Andreas Katsiotis

The chromosomal distributions of Ty1-copia group retrotransposable elements in higher plants and their implications for genome evolution

Retrotransposons make up a major fraction – sometimes more than 40% – of all plant genomes investigated so far. We have isolated the reverse transcriptase domains of the Ty1-copia group elements from several species, ranging in genome size from some 100 Mbp to 23 000 Mbp, and determined the distribution patterns of these retrotransposons on metaphase chromosomes and within interphase nuclei by DNA:DNA in situ hybridization. With some exceptions, the reverse transcriptase domains were distributed over the length of the chromosomes. Exclusion from rDNA sites and some centromeres (e.g., slash pine, 23 000 Mbp, or barley, 5500 Mbp) is frequent, whereas many species exclude retrotransposons from other sites of heterochromatin (e.g., intercalary and centromeric sites in broad bean). In contrast, in the plant Arabidopsis thaliana, widely used for plant molecular genetic studies because of its small genome (c. 100 Mbp), the Ty1-copia group reverse transcriptase gene domains are concentrated in the centromeric regions, collocalizing with the 180 bp satellite sequence pAL1. Unlike the pAL1 sequence, however, the Ty1-copia signal is also detectable as weaker, diffuse hybridization along the lengths of the chromosomes. Possible mechanisms for evolution of the contrasting distributions are discussed. Understanding the physical distribution of retrotransposons and comparisons of the distribution between species is critical to understanding their evolution and the significance for generation of the new patterns of variability and in speciation.

Genetic relationships among species and cultivars of Pistacia using RAPDs and AFLPs

The genus Pistacia (Anacardiaceae) includes 11 species divided into four sections, according to leaf characters and nut morphology. Recently two monophyletic groups have been proposed by using cpDNA,Lentiscus and Terebinthus,containing evergreen and deciduous species, respectively. In the present work molecular markers, derived from two different methods, RAPD and AFLP, are used to study the relationships of native and introducedPistacia species present in Greece. According to the cophenetic correlation coefficients best results for both methods were obtained by using the Jaccard algorithm and the UPGMA clustering method. However, phenograms were constructed using the NJ method (rcs= 0.987 for RAPDs and rcs= 0.982 for AFLP), since it is less sensitive against varying mutation rates. Correlation among the genetic similarity (GS) matrices for the two methods was high(r = 0.941). The AFLP and RAPD phenograms were comparable with minor clustering differences. According to our results, two main branches are obtained, one containing the evergreen species P. lentiscusand the resin producing P. lentiscuscv. Chia (cultivated only in the island of Chios), and the other branch containing the deciduous species P. terebinthus,P. palaestina and P. vera, P. chinensis was clustered either with the evergreen species (RAPD data) or with the deciduous species (AFLP data). P. palaestina is clustered to P. terebinthus and can be considered as a subspecies of P. terebinthus, since its GS values are close or smaller than GS values of entries within species (P. vera). The four female cultivars were found to have a narrow genetic basis, probably related to cultivar ‘Nazareth‘ with Syrian origin.

The origin of the C-genome and cytoplasm of Avena polyploids

The contribution of C-genome diploid species to the evolution of polyploid oats was studied using C-genome ITS-specific primers. SCAR analysis among Avena accessions confirmed the presence of C-genome ITS1-5.8S-ITS2 sequences in the genome of AACC and AACCDD polyploids. In situ hybridization and screening of more than a thousand rRNA clones in Avena polyploid species containing the C-genome revealed substantial C-genome rRNA sequence elimination. C-genome clones sequenced and Maximum Likelihood Parsimony analysis revealed close proximity to Avena ventricosa ITS1-5.8S-ITS2 sequences, providing strong evidence of the latter’s active role in the evolution of tetraploid and hexaploid oats. In addition, cloning and sequencing of the chloroplastic trnL intron among the most representative Avena species verified the maternal origin of A-genome for the AACC interspecific hybrid formation, which was the genetic bridge for the establishment of cultivated hexaploid oats.

Genomic and chromosomal organization of Ty1-copia-like sequences in Olea europaea and evolutionary relationships of Olea retroelements

Abstract.The Ty1-copia-like retrotransposon is one of the commonest class of transposable elements in the plant kingdom, often comprising several percent of the total DNA content. We aimed to study the evolutionary relationships of Olea retroelements, using part of the reverse transcriptase domain, as well as the genomic and chromosomal organization of these sequences in Olea europaea chromosomes and their transcription activity and copy number. Fourteen clones, that were isolated from four different species, were sequenced and a phylogenetic tree was constructed based on their predicted amino acids. Five clones derived from O. europaea were clustered together with a 87% nucleotide sequence homology and two Olea oleaster clones showed 98% sequence homology. The rest of the clones showed heterogeneity among them, leading to a common ancestral transposon that existed before the genus arose. The Ty1-copia-like sequences have a dispersed genomic organization, physically distributed on all chromosomes, showing minor clustering in some cases and low copy numbers in the smallest chromosome pair. The total copy number in the O. europaea genome was estimated by dot blotting to be 40,000 in a haploid nucleus, but a number of these are non-functional since the sequenced clones contained stop codons and frame-shifts. Some Ty1-copia-like copies, present in O. europaea, were found to be methylated, while no differences in methylation were observed between DNA isolated from young leaves and callus-suspension cultures.

Molecular and chromosomal organization of two repetitive DNA sequences with intercalary locations in sugar beet and other Beta species

Abstract In a search for repetitive DNA sequences in the sugar beet genome, two sequences with repeat unit lengths of 143 and 434 bp were isolated and characterized. The pSV family showed an unusual conservation of restriction sites reflecting homogenization of the analyzed repeats. Members of the family are organized as tandem repeats as revealed by PCR and sequencing of dimeric units. The pSV satellite occurs in large intercalary arrays which are present on all chromosome arms of sugar beet. The pSV sequence family is present in different abundance in the sections Beta, Corollinae and Nanae but is not detectable by Southern hybridization in the section Procumbentes. The pDRV family is characterized by an interspersed genomic organization. The sequence is detectable in all sections of the genus and is amplified in species of the section Beta but was also detected, although at lower abundance, in the remaining three sections. Fluorescent in situ hybridization has shown that the pDRV sequence family is dispersed over all chromosomes of the sugar beet complement with some regions of clustering and centromeric depletion.

Genetic diversity of Avena ventricosa populations along an ecogeographical transect in Cyprus is correlated to environmental variables

Avena ventricosa Balansa ex Coss. is considered the C-genome donor of the cultivated hexaploid oat and is a ‘priority’ species for conservation, since it has limited geographic distribution and the only recorded populations in Europe are present in Cyprus. The current study attempts to characterize the genetic structure and fragmentation of the species via the application of genotypic markers. It was revealed that the genetic variety was mainly allocated among the populations collected, since clustering obtained was according to the geographic origin of the samples and the habitat. Species distribution modeling showed that the most important climatic variable defining A. ventricosa distribution is the mean diurnal temperature. Furthermore, significant association of the genetic structure to environmental variables was detected; overall, a negative association to precipitation was confirmed, while significant correlations of genetic structure and the temperature at the time of anthesis and germination were established. The safeguarding of this valuable genetic resource is discussed.