Identification of sex-specific DNA markers in betel vine (Piper betle L.)

The Random Amplified Polymorphic DNA (RAPD) technique was used to amplify DNA segments, with the objective of finding markers linked to sex determination in male and female plants of Piper betle L. Two bulks of DNA were made drawing one each from male and female, by pooling an equal volume of DNA samples from each group of individual contributing to the bulk analysis. Fifty different random decamer primers were screened with the two bulks to identify markers associated with sex expression of which only four primers were found to be associated with sex expression. These four primers were then tested with individual plant DNA samples where sex-associated RAPD markers were identified. A ~1,400 and ~850 bp fragment from the primer OPA04 and OPN 02 respectively was found to be present in all the male individuals and absent in all the female plants. In another primer, a ~980 bp amplification product from the primer OPC 06 was present only in the female individuals. A common primer OPA 08 showed both male and female specific markers of 650 and 1,200 bp respectively. Thus, the three male- specific RAPD markers OPA041400, OPA08650 and OPN02850 and two female-specific markers OPA081200 and OPC06980 can reliably differentiate the male and female plants of P. betle L. Ploidy comparison also showed the differences in male and female plants.