Characterization of 12 microsatellite loci for Hypochaeris chillensis (Asteraceae) and cross-amplification in related species.

PREMISE OF THE STUDY: Hypochaeris is considered a biological model to understand evolutionary processes in the vascular flora of South America, particularly from the temperate portion of the continent. We report the development and characterization of microsatellite markers for H. chillensis to assess the genetic variability and patterns of population structure of the species. • METHODS AND RESULTS: Twelve microsatellite primers were isolated using a CT- and GT-enriched genomic library. PCR amplification detected one to five alleles, with 2.91 alleles per locus on average. Tested for cross-amplification, all primer pairs were successfully amplified in 10 South American species and in the putative ancestor of the group, H. angustifolia. • CONCLUSIONS: The microsatellites can be used to assess genetic diversity and population structure of H. chillensis. Application in other species will focus on the elucidation of adaptive radiation of the genus in South America.

Characterization, genomic organization and chromosomal distribution of Ty1-copia retrotransposons in species of Hypochaeris (Asteraceae).

This study aims to analyze the diversity of Ty1-copia retrotransposons in 18 taxa of Hypochaeris, including two Old World species H. maculata (2n=2x=10) and H. angustifolia (2n=2x=8), and representatives of the South American species (16 accessions of 15 species; all 2n=2x=8). Analysis of 380 PCR-amplified sequences, corresponding to a conserved domain of the subset of Ty1-copia reverse transcriptase (rt) gene amplifiable with degenerate standard primers, showed high levels of intra- and interspecific heterogeneity. Nucleotide diversity (Pi) of the copia fragments was high in all species and varied from 0.229 (H. angustifolia) to 0.412 (H. chillensis). Higher sequence heterogeneity correlates positively with larger genome size among analyzed species. Phylogenetic analyses of amplified fragments revealed different patterns of intraspecific heterogeneity within species, with most sequences forming one well-supported main clade while a few sequences fall into small clades or are left ungrouped. The combined analysis of all sequences revealed the presence of three main clades and showed that highly diverged species contain closely related Tyl-copia group retrotransposons. One of the main clades includes rt sequences of all South American species and three sequences of their putative ancestor, H. angustifolia, but no sequence of the Old World H. maculata. FISH with copia retrotransposons in four Hypochaeris species, including H. maculata and H. angustifolia and New World H. apargioides and H. spathulata, revealed differences in the chromosomal distribution between the two groups. In Old World species copia retroelements are distributed over the whole length of the chromosomes, excluding rDNA sites and some centromeres. In the South American species the two largest chromosome pairs are enriched in copia, while most of the long arms of the two small pairs of chromosomes are devoid of these elements. The patterns of heterogeneity and chromosomal distribution of Ty1-copia retrotransposons in Hypochaeris are discussed in the context of the origin, genome evolution and organization of the South American species.

Chromosomal organization and phylogenetic relationships in Hypochaeris species (Asteraceae) from Brazil

The association of cytogenetic and molecular techniques has contributed to the analysis of chromosome organization and phylogeny in plants. The fluorochrome GC-specific CMA3, fluorescent in situ hybridization (FISH) and RAPD (Random Amplified Polymorphic DNA) markers were used to investigate chromosome structure and genetic relationships in Hypochaeris (Asteraceae). Seven species native to South America, and two species introduced from Europe (H. glabra and Hypochaeris sp) were studied. FISH with rDNA probes identified one or two loci of 18S-5.8S-25S rDNA in the South American Hypochaeris species and one locus in the European species. Only one 5S rDNA locus was seen in all species studied. Blocks of GC-rich heterochromatin (CMA-positive bands) associated to 18S-5.8S-25SrDNA loci were detected in all species investigated. Co-location of 5S rDNA and CMA bands was also observed, except for three South American species and Hypochaeris sp. In two South American species, additional CMA bands not related to rDNA were observed on the long arm of chromosome 2, near to the centromere. Hypochaeris glabra exhibited additional CMA-positive signals distributed at pericentromeric regions, on the short arms of all chromosomes. A total of 122 RAPD markers were used to determine the genetic relationships among species. The level of polymorphism was very high, revealing two genetic groups comprising the South American and the European species, thus supporting a previous hypothesis of monophyly of the South American Hypochaeris species. The coefficients of genetic similarity between European and South American species were 0.35, on average. Polymorphism was also high within the two groups. The genetic associations observed with RAPD markers were consistent with chromosome characteristics. Species carrying similar distribution of 45S rDNA loci and CMA-positive signals were included in the same group revealed by RAPDs. Cytogenetic and molecular data support the view that not only chromosome rearrangements, but also changes in DNA sequence took place during the diversification of the South American Hypochaeris species.

Screening of Brazilian plants for the presence of peroxides

Chloroform or dichloromethane extracts of 357 southern Brazilian plant species were tested for the presence of peroxides by thin-layer chromatography, using the spray reagent from Huber & Fröhlke. >From the species tested, 71 (20 por cento) showed positive results and most of them (56 por cento) are Asteraceae species. The species tested were mainly from Asteraceae, but 55 more families were screened, in a total of 77 genera surveyed...