02467nas a2200205 4500008004100000245011500041210006900156260001900225300001200244490000700256520183400263653001502097653001502112653002302127653001602150653001602166100001902182700002102201856003902222 2019 eng d00aHairy Root Technology: Effect of Etiolation on Datura sp. Transgenic Root Induction and Hyoscyamine Production0 aHairy Root Technology Effect of Etiolation on Datura sp Transgen cSeptember 2019 a991-9950 v113 a

Introduction: Hyoscyamine, because of its medicinal properties, is an important tropane alkaloid. In order to implement a biotechnological process for its production, hairy roots of Datura species resulting from genetic transformation by Agrobacterium rhizogenes A4 strain have been the subject of this work. In the in vitro alkaloids production programs, optimization of the transformation is a crucial step for obtaining a large number of root lines allowing the selection of efficient lines. Methods: The conditions for hairy roots induction were explored by studying the effect of Datura sp. in vitro seedling etiolation on the genetic transformation. The first step was the establishment of Datura in vitro plantlet cultures followed by the hairy roots induction with A4 strain of A. rhizogenes. The confirmation of the genetic transformation was performed by polymerase chain reaction (PCR) analysis of rolB gene in the roots. After hyoscyamine extraction, it was analyzed (quantitatively and qualitatively) by gas chromatography. Results: 343 root lines were obtained in total, 96 root lines from non-etiolated Datura in vitro seedlings and 247 root lines from etiolated in vitro plantlets. After the selection of six transgenic root lines, tow root lines from each species (D. tramonium, D. tatula and D. innoxia), one from non-etiolated in vitro plantlet and one from etiolated in vitro plantlet, the most hyoscyamine content was 8.43 mg/g D.W. obtained with D. tatula etiolated in vitro seedling. Conclusion: The etiolated in vitro plantlets seem more favorable to hairy roots induction.

10aDatura sp.10aEtiolation10aGas chromatography10aHairy roots10aHyoscyamine1 aHARFI, Boualem1 aKHELIFI, Lakhdar uhttps://www.phcogj.com/article/977