Database Derived Microsatellite Markers (SSRs) of Stevia rebaudiana for Cross-transferability Testing Across Species in Family Asteraceae

Abstract

Aims: Simple sequence repeat markers derived from expressed sequences tags (ESTs) were used to carry out transferability studies across members of Asteraceae family. Study Design: NTSY Spc ver.2.0 was used to construct similarity matrix and dendrogram. Place and Duration of Study: Present study was undertaken in Department of Biotechnology, Dr Y S Parmar University of Horticulture and Forestry, Nauni, Solan, Himachal Pradesh, India -173230 from 2013-2015. Methodology: EST-SSRs were used for PCR amplification of genomic DNA of Asteraceae family members. The data was compiled in the form of per cent polymorphism depending on polymorphic and monomorphic bands. Similarity matrix was generated to find out per cent similarity between species and dendrogram represented visual phylogenetic tree of species used. Results: In this study we have studied the level of transferability of 45 Stevia EST-SSRs in 20 members of Asteraceae family, representing 20 genera of three subfamilies of Asteraceae. All selected 45 primers generated polymorphism. Transferability of the EST-SSRs ranged from 6.5% in Dimorphothica sinuata to 75.5% in Tagetes erecta, both of which belong to subfamily Asteroideae. Narrow base of studied material was depicted as similarity matrix values ranged from 0.00 to 0.30. Dendrogram was divided into two main clusters. Cluster ‘I’ contained only one genotype i.e. ‘Dimorphothica sinuata’, while Cluster ‘II’ consisted rest 20 species.  Conclusion: Present study targeted those important members of Asteraceae family which were not undertaken all together for molecular studies. As number of sequences in database repository for Asteraceae family is low, thus utilizing the repository from one crop to other members of same family is the only way to carry forward molecular research. Overall our findings suggested that transferability of Stevia EST-SSRs across Asteraceae genera is varied, yet valuable, thereby providing a good set of markers for genetic diversity, molecular mapping, gene tagging and comparative mapping studies.