Flow cytometric and karyological analyses of Calendula species from Iberian Peninsula

Abstract

Calendula L. (Asteraceae) is a taxonomically and cytologically complex genus due to its high morphological and karyological variation. To gather consistent cytological information aiming to consolidate the existing knowledge, sustain the taxonomic revision of the genus and explore the evolutionary relationships among species, the genome size and chromosome number of the Iberian Peninsula representatives of this genus were assessed. The study included 11 taxa that occur in the Iberian Peninsula, one in Madeira and two from Morocco. Chromosome counts were made using the squash technique in root tips and flower buds, while nuclear DNA contents were assessed using propidium iodide flow cytometry. The following chromosome numbers are reported: 2n = 44 for C. arvensis, 2n = 30 for C. tripterocarpa, and 2n = 32 for the remaining Iberian taxa. The genome size of Calendula species was assessed for the first time and ranged from 1.75 pg/2C in C. maroccana to 5.41 pg/2C in C. arvensis. Within the complex formed by C. incana and C. suffruticosa, a gradient of genome size values was obtained. Intraspecific variation in genome size was detected in some taxa. The obtained genome size values and their variation are discussed in the light of the theories proposed for the speciation of the genus, with events of hybridization, genome duplication and dysploidy being hypothesized to play a major role in the evolution of this genus.

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Acknowledgments

The authors are grateful to the ICNF (Instituto de Conservação da Natureza e das Florestas), namely Dr Luis Silva (UACI) and the Directors of several Departments (Departamento de Gestão de Áreas Classificadas—Zonas Húmidas, Departamento de Gestão de Áreas Classificadas do Litoral de Lisboa e Oeste, Departamento de Gestão de Áreas Classificadas Sul) for allowing the collection of samples on several nature reserves; to Miguel Sequeira, from the Biology Department of the University of Madeira, for providing Calendula incana ssp. maderensis material; to Abelardo Aparicio for methodological help in the chromosome counts. Although mostly not directly used for this study, but important to the general knowledge of the genus and planning of field work, we are grateful to the Directors of AVE, BC, BCN, BM, BR, C, COI, ELVE, G, GAT, GDA (GDAC), HAL, K, LD, MA, MAF, MGC, O, P, PO, RAB, SALA, SANT, SEV, TFMC, US herbaria for the loan of herbarium vouchers and/or for allowing us to visit them and study their collections on site and/or for making images of specimens available for study. Additionally, BH and HAL offered duplicates of some Calendula specimens. Portuguese Foundation for Science and Technology and European Social Fund financed the work of SC with the scholarship FCT/BPD/41200/2007, the work of HO with the scholarship SFRH/BPD/48853/2008 and the work of AG with the scholarship SFRH/BD/51464/2011.

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Nora, S., Castro, S., Loureiro, J. et al. Flow cytometric and karyological analyses of Calendula species from Iberian Peninsula. Plant Syst Evol 299, 853–864 (2013). https://doi.org/10.1007/s00606-013-0767-0

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Keywords

  • Calendula
  • Chromosome numbers
  • Flow cytometry
  • Genome size
  • Iberian Peninsula
  • Karyology
  • Nuclear DNA content