Assessment of genetic variations in EMS-exposed Petunia Tested for Salt in Vitro Tolerance Using RAPD

Marcelina Krupa-Małkiewicz

Anna Bienias

Abstract

Random Amplified Polymorphic DNA (RAPD) was used to assess the genetic variability within somaclones of Petunia cv. Prism Red exposed to 0.5 mM ethyl methane-sulphonate (EMS), and tested for salt tolerance in vitro. Twenty RAPD primers were utilized; 8 out of which amplified specific fragments generating a total of 39 alleles, with a mean of 3.9 alleles per locus. Three arbitrary oligonucleotide primers revealed polymorphisms between non-mutated and mutated plants. Whereas, six RAPD primers generated polymorphic products characteristic only for EMS putative mutants or for EMS putative mutants tested for salt tolerance. The extent of polymorphism indicated the existence of variability within DNA in induced mutated somaclones. Cluster analysis using the Nei’s similarity coefficient values and UPGMA algorithm detected genetic variation within non-mutated and mutated plants, as they are placed in different clusters/groups far from each other. Similarity matrices mostly ranged from 0.5 to 0.99. Results were indicative that induced mutation bears great potential in improving petunia for salinity resistance which can be considered as potential samples for further breeding programs.