Optimal inductive and cultural conditions of Polygonum multifl orum transgenic hairy roots mediated with Agrobacterium rhizogenes R1601 and an analysis of their anthraquinone constituents
Bing Huang1, Huanjie Lin1, Chuanyan Yan1★, Hongyan Qiu1, Lipeng Qiu2, Rongmin Yu2★ Corresponding author
- 1Department of Natural Medicinal Chemistry, College of Pharmacy, Guangdong Pharmaceutical University, China.
- 2Biotechnology Institute of Chinese Materia Medica, Jinan University, Guangzhou, China.
CORRESPONDENCE
Chuanyan Yan
Department of Natural Medicinal Chemistry, College of Pharmacy, Guangdong Pharmaceutical University, China.
Received: 18-12-2012; Revised: 17-01-2013.
Volume 10, Issue 37 · pp. 77–82 · PUBLISHED 7 February 2014 · DOI: 10.4103/0973-1296.126671
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ABSTRACT
Background: Polygonum multiflorum is an important medicinal plant. Hairy roots systems obtained by transforming plant tissues with the natural genetic engineer Agrobacterium rhizogenes can produce valuable biological active substances, which have immense potential in the pharmaceutical industry. Objective: To optimize the inductive and cultural conditions of P. multiflorum hairy roots and to identify the major active secondary metabolites in hairy roots. Materials and Methods: P. multiflorum hairy root were mediated with A. rhizogenes R1601 to induce hairy roots. Four combinations, including Murashige–Skoog (MS), 1/2 MS, B5, and White, were investigated to optimize the culture medium. MS medium was selected for the growth measurement. The qualitative and quantitative determinations of free anthraquinone in hairy roots were compared with the calli and aseptic plantlets using high-performance liquid chromatography. Results: The inductive rates of hairy roots by leaves were higher than for any other explants. The presence of agropine in the P. multifl orum hairy roots confi rmed that they were indeed transgenic. MS medium was the most suitable of the four media for hairy root growth. Meanwhile, the growth kinetics and nutrient consumption results showed that the hairy roots displayed a sigmoidal growth curve and that their optimal inoculation time was 18-21 days. The determination of the anthraquinone constituents indicated that the rhein content of the hairy roots reached 2.495 μg g−1 and was 2.55-fold higher than that of natural plants. Conclusion: Transgenic hairy roots of P. multifl orum could be one of the most potent materials for industrial-scale production of bioactive anthraquinone constituents.
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Huang, B., Lin, H., Yan, C., Qiu, H., Qiu, L., & Yu, R. (2014). Optimal inductive and cultural conditions of Polygonum multifl orum transgenic hairy roots mediated with Agrobacterium rhizogenes R1601 and an analysis of their anthraquinone constituents. Pharmacognosy Magazine, 10(37), 77–82. https://doi.org/10.4103/0973-1296.126671