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Genome size variation and morphological differentiation within Ranunculus parnassifolius group (Ranunculaceae) from calcareous screes in the Northwest of Spain

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Abstract

Ranunculus parnassifolius is an orophilous plant distributed throughout Central and Southwestern Europe (Alps, Pyrenees and Cantabrian Mountains). Its evolutionary history and taxonomy are often complicated, having been little studied before now. The purpose of this article is to present flow cytometry measurements and multivariate morphometric analyses to ascertain cytotype distribution patterns and the morphological differentiation of R. parnassifolius s.l. from calcareous screes in the Northwest of Spain. DNA ploidy level and morphometric analysis were determined for plants of R. parnassifolius s.l. using flow cytometry (112 individuals) and multivariate analysis (152 individuals). Specimens were collected in eight localities in the Northwest of the Iberian Peninsula. Different sample preservation methods (fresh, frozen, and herbarium specimens) were employed as well as the use of various buffers and internal standards, in order to test the reproducibility of DNA flow cytometry. Three ploidy levels were detected in the study area (diploid, tetraploid, and pentaploid), and mixed-cytotype populations were also found. The mean nuclear DNA content of the R. parnassifolius group ranged from 7.43 ± 0.185 to 7.63 ± 0.339 pg/2C in diploids and from 15.09 ± 0.161 to 15.85 ± 0.587 pg/2C in tetraploids. The analysis of the monoploid genome sizes (1Cx) did not reveal a clear difference among cytotypes. These results suggest low intraspecific variation, at least among the populations studied. In addition, a comparison of different DNA reference standards was conducted. A new value for the chicken genome size was used as internal reference standard (2C = 3.14 ± 0.155 pg), with similar results found using both animal and plant standards (Pisum sativum and Solanum lycopersicum). Finally, herbarium vouchers and frozen tissue were proved to be suitable for DNA ploidy level measurements. This study provided a first assessment of C values in the R. parnassifolius group using flow cytometry. The weak morphological distinction of the cytotypes and the existence of mixed-cytotype populations in the Northwest of Spain are reported here for the first time. The different distribution pattern of the two cytotypes is discussed.

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Acknowledgments

We acknowledge the technical assistance of Ana Salas and Marta Alonso Guervós (Flow Cytometry Area, Scientific-Technical Services, University of Oviedo). Seeds of tomato cv. ‘Stupicke’ and peas cv. ‘Ctirad’ were supplied by Dr. J. Doležel (Laboratory of Molecular Cytogenetics and Cytometry, Institute of Experimental Botany, Olomouc, Czech Republic). We express our gratitude to Anu Väinölä, Frantisek Krahulec, Elvira Hörandl, Eva M. Temsch, and Miguel de Cáceres for their encouragement and their helpful comments. We also express our gratitude to the curators and directors of all herbaria consulted. This research was funded by the Spanish Ministerio de Educación y Ciencia (CGL2006-11743). E. Cires is supported by a predoctoral grant by the University of Oviedo (UNOV-06-BECDOC-2).

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  1. Área de Botánica, Departamento de Biología de Organismos y Sistemas, Universidad de Oviedo, Catedrático Rodrigo Uría s/n, 33071, Oviedo, Spain

    Eduardo Cires & José Antonio Fernández Prieto

  2. Área de Fisiología Vegetal, Departamento de Biología de Organismos y Sistemas, Universidad de Oviedo, Catedrático Rodrigo Uría s/n, 33071, Oviedo, Spain

    Candela Cuesta, Elena L. Peredo & María Ángeles Revilla

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Cires, E., Cuesta, C., Peredo, E.L. et al. Genome size variation and morphological differentiation within Ranunculus parnassifolius group (Ranunculaceae) from calcareous screes in the Northwest of Spain. Plant Syst Evol 281, 193–208 (2009). https://doi.org/10.1007/s00606-009-0201-9

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