Hairy root induction and plant regeneration of tobacco (Nicotiana tabacum L.)

Abstract

The present study showed the morphogenic potential of an important aromatic and industrial tobacco plants for the development of transgenic plants through Agrobacterium rhizogenes (strain 15834) for rolA, rolB, and rolC genes. Leaf explants were inoculated with 1/25, 1/30, and 1/50 diluted bacterial culture (OD600=0.8) for 30 min. containing different acetosyringone doses 250 microM, 200 microM and 150 microM respectively. The highest number of hairy roots was obtained by 1/25 containing 250 microM acetosyringone was 100%. These hairy roots were then transferred to media containing different combinations of plant growth hormones 0.5, 1.0, and 2.0 mg/L of BAP and NAA. The nutrient medium consisting of MS vitamins supplemented with 2 mg/L BAP and 0 mg/L NAA 91.7 had the highest callus formation. However, the highest callus weight was detected in 0.5 mg/L BAP and 1 mg/L NAA
medium, while the highest shoot formation explant rate was 56.7% in 2 mg/L BAP and 1 mg/L NAA medium. Number of shoots per explant was detected in 1 mg/L BAP medium. Putative transgenic plants identified by PCR analyzez. As a result of the study, out of 54 putative transgenic plants 37 samples were confirmed by PCR for primers rolA, rolB, and rolC.