Induced mutations through EMS treatment and in vitro screening for salt tolerance plant of Petunia × atkinsiana D. Don

Abstract

In vitro culture is a practical plant breeding tool in developing plant resistant to different abiotic stresses such as salinity. In this study, the focus is on inducing mutations for salt tolerance using ethyl methane sulphonate (EMS) in calli of petunia (Petunia × atkinsiana D. Don 'Prism Red'), followed by cell line selection and subsequent plant regeneration. Callus cultures were initiated from leaf fragments soaked in mutagen solution in two concentrations 0.5 and 5.0 mM for 60, 120 and 180 min, respectively. Then, the callus was rinsed three times in deionized, sterile water and was transferred onto embryo formation medium Murashige and Skoog (MS) supplemented with 2.0 mg l-1 1-napthaleneaceticacid (NAA) and 5.0 mg l-1 (6-benzyloaminopurine) BAP. After 28 days, somatic embryos were transferred onto MS medium supplemented with 4.0 mg l-1 BAP for regeneration. Results of our studies showed that the callus immersion in the 5.0 mM EMS solution caused damage to the cells, and indicated by the change in callus colour into brown. The plants regenerated from the somatic embryos induced from callus treated with 0.5 mM solution of mutagen for 60 and 120 min were determined as M1 and M2 mutant variants, respectively. After the multiplication of the obtained plants, their tolerance to salinity stress was determined. The obtained results have showed that the M1 and M2 mutant lines were characterized by elevated tolerance to the stress factor in comparison to non-mutated plants.