Genetic diversity analysis in tuberose (Pollianthes tuberosa) genotypes through Randomly Amplified Polymorphic DNA

Abstract

The present investigation was undertaken for characterizing tuberose genotypes using DNA marker technology. Twenty tuberose genotypes comprising of both single- and double-petal types collected from different parts of India were selected for analysis. The DNA extraction and RAPD conditions were standardized. For RAPD analysis 20 ng DNA template, 2.5 mM MgCl2 and 1U TaqDNA polymerase was found effective. Out of 80 random decamer primers tested, 17 were selected based on high level of polymorphism. On the basis of primer resolving power and marker index, RAPD primers OPC-13 and OPD- 12 were identified as efficient primers for diversity analysis of tuberose. A total of 157 RAPD bands were generated by the 17 random decamer primers. The selected primers proved effective for DNA profiling in addition to diversity analysis. Genotypes Guwahati Double and Swarnrekha showed good morphological similarity revealing high similarity coefficient (0.90) suggesting them to be very closely related. The RAPD analysis also confirmed their relatedness as they grouped in the same cluster. The suitability of this technique for genotyping and diversity analysis was also established. Genotypes, Vaibhav and Pune Single were found to have least pair-wise similarity although they had a greater morphological similarity with each other.