Cires, E., Cuesta, C., Fernández, M.A., Nava, H.S. Flow cytometry analysis has been widely applied in the determination of nuclear DNA content and ploidy level in many organisms. Despite being the most appropriate method for DNA content measurement, flow cytometry also presents some limitations. A fairly common, but little-studied problem is the effect on measurements of the presence of secondary metabolites. A good example is the genus Viola, which is composed of 525-600 species distributed worldwide. These species have proved to be problematic for flow cytometric analyses due to the release of extremely mucilaginous compounds into the nuclear suspension. In this work, the genome size of 13 species of Viola using flow cytometry are presented for the first time. Despite obtaining histograms with high coefficients of variation, we here present an optimized protocol to remove cytoplasmic compounds, particularly mucilaginous ones, from plant nuclei that pave the way for its application to estimate the genome size of other species exhibiting similar problems. Statistical analyses revealed significant differences between sections Viola and Melanium, and within each section (P < 0.001). Furthermore, statistically significant differences were not detected among samples of the same species. ResumenCires, E., Cuesta, C., Fernández, M.A., Nava, H.S., Vázquez, V.M. & Fernández, J.A. 2011. Aislamiento de núcleos para citometría de flujo en plantas con alto contenido de compuestos mucilaginosos: un ejemplo en el género Viola L. (Violaceae). Ana les Jard. Bot. Madrid 68(2): 139-154 (en inglés).El análisis mediante citometría de flujo ha sido aplicado de modo general para determinar el contenido de ADN nuclear y el nivel de ploidía en muchos organismos. A pesar de ser el método más adecuado para medir la cantidad de ADN, esta técnica también presenta algunas limitaciones. Un problema bastante común, aunque poco estudiado, es el efecto de los metabolitos secundarios en los resultados obtenidos. Un ejemplo respecto a la presencia de estos compuestos se encuentra en el género Viola, compuesto por 525-600 especies distribuidas por todo el mundo. Las especies de este género ya han sido previamente descritas como problemáticas en los análisis de citometría de flujo debido a la presencia de compuestos extremadamente mucilaginosos en las suspensiones de núcleos. En el presente trabajo se analiza por primera vez el tamaño genómico de 13 especies del genero Viola mediante el empleo de citometría de flujo. A pesar de los altos valores mostrados en los coeficientes de variación de los histogramas, se presenta un protocolo optimizado para eliminar compuestos citoplasmáticos, y más concretamente mucilaginosos de las suspensiones nucleares, siendo de aplicación en la estimación del tamaño genómico de plantas con problemas similares. Los análisis estadísticos mostraron diferencias significativas entre las secciones Viola y Melanium, así como dentro de cada sección (P < 0,001). Además, no se encontraron diferencias significativas entre...

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The fluorescence properties of spores and pollen grains examined under ultraviolet incident light are used to assess the maturity of sedimentary organic matter and may have other applications in relation to recent sediments, in areas such as palaeoenvironmental research. In this study pollen grains and spores from 33 species common in peat ecosystems were mounted on a glass slide in accordance with standard palynologycal procedures for recent plants. The main objective of this work was to assess the variability of fluorescence spectra of pollens and spores within a single species or even within a single sample. A minimum of 10 spectra were recorded from each sample and were averaged to obtain a spectrum characteristic of each sample. Both the average scattering and the scattering in different spectral regions were calculated using the standard deviation (SD) and the coefficient of variation (CV). The effect of the preparation techniques was assessed on some samples of Ericaceae taxa. The results indicated similar spectra for alcohol-washed and distilled water-washed samples, whereas the application of an acetolysis solution caused an increase in intensity and a shift to longer wavelengths. The spectra corresponding to the Sphagnum spores had the lowest intensity of all the families studied and displayed their maxima at the lowest registered wavelengths. They often showed a peak in the red region of the spectra, causing a larger scatter in fluorescence in this region. This peak is probably the result of wax or cytoplasmic material attached to the exospore. A significant number of Ericaceae taxa had two fluorescing pollen populations: a blue one of high intensity andsmaller size a yellow-orange one of low intensity and larger size. This difference could be related to different degrees of maturity of the pollen grains. In the case of pollen grains of herbaceous, tree and bush plants the largest scatter was found in the tails of the spectra towards the blue and red regions. The decreasing trend of fluorescence intensity with the shift of the spectra towards red was not observed in the pollen and spores of fresh plants. A good correlation was found between the spectral maxima ( max ) and the red-green quotient (Q R/G ) regardless of the type of plant.

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FERNANDEZ PRIETO, J. A., VERA, M. L., ALVAREZ MARTINEZ, M. J., DIAZ, T. E., FERNANDEZ CASADO, M. A., FERNANDEZ‐CARVAJAL, M. C, GUTIERREZ VILLARIAS, M. I. & NAVA, H. S., 1993. Chromosome numbers and geographical distribution of Ulex europaeus subsp. europaeus (Leguminosae). Chromosome numbers are reported in Ulex europaeus subsp. europaeus (99 samples from northern Spain, France and Great Britain). Three levels of ploidy (diploid, tetraploid and hexaploid) have been detected; the diploid and tetraploid levels, hitherto unknown in this taxon, have been found exclusively in coastal areas of northern Spain and western France.