The next day, embryos were sliced long itudinally and inoculated with Agrobacterium suspension after
adjusting the OD600to 1–1.2 for 60 min. After 3–4 days of co-cultivation, the explants were washed thoroughly in distilled water and then in antibiotic solution to eliminate the Agrobacterium growth. Subsequently, the explants were transferred to shoot induction medium for 10 days and finally transferred to selective regeneration medium where the regenerated shoots were subjected to in-creased concentration of selection agent (PPT: 2.5, 5, 7.5 and 10 mg/L) every three weeks. In order to recover the putative trans-genic shoots for further molecular and functional analyses, the in vitro putative transgenic shoots were micrografted on to seedling root stock raised on soil substrate until
flowering and setting the T0 seeds. Leaf samples were collected
from successfully grafted and well grown plants for molecular
analysis