Abstract
The study reports a protocol for the regeneration of Glycyrrhiza glabra plantlet from the leaf explant. Notably, the chemical analysis of various developmental stages revealed presence of glycyrrhizin in the underground (7.0–29.8 µg/g) & aerial (7.3–23.4 µg/g) tissues of the in vitro regenerated plants, which was otherwise not detected in the aerial tissues of the field plant. Glycyrrhizin accumulation was reduced or undetected in the regenerated plants transferred to the glasshouse and subsequently under field conditions. Further, spatio-temporal relative gene expression analysis of aerial tissues of in-vitro regenerated G. glabra showed expression of all the known genes committed to glycyrrhizin pathway. In the shoot system, maximum expression of squalene epoxidase (7.9 fold), β-amyrin synthase (21.8 folds), Licorice β-amyrin 11-oxidase (5.9 folds) and UDP-glucosyltransferase (1.7 folds) was observed in different months. However, no expression was detected in the aerial tissues of the field grown plant. Also, a correlation was found between the expression patterns of Licorice β-amyrin 11-oxidase with glycyrrhizin accumulation. This is the first study demonstrating the presence of glycyrrhizin in the aerial tissues of the in-vitro regenerated Glycyrrhiza plant. This study will help in understanding glycyrrhizin regulation, precursor accumulation and their transport. Also, it will unlock the possibilities of understanding and enhancing glycyrrhizin biosynthesis and accumulation in the plant, under in-vitro conditions.
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Acknowledgements
MM, PP & PG acknowledges Council of Scientific and Industrial Research, Government of India & University Grant Commission, India for providing the fellowships. SG acknowledges the Science and Engineering Research Board, Department of Science and Technology, Government of India (SERB/SB/SO/PS/90/2013) for the grant to carry out the research work.
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MM & PG have performed all the experiments. PP, SK & AA have done the initial tissue culture optimisation; APG & PJ were involved in all the chemical analysis and validation. SG, PM & RV were involved in conceptualisation of the study and manuscript preparation. SG has contributed to supervision, investigation, project administration and SG & RV were involved in fund acquisition.
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Manzoor, M.M., Goyal, P., Gupta, A.P. et al. Chemical and real-time based analysis revealed active gene machinery of glycyrrhizin biosynthesis and its accumulation in the aerial tissues of in-vitro regenerated Glycyrrhiza glabra L.. Plant Growth Regul 92, 263–271 (2020). https://doi.org/10.1007/s10725-020-00635-y
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DOI: https://doi.org/10.1007/s10725-020-00635-y