PCR-based molecular markers in Asiatic hybrid lily

Abstract

PCR-based DNA markers in Asiatic hybrid lily were examined. Random primers with various lengths (10-, 12-, 15- and 20-base) were tested in randomly amplified polymorphic DNA (RAPD) reactions with total DNA from two cultivars, ‘Montreux’ and ‘Connecticut King’. Thirty-three out of 61 (54%) 15-base primers and 14 out of 21 (67%) 20-base primers generated polymorphic fragments whereas 17 out of 145 (12%) and 4 out of 24 (17%) of the 10- and 12-base primers, respectively, amplified them. The efficiency of the RAPD reaction increased with increasing primer length. We examined inter-simple sequence repeat (ISSR) reactions using 3′-anchored simple sequence repeat (ASSR) primers, and found 33 out of 63 (52%) primers that amplified polymorphic bands between the two cultivars. We repeated all RAPD and ISSR reactions at least two times and obtained the same DNA fingerprints, indicating high reproducibility obtained with these reactions. All polymorphic fragments were inherited into their F 1 hybrids, indicating that these DNA markers are genetically stable.