Gene-ecology of durum wheat HMW glutenin reflects their diffusion from the center of origin.

The production of many food items processed from wheat grain relies on the use of high gluten strength flours. As a result, about 80% of the allelic variability in the genes encoding the glutenin proteins has been lost in the shift from landraces to modern cultivars. Here, the allelic variability in the genes encoding the high molecular weight glutenin subunits (HMW-GSs) has been characterized in 152 durum wheat lines developed from a set of landraces. The allelic composition at the two Glu-1 loci (Glu-A1 and -B1) was obtained at both the protein and the DNA level. The former locus was represented by three alleles, of which the null allele Glu-A1c was the most common. The Glu-B1 locus was more variable, with fifteen alleles represented, of which Glu-B1b (HMW-GSs 7 + 8), -B1d (6 + 8) and -B1e (20 + 20) were the most frequently occurring. The composition of HMW-GSs has been used to make inferences regarding the diffusion and diversification of durum wheat. The relationships of these allelic frequencies with their geographical distribution within the Mediterranean basin is discussed in terms of gene-ecology.

Mutagen specificity in the induction of chromosomal aberrations in somatic cells of Drosophila melanogaster.

The distribution of chromosomal aberrations between and within chromosomes of male D, melanogaster somatic cells after treatment with UV has been analyzed. -- Distribution of the breaks between chromosomes was largely nonrandom since we found a higher aberration frequency than that expected on the Y chromosome. Moreover, within the chromosomes the aberrations are clustered in the pericentromeric heterochromatic regions. The above distribution is compared with that of the breaks induced by X rays and methyl-methane-sulphonate (MMS) which were distributed in a different pattern.

X-ray induction of chromatid interchanges is somatic cells of Drosophila melanogaster: variations through the cell cycle of the pattern of rejoining.

The relative frequencies of X-ray-induced symmetrical and asymmetrical chromatid interchanges were analysed as a function of the cell cycle in somatic cells of Drosophila melanogaster. In exchanges between autosomes, during the S phase, a clear prevalence of asymmetrical rejoining was observed. Later, this prevalence became reduced as the cell approached mitosis. On the other hand, in the exchanges between X-chromosomes in the females there were no significant variations in the type of rejoining through the cell cycle, there being a slight but consistent preference for symmetrical rejoining. These data are discussed in relation to the hypothesis proposed by Olivieri et al. [12] on the organization of the chromosomal sub-units in the interphase nucleus.

Identification of Lens culinaris ssp. culinaris chromosomes by physical mapping of repetitive DNA sequences.

We describe the characterisation and the chromosomal localisation of two repeated DNA sequences, named pLc30 (466 bp long, 64% AT residues) and pLc7 (408 bp long, 61% AT residues), isolated from lentil (Lens culinaris ssp. culinaris) genomic DNA. The pLc30 family is characterised by four internal repeats organised in a head-to-tail orientation, whereas the pLc7 contains many short direct subrepeats. The two families do not share significant sequence similarity. The distribution of these repetitive sequences in different Lens species and in other legumes was investigated. pLc30 is present in all Lens species investigated but absent from other genera examined. In contrast, pLc7 is present also in the genome of other legumes. As determined by FISH, the pLc30 sequence hybridises on six out of seven lentil chromosome pairs, while pLc7 hybridises on one only. The distribution of the nine different hybridisation sites of pLc30 allows the discrimination of all seven chromosome pairs and the construction of a karyotype of L. culinaris ssp. culinaris. Additionally, the combination of simultaneous and successive FISH with pLc7, 5S rRNA, 18S-5.8S-25S rRNA genes, and a telomeric sequence allowed the assembly of a physical map based on lentil karyotype.

Characterization of Dasypyrum villosum (L.) candargy chromosomal chromatin by means of in situ restriction endonucleases, fluorochromes, silver staining and C-banding.

The distribution of C-banded heterochromatin was determined in an inbred line of Dasypyrum villosum. Practically no difference in chromosome morphology or band distribution could be observed within the chromosomes of the same pair. Heterochromatin bands, revealed by Giemsa banding, were characterized by means of their differential reaction to fluorochromes, silver staining and in situ digestion with different restriction endonucleases. The results clearly indicate that in D. villosum two different classes of heterochromatin with different chromosomal local-ization exist: one is evidenced by both C-banding and DAPI staining and has mainly telomeric distribution, the other is evidenced only by C-banding and has mainly centromeric distribution.

Chromatin characterization by banding techniques, in situ hybridization, and nuclear DNA content in Cicer L. (Leguminosae).

