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Response surface modeling of natural alizarin production in hairy root cultures of Rubia tinctorum L. upon elicitation with fungal mycelia

Mehdi Ghorbani, Mansoor Omidi, Seyed Ali Peighambari, Karamatollah Rezaei


The roots of Rubia tinctorum L., the common madder contain natural red dye known as alizarin. In the current study, central composite design of response surface methodology was employed for modeling of fungal elicitor treatment on natural alizarin production in uniform hairy root cultures of common madder in liquid 1/2 B5 medium. Upon fungal elicitation assay, using two fungal mycelia elicitors (Aspergillus niger and Bipolaris maydis) at three different times (0, 12 and 24h), the production of alizarin was determined. According to the results, after 24h; modeling and optimization conditions, including combination of 2 % of both elicitors for alizarin production equal to 10.0 mg.g-1 DW was evaluated. Optimal process parameters have been determined by using a high desirability value of 1.00 in Design-Expert® software. Our results, altogether, offer a promising method regarding to the improvement of the alizarin production, as a pivotal natural dye in industrial applications.


Bahabadi, S.E., Sharifi, M., Safaie, N., Murata, J., Yamagaki, T., Satake, H., 2011. Increased lignan biosynthesis in the suspension cultures of Linum album by fungal extracts. Plant. Biotechnol. Report., 5, 367-373.

Banyai, P., Kuzovkina, I.N., Kursinszki, L., Szoke, E., 2006. HPLC analysis of alizarin and purpurin produced by Rubia tinctorum L. hairy root cultures. Chromatographia, 63, S111-S114.

Boka, K., Jakab, J., Kiraly, I., 2002. Comparison of the effect of different fungal elicitors on Rubia tinctorum L. suspension culture. Biol. Plantarum., 45, 281-290.

Ghorbani, M., Omidi, M., Peighambari, S.A., 2014. L-Arginine optimization medium for hairy root induction of madder (Rubia tinctorum L.) based on one factor model of response surface methodology. Sci. J. Biol. Sci., 3, 117-125

Guillon, S., Trémouillaux-Guiller, J., Pati, P.K., Rideau, M., Gantet, P., 2006. Hairy root research: recent scenario and exciting prospects. Curr. Opin. Plant. Biol., 9, 341-346.

Hu, Z-B., Du, M., 2006. Hairy root and its application in plant genetic engineering. J. Integr. Plant. Biol., 48, 121-127.

Imanishi, L., Vayssières, A., Franche, C., Bogusz, D., Wall, L., Svistoonoff, S., 2011. Transformed hairy roots of discaria trinervis: A valuable tool for studying actinorhizal symbiosis in the context of intercellular infection. Mol. Plant. Microbe. Interact., 24, 1317-1324.

Khan, S., Qureshi, M.I., Alam, T., Abdin, M.Z., 2007. Protocol for isolation of genomic DNA from dry and fresh roots of medicinal plants suitable for RAPD and restriction digestion. Afr. J. Biotechnol., 6, 175-178.

Nartop, P., Akay, Ş., Gürel, A., 2013. Immobilization of Rubia tinctorum L. suspension cultures and its effects on alizarin and purpurin accumulation and biomass production. Plant Cell, Tissue and Organ Culture (PCTOC). 112, 123-128.

Navaei, M.N., Mirza, M., Dini, M., 2006. Chemical composition of the essential oils of Rubia tinctorum L. aerial parts from Iran. Flavour. Fragr. J., 21, 519-520.

Oksman-Caldentey, K.M., 2002. Agrobacterium rhizogenes‐mediated transformation: Root cultures as a source of alkaloids. Plant. Med., 68, 859-868.

Ono, N.N., Tian, L., 2011. The multiplicity of hairy root cultures: Prolific possibilities. Plant. Sci., 180, 439-446.

Orbán, N., Boldizsár, I., Szűcs, Z., Dános, B., 2008. Influence of different elicitors on the synthesis of anthraquinone derivatives in Rubia tinctorum L. cell suspension cultures. Dyes and Pigments, 77, 249-257.

Ozen, E., Yeniocak, M., Goktas, O., Alma, M.H., Yilmaz, F., 2014. Antimicrobial and antifungal properties of Madder Root (Rubia tinctorum) colorant used as an environmentally-friendly wood preservative. Bio. Resour., 9, 1998-2009.

Perassolo, M., Quevedo, C.V., Busto, V.D., Giulietti, A.M., Talou, J.R., 2011a. Role of reactive oxygen species and proline cycle in anthraquinone accumulation in Rubia tinctorum cell suspension cultures subjected to methyl jasmonate elicitation. Plant. Physiol. Biochem., 49, 758-763.

Perassolo, M., Quevedo, C.V., Giulietti, A.M., Talou, J.R., 2011b. Stimulation of the proline cycle and anthraquinone accumulation in Rubia tinctorum cell suspension cultures in the presence of glutamate and two proline analogs. Plant Cell, Tissue and Organ Culture (PCTOC). 106, 153-159.

Vasconsuelo, A., Giulietti, A.M., Boland, R., 2004. Signal transduction events mediating chitosan stimulation of anthraquinone synthesis in Rubia tinctorum. Plant. Sci., 166, 405-413.

Zargari, A., 1992. Medicinal Plants vol 2. University of Tehran Press, Tehran.

Zhou, M.L., Zhu, X.M., Shao, J.R., Tang, Y.X., Wu, Y.M., 2011. Production and metabolic engineering of bioactive substances in plant hairy root culture. Appl. Microbiol. Biotechnol., 90, 1229-1239.

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