BSA and molecular markers screening for salt stress tolerant mutant of Petunia obtained in in vitro culture

Abstract

In this study, we performed BSA to identify genetic markers linked to salt tolerance. We tested the genetic diversity among four bulked DNA samples of EMS induced mutant clones and one bulked DNA sample of non-mutated clone of Petunia for salt tolerance in in vitro callus cultures using RAPD and ISSR markers. Out of the 36 RAPD and 16 ISSR primers identified, 25 and 13 were effectively used to amplify genomic DNA of all the five bulked samples, respectively. In total, 114 RAPD amplifications products were obtained, of which 28% were polymorphic and 2% were genotype-specific bands. Out of the 64 ISSR amplification products obtained, 51% were polymorphic and 1% was genotype-specific bands. Results of this study indicated the existence of two patterns of distorted segregation among the studied markers. The first one indicates the differences between non-mutated clones of Petunia and its putative mutants. The second one was observed only between putative mutants and putative mutants tested for salt tolerance in in vitro culture. Both RAPD and ISSR analysis successfully detected the association with changes induced by chemical mutagenesis and salinity. Furthermore, our results indicate that BSA method can be useful in the rapid detection of molecular markers for further marker-assisted selection.

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“…Also, various researchers indicated that this technique based on the PCR reaction is a fantastic tool [64] for genetic improvement of crop plants to tolerate various environmental stresses [65,66,67]. Also, different researches performed association analysis to find the ISSR markers which are linked with salinity tolerance [64,68,69,70].…”

“…Considering the high percentage of polymorphism (80%), it could be expected that these markers may have acted as a powerful tool in identifying and distinguishing the treatments [71]. In their study, Krupa-Małkiewicz and Bienias [68] found that both ISSR and Random Amplification of Polymorphic DNA (RAPD) primers successfully detected the association with changes induced by chemical mutagenesis and salinity. Like this study, they concluded that the bulked segregant analysis (BSA) technique using an ISSR marker is a rapid tool for detecting salt-tolerant genotypes.…”