Die Ergebnisse elektronenmikroskopischer Untersuchungen an somatischen Chromosomen des Menschen

Zusammenfassung Die Ergebnisse elektronenmikroskopischer Untersuchungen an somatischen Chromosomen werden zusammenfassend dargestellt und diskutiert. Es scheint kein Zweifel zu bestehen, daß das wesentliche Bauelement der Chromosomen Fibrillen sind, die als Elementarfibrillen bezeichnet werden. Die Elementarfibrillen haben einen Durchmesser von 100–150 Å in sofort fixiertem Material und 150–300 Å in Material, das vor der Fixierung einer hypotonen Behandlung ausgesetzt worden war. Dies gilt sowohl für Schnittpräparate wie für mit verschiedenen Techniken gewonnene Totalpräparate. Die Elementarfibrille entspricht dem DNS-Histon-Komplex, sie besteht wahrscheinlich aus einer in Falten gelegten feinen Fibrille von 20 Å Dicke (vermutlich ein DNS-Doppelschraubenmolekül) mit einer Histonhülle. Wie viele Elementarfibrillen ein Chromatid aufbauen, ist nicht sicher anzugeben, doch spricht nichts gegen die Annahme, daß ein Chromatid aus einer einzigen durchgehenden, in mehr oder weniger unregelmäßige Falten gelegten Elementarfibrille, und damit aus einem DNS-Molekül besteht. Menschliche Chromosomen lassen keine Subchromatidstrukturen erkennen.

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The results of electron microscopy on somatic chromosomes of man

Summary The results of electron microscopic studies on somatic chromosomes are reviewed and discussed. There seems to be no doubt that the main structural components of the chromosomes are fibrils; these are termed elementary fibrils. The diameter of these elementary fibrils is 100–150 Å in immediately-fixed material and 150–300 Å in material which before fixing has been subjected to hypotonic treatment. This applies equally to section preparations and to total preparations obtained using various techniques. The elementary fibril corresponds to the DNA-histone complex and is probably composed of a fine, multifolded fibril 20 Å thick, presumably a DNA double-helix molecule, possessing a histone sheath. It cannot be stated with certainty how many elementary fibrils form a chromatid, but nothing contradicts the assumption that a chromatid is formed of a single, continuous, irregularly-folded elementary fibril, i.e. of one DNA molecule. No subchromatids can be detected in human chromosomes.

     

Discrimination between closely related Triticeae species using genomic DNA as a probe

Summary Labelled total genomic DNA was used as a probe in combination with blocking DNA to discriminate between taxonomically closely related species in the genera Hordeum and Secale. Discrimination was possible both by Southern hybridization to size-fractionated restriction enzyme digests of genomic DNA and by in situ hybridization to chromosome preparations. To distinguish between two species (e.g. H. vulgare and H. bulbosum), genomic DNA from one species was used as the labelled probe, while unlabelled DNA from the other species was applied at a much higher concentration as a block. The blocking DNA presumably hybridized to sequences in common between the block and the labelled probe, and between the block and DNA sequences on the membrane or chromosomes in situ. If so, mainly species-specific sequences would remain as sites for probe hybridization. These species-specific sequences are dispersed and represent a substantial proportion of the genome (unlike many cloned, species-specific sequences). Consequently, rapid nonradioactive methods detected probe hybridization sites satisfactorily. The method was able to confirm the parentage of hybrid plants. It has potentially wide application in plant breeding for the detection of alien DNA transfer, and it can be easily adapted to many species.     

Characterization of progenies of Triticum aestivum- Psathyrostachys juncea derivatives by using genomic in-situ hybridization

Using genomicin-situ hybridization (GISH) technique, 7 translocation-addition lines, 6 translocation and translocation-addition lines, 2 ditelosomic addition lines and 1 translocation line were identified fromTriticum aestivum L. -Psathyrostachys juncea (Fisch.) Nevski intergeneric hybrids, of which translocation-addition and translocation and translocation-addition lines were not found in other reports. No substitutions and disornic additions were detected in the, hybrids and breakages occurred in allP. juncea chromosomes studied. Results have shown that the improved GISH technique is a rapid and economical method for use in this field.     

Parental genomes are separated throughout the cell cycle in a plant hybrid

The positions of the genomes originating from each parent were analysed in root-tip nuclei of the mature, sexual F₁ hybrid plant Hordeum vulgare (barley) x Secale africanum (a wild rye). The two genomes of the hybrid were identified in both spread and sectioned material by non-radioactive DNA:DNA in situ hybridization using labelled total genomic DNA from one parent as a proble and unlabelled total genomic DNA from the other parent to block non-specific hybridization. Complete three-dimensional reconstructions of sets of labelled sections enabled detailed analysis of the position of the genomes at interphase. The parental genomes lay in various non-intermixed configurations, including lateral and concentric arrangements. On spread preparations, the two parental genomes were found to be spatially separated throughout the cell cycle; the genome originating from H. vulgare tended to be located more centrally than that from S. africanum. This results show that the nucleus is spatially organized above the level of the DNA and chromosome at the genome level.by M.F. Trendelenburg     

Genomic in situ hybridization to identify alien chromosomes and chromosome segments in wheat

