Determination of Genetic Variation in Piper Species Using 4C Nuclear DNA and RAPD Markers

The present study utilized dominant markers such as Random Amplified Polymorphic DNA (RAPD) for the evaluation of relatedness among 6 Piper species were included in this study and to ensure such technique for claiming intellectual property right (IPR) by the plant breeder. Further, the study also aimed at estimation of the genetic relatedness of these 6 morpho-agronomically contrasting species. Six Piper species were screened using RAPD with decamer primer of arbitrary sequence. Out of 100 primers screened 12 were selected which gave clear and bright fragments. DNA banding patterns generated by RAPD were recorded as ‘1’ for presence of the RAPD marker and ‘0’ for absence. Genetic distance between these 6 species was calculated based on the RAPD data set as per Squared Euclidean distances. Based on the number of bands all the species were grouped into 3 clusters and the dendrogram revealed maximum similarity between P. betel and P. longum and also in between P. nigrum and P. mullesua species, altogether forming one cluster. The standardized method can identify any of the single hybrid or species tested.