The karyotypes of three accessions, one each from three annual species of the genus Cicer, namely Cicer arietinum, Cicer reticulation, and Cicer echinospermum, were examined and compared using C-banding, the fluorochromes chromomycin A3, DAPI, and Hoechst 33258, in situ hybridization of the 18S-5.8S-25S and 5S rDNA sequences, and silver staining. The nuclear DNA content of the three species and the amount of heterochromatin were also determined. The results suggest an evolutionary pathway in which C. reticulatum is the ancestral species from which both C. arietinum and C. echinospermum are derived with the loss of one pair of satellites; subsequently, C. echinospermum further differentiated by the accumulation of chromosomal rearrangement(s) that gave rise to a hybrid sterility barrier. Key words : Cicer, C-banding, fluorochromes, Ag staining, rRNA genes.

Genomic organization and phylogenetic relationships in the genus Dasypyrum analysed by southern and in situ hybridization of total genomic and cloned DNA probes.

Molecular cytogenetic methods have been used to study the controversial phylogenetic relationships between the species Dasypyrum villosum (L.) Candargy (2n=2x=14) and D. breviaristatum (Lindb. f.) Frederiksen (2n=4x=28). Using total genomic DNA from the two species as probes for in situ hybridization to chromosomes, we found that the pericentromeric regions of the chromosome arms of both species are similar, while distal regions show substantial differences. Two dispersed repetitive DNA sequences were isolated: pDbKB45 is distributed along the chromosomes but amplified in the subtelomeric regions of D. breviaristatum chromosomes, while pDbKB49, in both species, is less amplified in terminal regions. Size-separated restriction enzyme digests of DNA showed many repetitive fragments, but few in common between the two species. After probing Southern transfers with D. breviaristatum genomic DNA, all lanes showed similar hybridization patterns although one extra small band was evident in the D. breviaristatum lanes. In contrast, probing with D. villosum DNA showed very substantial differences between the two species. Genomic in situ hybridization to meiotic metaphases from an interspecific hybrid showed seven bivalents of D. breviaristatum origin and seven univalents from D. villosum. We also analysed the physical organization of 5S rDNA, 18S-25S rDNA and a tandemly repeated sequence from rye. Our data support an autotetraploid origin for D. breviaristatum, but its genome and that of D. villosum show extensive differences, so the tetraploid is unlikely to be directly derived from D. villosum.

The molecular cytogenetics of Vigna unguiculata (L.) Walp: the physical organization and characterization of 18s-5.8s-25s rRNA genes, 5s rRNA genes, telomere-like sequences, and a family of centromeric repetitive DNA sequences.

A knowledge of genome organization is important for understanding how genomes function and evolve, and provide information likely to be useful in plant breeding programmes involving hybridization and genetic manipulation. Molecular techniques, including in situ hybridization, molecular cloning and DNA sequencing, are proving valuable tools to investigate the structure, organization, and diversity of chromosomes in agricultural crops. Heterologous labelled 18 s-5.8 s-25 s (pTa71) and 5 s rDNAs (pTa794) were used for in situ hybridization on Vigna unguiculata (L.) Walp. chromosomes. Hybridization with 18 s-5.8 s-25 s rRNA gene probes occurred at the same chromosomal sites which were positive to the CMA fluorochrome. Silver staining of nucleolar-organizing regions indicated that all the rDNA sites detected using the 18 s-5.8 s-25 s rRNA gene probe possessed active genes. Degenerate telomeric repeats gave hybridization signals at the telomeres of most chromosomes and no intercalary sites were detected at metaphase; the sequences appear to have no preferential distribution in interphase nuclei. A repetitive DraI family from V. unguiculata was cloned (pVuKB1) and characterized. The DraI repeat is 488 nucleotides long, AT rich (74%), and hybridized on all chromosomes in the centromeric areas. The presence of this sequence family was investigated by Southern hybridization in different Vigna species and other Leguminoseae. It was only detected in V. unguiculata, and hence represents a species-specific DNA sequence.

S1 nuclease removes DNA from eukaryotic metaphase chromosomes: cytological evidence.

Fixed and unfixed human chromosomes, as well as fixed rye chromosomes were treated with S1 nuclease, which specifically cleaves single stranded DNA. Subsequent staining with either acridine orange, ethidium bromide or Giemsa revealed that, contrary to what has previously been reported, S1 digestion extensively altered chromosomal morphology and staining intensity, although the alteration was more pronounced in fixed as compared to unfixed metaphases. A number of mechanisms, which may account for our findings, have been invoked: a) the presence in metaphase chromatin of B-DNA/Z-DNA transitional junctions, b) the induction, by alcohol: acid fixation procedure, of nicks within regular B-DNA conformation and c) the induction of sites available to S1 by torsional stress due to metaphase high condensation degree.