Majallah-i ̒Ilmī-i Shīlāt-i Īrān (Jan 2008)
INTRODUCING MOLECULAR MARKERS FOR LERNAEA CYPRINACEA AND LERNAEA CTENOPHARYNGODONI USING RAPD TECHNIQUE
Abstract
Molecular comparison of two parasites Lernaea cyprinacea and Lernaea ctenopharyngodoni was carried out using RAPD (Random Amplified Polymorphic DNA) technique. A total of 43 Lernaea specimens belonging to the two species were collected from the Guilan and Khouzestan Provinces. DNA was extracted using the Phenol-chloroform method. The quality and quantity of DNA was assessed using 1% Agarose gel electrophoresis and spectrophotometer. Polymerase Chain Reaction (PCR) was conducted on the target DNA under specific conditions and PCR products were subjected to electrophoresis on polyacrylamide gels (6%). Polyacrylamide gels were stained using silver nitrate and DNA bands were analyzed with BioCapt software. The genetic analysis was conducted using POP GEN 32 software. Forty two primers, 10 nucleotides each were used for PCR reaction. Totally, 397 RAPD loci were counted on polyacrylamide gel where 349 identical loci were polymorphic of which some bands may be used as genetic markers for the identification of both Lernaea species. Data analysis on PCR products showed higher genetic variation (1.15%) of Lernaea ctenopharyngodoni in the Guilan Province as compared to that of the Khouzestan (0.0%).However, genetic variation (27.46%) of Lernaea cyprinacea in the Khouzestan province was 7.26 times higher than that of the Guilan province (3.78%). The two species showed a genetic differentiation of approximately 88%. Based on the observed molecular differences, we state that Lernaea ctenopharyngodoni is a genetically independent species from Lernaea cyprinacea.
