 in the series Studies inHeterobasidiomycetes from the Botanical Institute, University of Tübingen.     

In vitro culture of Dianthus zeyheri subsp. natalensis,a South African carnation

A South African carnation species Dianthus zeyheri subsp. natalensis was cultured in vitro using techniques similar to those developed for the cut carnation (Dianthus caryophyllus L.). The ease of callus and suspension culture establishment makes this species a useful tool for fundamental biochemical studies.Abbreviations 2,4-d 2,4-dichlorophenoxyacetic acid - NAA -naphthaleneacetic acid - KIN kinetin - pCPA p-chlorophenoxyacetic acid     

sll1722, an unassigned open reading frame of<Emphasis Type="Italic"> Synechocystis</Emphasis> PCC 6803, codes for <Emphasis Type="SmallCaps">l</Emphasis>-<Emphasis Type="Italic">myo</Emphasis>-inositol 1-phosphate synthase

l-myo-Inositol 1-phosphate synthase (EC 5.5.1.4; MIPS) catalyzes conversion of glucose 6-phosphate to l-myo-inositol 1-phosphate, the first and the rate-limiting step in the production of inositol, and has been reported from evolutionarily diverse organisms. Two forms of the enzyme have been characterized from higher plants, viz. cytosolic and chloroplastic, and the presence of MIPS has been earlier reported from the cyanobacteria (e.g. Spirulina sp.), the presumed chloroplast progenitors. The present study demonstrates possible multiple forms of MIPS and identifies the gene for one of them in the cyanobacterium Synechocystis sp. PCC 6803. Following detection of at least two immunologically cross-reactive MIPS forms, we have been able to identify from the fully sequenced Synechocystis genome an as yet unassigned open reading frame (ORF), sll1722, coding for the approx. 50-kDa MIPS protein, by using biochemical, molecular and bioinformatics tools. The DNA fragment corresponding to sll1722 was PCR-amplified and functional identity of the gene was confirmed by a complementation assay in Saccharomyces cerevisiae mutants containing a disrupted INO1 gene for the yeast MIPS. The sll1722 PCR product was cloned in Escherichia coli expression vector pET20b and the isopropyl -d-thiogalactopyranoside (IPTG)-induced overexpressed protein product was characterized following complete purification. Comparison of the sll1722 sequences with other MIPS sequences and its phylogenetic analysis revealed that the Synechocystis MIPS gene is quite divergent from the others.Abbreviations CBB Coomassie Brilliant Blue - EST Expressed sequence tag - G6P d-Glucose 6-phosphate - IPTG Isopropyl -d-thiogalactopyranoside - MIPS l–myo-Inositol 1-phosphate synthase - ORF Open reading frame     

Characterization of a Na+/glucose cotransporter cloned from rabbit small intestine

Summary The Na⁺/glucose cotransporter from rabbit intestinal brush border membranes has been cloned, sequenced, and expressed inXenopus oocytes. Injection of cloned RNA into oocytes increased Na⁺/sugar cotransport by three orders of magnitude. In this study, we have compared and contrasted the transport properties of this cloned protein expressed inXenopus oocytes with the native transporter present in rabbit intestinal brush borders. Initial rates of¹⁴C--methyl-d-glucopyranoside uptake into brush border membrane vesicles andXenopus oocytes were measured as a function of the external sodium, sugar, and phlorizin concentrations. Sugar uptake into oocytes and brush borders was Na⁺-dependent (Hill coefficient 1.5 and 1.7), phlorizin inhibitable (K _i 6 and 9 m), and saturable (-methyl-d-glucopyranosideK _m 110 and 570 m). The sugar specificity was examined by competition experiments, and in both cases the selectivity wasd-glucose>-methyl-d-glucopyranoside>d-galactose>3-O-methyl-d-glucoside. In view of the close similarity between the properties of the cloned protein expressed in oocytes and the native brush border transporter, we conclude that we have cloned the classical Na⁺/glucose cotransporter.