TY - JOUR
T1 - A hairy-root transformation protocol for Trigonella foenum-graecum L. as a tool for metabolic engineering and specialised metabolite pathway elucidation
AU - Garagounis, Constantine
AU - Beritza, Konstantina
AU - Georgopoulou, Maria-Eleni
AU - Sonawane, Prashant
AU - Haralampidis, Kosmas
AU - Goossens, Alain
AU - Aharoni, Asaph
AU - Papadopoulou, Kalliope K.
N1 - We would like to thank Irene Chionidou and Fotini Dimakou for assistance with fenugreek and Lotus hairy root transformations. We acknowledge support of this work by the project: “Synthetic Biology: from omics technologies to genomic engineering” (OMIC-ENGINE) (MIS 5002636), which is implemented under the Action Reinforcement of the Research and Innovation Infrastructure, funded by the Operational Programme Competitiveness, Entrepreneurship, and Innovation (NSRF 2014–2020) and co-financed by Greece and the European Union (European Regional Development Fund) (to KKP), and by the European Community's Seventh Framework Program (FP7/2007–2013) under grant agreement 613692 (TriForC) (AG, AA, KKP). CG was supported by an EMBO short-term fellowship, number 8090. CG and KKP conceived the study and designed and coordinated experimental efforts. CG, KB, MEG conducted experiments, analysed data and prepared figures. PDS provided technical expertise and carried out metabolite analysis. KH provided resources and helped conceive experiments. AG provided TSAR clones and facilitated analysis of samples from Lotus. AA provided analytical equipment for fenugreek metabolite analysis and resources. CG and KKP wrote the paper. All authors received and approved the manuscript.
PY - 2020/9
Y1 - 2020/9
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85087699044&partnerID=8YFLogxK
U2 - 10.1016/j.plaphy.2020.06.011
DO - 10.1016/j.plaphy.2020.06.011
M3 - مقالة
SN - 0981-9428
VL - 154
SP - 451
EP - 462
JO - Plant Physiology and Biochemistry
JF - Plant Physiology and Biochemistry
ER -