OPTIMIZATION OF IN-PLANTA TRANSFORMATION EFFICIENCY IN MUNGBEAN (VIGNA RADIATA L.) USING AN AGROBACTERIUM-MEDIATED APPROACH

Abstract

In planta transformation is a promising technique for genetic modification in mungbean (Vigna radiate L.), yet its efficiency is influenced by various factors. This study aimed to assess the impact of genotype, Agrobacterium culture concentration, and floral bud stage on transformation efficiency. The transformation process was evaluated using β-glucuronidase (GUS) staining, wherein successfully transformed embryos exhibited a blue coloration. The expression of the GUS gene was confirmed through visual examination and microscopic analysis. Results indicated that genotype significantly influenced transformation efficiency, with NM-19 exhibiting the highest mean transformation rate (6.0758%), whereas NCM-2013 had the lowest (0.050%). Similarly, Agrobacterium culture concentration played a crucial role, where an optical density (OD) of 1.0 at the late floral bud stage yielded the highest transformation efficiency (5.3483%). Conversely, lower transformation rates were observed at an OD of 0.5 during the early floral bud stage (2.9258%). The interaction between genotype and culture concentration was also significant, with NM-19 at OD 1.0 achieving the highest transformation efficiency (6.5500%). Additionally, the late floral bud stage resulted in significantly higher transformation efficiency compared to the early floral bud stage. Statistical analysis using ANOVA confirmed the significant effects of genotype, culture concentration, and floral bud stage on transformation efficiency. The findings suggest that selecting an appropriate genotype, optimizing bacterial culture concentration, and targeting the late floral bud stage can enhance transformation success in mungbean. These insights contribute to improving genetic transformation protocols for mungbean and may facilitate advancements in legume biotechnology.