Establishment of the hairy root culture of Echium plantagineum L. and its shikonin production
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ORIGINAL ARTICLE
Establishment of the hairy root culture of Echium plantagineum L. and its shikonin production Jiang‑Yan Fu1 · Hua Zhao1 · Jia‑Xin Bao1 · Zhong‑Ling Wen1 · Rong‑Jun Fang1 · Aliya Fazal1 · Min‑Kai Yang1 · Bao Liu2 · Tong‑Ming Yin3 · Yan‑Jun Pang1,3 · Gui‑Hua Lu1,3,4 · Jin‑Liang Qi1,3 · Yong‑Hua Yang1,3 Received: 6 March 2020 / Accepted: 31 August 2020 © King Abdulaziz City for Science and Technology 2020
Abstract Echium plantagineum L. (Boraginaceae) is an invasive species in Australia and contains medicinal shikonins in its roots. In this study, the hairy root lines of E. plantagineum were established using Agrobacterium rhizogenes strain ATCC15834 and confirmed by the amplification of the rolB gene. Results showed significant difference in shikonin production between the hairy root lines in the 1/2B5 and M9 media. The biomass of the lines in the 1/2B5 medium was fivefold of that in the M9 medium. However, the components of detected shikonins were similar in these two liquid media. By contrast, different accumulation profiles appeared in the hairy root lines. HPLC analysis revealed the presence of nine possible related compounds, including shikonins, and acetylshikonin was the most abundant shikonin derivative. The content of acetylshikonin in the 1/2B5 medium (36.25 mg/L on average) was twofold of that in the M9 medium. Our results showed that the hairy root cultures of E. plantagineum can be used in enhancing the production of potential pharmaceutical compounds, such as acetylshikonin. Keywords Culture medium · Echium plantagineum L. · Hairy root · In vitro biosynthesis · Shikonins
Introduction Plants can produce a range of secondary metabolites at different developmental stages or in specific tissues. These natural products contribute to the interaction of plants with the Jiang-Yan Fu and Hua Zhao contributed equally to this work. * Gui‑Hua Lu [email protected] * Jin‑Liang Qi [email protected] * Yong‑Hua Yang [email protected] 1
Institute for Plant Molecular Biology, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China
2
Key Laboratory of Molecular Epigenetics of the Ministry of Education (MOE), Northeast Normal University, Changchun 130024, China
3
Co‑Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China
4
School of Life Sciences, Huaiyin Normal University, Huaian 223300, China
environment and serve as pharmaceuticals, cosmetics, and dyes (Verpoorte and Alfermann 2000). Naphthoquinones constitute a class of secondary metabolites with a wide distribution among higher plant families. Boraginaceae has a broad distribution, with more than 2000 species in 146 genera, including shrubs, trees, and herbs (Babula et al. 2009). Shikonins are the most important naphthoquinones detected in this family. These red pigments are abundant in the roots of some genera, such as Lithospermum, Arnebia, Echium, and Onosma (Malik et al. 2016). Shikonin biosynthesis starts fr
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