Plant Biotechnology

Molecular Biology and Genetics

Electronic Journal of Biotechnology ISSN: 0717-3458 Vol. 13 No. 2, Issue of March 15, 2010
© 2010 by Pontificia Universidad Católica de Valparaíso -- Chile Received July 14, 2009 / Accepted October 23, 2009
DOI: 10.2225/vol13-issue2-fulltext-3

Development and application of high-throughput amplified fragment length polymorphism technique in Calluna vulgaris (Ericaceae)

Thomas Borchert*
Leibniz-Institute of Vegetable and Ornamental Crops
Department of Plant Propagation
Kuehnhaeuser Str. 101
Erfurt, Germany

Inka Gawenda
Crop Plant Diversity and Breeding Informatics Group
Institute of Plant Breeding, Seed Science and Population Genetics
University of Hohenheim
Stuttgart, Germany 


*Corresponding author

Financial support: The results were obtained within a third-party funded enterprise (German Federal Ministry of Education and Research, grant PGI-06.01-28-1-43.038-07). The experiments were conducted at the IGZ.

Keywords: DNA content, mapping, marker-assisted selection, ornamental, plant tissue.


AFLP®: amplified fragment length polymorphisms
htp: high-throughput
IRDye®: near infrared dye (LI-COR Biosciences GmbH)
ISSR: inter simple sequence repeat
RAPD: randomly amplified polymorphic DNA

Abstract   Full Text

Calluna vulgaris is an important ornamental crop of the horticultural industry in Europe. In order to improve breeding of this species, especially of the most important trait of ‘bud-flowering’, the implementation of molecular techniques that allow rapid, reproducible and efficient screening of whole segregating populations e.g. for molecular marker and mapping approaches is a requirement. We therefore aimed to introduce the powerful tool of amplified fragment length polymorphisms (AFLP®), a widely and successfully applied method, into our methodological assortment. As an essential prerequisite, the isolated DNA should be of adequate quality which is a common obstacle when dealing with woody species and their interfering secondary components/metabolites. The results of screening different and modified DNA isolation protocols are described. As the outcome of our evaluations of reaction conditions during the AFLP® procedure, we circumstantiate a functional protocol ranging from DNA extraction to visualization of AFLP® banding patterns for the woody crop C. vulgaris. This method is suitable for high throughput genetic applications and may even be transferable to other species. In addition, costs are reduced by reasonable reagents and multiplexing assays.

Supported by UNESCO / MIRCEN network