Genetic Diversity Analysis of Sodium Azide-Induced Mutants of Ornamental Banana (Musa ornata Roxb.) Using ISSR Markers
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Chemical mutagenesis is widely used to generate genetic variation in clonally propagated crops where conventional hybridization is limited. This study evaluated the mutagenic effects of sodium azide (NaN3) on Musa ornata Roxb. and assessed the resulting genetic diversity using inter simple sequence repeat (ISSR) markers. In vitro–propagated plantlets were treated with NaN3 concentrations ranging from 0.1 to 4.0 mM, and survival responses were used to determine optimal mutagenic doses. Plant survival decreased linearly with increasing NaN3 concentration (y = −20.92x + 93.59, R² = 0.938), with estimated LD30 and LD50 values of 1.13 mM and 2.08 mM, respectively. Sublethal NaN3 treatments induced measurable morphological variation in regenerated plantlets, including changes in fresh weight, leaf number, leaf length, and root development. Molecular analysis using ten ISSR primers generated 81 scorable ISSR bands, of which 36 were polymorphic (43.02%). Genetic diversity indices indicated moderate variation among mutants (PIC = 0.28, Nei’s gene diversity = 0.24, Shannon’s index = 0.34). Cluster analysis based on UPGMA grouped the plantlets into three major genetic clusters, with pairwise genetic dissimilarities ranging from 34.7% to 68.4%. The clustering pattern showed partial correspondence with observed morphological variation, suggesting that NaN3-induced genomic changes contributed to phenotypic diversification. These findings demonstrate that the combination of in vitro sodium azide mutagenesis and ISSR marker analysis is an effective approach for generating and identifying novel variation in M. ornata. The resulting lines represent valuable genetic resources for the improvement of ornamental bananas through mutation breeding.
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