Elsevier

Industrial Crops and Products

Volume 114, April 2018, Pages 173-179
Industrial Crops and Products

Elicitor mediated adventitious root culture for the large-scale production of anthraquinones from Oldenlandia umbellata L.

https://doi.org/10.1016/j.indcrop.2018.01.069Get rights and content

Highlights

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    Production of anthraquinones (AQ) from root cultures of Oldenlandia umbellata L.

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    Pectin (50 mg L−1) elicited cultures produced 24.33 mg g−1 dry weight of AQ.

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    13.52 mg g−1 dry weight alizarin was achieved in pectin elicited root cultures.

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    AQ production enhanced to 2.19 fold over control.

Abstract

Anthraquinones (AQs) have an increasing global demand due to its importance in both industrial and pharmaceutical fields. Increased demand can be met with large-scale production of the compound. Enhanced production of AQs was achieved through elicitor mediated adventitious root cultures of Oldenlandia umbellata L. Adventitious root cultures were raised by incorporating different auxins into the half strength MS liquid medium. The root cultures were incubated for a period of 60 days with continuous monitoring at 10 days interval. Among the different auxins used, 1 μM indole 3 acetic acid (IAA) produced highest amount of total AQ (11.09 mg g−1 dry weight). Later, for enhanced production of AQs, elicitors viz., yeast extract, pectin, xylan, or precursors like α-keto glutaric acid and L- phenylalanine and an AQ inhibitor, piroxicam were added at a concentration of 10, 25 and 50 mg L−1 to the medium on 35th day of incubation and roots were harvested at 40th day of incubation. Addition of 50 mg L−1 pectin resulted highest (24.33 mg g−1 dry weight) elicitation of AQs from adventitious roots. HPLC analysis of samples from adventitious roots confirmed the presence of alizarin in the extract. By adopting adventitious root culture through pectin (50 mg L−1) as elicitor, 2.19 fold enhanced AQ production over control was achieved in O. umbellata.

Introduction

Anthraquinones (AQs) are the largest group of dyes that positions second among textile dyes and were used for colouring fabrics since ancient times. Their resistance to biodegradation helps to retain colours in fabrics for a long period which makes them preferable natural dyestuff (Banat et al., 1996; Siddiqui et al., 2010). Apart from its industrial importance, AQs also contribute to therapeutic fields such as antimicrobial, analgesic, hypotensive, anti-inflammatory, anti-genotoxic, antitumor and immune enhancing agents (Wang et al., 2002; Treetip et al., 2008; Sreeranjini and Siril, 2013). Nowadays, Chinese AQ industry is the prime producers and exporters of AQs worldwide. Most of the AQ producers in China adopt synthetic methods for AQ production, viz., from anthracene oil, crude anthracene, phthalic anhydride, etc… Most of these raw materials originate from coal and petrochemical industry. The price fluctuations in these industries significantly affect the cost of production and market demand of AQ. Plant originated AQs on the other hand have a stable global demand in diverse industries such as pharmaceutics, textile dying, cosmetics etc….The demand for natural AQ however, cannot met by conventional extraction based on plants growing at natural stands.

Oldenlandia umbellata L. (Chay root), is an important source of AQs (Siva, 2007, Siva, 2010; Siva et al., 2012) belongs to family Rubiaceae. In ancient period, O. umbellata was cultivated to extract AQ based natural dye from its roots (Siva, 2007). Chemical profiling of AQ derived from O. umbellata (Siva et al., 2009) revealed the presence of different AQ derivatives including alizarin and purpurin.

Phytochemical profile of O. umbellata suggests that it can be used as an alternative source of AQs over well-established sources such as Rubia cordifolia and Morinda citrifolia and can be used for the large-scale production of the natural dyes. In view of this an investigation to test a hypothesis was formulated that whether adventitious root cultures can be used as a method for the in vitro production of anthraquinones or not. In the present study, we examined effectiveness of various elicitors, precursor, or inhibitors to overproduce anthraquinones from adventitious root cultures of O. umbellata.

Section snippets

Induction of adventitious root culture

Oldenlandia umbellata shoots were developed in vitro, as per the protocol of Krishnan and Siril (2016a). The adventitious roots were induced (Fig. 1A) from in vitro shoots on agar gelled MS medium supplemented with 7.5 μM indole-3-butyric acid (IBA). Approximately 50 mg of roots were collected and then transferred to a 100 mL Erlenmeyer flask containing 50 mL half strength liquid MS medium enriched with 3% sucrose, and 1 μM Indole-3-Acetic Acid (IAA). Prior to autoclaving at 121 °C and 108 kPa

Standardization of adventitious root cultures

Adventitious root cultures were developed on half strength MS liquid medium supplemented with 1 μM IAA was further sub-cultured in the same medium with different concentrations (1.0, 2.5, 5.0 and 10.0 μM) of auxins; viz., NAA, IBA or IAA. Fresh weight of adventitious roots developed in medium supplemented with different types and concentrations of auxin showed an overall significant (P < 0.001) effect with regard to varying period of incubation (Table 1). Two-way ANOVA revealed significant

Conclusion

The present study put forward an initial step towards the large-scale production of AQs through adventitious root cultures of O. umbellata. There were previous reports on anthraquinone production from cell suspension cultures and hairy root culture of O. umbellata. However, the present protocol yields higher amount of AQ, especially alizarin than the previous reports. Compared to hairy root cultures, adventitious root cultures are genetically stable and easy to establish through auxin added

Acknowledgements

The authors acknowledge Dr. Suhara Beevy S., Associate Professor and Head, Department of Botany University of Kerala for facilities provided. The present work was funded by University of Kerala, Thiruvananthapuram, India in the form of research grant to SKSR (Ac E1B1/43700/2011 dt. 26/12/11).

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