Optimization of sonication-assisted Agrobacterium-mediated transformation for enhanced explant recovery and regeneration in soybean (Glycine Max L.)

Abstract:

This study optimizes a sonication-assisted Agrobacterium-mediated transformation protocol for the soybean genotype PDKV Amba. The research aimed to efficiently deliver the pDIRECT_22A vector, which targets the β-conglycinin allergen using CRISPR/Cas9 technology. Evaluated co-cultivation durations of 2, 4, and 6 days following a 30-second sonication treatment at 50Hz. While the 2 day period showed the highest initial germination (92%) and the 6 day period yielded maximum survival on selection media (90.47%), the 4 day co-cultivation proved optimal. It achieved the highest overall recovery rate (39.07%) and produced the most viable plants, successfully balancing Agrobacterium infection with explant viability.