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Kashani K, Jalali Javaran M, Moieni A, Mohebodini M. The Optimization of One and Two-Step Adventitious Organogenesis for Agrobacterium- Mediated Transformation in (Solanum tuberosum L.). NCMBJ 2014; 4 (14) :115-127
URL: http://ncmbjpiau.ir/article-1-531-en.html

Materials and Methods: 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. 

Results: 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). 

Conclusion: 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.