Enhanced production of phenolic compounds in hairy root cultures of Polygonum multiflorum and its metabolite discrimination using HPLC and FT-IR methods
- 270 Downloads
Hairy root cultures (HRCs) are characterized by fast and unlimited root growth, and they have greater genetic stability than other cultivation methods. HRCs are known to accumulate phytochemical levels comparable to those of intact plant. In this study, HRCs of Polygonum multiflorum were established from leaf explants infected with Agrobacterium rhizogenes strain KCCM 11879. Over 60% of the explants showed hairy root induction after 21 days of cultivation on hormone-free MS (Murashige and Skoog Physiol Plant 15:473–479, 1962) medium; induced roots were confirmed by PCR using a rolC-specific primer. Of the six lines of HRCs selected for further analysis, line HR-01 performed best, producing a root biomass (105.2 g L−1 of FW, 9.7 g L−1 of DW), which is 10-fold higher than that of non-transgenic roots. The HR-01 line also showed a significant increase in its total phenolic content (26.64 mg g−1 DW), while non-transgenic roots accumulated 8.36 mg g−1 DW of total phenolic. The levels of phenolic compounds in the HRCs increased more than 2.5-fold following exposure to 50 μM methyl jasmonate for 5 days. Fourier transform infrared (FT-IR) spectroscopic analysis of bioactive accumulation in P. multiflorum enabled discrimination between hairy root and adventitious root cultures. Thus, it is evident from this study that HRCs could be an attractive proposition for large-scale production of root biomass and secondary metabolites of P. multiflorum in bioreactors.
KeywordsFourier transform infrared spectroscopy Hairy root culture Methyl jasmonate Phenolic compounds Polygonum multiflorum
This work was supported by the Korean Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry and Fisheries (IPET) through the Advanced Production Technology Development Program, funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA) (Grant number 315013-4).
Thanh-Tam Ho, Jong-Du Lee, and Myung-Suk Ahn contributed to data acquisition and manuscript writing. Suk-Weon Kim participated in the interpretation of data and revision for important intellectual content. So-Young Park made substantial contributions to the conception and design of the study.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflicts of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Ahn MS, Min SR, Jie EY, So EJ, Choi SY, Moon BC, Kang YM, Park SY, Kim SW (2015) Rapid comparison of metabolic equivalence of standard medicinal parts from medicinal plants and their in vitro-generated adventitious roots using FT-IR spectroscopy. Korean J Plant Biotechnol 42:257–264CrossRefGoogle Scholar
- McCoy E, O’Connor S (2008) Natural products from plant cell cultures. Prog Drug Res 65:330–370Google Scholar
- Palazón J, Cusidó R, Roig C, Piñol MT (1997) Effect of rol genes from Agrobacterium rhizogenes TL-DNA on nicotine production in tobacco root cultures. Plant Physiol Biochem 35:155–162Google Scholar
- Rogers SO, Bendich AJ, Gelvin SB (1988) Plant molecular biology manual. Springer, New YorkGoogle Scholar
- Sivanandhan G, Dev KG, Jeyaraj M, Rajesh M, Arjunan A, Muthuselvam M, Manickavasagam M, Ganapathi A (2013) Increased production of withanolide A, withanone and withaferin A in hairy root cultures of Withania somnifera (L.) Dunal elicited with methyl jasmonate and salicylic acid. Plant Cell Tissue Organ Cult 114:121–129CrossRefGoogle Scholar
- Trinh TH, Hieu T, Tam HT, Hoang NH, Hien VT, Cuong LK, Nam NB, Huy NP, Luan VQ, Vinh BT, Phuong TD, Luan TC, Ha CH, Nhut DT (2014) The effects of some elicitors on growth and ginsenoside production of secondary roots of Panax vietnamensis Ha et Grushv. cultured in vitro. Vietnam J Biotechnol 12:477–487Google Scholar
- Vereshchagina YV, Bulgakov VP, Grigorchuk VP, Rybin VG, Veremeichik GN, Tchernoded GK, Gorpenchenko TY, Koren OG, Phan NHT, Minh NT, Chau LT, Zhuravlev YN (2014) The rolC gene increases caffeoylquinic acid production in transformed artichoke cells. Appl Microbiol Biotechnol 98:7773–7780CrossRefGoogle Scholar