Molecular characterization of citrus butterfly, Papilio demoleus Linnaeus (Lepidoptera: Papilionidae) by using PCR based Randomly Amplified Polymorphic DNA (RAPD)
2016
Jahnavi, M. | Rao, A Ramakrishna | Sarada, G.
Random Amplified Polymorphic DNA (RAPD) is a rapid and sensitive technique in genetic diversity analysis of insect pests. Fifteen arbitraryprimers(OPA01, 05, 07, 13, 14, 15 OPC 08, 15 OPE 02, 06, 08 OPAA16 OPAB11, 12 and 18) were used for RAPD analysis and generated scorable PCR products by amplifying the template DNA with Taq polymerase. The RAPD products generated a total of 162 fragments with an average of 10.8 fragments per primer and the fragments were found 100% polymorphic. The maximum number of polymorphic bands (12) were obtained with primer OPE-02, OPE-08 and minimum number (6) were obtained with primer OPAB-18 and OPA-14. OPA-01, 05, 07, 13, 15; OPC 15; OPAB 11, 12, 18 showed 100 per cent polymorphism, OPC 15 exhibited 92.30 per cent polymorphism, OPC 08 depicted 90.90 per cent polymorphism, OPA 14 and OPE 08 displayed 85.71 per cent polymorphism, OPE 06 showed 80 per cent polymorphism and OPAA 16 exhibited 70 per cent polymorphism. The similarity index values obtained for each pair wise comparision among the 9 citrus butterfly population (C1, C2, C3, N1, N2, N3, K1, K2 and K3) based on 162 RAPD fragments ranged from 250bp to 3.0kb. The maximum genetic similarity value was 0.730 between K3 and K1 followed by 0.679 between C3 and C2. The lowest genetic similarity value of 0.318 was found between citrus population of K2 and N2. Generation of higher number of markers or alleles in the present study indicated higher amount of genetic diversity among different populations of citrus butterfly.
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