African Journal of Biotechnology Vol. 5 (8), pp. 566-569, 18 April 2006   

ISSN 1684–5315 © 2006 Academic Journals        

Full Length Research Paper

DGGE-RAPD analysis as a useful tool for cultivar identification

A. Bahieldin^1,2*, I.A. Ahmed³, Gh.A. Gad El-Karim¹, H.F. Eissa¹, H.T. Mahfouz¹ and O.M. Saleh⁴

¹Agricultural Genetic Engineering Research Institute (AGERI), ARC, Giza, Egypt.

²Department of Genetics, Faculty of Agriculture, Ain Shams University, Cairo, Egypt.

³Barley Department, Field Crops Research Institute, ARC, Giza, Egypt.

⁴National Center of Radiation Research and Technology, Egypt.

*Corresponding author. E-Mail: bahieldin@hotmail.com, Tel: 202-624-3233, Fax: 202-444-4460.

Accepted 16 May, 2005

Denaturing gradient gel electrophoresis-random amplified polymorphic DNA (DGGE-RAPD) was used to overcome the main drawbacks of RAPD (i.e., the low levels of reproducibility and polymorphism). As a model, six barley cultivars of known origin were tested for RAPD markers using DGGE methodology with 29 arbitrary primers. Among a total of 418 bands observed, as high as 99 were polymorphic. Comparison between agarose-RAPD and DGGE-RAPD revealed that the latter was highly reproducible and gave higher level of polymorphism and consequently more markers. The relationships among barley cultivars derived from this study based on DGGE-RAPD are consistent with the known lineage of these cultivars. In conclusion, we recommend the use of DGGE-RAPD as an alternative tool to the more costly DNA-based analysis in cultivar identification in laboratories with limited funds.

Key words:Denaturing gradient gel electrophoresis-random amplified polymorphic DNA, artificial heteroduplex, dendogram, lineage.