Abstract
Hairy root cultures are widely used for the production of different secondary metabolites, since their level of synthesis in transgenic roots is often higher than that of non-transgenic tissues. However, the mechanism of influence of T-DNA genes on plant metabolism is not studied well. We offer a diagnostic panel for Fluidigm BioMark for investigation of the expression of genes associated with the synthesis of alkaloids in tobacco hairy roots and related PCR protocol. The list of investigated genes includes genes of alkaloid biosynthesis, genes of transcription factors participating in the regulation of alkaloid content, rol genes, and reference genes.
Keywords
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Dewey R, Xie J (2013) Molecular genetics of alkaloid biosynthesis in Nicotiana tabacum. Phytochemistry 94:10–27. https://doi.org/10.1016/j.phytochem.2013.06.002
Mehrotra S, Srivastava V, Rahman LU, Kukreja AK (2015) Hairy root biotechnology—indicative timeline to understand missing links and future outlook. Protoplasma 252:1189–1201. https://doi.org/10.1007/s00709-015-0761-1
Omote H et al (2006) The MATE proteins as fundamental transporters of metabolic and xenobiotic organic cations. Trends Pharmacol Sci 27(11):587–593. https://doi.org/10.1016/j.tips.2006.09.00
Pistelli L, Giovannini A, Ruffoni B, Bertoli A, Pistelli L (2010) Hairy root cultures for secondary metabolites production. Adv Exp Med Biol 698:167–184
Real-Time PCR Analysis User Guide (PN 68000088 N1). https://www.fluidigm.com/binaries/content/documents/fluidigm/resources/real-time-pcr-analysis-ug-68000088/real-time-pcr-analysis-ug-68000088/fluidigm%3Afile. Accessed 10.06.2019
Shoji T, Inai K, Yazaki Y, Sato Y, Takase H, Shitan N, Yazaki K, Goto Y, Toyooka K, Matsuoka K et al (2009) Multidrug and toxic compound extrusion-type transporters implicated in vacuolar sequestration of nicotine in tobacco roots. Plant Physiol 149:708–718
Shoji T, Kajikawa M, Hashimoto T (2010) Clustered transcription factor genes regulate nicotine biosynthesis in tobacco. Plant Cell. https://doi.org/10.1105/tpc.110.078543
Srivastava V, Mehrotra S, Mishra S (2016) Biotransformation through hairy roots: perspectives, outcomes, and major challenges. In: Jha S (ed) Transgenesis and secondary metabolism, Reference series in Phytochemistry. Springer, Cham, pp 1–24
Zenkner FF, Margis-Pinheiro M, Cagliari A (2019) Nicotine biosynthesis in Nicotiana: a metabolic overview. Tobacco Sci 56:1–9
Acknowledgment
The authors are grateful to RC Development of Molecular and Cellular Technologies of Saint Petersburg State University for providing the necessary facilities to carry out the work. Grant 18-016-00118 provided by Russian Foundation for Basic research for TM is acknowledged.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Nature Singapore Pte Ltd.
About this chapter
Cite this chapter
Matveeva, T., Amini, G. (2020). Fluidigm BioMark Diagnostic Panel for Analysis of the Expression of Nicotiana tabacum Genes, Associated with Alkaloid Synthesis. In: Srivastava, V., Mehrotra, S., Mishra, S. (eds) Hairy Root Cultures Based Applications. Rhizosphere Biology. Springer, Singapore. https://doi.org/10.1007/978-981-15-4055-4_15
Download citation
DOI: https://doi.org/10.1007/978-981-15-4055-4_15
Published:
Publisher Name: Springer, Singapore
Print ISBN: 978-981-15-4054-7
Online ISBN: 978-981-15-4055-4
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)