Fenugreek (Trigonella foenum-graecum L.) is highly esteemed for its therapeutic properties and is widely used in traditional medicine and modern pharmacology. Enhancing its genetic traits and phytochemical profile, particularly its trigonelline content, can significantly increase its medicinal and agricultural value. This study aims to investigate the effects of gamma rays and Ethyl Methane Sulphonate (EMS) as mutagenic agents on the genetic and phytochemical characteristics of the M2 generation of fenugreek, focusing on genetic diversity and desirable trait enhancement. To achieve this, various concentrations of EMS and gamma rays were administered to fenugreek seeds, and 27 traits were assessed in the resulting M2 generation. These traits were analyzed for variance, mean values, and correlations. The genetic diversity of 23 M2 offspring was investigated using nine Start Codon Targeted (SCoT) markers. The genetic diversity assessment involved Principal Coordinate Analysis (PCoA) and cluster analysis, utilizing the Dice similarity coefficients and the Unweighted Pair Group Method with Arithmetic Mean (UPGMA). A Bayesian model provided deeper insights into the genetic structure. Results revealed that lower doses of gamma rays (100Gy) and EMS (0.2%) positively impacted specific traits. In comparison, higher doses (200Gy and 0.4% EMS) increased seed trigonelline content to 0.71mg/g dry weight. Among the SCoT markers, SCoT-9 was the most efficient, segregating the populations into three clusters. The first three principal components in the PCoA explained 20% of the total variance, leading to seven subgroup populations distinction. These findings underscore the potential of induced mutagenesis in enhancing desirable traits in fenugreek.
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