Diversity of satDNA fraction in Cestrum, the Genus with the Largest Genomes within Solanaceae

Cestrum species present large genomes (~24 pg), a high occurrence of B chromosomes, and great diversity in heterochromatin bands. Despite this, there is maintenance of chromosome shape and karyotype symmetry. To deepen our knowledge on Cestrum genome composition, low coverage sequencing data of C. strigilatum and C. elegans were compared. Bioinformatics analyses showed retrotransposons comprising more than 70% of the repetitive fraction, followed by transposons (~18%). The four satDNA families that accumulated the most in the datasets were used as probes in FISH assays, and showed different distribution profiles along chromosomes. Most hybridization signals were located in the C-CMA/DAPI banding sites, including those related to AT-rich Cold-Sensitive Regions (CSRs) and heterochromatin. Although satellite probes hybridized in all tested species, a satDNA family named CsSat49 was highlighted as it predominates in centromeric regions. Data suggest that the satDNA fraction is still conserved in the genus, although there is variation in the number of FISH signals between karyotypes, as well as in the B chromosomes. This study brings an important advance in the knowledge on genome organization and heterochromatin composition in Cestrum, especially on the distribution and differentiation mechanisms of satellite fraction between species of a genus of Solanaceae with large genomes.

Integrative rDNAomics—Importance of the Oldest Repetitive Fraction of the Eukaryote Genome

Nuclear ribosomal RNA (rRNA) genes represent the oldest repetitive fraction universal to all eukaryotic genomes. Their deeply anchored universality and omnipresence during eukaryotic evolution reflects in multiple roles and functions reaching far beyond ribosomal synthesis. Merely the copy number of non-transcribed rRNA genes is involved in mechanisms governing e.g., maintenance of genome integrity and control of cellular aging. Their copy number can vary in response to environmental cues, in cellular stress sensing, in development of cancer and other diseases. While reaching hundreds of copies in humans, there are records of up to 20,000 copies in fish and frogs and even 400,000 copies in ciliates forming thus a literal subgenome or an rDNAome within the genome. From the compositional and evolutionary dynamics viewpoint, the precursor 45S rDNA represents universally GC-enriched, highly recombining and homogenized regions. Hence, it is not accidental that both rDNA sequence and the corresponding rRNA secondary structure belong to established phylogenetic markers broadly used to infer phylogeny on multiple taxonomical levels including species delimitation. However, these multiple roles of rDNAs have been treated and discussed as being separate and independent from each other. Here, I aim to address nuclear rDNAs in an integrative approach to better assess the complexity of rDNA importance in the evolutionary context.

Tandemly repeated C-C-C-C-A-A hexanucleotide of Tetrahymena rDNA is present elsewhere in the genome and may be related to the alteration of the somatic genome.

The ribosomal RNA genes of the Tetrahymena macronucleus exist as extrachromosomal, linear molecules. The termini of these molecules have been shown to contain the tandemly repeated hexanucleotide (C-C-C-C-A-A)n. In this study the same or related sequences were found in other locations of the genome. Using the depurination method, we showed that macronuclear DNA contained this sequence even after rDNA had been removed. The sequence was found mainly in the repetitive fraction of the DNA. The presence of this sequence in both the macronucleus and the micronucleus was also shown by Southern hybridization using C-C-C-C-A-A repeat as a probe. Comparison between the hybridization patterns of macronuclei and micronuclei reveals interesting differences. Whereas the two nuclei share the same genetic origin, the majority of the restriction enzyme digestion sites flanking the C-C-C-C-A-A repeat appear to be different. Such a difference was found to be specific for this sequence, because it was not detected when other sequences were used for hybridization. These results suggest that some kind of alteration has occurred in the genome during the formation of the macronucleus, and that the C-C-C-C-A-A repeat may be related to this process.

Analysis of the genetic complexity of a hybrid plant

Nuclear DNA from wheat, rye, and Triticale plantlets was isolated and purified. Purity of the samples was tested spectrophotometrically. Tm values and hyperchromicities were determined for the three DNA samples. The DNA(s) were denaturated and annealed under controlled conditions. Cot curves were carried out for total DNA on the three samples and for the repetitive DNA fractions.The analysis of those curves indicated that the genomes of wheat and rye are made of highly repetitive, intermediate, and unique DNA fractions. The highly repetitive fraction (107) is suggested as corresponding to palindrome areas in the genome.The Triticale genome showed a similar pattern to the parents, except for the highly repetitive DNA fraction which showed different orders of repetitiveness.