Reza FARJAMINEZHAD, Nasser ZARE, Rasool ASGHARI-ZAKARIA, Manoochehr FARJAMINEZHAD

Establishment and optimization of cell growth in suspension culture of Papaver bracteatum: a biotechnology approach for thebaine production

Iranian poppy (Papaver bracteatum) is an important medicinal plant that is the main source of the opium alkaloids codeine, morphine, and thebaine. To establish an efficient protocol for cell suspension culture and growth, the effects of different plant growth regulators (2,4-D, NAA, BAP, and kinetin) on callus induction and cell suspension culture of Iranian poppy were evaluated. The maximum percentage of callus induction (86.67%) and fresh weight of callus were obtained in MS medium supplemented with 1 or 2 mg/L 2,4-D, 0.1 or 0.2 mg/L kinetin and 15 mg/L ascorbic acid. The addition of 15 mg/L ascorbic acid to the medium was effective in reducing browning of explants and calli. In cell suspension cultures, MS medium supplemented with 1 mg/L NAA, 1 mg/L BAP, and 15 mg/L ascorbic acid gave maximum cell growth and proliferation compared to other treatments evaluated. Among the auxins, NAA was more effective than 2,4-D for growth of P. bracteatum cell suspension cultures. However, there was no statistically significant difference between kinetin and BAP. The thebaine yield of P. bracteatum cell suspension at 12 days after culture was 4.67 µg/g dry weight, which was significantly lower than that at 16 days (10.76 µg/g dry weight).

Anahtar Kelimeler:

Key words: Cell suspension culture, Papaver bracteatum, secondary metabolites, thebaine

Establishment and optimization of cell growth in suspension culture of Papaver bracteatum: a biotechnology approach for thebaine production

Keywords:

Key words: Cell suspension culture, Papaver bracteatum, secondary metabolites, thebaine,

