Biologia plantarum 2014, 58:601-610 | DOI: 10.1007/s10535-014-0434-5

In vitro culture of sweet basil: gas exchanges, growth, and rosmarinic acid production

C. Kiferle1, M. Lucchesini1,*, R. Maggini1, A. Pardossi1, A. Mensuali-Sodi2
1 Department of Agriculture, Food and Environment, University of Pisa, Pisa, Italy
2 Life Science Institute, Scuola Superiore Sant'Anna, Pisa, Italy

Five in vitro culture systems with different ventilation rates were used to investigate the influence of vessel environment on photosynthesis, dark respiration, ethylene evolution, and rosmarinic acid (RA) production in sweet basil (Ocimum basilicum L.) micropropagated shoots. The systems under comparison were two bioreactors with either temporary (RITA™) or stationary (Growtek™) immersion, and three types of vessels (Magenta™, Microbox ECO 2 ™, and PCCV25™) that are largely used for plant micropropagation. Shoots of green-leaved cv. Genovese and purple-leaved cv. Dark Opal were cultured on a modified Murashige and Skoog medium containing 0.25 mg dm-3 6-benzylaminopurine. The instantaneous rates of photosynthesis, dark respiration, and ethylene production were determined by gas chromatography measuring CO2 and ethylene concentrations in vessel headspaces. The tissue RA content was determined by HPLC in HCl-methanol extracts. The explant growth and morphology were significantly affected by culture conditions and cultivars. The largest biomass production was observed under the photomixotrophic culture conditions provided by Growtek™, whereas the highest RA content in shoot tissues was found in the RITA™ photomixotrophic system, where ethylene accumulated to the greatest extent.

Keywords: bioreactors; ethylene; mixotrophic culture; photosynthetetic rate; respiration rate; secondary metabolism
Subjects: in vitro culture; gas exchange; rosmarinic acid; ethylene; net photosynthetic rate; respiration rate; growth analysis; sweet basil
Species: Ocimum basilicum

Received: November 13, 2013; Revised: February 1, 2014; Accepted: March 20, 2014; Published: December 1, 2014Show citation

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Kiferle, C., Lucchesini, M., Maggini, R., Pardossi, A., & Mensuali-Sodi, A. (2014). In vitro culture of sweet basil: gas exchanges, growth, and rosmarinic acid production. Biologia plantarum58(4), 601-610. doi: 10.1007/s10535-014-0434-5.
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References

