ISSN 2736-1756
Advanced Journal of Microbiology Research Vol. 2006
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2006 International Scholars Journals
Full Length Research Paper
Optimization of ISSR marker for African edible-seeded Cucurbitaceae species’ genetic diversity analysis
Djè Y1, Tahi GC3, Zoro Bi IA1, Malice M2, Baudoin JP2 and Bertin P3
1Laboratoire de génétique, UFR des Sciences de la nature, Université d’abobo-adjamé, 02 BP 801, Abidjan 02, Côte d’Ivoire.
2Unité d’horticulture et phytotechnie tropicale, Faculté des Sciences Agronomiques de Gembloux, Belgique.
3Laboratoire d’écologie et amélioration végétale, Université catholique de Louvain-la-neuve, Belgique.
Accepted 14 October, 2005
Abstract
We applied a molecular approach using inter-simple sequence repeat (ISSR) markers on three African edible-seeded cucurbits (Citrullus lanatus L. Matsumura and Nakai, Cucumeropsis mannii L. Naudin and Cucumis melo var. agrestis L. Naudin). To obtain clear and reproducible bands on 1.5% agarose gels, we screened 21 ISSR primers and three parameters (annealing temperature, gel tray, and voltage and running time). The resolution of 11 ISSR markers was performed, with optimal annealing temperature (Ta) varying from 50 to 52°C. The best combination to obtain clear and well-distinguished band patterns was 1.5% agarose gel with a 20-lanes tray (6 mm width) at 80 V for 5 h. Applying the 11 ISSR primers on DNA extracted from an accession of C. lanatus, 66 bands with 4 to 11 bands per primer was observed.
Key words: ISSR, Cucurbits, genetic diversity, molecular markers, optimization.