AFLP and MS-AFLP analysis of the variation within saffron crocus (Crocus sativus L.) germplasm

PLoS One. 2015 Apr 17;10(4):e0123434. doi: 10.1371/journal.pone.0123434. eCollection 2015.

Abstract

The presence and extent of genetic variation in saffron crocus are still debated, as testified by several contradictory articles providing contrasting results about the monomorphism or less of the species. Remarkably, phenotypic variations have been frequently observed in the field, such variations are usually unstable and can change from one growing season to another. Considering that gene expression can be influenced both by genetic and epigenetic changes, epigenetics could be a plausible cause of the alternative phenotypes. In order to obtain new insights into this issue, we carried out a molecular marker analysis of 112 accessions from the World Saffron and Crocus Collection. The accessions were grown for at least three years in the same open field conditions. The same samples were analysed using Amplified Fragment Length Polymorphism (AFLP) and Methyl Sensitive AFLP in order to search for variation at the genetic (DNA sequence) and epigenetic (cytosine methylation) level. While the genetic variability was low (4.23% polymorphic peaks and twelve (12) effective different genotypes), the methyl sensitive analysis showed the presence of high epigenetic variability (33.57% polymorphic peaks and twenty eight (28) different effective epigenotypes). The pattern obtained by Factorial Correspondence Analysis of AFLP and, in particular, of MS-AFLP data was consistent with the geographical provenance of the accessions. Very interestingly, by focusing on Spanish accessions, it was observed that the distribution of the accessions in the Factorial Correspondence Analysis is not random but tends to reflect the geographical origin. Two clearly defined clusters grouping accessions from the West (Toledo and Ciudad Real) and accessions from the East (Cuenca and Teruel) were clearly recognised.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amplified Fragment Length Polymorphism Analysis
  • Crocus / genetics*
  • Genetic Variation*
  • Genotype*
  • Phenotype*
  • Phylogeny

Grants and funding

This work was supported by: 1) Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (ES); Creación de la colección española de azafrán y especies afines, grant number RF2004-00032; URL: http://www.inia.es/IniaPortal/verPresentacion.action; grant holder MDM; 2) Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (ES); Colección española de germoplasma de azafrán y especies afines, grant number RF2008-00012; URL: http://www.inia.es/IniaPortal/verPresentacion.action; grant holder MDM; 3) Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (ES); Avances en la estrategia de conservación, multiplicación y caracterización de la colección española de germoplasma de azafrán y especies afines, grant number RF2011-00005; URL: http://www.inia.es/IniaPortal/verPresentacion.action; grant holder MDM; 4) European commission; Genetic resources of saffron and allies (crocus spp.): Crocusbank, grant number AGRI-2006-0265; URL: http://ec.europa.eu/; grant holder JAF; 5) European Cooperation in Science and Technology; Omics technologies for crop improvement, traceability, determination of authenticity, adulteration and origin in saffron, COST action FA1101; URL: http://www.cost.eu/; grant holder MR.