Development and characterization of nuclear microsatellite markers for Eremanthus erythropappus and their transferability across related species

BACKGROUND: We developed simple sequence repeats (SSR) for Eremanthus erythropappus (DC.) MacLeish, an endangered tree species endemic to the Brazilian Savanna and Atlantic Forest biomes, and tested their transferability to two closely related Eremanthus species. RESULTS: Using a genomic library enriched with tandem repeat motifs, we identified 16 primer pairs, and characterized them in two populations. Nine primers amplified the expected size fragments and seven SSRs were polymorphic, providing a total of 38 alleles and an average of 4.22 alleles per marker. The polymorphic information content (PIC) ranged from 0.44 to 0.94 with an average of 0.65. The average observed heterozygosity across all loci varied from 0.61 to 1.00. The observed ( HO ) and expected ( HE ) heterozygosity within the two populations varied from 0.65 to 1.00 and from 0.31 to 1.00, respectively. CONCLUSIONS: These newly developed SSR markers are a powerful tool for population genetic analyses and may be useful in studies on species ecology, evolution, and taxonomy.

Inferring possible population divergence in Espeletia pycnophylla (Asteraceae) through morphometric and paleogeographic approaches

The phenotypic structure within and between plant populations is generally influenced by their distribution patterns in space and time; therefore, the study of their divergence is a central issue for the understanding of their microevolutive processes. We boarded the hypothesis that three populations of Espeletia pycnophylla show phenotypic divergence as one of the possible implications of their geographic isolation in the Southern Colombian Andes. We used the Elliptic Fourier Descriptors (leaf shape) and traditional leaf morphometry (leaf size) of 347 leaves to measure inter and intra-population variation and a comparison between a paleogeographic reconstruction with an actual estimate of the distribution areas of E. pycnophylla in order to identify their main changes during the last 14 000 years. The three populations showed significant differences in leaf morphometry and a positive correlation between the matrices of morphometric and geographic dissimilarities, indicating that the inter-population divergence increases between further populations, so that the morphometric structure reflects their spatial distribution. The geographical and paleogeographical estimates evidenced a conspicuous process of reduction and fragmentation of the distribution area of E. pycnophylla since the Late-Glacial until the Holocene. We suggest that these results support possible scenarios of vicariance events, which allow us to approach the divergence of these populations in terms of their historic biogeographic relations. However, genetic analyses are still needed to support these results. Rev. Biol. Trop. 58 (4): 1261-1270. Epub 2010 December 01.

La estructuración fenotípica entre y dentro de poblaciones vegetales responde generalmente a sus patrones de distribución espacio-temporales, por lo tanto, el estudio de su divergencia es un tema central dentro de sus procesos microevolutivos. En esta investigación abordamos la hipótesis que tres poblaciones de Espeletia pycnophylla presentan divergencia fenotípica como uno de los posibles efectos de su aislamiento geográfico en los Paramos del suroeste de Colombia. Utilizamos los Descriptores Elípticos de Fourier (forma foliar) de 117 hojas y algunas medidas morfológicas tradicionales (tamaño foliar) como indicadores de la variación intra e inter-poblacional, además de una comparación entre una reconstrucción paleogeográfica con una estimación del área de distribución actual de la especie para identificar sus principales cambios durante los últimos 14 000 años. Todas las poblaciones mostraron diferencias significativas y además, existió una correlación positiva entre las matrices de disimilitud morfométrica y geográfica, indicando que la divergencia incrementa entre poblaciones lejanas. Paralelamente, las estimaciones geográficas y paleogeográficas evidenciaron un proceso de reducción y fragmentación del área de distribución de E. pycnophylla desde el Tardiglacial hasta el Holoceno. Sugerimos que ambos resultados apoyan escenarios de posibles series de eventos de vicarianza para las poblaciones estudiadas y nos permiten entender su divergencia en términos de sus relaciones biogeográficas históricas.

Contrasting genetic diversity and differentiation of populations of two successional stages in a neotropical pioneer tree (Eremanthus erythropappus, Asteraceae)

Eremanthus erythropappus, commonly known as “candeia”, is an abundant pioneer tree species, forming dense populations known as “candeial”, but it is also found in forests at middle stages of succession. Trees from forests are bigger and occur in lower density than in the “candeial”. The objectives of the present study were to investigate if the decrease in population density during successional process is accompanied by 1) changes in within-population genetic diversity, and 2) differentiation of populations. Eight populations, four of early successional stage (“candeial”) and four of middle successional stages (forest), were analyzed with RAPD markers. The genetic diversity found was high compared to other tree species analyzed with RAPD markers. AMOVA revealed that most of the genetic variations of E. erythropappus were found within populations (85.7%), suggesting that this species is predominantly outcrossing. The relatively low differentiation among the populations can be attributed to small distances among the populations analyzed (0.2 to 10.8 km). No indication that populations from middle successional habitats show lower genetic variation than populations from early successional stages was found. The percentage of polymorphic fragments (82.8 and 84.8%) and the Shannon indexes (0.442 and 0.455) were similar in “candeial” and forest, respectively. These results suggest that if an increase in selection intensity occurred during succession, it did not result in a decrease in genetic diversity or that the selection effect was balanced by other factors, such as gene flow. Higher significant differentiation among E. erythropappus populations from “candeial” in relation to that among populations from forest was also not detected.

Effects of Pterocaulon polystachyum DC. (Asteraceae) on onion (Allium cepa) root-tip cells

In Brazil leaf infusions of the plant Pterocaulon polystachyum DC (Asteraceae) are used to treat digestive problems. We used plants from six P. polystachyum populations to prepare fresh aqueous leaf infusions at the concentration normally used in Brazil (2.5 g L-1) and at twice (5 g L-1) and four times (10 g L-1) this concentration. We rooted onion bulbs in a water control or one of the infusions for 24 h and used the in vivo onion (Allium cepa) root-tip cell test to investigate the potential mutagenicity of the infusions by calculating the mitotic index for the control and the experimental treatments and comparing them using the Chi-squared test (chi2, p = 0.05). There was a highly significant decrease in the mitotic index of root-tip cells treated with infusion as compared to those exposed to water only. These results indicate that P. polystachyum infusions present cytotoxic and anti-proliferative activity and therefore have therapeutic potential.

Cytogenetic study of the genus Cousinia ( Asteraceae, section Serratuloideae ) in Iran

Meiotic studies of ploidy level, chromosome paring and chiasma frequency were performed on 11 Cousinia (Asteraceae) species of the section Serratuloideae. The diploid number of the species studied was 2n = 2x = 24 and 26 so these species possess two different basic numbers (x = 12 or 13), a phenomenon common to other sections of the genus. The chromosome numbers of 9 species are reported here for the first time. When the 2n = 24 and 2n = 26 species were subjected to cluster analysis based on relative meiotic characters two different clusters were formed indicating their distinctness. Our data support the results obtained from morphometry, anatomy, pollen morphology and molecular studies of the genus Cousinia.