Easy determination of ploidy level in Arabidopsis thaliana plants by means of pollen size measurement

Summary Cytogenetic examination of transgenic Arabidopsis thaliana (L.) Heynh. plants obtained by Agrobacterium-mediated gene transfer to cotyledon- and root-explants or by direct gene transfer into protoplasts revealed a high percentage of tetraploid or aneuploid transformants. Depending on the transformation procedure used, 13% (root explant transformation), 33% (cotyledon explant transformation), or 38% (direct gene transfer) of the transformants showed aberrant ploidy levels. A good correlation between the ploidy level of a plant and the size of its pollen grains was observed. This allows quick and simple testing of the ploidy level of transgenic Arabidopsis plants.Abbreviations AM Arabidopsis medium - ANOVA analysis of variance - DAPI 4,6-Diamidino-2-phenylindole - PEG polyethyleneglycol     

Purification and properties of poly(3-hydroxyvaleric acid) depolymerase from Pseudomonas lemoignei

During growth on poly(3-hydroxyvaleric acid), P(3HV), or valerate Pseudomonas lemoignei secretes a P(3HV) depolymerase. This P(3HV) depolymerase was purified from the culture medium of valerate-grown cells by ammonium sulphate precipitation, chromatography on DEAe-sephacel and CM-Sepharose CL 6B. The relative molecular masses of the native as well as the sodium dodecyl sulphate (SDS)-treated enzyme were 53 000 or 54 000, respectively. In contrast to the poly(3-hydroxybutyric acid), P(3HB), depolymerase of Comamonas sp. and P(3HB) depolymerases A and B of P. lemoignei, which are specific for the hydrolysis of P(3HB), the purified P(3HV) depolymerase hydrolysed P(3HB), P(3HV) and co-polymers of 3-hydroxybutyric acid and 3-hydroxyvaleric acid at similar rates. Poly(hydroxyalkanoic acids), consisting of monomers with six and more carbon atoms or substrates characteristic for lipases such as Tween 80 or triolein were not hydrolysed. Maximum activities were measured in 50mm TRIS-HCl buffer, pH 8.0, at 55° C. The apparent K _m values of the purified P(3HV) depolymerase for P(3HB) and P(3HV) were 77 and 65 g polyester/ml, respectively. As the main product of enzymatic hydrolysis of P(3HV), 3-hydroxyvalerate was identified. The depolymerase was insensitive to p-hydroxymercuribenzoate but sensitive to dithioerythritol and phenylmethylsulphonyl fluoride, indicating the absence of active reduced sulphur groups and the presence of essential disulphide bonds and serine residues. Correspondence to: D. Jendrossek     

Evidence for the Existence of Differential O-Glycosylated α5-Subunits of the γ-Aminobutyric AcidA Receptor in the Rat Brain

Abstract: Polyclonal antibodies were raised to synthetic peptides having amino acid sequences corresponding with the N- or C-terminal part of the γ-aminobutyric acid_A (GABA_A) receptor α₅-subunit. These anti-peptide α₅(2–10) or anti-peptide α₅(427–433) antibodies reacted specifically with GABA_A receptors purified from the brains of 5–10-day-old rats in an enzyme-linked immunosorbent assay and were able to dose-dependently immunoprecipitate up to 6.3 or 13.1% of the GABA_A receptors present in the incubation, respectively. In immunoblots, each of these antibodies reacted with the same two protein bands with apparent molecular mass of 53 or 57 kDa. After exhaustive treatment of purified GABA_A receptors with N -Glycanase, each of these antibodies identified two proteins with apparent molecular masses of 46 and 48 kDa. Additional treatment of GABA_A receptors with neuraminidase and O -Glycanase resulted in an apparently single protein with molecular mass of 47 kDa, which again was identified by both the anti-peptide α₅(2–10) and the anti-peptide α₅(427–433) antibody. These results indicate the existence of at least two different α₅-sub-units of the GABA_A receptor that differ in their carbohydrate content. In contrast to other α- or β-subunits of GABA_A receptors so far investigated, at least one of these two α₅-subunits contains O-linked carbohydrates.     

GENOTYPIC VARIATION AND ALTERNATING DNA LEVELS AT CONSTANT CHROMOSOME NUMBERS IN THE LIFE HISTORY OF THE BROWN ALGA HAPLOSPORA GLOBOSA (TILOPTERIDALES)1

The brown algal order Tilopteridales contains three monospecific genera with reduced life histories, Which are assumed to have been derived form ancestors with oogamous reproduction and alternation of generations. The Newfoundland population of Haplospora globosa Kjellman still shows an alternation of gametophytes and sporophytes, but the chromosome Numbers remain equal because of parthenogenesis and apomeiosis, However, DNA fluorometry showed that the DNA level is twice as high in the Sporophytes as in the gametophytes, The DNA variation at constant chromosome numbers is presumably due to endomitosis combined with a law degree of polyteny. A genotypic variant of Haplospora is represented by the population at Helgoland (F.R.G.) where only sporophytes exist, Spores develop into sporophytes instead of gametophytes, and the plants have reduced chromosome number but the same DNA level as the Newfoundland sporophytes     

