The Optimization of One and Two-Step Adventitious Organogenesis for Agrobacterium- Mediated Transformation in (Solanum tuberosum L.)

Aim and Plant based systems offer the potential for safe, economical and high-capacity production for many biopharmaceuticals. Potato (Solanum tuberosum L.) is a proper bioreactor and a very good candidate for genetic manipulation. After gene transformation to the explants, shoot regeneration is the first and the most remarkable necessity. After investigation of various one and two-step culture media for shoot regeneration of Desiree, Agria and Marfona internodal explants and calculation of shoot regeneration percentage, the best one and two-step shoot regeneration media is introduced for adventitious organogenesis in each of these varieties. Human proinsulin gene is transferred into nuclear genome of potato plant via Agrobacterium tumefaciens-mediated transformation method. The appropriate amounts of Zeatin Riboside, Naphthalene Acetic Acid and Gibberellic Acid hormones needed for maximum regeneration in two-step culture media for shoot regeneration of Desiree, Agria and Marfona internodal explants in the first stage were (3, 0.1, 0.02), (3, 0.1, 0.02) and (3.5, 0.1, 0.02) mg L-1 respectively. These amounts in one- step culture media were (4, 0.02, 0.05), (3, 0.02, 0.05) and (4, 0.02, 0.05) mg L-1 in that order. In the second stage of two- step culture media, zeatin riboside and naphthalene acetic acid levels were reduced by 20% and the factor of 10 respectively. Assessment of transgenic plants was carried out in 3 levels (DNA, RNA and protein). The optimization of adventitious organogenesis in Desiree, Agria and Marfona cultivars will be helpful for other researchers who want to study the transformation of these cultivars in future.