Abstract
An efficient and reproducible protocol for Agrobacterium rhizogenes-mediated hairy root were efficiently induced from in vitro grown leaf explants of H. enneaspermus. Among the diverse factors analysed, acetosyringone (100 µM), sonication (40 s), and vacuum infiltration (2 min) followed by 2 days co-cultivation period revealed maximal percentage of hairy root induction using A4 strain (29.6%). PCR analysis were confirmed the transgenic status of hairy root by using rol B gene specific primer. Further, the highest amount of fresh weight (17.71 g) and dry weight (2.14 g) was noted at 40-day old A. rhizogenes induced hairy root cultures. Precursor L-tyrosine (100 µM) treated hairy roots produced higher L-Dopa (32.71 mg/g DW) content when compared with control culture. The transformation procedure designed in the current research could be valuable to transferring the gene of interest for secondary metabolism that was introduced into hairy roots of Hybanthus, and it could provide a feasible and sustained product platform for naturally derived, high quality and potential phytochemicals.
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Abbreviations
- FW:
-
Fresh weight
- DW:
-
Dry weight
- AS:
-
Acetosyringone
- PCR:
-
Polymerase chain reaction
- LB:
-
Luria-Bertani
- OD:
-
Optical Density
- MS:
-
Murashige and Skoog
- CTAB:
-
Cetyl Trimethyl Ammonium Bromide
- N:
-
Normality
- HPLC:
-
High Performance Liquid Chromatography
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The authors thank to “RUSA, 2.0 - Biological Sciences, Bharathidasan University” for providing financial assistance.
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SS performed experiments, analyzed, assembled the data and prepared manuscript. VV assisted to perform HPLC analysis. SK and GP contributed to results analysis. RS evaluated the data. MM supervised the findings of this research and checked the manuscript.
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Sathish, S., Vasudevan, V., Karthik, S. et al. Precursor feeding enhances L-Dopa production in hairy root culture of Hybanthus enneaspermus (L.) F.Muell. Biologia 78, 913–923 (2023). https://doi.org/10.1007/s11756-022-01308-z
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DOI: https://doi.org/10.1007/s11756-022-01308-z