Using AFLP markers for species differentiation and assessment of genetic variability of in vitro-cultured Papaver bracteatum (section Oxytona)

Abstract

SummaryAmplified fragment length polymorphism (AFLP) markers were employed to detect genetic variation among species of Papaver (section Oxytona ) and assess genetic fidelity between in vitro cell lines of Papaver bracteatum and mature plants derived from the propagation of their callus cultures. Regenerated plants exhibited morphological and phytochemical characteristics dissimilar to those of their source material. Thebaine, the dominant alkaloid produced by Papaver bracteatum, was not detected in capsules from mature regenerated accessions, indicating that there may have been a loss of genetic uniformity. Instead, the dominant alkaloid produced by the regenerated plant was shown to be isothebaine (by TLC and GC/MS), a metabolic characteristic of P. pseudo-orientale. A Neighbor-Joining tree constructed from AFLP fingerprints distinctly separates the three species of Oxytona while firmly grouping the in vitro-cultured plants with P. pseudo-orientale. Additionally, phytochemical data and chromosome counts indicate that the seed used to initiate cultures was of hybrid origin and that the loss in genetic uniformity was not due to somaclonal variation occurring during the in vitro culture process. AFLP fingerprinting was therefore able to differentiate Oxytona species and investigate allopolyploidy in closely related Papaver species.

Molecular characterization of the genus Papaver section Oxytona using ISSR markers

“…It has been reported that regenerated plants showed different morphological and phytochemical characteristics from those of their source material. In addition, phytochemical chromosome data indicated that the seed used in cultures was of hybrid origin and that the loss in genetic uniformity was not due to somaclonal variation occurring during the in vitro culture process (Carolan et al, 2002). In our study, P. pseudo-orientale and P. orientale had higher genetic variation than P. bracteatum.…”

Phylogenetics of Miscanthus , Saccharum and related genera (Saccharinae, Andropogoneae, Poaceae) based on DNA sequences from ITS nuclear ribosomal DNA and plastid trnL intron and trnL-F intergenic spacers

“…Amplified fragment length polymorphism (AFLP) DNA fingerprinting (Vos et al 1995;Reeves et al 1998;Carolan et al 2002) data showed that M. nepalensis was clearly separate from other Miscanthus species (Hodkinson et al 2002b). Species of section Diandra resemble Miscanthus s.s. on morphological grounds, but possess two anthers, instead of the usual three, and a basic chromosome number of five or ten instead of 19 (Mehra and Sharma 1975).…”

Assessment of Genetic Stability Among In Vitro Plants of Arachis retusa Using RAPD and AFLP Markers for Germplasm Preservation

“…More recently, the amplified fragment length polymorphism (AFLP) technique has proven to be a highly efficient tool for characterizing somaclonal variation (Carolan et al 2002;Popescu et al 2002), in addition to cultivar identification (Ferriol et al 2003) or redundancy reduction in germplasm collection (van Treuren et al 2004). In Arachis, this technique was used to detect DNA polymorphism in the cultivated peanut (Herselman 2003) and to establish genetic relationships among species (Gimenes et al 2002).…”

“…However, there are still comparatively few reports on the use of AFLP in monitoring the genetic stability of in vitro plants obtained through different regeneration pathways. These studies were performed, for example, in plants of pecan (Vendrame et al 1999), cork oak (Hornero et al 2001), Arabidopsis thaliana (Polanco et al 2002), grapevines (Popescu et al 2002), and among in vitro cell lines of Papaver bracteatum (Carolan et al 2002).…”