Application possibilities of AFLP fingerprinting technique in maize DNA profiling and plant variety protection

Abstract

As a contribution to DUS testing within the system of protection of plant breeders' rights (PBR), the AFLP molecular system has been used in this study to produce DNA fingerprinting profiles. DNA polymorphism and genetic distance of nine agronomicaly important maize genotypes has been investigated using the AFLP technique. Two specific adapters, two preselective primers and twenty selective primers were utilized for DNA amplification. The selective primers were GC rich, each having a 3-mer selective sequence at 3' termini. Ten double stranded primer combinations were made out of the twenty primers but only five of them turned out to be reliable. Out of 253 amplified DNA fragments, 177 were polymorphic (70%). The CGA/GAG (B) primer combination has proved to be the most polymorphic (44 polymorphic fragments have been recorded) revealing the polymorphism rate of 81.5%. Genotypes g1 and g7 were most distinct (GD=55% and GD=79%, respectively) and genotypes g1. g4 and g8 were closest (GD=55% in all cases). The paper discusses possible uses of AFLP DNA profiling technique to achieve a unique fingerprinting pattern of agronomicaly important maize genotypes.

U ovom radu korišćen je AFLP molekularni sistem u cilju ispitivanja mogućnosti njegove primene u karakterizaciji fingerprinting profila devet agronomski značajnih genotipova kukuruza. Za umnožavanje specifičnih DNK fragmenata korišćena su dva specifična adaptera, dva preselektivna prajmera i dvadeset selektivnih prajmera. Od ukupno 10 testiranih prajmer kombinacija, 5 je pokazalo značajnu polimorfnost. Od 253 umnožena fragmenta, 177 su bila polimorfna (70%). Prajmer kombinacija B (CGA/GAG) bila je najpolimorfnija (81,5%) a genotipovi gl(GD=55%) i g7 (79%) pokazali su najveću genetičku udaljenost. Na osnovu dobijenih rezultata zaključili smo daje AFLP molekularni sistem pogodna i pouzdana metoda za određivanje lične karte agronomski značajnih genotipova kukuruza i kao takva primenljiva u zaštiti autorskih prava.