Effect of Some Phytohormones on Podophyllotoxin Production in Cell and Plantlets Cultures of Linum album


The natural lignan podophyllotoxin, a dimerized product of two phenylpropanoid moieties which occurs in a few plant species, is a pharmacologically important compound for its anticancer activities. Optimization of the in vitro cultures might lead to a renewable source of lignans for medicinal uses. In order to maximize efficiency of podophyllotoxin accumulation via in vitro cultures, the effect of different concentrations of α-Naphthalene acetic acid (NAA) (0, 0.4 and 1 mg/l), Kinetin (Kin) (0, 0.2 and 0.4 mg/l) and combinations of NAA and Kin [(0.4 and 0.2), (0.4 and 0.4), (1 and 0.2) and (1 and 0.4) mg/l] on growth and accumulation of podophyllotoxin in callus, cell culture and in vitro propagated plantlets of Linum album was evaluated. The highest growth and accumulation of podophyllotoxin in callus culture was obtained when it grown under dark condition in comparison to photoperiod. The biomass of cell and callus do not changed under various hormone concentrations. Content of podophyllotoxin was enhanced in cell and callus cultures supplemented with NAA 1 mg/l + Kin 0.4 mg/l. The experiments demonstrated Kin 0.2 or 0.4 mg/l stimulated multiplication and production of podophyllotoxin in shoot culture.


1. Schmitt J, Petersen M. Pinoresinol and matairesinol accumulation in a Forsythia × intermedia cell suspension culture. PCTOC 2002;68:91–98.

2. Farkya S, Bisaria VS, Sirvastava AK. Biotechnological aspects of the production of the anticancer drug podophyllotoxin. Appl Microbiol Biotechnol. 2004;65:504-519.

3. Fuss E. Lignans in plant cell and organ cultures: An overview. Phytochem Rev. 2003;2:307-320.

4. Yousefzadi M, Sharifi M, Ahmadian Chashmi N, Behmanesh M, Ghasempour, A. Optimization of podophyllotoxin extraction method from Linum album cell cultures.Pharm Biol. 2010a;12:1421-1425.

5.Smollny T, Wichers H, Kalenberg S, Shahsavari A, Petersen M, Alfermann, AW. Accumulation of podophyllotoxin and related lignans in cell suspension cultures of Linum album. Phytochemistry 1998;6:975-979.

6. Yousefzadi M, Sharifi M, Behmanesh M, Ghasempour A, Moyano E, Palazon J. Salicylic acid improves podophyllotoxin production in cell cultures of Linum album by increasing the expression of genes related with its biosynthesis. Biotechnol Lett. 2010b;32:1739-1743.

7. Esmaeilzadeh-Bahabadi S, Sharifi M, Behmanesh M, Safaie N, Murata J, Araki R, Yamagaki T, Satake H. Time-course changes in fungal elicitor-induced lignan synthesis and expression of the relevant genes in cell cultures of Linum album. J Plant Physiol. 2011;169:487- 491.

8. Rao SR, Ravishankar GA. Plant cell cultures: Chemical factories of secondary metabolites. Biotechnol Adv. 2002;20,101-153.

9. Kim YS, Yeung EC, Hahn EJ, Paek KY. Combined effects of phytohormone, indole-3-butyric acid, and methyl jasmonate on root growth and ginsenoside production in adventitious root cultures of Panax ginseng C.A. Meyer. Biotechnol Lett. 2007;29:1789–1792.

10. Zhong JJ, Bai Y, Wang SJ. Effects of plant growth regulators on cell growth and ginsenoside saponin production by suspension cultures of Panax quinquefolium. J Biotechnol. 1996;45:227-234.

11. Khan T, Krupadanam D, Anwar SY. The role of phytohormone on the production of berberine in the calli cultures of an endangered medicinal plant, turmeric (Coscinium fenestratum). AJB 2008;18:3244-3246.

12. Murashige T, Skoog F. A revised medium for rapid growth and bioassay of tobacco tissue culture. Physiol Plantarum. 1962;15:473-497.

13. Salehi H, Salehi M, Ashiri F. Some conditions for the best induction in common Bermudagrass (Cynodon dactylon (L.) Pers. (California origin). Amer-Euroasian J Agric Environ Sci. 2008;3:409-413.

14. Summart J, Panichajaku S, Prathepha  P, Thanonkeo P. Callus Induction and Influence of Culture Condition and Culture Medium on Growth of Thai Aromatic Rice, Khao Dawk Mali 105, Cell Culture. WASJ 2008;2:246-251.

15. Suzuki S, Miki O, Masaru N. Embryogenic callus induction from leaf explants of the Liliaceous ornamental plant, Agapanthus praecox sp. orientalis. Histological study and response to selective agents. Sci Hort. 2002;95:123-132.