  1. Afreen, F., Zobayed, S.M.A., Kozai, T.: Photoautotrophic culture of Coffea arabusta somatic embryos: development of a bioreactor for large-scale plantlet conversion from cotyledonary embryos. - Ann. Bot. 90: 21-29, 2002. Go to original source...
  2. Begum, F., Amin, M.N., Azad, M.A.K.: In vitro rapid clonal propagation of Ocimum basilicum L.. - Plant Cell Tissue Organ Cult. 12: 27-35, 2002.
  3. Bertoli, A., Lucchesini, M., Mensuali-Sodi, A., Leonardi, M., Doveri, S., Magnabosco, A., Pistelli L.: Aroma characterisation and UV elicitation of purple basil from different plant tissue cultures. - Food Chem. 141: 776-787, 2013. Go to original source...
  4. Biddington, N.L.: The influence of ethylene in plant tissue culture. - Plant Growth Regul. 11: 173-187, 1992. Go to original source...
  5. Bourgaud, F., Gravot, A., Milesi, S., Gontier, E.: Production of plant secondary metabolites: a historical perspective. - Plant Sci. 161: 839-851, 2001. Go to original source...
  6. Chakrabarty, D., Park, S.Y., Ali, M.B., Shin, K.S., Paek, K.Y.: Hyperhydricity in apple: ultrastuctural and physiological aspects. - Tree Physiol. 26: 377-388, 2005.
  7. Chanemougasoundharam, A., Sarkar, D., Pandey, S.K., Al-Biski, F., Helali, O., Minhas, J.S.: Culture tube closure-type affects potato plantlets growth and chlorophyll contents. - Biol. Plant. 48: 7-11, 2004. Go to original source...
  8. Cho, G.H., Kim, D.I., Pedersen, H., Chin, C.K.: Ethephon enhancement of secondary metabolite synthesis in plant cell cultures. - Biotech. Progr. 4: 184-188, 1988. Go to original source...
  9. Deng, Z., Chu, J., Wang, Q., Wang, L.: Effect of different carbon sources on the accumulation of carbohydrate, nutrient absorption and the survival rate of Chinese Ash (Fraxinus mandshurica) explants in vitro. - Afr. J. agr. Res. 7: 3111-3119, 2012.
  10. Dey, S.: Cost-effective mass cloning of plants in liquid media using a novel Growtek bioreactor. - In: Hvoslef-Eide, A.K., Preil W. (ed.): Liquid Culture Systems for in Vitro Plant Propagation. Pp. 127-141. Springer, Dordrecht 2005.
  11. Diarra, S.T., He, J., Wang, J., Li J.: Ethylene treatment improves diosgenin accumulation in in vitro cultures of Dioscorea zingiberensis via up-regulation of CAS and HMGR gene expression. - Electron. J. Biotechn. 16: http://dx.doi.org/10.2225/vol16-issue5-fulltext-9, 2013. Go to original source...
  12. Dias, M.C., Pinto, G.C., Correia, M., Moutinho-Pereira, J., Silva, S., Santos, C.: Photosynthetic parameters of Ulmus minor plantlets affected by irradiance during acclimatization. - Biol. Plant. 57: 33-40, 2013. Go to original source...
  13. Fujiwara, K., Kozai, T., Watanabe, I.: Measurements of carbon dioxide gas concentration in stoppered vessels containing tissue cultured plantlets and estimates of net photosynthetic rate of the plantlets. - J. Agr. Meteorol. 43: 21-30, 1987. Go to original source...
  14. Gaspar, T., Franck, T., Bisbis, B., Kevers, C., Jouve, L., Haussman, J.F., Dommes, J.: Concepts in plant stress physiology. Application to plant tissue cultures. - Plant Growth Regul. 37: 263-285, 2002. Go to original source...
  15. Georgiev, M.I., Weber, J., Maciuk, A.: Bioprocessing of plant cell cultures for mass production of targeted compounds. - Appl. Microbiol. Biotechnol. 83: 809-823, 2009. Go to original source...
  16. Hakkim, F.L., Shankar, C.G., Girua, S.: Chemical composition and antioxidant property of holy basil (Ocimum sanctum L.) leaves, stems, and inflorescence and their in vitro callus cultures. - J. Agr. Food Chem. 55: 9109-9117, 2007. Go to original source...
  17. Hazarika, B.N.: Morpho-physiological disorders in in vitro culture of plants. - Sci. Hort. 108: 105-120, 2006. Go to original source...
  18. Ikemeyer, D., Barz, W.: Comparison of secondary product accumulation in photoautotrophic, photomixitrophic and heterotrophic Nicotiana tabacum cell suspension cultures. - Plant Cell Rep. 8: 479-482, 1989. Go to original source...
  19. Jackson, M. B.: Aeration stress in plant tissue cultures. - Bulg. J. Plant Physiol. 96(Special Issue): 109, 2003.
  20. Jayasinghe, C., Gotoh, N., Aoki, T., Wada, S.: Phenolics composition and antioxidant activity of sweet basil (Ocimum basilicum L.). - J. Agr. Food Chem. 51: 4442-4449, 2003. Go to original source...
  21. Juliani, H.R., Koroch, A.R., Simon, J.E.: Basil: A new source of rosmarinic acid - In: Ho, C.T., Simon, J.E., Shahidi, F., Shao, Y. (ed.): Dietary Supplements. Pp. 129-143. American Chemical Society, Washington 2008.
