A hairy-root transformation protocol for Trigonella foenum-graecum L. as a tool for metabolic engineering and specialised metabolite pathway elucidation

Constantine Garagounis, Konstantina Beritza, Maria-Eleni Georgopoulou, Prashant Sonawane, Kosmas Haralampidis, Alain Goossens, Asaph Aharoni, Kalliope K. Papadopoulou

Research output: Contribution to journalArticlepeer-review

Abstract

The development of genetic transformation methods is critical for enabling the thorough characterization of an organism and is a key step in exploiting any species as a platform for synthetic biology and metabolic engineering approaches. In this work we describe the development of an Agrobacterium rhizogenes-mediated hairy root transformation protocol for the crop and medicinal legume fenugreek (Trigonella foenum-graecum). Fenugreek has a rich and diverse content in bioactive specialised metabolites, notably diosgenin, which is a common precursor for synthetic human hormone production. This makes fenugreek a prime target for identification and engineering of specific biosynthetic pathways for the production of triterpene and steroidal saponins, phenolics, and galactomanans. Through this transformation protocol, we identified a suitable promoter for robust transgene expression in fenugreek. Finally, we establish the proof of principle for the utility of the fenugreek system for metabolic engineering programs, by heterologous expression of known triterpene saponin biosynthesis regulators from the related legume Medicago truncatula in fenugreek hairy roots.

Original languageEnglish
Pages (from-to)451-462
Number of pages12
JournalPlant Physiology and Biochemistry
Volume154
Early online date3 Jul 2020
DOIs
StatePublished - Sep 2020

All Science Journal Classification (ASJC) codes

  • Physiology
  • Genetics
  • Plant Science

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