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In the present study, the experimental validation of Genic-SSR markers developed from transcriptome data of Selaginella bryopteris was done, and their cross-species transferability was examined across genus and species of pteridophytes. Twenty primer pairs were designed to amplify the various repeats of SSR markers, and first validated with complementary DNA (cDNA) of S. bryopteris. Out of twenty primer pair, fourteen primer pairs (70%) could amplify the DNA of S. bryopteris. Among Azolla spp., four SSR markers (SbSSR4, SbSSR12, SbSSR14 and SbSSR18) were found to be 100% transferable, while others SbSSR13 (80%), SbSSR3 and SbSSR20 (50%) and four markers SbSSR6, SbSSR10, SbSSR15 and SbSSR19 (10%). Out of twenty validated primers, 11 primers were found to be polymorphic. The value of polymorphism information content (PIC) ranged from 0.47 to 0.86 with an average of 0.49. The Jaccard’s similarity coefficient ranged from 0.10 to 0.93 which revealed a wide range of genetic identity. The similarity between A. fuliculoids and A. maxicana (0.93), and A. pinnata and A. rubra (0.93) was high and while low similarity coefficient of S. bryopteris with P. vittata (0.18), local fern (0.20) and with Azolla spp. (0.10- 0.25). Two major Clusters formed based on UPGMA i.e. S. bryopteris, P. vittata, local fern in Cluster I and Azolla spp. in Cluster II, a clear divergence between S. bryopteris and other pteridophytes. BAUS1 was found to be more genetically divergent followed by BAUS2, BAUS1 and BAUS3 in comparison to known species of Azolla. The transferability of Genic- SSR markers of S. bryopteris demonstrated the utility for elucidating genetic relationship among selected pteridophytes.

Transferability of Genic-SSR markers developed from the transcriptome data of Selaginella bryopteris (L.) Baker