16. Shabbir K, Ahmad T, Ahmad Hafiz I, Hussain A, Akhtar Abbasi N, Ahmad J. In vitroregeneration of Gerbera jamesonii cv. Sunglow. AJB 2012;42:9975-9984.

17. Nahid JS, Saha S, Hattori K. High frequency shoot regeneration from petal explants of Chrysanthemum morifolium Ramat. in vitro. Pak J Biol Sci. 2007;10:3356-3361.

18. Ahmad R, Sharma VK, Rai AK, Shivananda RD, Shivananda BG. Production of lignans in callus culture of Podophyllum hexandrum. TJPR 2007;4:803-808.

19. Ionkova I, Antonova I, Momekov G, and Fuss E. Production of podophyllotoxin in Linum linearifolium in vitro cultures.Phcog Mag. 2010;23:180-185.

20. Caretto S, Quarta A, Durante M, Nisi R, De Paolis A, Blando F, Mita, G. Methyl jasmonate and miconazole differently affect arteminisin production and gene expression in Artemisia annua suspension cultures. Plant Biol. 2011;1:51-58.

21. Hobbs MC, Yeoman MM.Effect of light on alkaloid accumulation in cell cultures of Nicotiana species. J Exp Bot. 1991;42:1371-1378.

22. Chattopadhyay S, Srivastava AK, Bhojwani SS, Bisaria VS. Production of podophyllotoxin by plant cell cultures of Podophyllum hexandrum in bioreactor. J Biosci Bioeng. 2002;93:215-220.

23. Duangporn P, Siripong P. Effect of auxin and cytokinin on phyllanthusol a production by callus cultures of Phyllanthus acidus skeels. Amer-Eurasian J Agri Environ Sci. 2009;2:258-263.

24. Kadkade PG. Growth and podophyllotoxin production in callus tissues of Podophyllum peltatum. Plant Sci Lett. 1982;25:107-115.

25. Zia M, Mannan A, Chaudhary MF. Effect of growth regulators and amino acids on artemisinin production in the callus of Artemisia absinthium. Pakistan J Bot. 2007;2:799-805.

26. Borroto J, Coll J, Rivas M, Blanco M, Concepcio O, Tandron Y, Hernandez M, Trujillo R. Anthraquinones from in vitro root culture of Morinda royoc L. PCTOC 2008;94:181-187.

27. Abd Alhady MRA. Micropropagation of Stevia rebaudiana Bertoni. A new sweetening crop in Egypt.GJBBR 2011;6:178-182.

28. Komalavalli N, Rao MV. In vitro micropropagation of Gymnema sylvestre- a multipurpose medicinal plant. PCTOC 2000;61:97-105.

29. Raha, S, Roy SC. In vitro plant regeneration in Holarrhena antidysenterica Wall. Though high frequency axillary shoot proliferation. In vitro Cell Dev Biol Plant. 2001;37:232-236.

30. Nikam TD, Savant RS. Multiple shoot regeneration and alkaloid cerpegin accumulation in callus culture of Ceropegia juncea Roxb. Physiol Mol Biol Plants. 2009;15:71-77.

31. Lakshmi SR, Franklin BJH, Senthil KT, Murthy GVS, Rao MV. In vitro propagation of Hoya wightii ssp. palniensis (Mathew KT Ed), a highly vulnerable and endemic species of Western Ghats of Tamil Nadu, India. AJB 2010;9:620-627.

32. Zare AR, Solouki M, Omidi M, Irvani N, Mahdi Nezad N, Rezazadeh SH. Callus induction and plant regeneration in Ferula assa-foetida L. (Asafetida), and endangered medicinal plant. TJS 2010;1:11-18.

33. Khatun MM, Hossain MS, Haque1 MA, Khalekuzzaman M. In vitropropagation of Citrullus lanatus Thumb. from nodal explants culture. J. Bangladesh Agric Univ. 2010;8:203-206.

34. SilveiraDG, Souza FVD, Pelacani CR, Souza AS, Ledo CAS, Santana JRF. Micropropagation and in vitro conservation of Neoglaziovia variegata (Arr. Cam.) mez, a fiber producing bromeliad from Brazil. Braz Arch Biol Technol. 2009;52:923-932.

35. Kovacevic N, Grubisic D. In Vitro. Cultures of plants from the rhamnaceae: shoot propagation and anthraquinones production. Pharm Biol. 2005;43:420-424.

36. Gadzovska S, Pavlova V, Dimovska K, Spasenoski M. Phenylpropanoid productions in in vitro shoot cultures of St. John’s Wort (Hypericum perforatum L.). Proceedings of the III Congress of Ecologists of the Republic of Macedonia with International Participation, 06-09.10.2007, Struga. Special Issues of Macedonian Ecological Society. 2008;8:47-53.

37. Vasilev N, Ionkova I. Lignan accumulation in cell cultures of Linum strictum ssp. strictum L. Acta Pharm. 2004;54:347–351.