Abstract
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.
Highlights
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
The first step was the establishment of Datura in vitro plantlet cultures followed by the hairy roots induction with A4 strain of A. rhizogenes
Summary
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. 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. The wild population of Datura species (D. stramonium, D. tatula and D. innoxia) that grows in Algeria is rich in hyoscyamine This last one is a tropane alkaloid used in medicine as useful natural product.[1,2]. In vitro plant cultures drawn an interest and cost-effective alternative to classical approaches to plant secondary metabolite production. They constitute sustainable and eco-friendly system to obtain complex chemical structures biosynthesized by concerned plant species. “Plant Cell Factory” concept defined as a biotechnological systems developed to improve the production of plantderived compounds with biological activity.[3]
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