  22. Karuppusamy, S.: A review on trends in production of secondary metabolites from higher plants by in vitro tissue, organ and cell cultures. - J. med. Plants Res. 3: 1222-1239, 2009.
  23. Kiferle, C., Lucchesini, M., Mensuali-Sodi, A., Maggini, R., Raffaelli, A., Pardossi A.: Rosmarinic acid content in basil plants grown in vitro and in hydroponic. - Cent. Eur. J. Biol. 6: 946-957, 2011. Go to original source...
  24. Kiferle, C., Maggini, R., Pardossi, A. Influence of nitrogen nutrition on growth and accumulation of rosmarinic acid in sweet basil (Ocimum basilicum L.) grown in hydroponic culture. - Aust. J. Crop Sci. 7: 321-327, 2013.
  25. Kintzios, S., Kollias, H., Straitouris, E., Makri, O.: Scale-up micropropagation of sweet basil (Ocimum basilicum L.) in an airlift bioreactor and accumulation of rosmarinic acid. - Biotechnol. Lett. 26: 521-523, 2004. Go to original source...
  26. Kintzios, S., Makri, O., Panagiotopoulos, E., Scapeti, M.: In vitro rosmarinic acid accumulation in sweet basil (Ocimum basilicum L.). - Biotechnol. Lett. 25: 405-408, 2003. Go to original source...
  27. Kobayashi, Y., Fukui, H., Tabata, M.: Effect of carbon dioxide and ethylene on berberine production and cell browning in Thalictrum minus cell cultures. - Plant Cell Rep. 9: 496-499, 1991a. Go to original source...
  28. Kobayashi, Y., Hara, M., Fukui, H., Tabata, M.: The role of ethylene in berberine production by Thalictrum minus cell suspension cultures. - Phytochemistry 30: 3605-3609, 1991b. Go to original source...
  29. Kozai, T.: Autotrophic micropropagation. - In: Bajaj, Y.P.S. (ed.): Biotechnology in Agriculture and Forestry: High Tech and Micropropagation I. Pp. 313-343. Springer-Verlag, Berlin - Heidelberg 1991.
  30. Kozai, T., Kubota, C.: Concepts, definitions, ventilation methods, advantages and disadvantages. - In: Kozai T., Afreen, F., Zobayed, S.M.A. (ed.): Photoautotrophic (Sugar-Free Medium) Micropropagation as a New Propagation and Tansplant Production System. Pp. 9-30. Springer, Berlin 2005.
  31. Kozai, T., Kubota, C., Chun, C., Afreen, F., Ohyama, K.: Necessity and concept of the closed transplant production system. - In: Kubota, C., Chun, C. (ed.): Proceedings for the International Symposium on Transplant Production in Closed System. Pp. 3-19. Kluwer Academic Publisher, Dordrecht 2000.
  32. Lucchesini, M., Bertoli, A., Mensuali-Sodi, A., Pistelli, L.: Establishment of in vitro tissue cultures from Echinacea angustifolia D.C. adult plants for the production of phytochemical compounds. - Sci. Hort. 122: 484-490, 2009. Go to original source...
  33. Lucchesini, M., Mensuali-Sodi, A.: Plant tissue culture-an opportunity for the production of nutraceuticals. - In: Giardi, M.T., Rea, G., Berra, B. (ed.): Bio-Farms for Nutraceuticals: Functional Food and Safety Control by Biosensor. Pp: 185-200. Springer, New York 2010.
  34. Lucchesini, M., Mensuali-Sodi, A., Massai, R., Gucci, R.: Development of autotropy and tolerance to acclimatization of Myrtus communis transplant cultured in vitro under different aeration. - Biol. Plant. 44: 167-174, 2001. Go to original source...
  35. Lucchesini, M., Monteforti, G., Mensuali-Sodi, A., Serra, G.: Leaf ultrastructure, photosynthetic rate and growth of myrtle plantlets under different in vitro culture condictions. - Biol. Plant. 50: 161-168, 2006. Go to original source...
  36. Makri, O., Kintzios, S.: Ocimum sp. (basil): botany, cultivation, pharmaceutical properties, and biotechnology. - J. Herbs Spices med. Plants 13: 123-150, 2007.
  37. Martre, P., Lacan, D., Just, D., Teisson, C.: Physiological effects of temporary immersion on Hevea brasiliensis callus. - Plant Cell Tissue Organ Cult. 67: 25-35, 2001. Go to original source...
  38. Matkowski, A.: Plant in vitro for the production of antioxidants, a review. - Biotech. Adv. 26: 548-560, 2008. Go to original source...
  39. Mensuali-Sodi, A., Lucchesini, M., Maltinti, S., Serra, G., Tognoni, F.: Leaf senescence in tissue culture of Passiflora incarnata L.: the role of ethylene. - In: Ramina, A., Chang, C., Giovannoni, J., Klee, H., Perata, P., Woltering, E. (ed.): Advances in Plant Ethylene Research. Pp. 151-152. Springer, Dordrecht 2007.
  40. Mensuali-Sodi, A., Panizza, M., Tognoni, F.: Studies on lavandin callus cultures: ethylene production in relation to the growth. - Biol. Plant. 31: 247-253, 1989. Go to original source...
