Usage of RAPD-PCR Analysis of DNA to Differentiate Lady Bird Beetles Among Family Coccinelidae

Document Type : Original Article


1 Biological Control Res. Depart., Plant Protection Res. Institute., Agricult. Res. Center

2 Biological Control Res. Depart., Plant Protection Res. Institute., Agricult. Res. Center.


Random amplified polymorphic DNA (RAPD) analysis, analysed by polymerase chain reaction (RAPD-PCR), as a tool to differentiate between six coccinellid beetles (Coccinella undecimpunctata L. 1758; C. Novemnotata Herbst 1793; Hippodamia tredecimpunctata L. 1758; Cheilomenes propinqua vicina (Mulsant, 1850); Scymnus (Pullus) subvillosus (Goeze, 1777) and Cydonia vicina nilotica Mulsant 1850) evaluate the variation and the genetic structure of the different species with three decamer oligonucleotide primers (MWG1, MWG2, and MWG3). MWG1 primer was successfully used to evaluate the similarity and compare genetic variation among the six insects, the total number of amplicons ranged from 10 to 16 per primer with an average of 14 amplicons.  Amplicons size (bp) were ranged from 100 bp to 1311 bp. Polymorphic amplicons were ranged from 9 to 16.  The maximum of polymorphic bands belonged to MWG2 primer 16 bands with 100% polymorphism, and the minimum of polymorphic bands belonged to MWG3 primer (9bands) with 90% polymorphism.  In addition, MWG1 primer was produced (14 bands) with 100% polymorphism. The RAPD analysis data grouped into two main clusters and an independent one, the first cluster was C. vicina nilotica, and C. novemnotata classified as the most related species with 72%, and they close to H. tradecimpunctata with 55%. The second cluster was C. undecimpunctata, and S. subvillosus were closed with 58% while, C. vicina is relatively distant from the other five species, and meet with them at common ancestor with 45% using   RAPD-PCR analysis and the results were cleared genetic variations between the six coccinellid spp < em>. The polymorphic percentage ranged between 90% and 100%, and the similarity percentage ranged between 0.32 and 0.71.