Summary Genomic in situ hybridization was used to identify alien chromatin in chromosome spreads of wheat, Triticum aestivum L., lines incorporating chromosomes from Leymus multicaulis (Kar. and Kir.) Tzvelev and Thinopyrum bessarabicum (Savul. and Rayss) Löve, and chromosome arms from Hordeum chilense Roem. and Schult, H. vulgare L. and Secale cereale L. Total genomic DNA from the introgressed alien species was used as a probe, together with excess amounts of unlabelled blocking DNA from wheat, for DNA:DNA in-situ hybridization. The method labelled the alien chromatin yellow-green, while the wheat chromosomes showed only the orange-red fluorescence of the DNA counterstain. Nuclei were screened from seedling root-tips (including those from half-grains) and anther wall tissue. The genomic probing method identified alien chromosomes and chromosome arms and allowed counting in nuclei at all stages of the cell cycle, so complete metaphases were not needed. At prophase or interphase, two labelled domains were visible in most nuclei from disomic lines, while only one labelled domain was visible in monosomic lines. At metaphase, direct visualization of the morphology of the alien chromosome or chromosome segment was possible and allowed identification of the relationship of the alien chromatin to the wheat chromosomes. The genomic in-situ hybridization method is fast, sensitive, accurate and informative. Hence it is likely to be of great value for both cytogenetic analysis and in plant breeding programmes.     

Nucleolar changes in human phytohaemagglutinin-stimulated lymphocytes

Summary The nucleoli of lymphocytes from circulating peripheral blood and from phytohaemagglutinin (PHA)-stimulated cultures (from 2 h–96 h) were studied using a silver method, RNA-specific fluorescent staining, and electron microscopy of ultrathin sections. In peripheral blood about 75% of the lymphocytes have one ring-shaped nucleolus composed of a distinct fibrillar centre surrounded by a dense pars fibrillaris and little granular material; the remaining lymphocytes showing two or more small ring-shaped nucleoli. With PHA stimulation, the number of cells with several nucleoli increases first (from 2 h–12 h). Next, cells containing one or, at most, two large nucleoli with nucleolonema devoid of fibrillar centers are seen (from 4 h on). 34 h after PHA, nucleoli of the compact type containing one or more fibrillar centres appear and comprise about 60% of the cells after 72 h. The appearance of more than one nucleolus per cell shortly after PHA administration suggests an activation of additional nucleolar organizer regions (NOR), which fuse to form one or two large nucleoli with nucleolonema. These are then transformed into compact nucleoli. The fibrillar centers stain preferentially with silver. They contain nonchromosomal proteins and may serve as stores for nucleolar proteins. The fusion of activated NORs during the first cell cycle explains the relatively high frequency of satellite associations in first mitoses compared to later mitoses after stimulation.     

High levels of genetic diversity throughout the range of the Portuguese wheat landrace 'Barbela'

BACKGROUND AND AIMS: Landrace populations represent an important intra-crop reservoir of biodiversity and source of novel gene alleles for use in breeding programmes. Here the aim was to measure the diversity of a wheat landrace, 'Barbela', from the north of Portugal. METHODS: DNA was extracted from 59 accessions of Barbela collected across its geographical range. Diversity was measured by microsatellite length polymorphisms using 27 primer pairs amplifying 34 polymorphic microsatellite loci. KEY RESULTS: High levels of polymorphism were found, with an average polymorphism information content of 0.52; an average of 4.77 alleles (range 2-11) were present at each locus, and half of these loci showed an additional allele in the reference variety 'Chinese Spring'. CONCLUSIONS: 'Barbela' is maintained from seeds collected by farmers, but it maintains high allelic variation, and no groupings of accessions were detected when analysed by geographical region, farm or climate, indicating that the wheat landrace is a homogeneous entity. The diversity within the farmer-maintained landrace demonstrates the importance of characterization and maintenance of landrace collections before valuable genetic combinations are lost as uniform commercial crops are introduced.     

Characterisation of mildew resistant wheat-rye substitution lines and identification of an inverted chromosome by fluorescent in situ hybridisation

Seven different mildew resistant wheat lines derived from crosses between triticale and bread wheat were examined by molecular cytogenetics and chromosome C-banding in order to determine their chromosomal composition. Genomic in situ hybridisation (GISH) showed the presence of rye germplasm in all the lines and identified three substitution lines, three double substitution lines and one addition-substitution line. C-banding identified rye chromosomes 1R and 4R in the addition-substitution line, rye chromosomes 1R and 6R in two substitution lines and 1R and 2R in the third line, and rye chromosome 1R in the three substitution lines. Two of the latter lines (7-102 and 7-169) contained a modified form of the chromosome; fluorescent in situ hybridisation (FISH) using five different repetitive DNA-probes showed a pericentric inversion of 1R in both lines. The breakpoints of the 1R inversion were between (1) the 5S rDNA site and the NOR-region on the satellite of the short arm, and (2) between two AAC(5) sites close to the centromere on the long arm. The role of the rye chromosomes in the mildew resistance, the utilisation of the inverted 1R and the significance of the lines in wheat breeding are discussed.