Validation of DNA fingerprinting techniques in characterization of entomopathogenic nematodes
Nour El-Deen AH
Three isolates of entomopathogenic nematodes (EPNs) Steinernema feltiae, Heterorhabditis bacteriophora and Steinernema sp. isolated from Egypt and Kingdom of Saudi Arabia have been, examined and evaluated for their molecular characterization. Genetic diversity evaluation among three EPNs genotypes were determined via RAPD and ISSR markers. The highest degree of polymorphism was selected from RAPD and ISSR primers. A total of 45 bands of polymorphic ISSR bands were revealed from 61 bands, with an average of 4.5 polymorphic fragments per primer. The number of amplified fragments with ISSR primers ranged from 2 to 11, with the size of amplicons ranging from 93 to 2043 bp. The polymorphism ranged from zero to 100.0, with an average of 73%. The three RAPD primers produced 12 bands across 3 genotypes, of which 10 were polymorphic, with an average of 3.3 polymorphic fragments per primer. The number of amplified bands varied from 3 to 5, the amplicons size ranging from 188 to 295 bp. The percentage of polymorphism using RAPD primers ranged from 33.3 to 100 with an average of 77.7%. The results of molecular characterization of isolated EPNs represent that S. feltiae and S. sp. are relatives more than H. bacteriophora.