Hairy root transformation of Brassica rapa with bacterial halogenase genes and regeneration to adult plants to modify production of indolic compounds.

Autor: Neumann M; Faculty of Biology, Institute of Botany, Technische Universität Dresden, 01062, Dresden, Germany., Prahl S; Vita34 AG, 04103, Leipzig, Germany., Caputi L; Department of Natural Product Synthesis, Max Planck Institute for Chemical Ecology, 07745, Jena, Germany., Hill L; John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, United Kingdom., Kular B; John Innes Centre, Norwich Research Park, Norwich, NR4 7UH, United Kingdom., Walter A; Faculty of Biology, Institute of Botany, Technische Universität Dresden, 01062, Dresden, Germany., Patallo EP; Biochemistry, Technische Universität Dresden, 01062, Dresden, Germany., Milbredt D; Biochemistry, Technische Universität Dresden, 01062, Dresden, Germany., Aires A; Centre for the Research and Technology for Agro-Environment and Biological Sciences, CITAB, University of Trás-os-Montes e Alto Douro, 5001-801, Vila Real, Portugal., Schöpe M; Vita34 AG, 04103, Leipzig, Germany., O'Connor S; Department of Natural Product Synthesis, Max Planck Institute for Chemical Ecology, 07745, Jena, Germany., van Pée KH; Biochemistry, Technische Universität Dresden, 01062, Dresden, Germany., Ludwig-Müller J; Faculty of Biology, Institute of Botany, Technische Universität Dresden, 01062, Dresden, Germany. Electronic address: Jutta.Ludwig-Mueller@tu-dresden.de.
Jazyk: angličtina
Zdroj: Phytochemistry [Phytochemistry] 2020 Jul; Vol. 175, pp. 112371. Date of Electronic Publication: 2020 Apr 10.
DOI: 10.1016/j.phytochem.2020.112371
Abstrakt: During the last years halogenated compounds have drawn a lot of attention. Metabolites with one or more halogen atoms are often more active than their non-halogenated derivatives like indole-3-acetic acid (IAA) and 4-Cl-IAA. Within this work, bacterial flavin-dependent tryptophan halogenase genes were inserted into Brassica rapa ssp. pekinensis (Chinese cabbage) with the aim to produce novel halogenated indole compounds. It was investigated which tryptophan-derived indole metabolites, such as indole glucosinolates or potential degradation products can be synthesized by the transgenic root cultures. In vivo and in vitro activity of halogenases heterologously produced was shown and the production of chlorinated tryptophan in transgenic root lines was confirmed. Furthermore, chlorinated indole-3-acetonitrile (Cl-IAN) was detected. Other tryptophan-derived indole metabolites, such as IAA or indole glucosinolates were not found in the transgenic roots in a chlorinated form. The influence of altered growth conditions on the amount of produced chlorinated compounds was evaluated. We found an increase in Cl-IAN production at low temperatures (8 °C), but otherwise no significant changes were observed. Furthermore, we were able to regenerate the wild type and transgenic root cultures to adult plants, of which the latter still produced chlorinated metabolites. Therefore, we conclude that the genetic information had been stably integrated. The transgenic plants showed a slightly altered phenotype compared to plants grown from seeds since they also still expressed the rol genes. By this approach we were able to generate various stably transformed plant materials from which it was possible to isolate chlorinated tryptophan and Cl-IAN.
Competing Interests: Declaration of competing interest The authors declare no conflict of interest.
(Copyright © 2020 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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