PDF

___

Bais HP, Walker TS, McGrew JJ, Vivanco JM (2002). Factors affecting growth of cell suspension cultures of Hypericum perforatum L. (St. John’s wort) and production of hypericin. In Vitro Cell Dev-Pl 38: 58–65.
Cline SD, Coscia CJ (1988). Stimulation of sanguinarine production by combined fungal elicitation and hormonal deprivation in cell suspension cultures of Papaver bracteatum. Plant Physiol 86: 161–165.
Daud Z, Keng CL (2006). Effects of plant growth regulators on the biomass of embryogenic cells of Cyperus aromaticus (Ridly) Mattf and Kukenth. Biotechnol 5: 75–78.
Falco MC, Mendes BMJ, Neto AT (1996). Cell suspension culture of sugarcane: growth, management, and plant regeneration. R Bras Fisiol Veg 8: 1–6.
Gadzovska S, Maury S, Delaunay A, Spasenoski M, Joseph C, Hagège D (2007). Jasmonic acid elicitation of Hypericum perforatum L. cell suspensions and effects on the production of phenylpropanoides and naphtodianthrones. Plant Cell Tiss Org 89: 1–13.
Godoy-Hernández G, Vázquez-Flota FA (2006). Growth measurements: estimation of cell division and cell expansion. In: Loyola-Vargas VM, Vazquez-Flota F, editors. Plant Cell Culture Protocols, 2nd ed. Totowa, NJ, USA: Humana Press Inc., pp. 51–58.
Hamill JD, Rhodes MJC (1993). Manipulating secondary metabolism in culture. In: Grierson D, editor. Plant Biotechnology, Volume III: Biosynthesis and Manipulation of Plant Products. London: Blackie Academic and Professional (Chapman and Hall), pp. 178–209.
Hossain MJ, Raman M, Bari MA (2007). Establishment of cell suspension culture and plantlet regeneration of Brinjal (Solanum melongena L.). J Plant Sci 2: 407–415.
Ilahi I, Ghauri EG (1994). Regeneration in cultures of Papaver bracteatum as influenced by growth hormones and temperature. Plant Cell Tiss Org 38: 81–83.
Jayanthi M, Mandal, PK (2001). Plant regeneration through somatic embryogenesis and rapid analysis of regenerated plants in Tylophora indica (Burm. F. Merrill.). In Vitro Cell Dev-Pl 37: 576–580.
Kamimura S, Akutsu M (1976). Cultural conditions on growth of cell culture of Papaver bracteatum. Agr Biol Chem Tokyo 40: 899–906.
Kamimura S, Akutsu M, Nishikawa M (1976). Formation of thebaine in the suspension culture of Papaver bracteatum. Agr Biol Chem Tokyo 40: 913–919.
Machakova I, Zazimalova E, George EF (2008). Plant growth regulators I: Introduction; auxins, their analogues and inhibitors. In: George EF, Hall MA, de Klerk G-J, editors. Plant Propagation by Tissue Culture. Vol. 1. The Background, 3rd ed. Dordrecht: Springer, pp. 175–204.
Milo J, Levy A, Palevitch D (2006). An alternative raw—the cultivation and breeding of Papaver bracteatum. In: Bernáth J, editor. Poppy: the Genus Papaver. Amsterdam: Harwood Academic Publishers, pp. 279–289.
Murashige T, Skoog F (1962). A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plantarum 15: 473–497.
O’Dowd NA, McCauley PG, Richardson DHS, Wilson G (1993). Callus production, suspension culture and in vitro alkaloid yields of Ephedra. Plant Cell Tiss Org 34: 149–155.
Oksman-Caldentey KM, Inze D (2004). Plant cell factories in the postgenomic era: new ways to produce designer secondary metabolites. Trends Plant Sci 9: 433–440.
Okumuş V, Pirinç V, Onay A, Başaran D (2011). In vitro propagation of Diyarbakır watermelons and comparison of direct-seeded and transplanted watermelon. Turk J Biol 35: 601–610.
Pan MJ, Van Staden J (1998). The use of charcoal in in vitro culture: a review. Plant Growth Regul 26: 155–163.
Phillipson JD (1990). Plants as source of valuable products. In: Charlwood BV, Rhodes MJC, editors. Secondary Products from Plant Tissue Culture. Oxford: Clarendon Press, pp. 1–21.
Ramachandra Rao S, Ravishankar GA (2002). Plant cell cultures: chemical factories of secondary metabolites. Biotechnol Adv 20: 101–153.
Ravishankar GA, Bhyalakshmi N, Ramachandra Rao S (1999). Production of food additives. In: Ramawat KG, Mérillon JM, editors. Biotechnology: Secondary Metabolites. New Delhi, Oxford: IBH, pp. 89–110.
Richard D, Lescot M, Inze D, De Veylder L (2002). Effect of auxin, cytokinin, and sucrose on cell cycle gene expression in Arabidopsis thaliana cell suspension cultures. Plant Cell Tiss Org 69: 167–176.
Roberts MF (1988). Isoquinolines (Papaver alkaloids). In: Constabel F, Vasil IK, editors. Cell Culture and Somatic Cell Genetics of Plants. San Diego: Academic Press Inc., pp. 315–334.
Rostampour S, Sohi HH, Dehestani A (2010). In vitro regeneration of Persian poppy (Papaver bracteatum). Biologia 65: 647–652.
Rostampour S, Sohi HH, Jourabchi E, Ansari E (2009). Influence of Agrobacterium rhizogenes on induction of hairy roots and benzylisoquinoline alkaloids production in Persian poppy (Papaver bracteatum Lindl.): preliminary report. World J Microb Biot 25: 1807–1814.
Ryu DDY, Lee SO, Romani RJ (1990). Determination of growth rate for plant cell cultures: comparative studies. Biotechnol Bioeng 35: 305–311.
Shibli RA, Smith MAL, Sharnawi MA (1999). Pigment recovery from encapsulated–dehydrated Vaccinium pahalae (Ohelo) cryopreserved cells. Plant Cell Tiss Org 55: 119–123.
Silveira V, Floh EIS, Handro W, Pedro Guerra M (2004). Effect of plant growth regulators on the cellular growth and levels of intracellular protein, starch, and polyamines in embryogenic suspension cultures of Pinus taeda. Plant Cell Tiss Org 76: 53–
Stals H, Inze D (2001). When plant cells decide to divide. Trends Plant Sci 8: 359–364.
Tisserat B, Berhow M (2009). Production of pharmaceuticals from Papaver cultivars in vitro. Eng Life Sci 9: 190–196.
Toering A, Pullman GS (2005). Modeling available 2,4-dichlorophenoxyacetic acid in a tissue culture medium containing activated carbon. Plant Cell Tiss Org 82: 179–188.
Van Winkle SC, Pullman GS (2005). Achieving desired plant growth regulator levels in liquid plant tissue culture media that include activated carbon. Plant Cell Rep 24: 201–208.
Venkatachalam P, Jiayabalan N (1997). Effect of auxins and cytokinins on efficient plant regeneration and multiple-shoot formation from cotyledons and cotyledonary-node explants of groundnut (Arachis hypogaea L.) by in vitro culture technology. Appl Biochem Biotech 67: 237–247.
Verma SK, Yücesan BB, Gürel S, Gürel E (2011). Indirect somatic embryogenesis and shoot organogenesis from cotyledonary leaf segments of Digitalis lamarckii Ivan., an endemic medicinal species. Turk J Biol 35: 743–750.
Verpoorte R, Contin A, Memelink J (2002). Biotechnology for the production of plant secondary metabolites. Phytochem Rev 1: 13–
Wan Y, Sorensen EL, Liang GH (1988). Genetic control of in vitro regeneration in alfalfa (Medicago sativa L). Euphytica 39: 3–9.
Wickremesinhe ERM, Arteca RN (1993). Establishment of fastgrowing callus and root cultures of Cephalotaxus harringtonia. Plant Cell Rep 12: 80–83.
Zhong JJ, Bai Y, Wang SJ (1996). Effects of plant growth regulators on cell growth and ginsenoside saponin production by suspension cultures of Panax quinquefolium. J Biotechnol 45: 227–234.