A novel type of meso-diaminopimelic acid-based peptidoglycan and novel poly (erythritol phosphate) teichoic acids in cell walls of two coryneform isolates from the surface flora of French cooked cheeses

The primary structure of the peptidoglycan and the teichoic acids of two coryneform isolates from the surface flora of French cooked cheeses, CNRZ 925 and CNRZ 926, have been determined. In the peptidoglycan, meso-diaminopimelic acid was localized in position three of the peptide subunit. It contained an d-glutamyl-d-aspartyl interpeptide bridge, connecting meso-diaminopimelic acid and d-alanine residues of adjacent peptide subunits. The -carboxyl group of d-glutamic acid in position two of peptide subunits was substituted with glycine amide. The teichoic acid pattern and composition differed between the strains: both contained an erythritol teichoic acid and strain CNRZ 925 also contained an N-acetylglucosaminylphosphate polymer. The erythritol teichoic acids differed in terms of the quality and quantity of substituents, but they both had N,N-diacetyl-2,3-diamino-2,3-dideoxyglucuronic acid in common.Abbreviations DNP dinitrophenyl - Ery erythritol - Gal galactose - GlcN glucosamine - GlcNAc N-acetylglucosamine - GlcUANAc₂ N,N-diacetyl-2,3-diamino-2,3-dideoxyglucuronic acid - Hex UANAc₂ N,N-diacetyl-2,3-diamino-2,3-dideoxyhexuronic - acid m-Dpm, meso-diaminopimelic acid - Mur muramic acid - MurNAc N-acetylmuramic acid     

Age-related nonrandom chromosomal abnormalities in human low-grade astrocytomas

We report a cytogenetic investigation of 55 low-grade astrocytomas in 52 patients, 15 children and 37 adults. In addition to numerical aberrations such as trisomy 7 and gonosomal losses, we found structural and/or numerical aberrations of chromosome 1 in eight astrocytomas. There was a striking difference between the rearranged chromosomes in pediatric and adult patients. Whereas the pediatric tumors revealed monosomies 1p with accompanying trisomy 1q, the astrocytomas in adults showed partial or complete monosomies 1q.     

Retrospective study of the parental origin of the extra chromosome in trisomy 18 (Edwards syndrome)

The parental origin of the extra chromosome in trisomy 18 was traced in 30 informative families using highly polymorphic (CA) repeats mapped on the long arm of chromosome 18. Proband DNA was recovered from slides of chromosome preparations in 28 cases and from paraffin-embedded tissues in two cases. The extra chromosome was found to be of maternal origin in 26 cases (86.7%), and paternal origin in 4 cases (13.3%).     

Elevated CO2 and the mineral content of herbaceous and woody plants

The CO₂ enrichment effects (300–650 µmol mol^-1) on mineral concentration (N, P, K, Ca, Mg, Mn, Fe, Zn), absolute total mineral contents per individual and of whole stands of four herbaceous (Trifolium repens L.,Trifolium pratense L.,Lolium perenne L.,Festuca pratensis HUDS.) and two woody species (Acer pseudo-platanus L.,Fagus sylvatica L.) were investigated.In general, the mineral concentration of the plant tissues decreased (all six species: N>Ca>K>Mg) with the exception of P. Mn and Fe were only determined for the tree species. Both decreased in concentration (Mn>Fe). Zn was only analysed forTrifolium pratense andFestuca pratensis and decreased significantly in the grass.Despite of decreases in concentrations of as much as 20% in some cases there were increases in absolute amounts per individual and, therefore, in the whole vegetation up to 25% because of the enhanced dry matter accumulation at elevated CO₂ supply.Dedicated to Prof. Dr. R. Bornkamm, TU-Berlin, on behalf of his 60th birthday     

Über schlauchförmige Ausstülpungen an der Zellmembran der Makrogametocyten von Eimeria perforans

Summary The surface of the gametocytes and gametes of Eimeria perforans reveals tube-like extrusions which have not been discovered so far in coccidia. These slender tubes are 650 Å in width and at least 1,3 in length. Probably they represent resorbing organelles. The tubes occur only at the surface of older female gametocytes and gametes and have not been observed in merozoites, schizonts, microgametocytes and male gametes. In transversal sections the tubes look like vesicles and are bordered by a membrane the layers of which are not sharply defined. The interior of the tubes seems to be empty after electron microscope observations. In longitudinal sections the membrane of the tubes is striated. The repeating unit of the dark and light bands amounts to about 165 Å. This appearance cannot be explained so far.

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Für beratende Hilfe sind wir Herrn Prof. Dr. R. Danneel und Herrn Prof. Dr. K. E. Wohlfarth-Bottermann zu Dank verpflichtet, für technische Unterstützung Fräulein cand. rer. nat. B. Volkmann. Herrn Dr. D. Spiecker von der Forschungsstelle für Jagdkunde und Wildschadenverhütung, Beuel, danken wir für die Durchführung der Infektionen. Die Mittel für die Untersuchungen stellte uns die Deutsche Forschungsgemeinschaft zur Verfügung.