  41. Mensuali-Sodi, A., Panizza, M., Tognoni, F.: Quantification of ethylene losses in different container-seal systems and comparison of biotic and abiotic contributions to ethylene accumulation in cultured tissues. - Physiol. Plant. 84: 472-476, 1992. Go to original source...
  42. Mingozzi M, Morini S., Lucchesini M., Mensuali-Sodi A.: Effects of leaf soluble sugars content and net photosynthetic rate of quince donor shoots on subsequent morphogenesis in leaf explants. - Biol. Plant. 55: 237-242, 2011. Go to original source...
  43. Morgan, P.W., Drew, M.C.: Ethylene and plant responses to stress. - Physiol. Plant. 100: 620-630, 1997. Go to original source...
  44. Mosaleeyanon, K., Zobayed, S.M.A., Afreen, F., Kozai, T.: Relationships between net photosynthetic rate and secondary metabolite contents in St. John's wort. - Plant. Sci. 169: 523-537, 2005. Go to original source...
  45. Moschopoulou, G., Kintzios, S.: Achievement of thousand-fold accumulation of rosmarinic acid in immobilized cells of sweet basil (Ocimum basilicum L.) by ten-fold increase of the volume of the immobilization matrix. - J. biol. Res. Thessalon. 15: 59-65, 2011.
  46. Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures. - Physiol. Plant. 15: 473-493, 1962. Go to original source...
  47. Nguyen, P.M., Kwee, E.M., Niemeyer, E.D.: Potassium rate alters the antioxidant capacity and phenolic concentration of basil (Ocimum basilicum L.) leaves. - Food Chem. 23: 1235-1241, 2010. Go to original source...
  48. Panizza, M., Mensuali-Sodi, A., Tognoni, F.: Role of ethylene in axillary shoot proliferation of lavandin-interaction with benzyladenine and polyamines. - J. exp. Bot. 44: 387-394, 1993. Go to original source...
  49. Park, S.U., Uddin, M.R., Xu, H., Kim, Y.K., Lee, S.Y.: Biotechnological applications for rosmarinic acid production in plant. - Afr. J. Biotechnol. 7: 4959-4965, 2008.
  50. Patnaik, J., Sahoo, S., Debata, B.K.: Somaclonal variation in cell suspension culture-derived regenerants of Cymbopogon martini (Roxb.) Wats var. motia. - Plant Breed. 118: 351-354, 1999. Go to original source...
  51. Petersen, M., Simmonds, M.S.J.: Molecules of interest: rosmarinic acid. - Phytochemistry 62: 121-125, 2003. Go to original source...
  52. Pospíšilová, J., Haisel, D., Synková, H., Čatský, J., Wilhelmová, N., Plzáková, Š., Procházková, D., Šrámek, F.: Photosynthetic pigments and gas exchange during ex vitro acclimation of tobacco plants as affected by CO2 supply and abscisic acid. - Plant Cell Tissue Organ Cult. 61: 125-133, 2000.
  53. Rady, M.R., Nazif, N.M.: Rosmarinic acid content and RAPD analysis of in vitro regenerated basil (Ocimum basilicum L.) plants. - Fitoterapia 6: 525-533, 2005. Go to original source...
  54. Saher, S., Piqueras, A., Hellin, E., Olmos, E.: Hyperhydricity in micropropagated carnation shoots: the role of oxidative stress. - Physiol. Plant. 120: 152-161, 2004. Go to original source...
  55. Sahoo, Y., Patfnaik, S.K., Chand, P.K.: In vitro propagation of an aromatic medicinal herb Ocimum basilicum L. (sweet basil) by axillary shoot proliferation. - In vitro cell. dev. Biol. Plant. 33: 293-296, 1997. Go to original source...
  56. Savio, L.E. B., Astarita, L.V., Santarém, E.R.: Secondary metabolism in micropropagated Hypericum perforatum L. grown in non-aerated liquid medium. - Plant Cell Tissue Organ Cult. 108: 465-472, 2012. Go to original source...
  57. Steyn, W.J., Wand, S.J.E., Holcroft, D.M., Jacobs, G.: Anthocyanins in vegetative tissues: a proposed unified function in photoprotection. - New Phytol. 55: 349-361, 2002. Go to original source...
  58. Xiao, Y., Niu, G., Kozai, T.: Development and application of photoautotrophic micropropagation plant system. - Plant Cell Tissue Organ Cult. 105: 149-158, 2011. Go to original source...
  59. Yasmin, S., Mensuali-Sodi, A., Perata, P., Pucciariello, C.: Ethylene influences in vitro regeneration frequency in the FR13A rice harbouring the SUB1A gene. - Plant Growth Regul. 72: 97-103, 2014. Go to original source...
  60. Zhao, J., Davis, L.C., Verpoorte, R.: Elicitor signal transduction leading to production of plant secondary metabolites. - Biotechnol. Adv. 23: 283-333, 2005. Go to original source...
  61. Ziv, M.: Bioreactor technoloy for plant micropropagation. - Hort. Rev. 24: 1-30, 2000.
  62. Zobayed, S.M.A., Saxena, P.K.: Production of St. John's wort plants under controlled environment for maximizing biomass and secondary metabolites. - In vitro cell. dev. Biol. Plant. 40: 108-114, 2004. Go to original source...