Induction of morphological, anatomical, and genomic variability in in vitro Moringa concanensis Nimmo. via ethyl methylsulfonate and sodium azide mutagenesis
摘要
The current study explores the in vitro growth of Moringa concanensis Nimmo. coupled with in vitro mutagenesis. In vitro regenerated shoots were sub-cultured and grown in chemical mutagens, including EMS and SA, fortified media with varying concentrations. All the EMS-treated shoots survived with a 100% survival rate. SA was most effective at its lower concentration of 0.1 mg L−1, although shoot survival declined with increasing concentrations. Anatomical analysis revealed structural changes in mutagen-treated shoots, including a multilayered epidermis, elongated cortical cells, enhanced pericycle tissue, and increased vascular tissue content, particularly in samples treated with SA. These anatomical improvements contributed to a 70% survival rate of the plantlets. CBDP (CAAT Box Derived Polymorphism), SCoT (Start Codon Targeted) gene-based, and ISSR (Inter Simple Sequence Repeat) intergenic sequence-based markers evaluated the mutagen-derived variations and genomic instability in treated shoots. Marker analysis of mutagen-treated shoots revealed high levels of polymorphism and corresponding PIC (Polymorphic Information Content) values: 82.35% and 0.24 (CBDP), 88.46% and 0.26 (ISSR), and 71.42% and 0.26 (SCoT). The observed polymorphism in genomic sequences demonstrates the effectiveness of chemical mutagens in generating variations in in vitro regenerated shoots of M. concanensis. These findings highlighted the application of EMS and SA mutagenesis could be effective in generating mutant variants that can contribute significantly to both breeding programs and the conservation of M. concanensis.