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Transformed Root Culture: From Genetic Transformation to NMR-Based Metabolomics

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Plant Cell Culture Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1815))

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Abstract

Hairy root (HR) culture is considered as “green factory” for mass production of bioactive molecules with pharmaceutical relevance. As such, HR culture has an immense potential as a valuable platform to elucidate biosynthetic pathways and physiological processes, generate recombinant therapeutic proteins, assist molecular breeding, and enhance phytoremediation efforts. However, some plant species appear recalcitrant to the classical Agrobacterium rhizogenes transformation techniques. Sonication-assisted Agrobacterium-mediated transformation (SAArT) is a highly effective method to deliver bacteria to target plant tissues that includes exposure of the explants to short periods of ultrasound in the presence of the bacteria.

Nuclear magnetic resonance (NMR)-based metabolomics is one of the most powerful and suitable platforms for identifying and obtaining structural information on a wide range of compounds with a high analytical precision. In terms of plant science, NMR metabolomics is used to determine the phytochemical variations of medicinal plants or commercial cultivars in certain environments and conditions, including biotic stress and plant biotic interaction, structural determination of natural products, quality control of herbal drugs or dietary supplements, and comparison of metabolite differences between plants and their respective in vitro cultures.

In this chapter, we attempt to summarize our knowledge and expertise in induction of hairy roots from rare and recalcitrant plant species by SAArT technique and further methodology for extraction of secondary metabolites of moderate to high polarity and their identification by using NMR-based metabolomics.

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Acknowledgment

This work has been supported by a grant from NSF of Bulgaria and DAAD Germany (Contract Number DNTS/Germany 01/8).

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Author notes

  1. Andrey S. Marchev and Zhenya P. Yordanova are contributed equally to this work.

Authors and Affiliations

  1. Group of Plant Cell Biotechnology and Metabolomics, The Stephan Angeloff Institute of Microbiology, Bulgarian Academy of Sciences, 139 Ruski Blvd., 4000, Plovdiv, Bulgaria

    Andrey S. Marchev & Milen I. Georgiev

  2. Department of Plant Physiology, Faculty of Biology, Sofia University “St. Kliment Ohridski”, 8 Dragan Tsankov Blvd., Sofia, 1164, Bulgaria

    Zhenya P. Yordanova

Authors
  1. Andrey S. Marchev
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  2. Zhenya P. Yordanova
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  3. Milen I. Georgiev
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Corresponding author

Correspondence to Milen I. Georgiev .

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Editors and Affiliations

  1. Centro de Investigación Científica de Yucatán, Unidad de Bioquímica y Biología, Molecular de Plantas, Mérida, Yucatán, Mexico

    Víctor M. Loyola-Vargas

  2. Departamento de Ingeniería Genética, Unidad Irapuato, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, Irapuato, Guanajuato, Mexico

    Neftalí Ochoa-Alejo

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Marchev, A.S., Yordanova, Z.P., Georgiev, M.I. (2018). Transformed Root Culture: From Genetic Transformation to NMR-Based Metabolomics. In: Loyola-Vargas, V., Ochoa-Alejo, N. (eds) Plant Cell Culture Protocols. Methods in Molecular Biology, vol 1815. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8594-4_32

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  • DOI: https://doi.org/10.1007/978-1-4939-8594-4_32

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  • Publisher Name: Humana Press, New York, NY

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