Optimization of Callus Induction and Cell Suspension in Catharanthus roseus

Document Type : Research Paper

Authors

Abstract

Periwinkle (Catharanthus roseus) has been considered as an important medicinal plant because it contains many alkaloids such as vincristine and vinblastine. In vitro culture of periwinkle provides new tissue sources such as callus, cell suspension and seedlings to produce secondary metabolites. As for the mass production of callus and cell suspension in order to optimize the extraction of secondary metabolites have been few studies, the present study describes two callus production optimization procedures and one cell suspension experiment. The first trail was a factorial experiment with three explants (roots, shoots and leaves) on Murashige and Skoog (MS) medium supplemented with different concentrations of BAP and 2,4-D. The second trial was a factorial experiment with six explants (roots, shoots, in-vitro grown leaves, ex-vitro grown leaves, seeds and nodes) and ten different hormonal combinations. The cell suspension experiment was conducted with six hormonal and one control (without hormone) treatments based on completely randomized design. Comparison of means showed that the maximum callus production was obtained from leaf explants, the root and shoot explants were in the second orders. In overall, 0.5 mg /l BAP + 1 mg/l 2,4-D and 0.5 mg /l KN + 1 mg/l 2,4-D concentrations proved to be optimal for the production of maximum callus. The best combination for suspension culture was 1 mg/l 2,4-D and/or 2 mg/l 2,4-D+1 mg /l BAP according to the dried cell weight

Keywords


periwinkle, callus. Suspension culture, In vitro

Chen LJ, Zhu XY, Gu L, Wu J (2005). Efficient callus induction and plant regeneration from anther of Chinese narcissus Narcissus tazetta L. var. chinensis Roem. Plant Cell Reports 24: 401-407.
Dutta A, Singh S, Kumar S, Sen J (2007). Transcript profiling of terpenoid indole alkaloid pathway genes and regulators reveals strong expression of repressors in Catharanthus roseus cell cultures. Plant Cell Reports 26: 907-915.
Gurel S, Gurel E, Kaya Z (2001). Callus development and indirect shoot regeneration from seedling of sugar beet (Beta vulgaris L.) cultured in vitro. Turkish Journal of Botany 25: 25-33.
Hussain Z, Haroon M, Bano R, Rashid H, Chaudhry Z (2010). Protocol optimization for efficient callus induction and regeneration in three Pakistani rice cultivars. Pakistan Journal of Botany 42: 879-887.
Kwon TH, Abe T, Sasahara T (1993). Efficient callus induction and plant regeneration in Sesamum species. Plant Tissue Culture Letters 10: 260-266.
Lee E, Mobin M, Hahn EJ, Paek KY (2006). Effects of sucrose, [noculum density, auxins, and aeration volume on cell growth of Gymnema sylvestre. Journal of Plant Biology 49: 427-431.
Mujib A, Bandhyopadhyay S, Ghosh PD (2000). Tissue culture derived plantlet variation in Caladium an important ornamental. Plant Cell, Tissue and Organ Culture 10: 149-155.
Mungole A, Awati R, Dey S, Chaturvedi A, Zanwar P (2009). In-vitro callus induction and shoot regeneration in Ipomoea obscura (L.): potent Indian medicinal plant. Indian Journal of Science and Technology 2: 24-26.
Ozgen M, Türet M, Altinok S, Sancak C (1998). Efficient callus induction and plant regeneration from mature embryo culture of winter wheat (Triticum aestivum L.) genotypes. Plant Cell Reports 18: 331-335.
Phillips GC, Hubstenberger JF, Hansen EE (1995). Plant regeneration by organogenesis from callus and cell suspension cultures. In: Gamborg OL, Phillips GC (eds), Plant Cell, Tissue and Organ Culture, pp. 67-78. Heidelberg: Springer and Verla.
Pietrosiuk A, Furmanowa M, Lata B (2007). Catharanthus roseus: micropropagation and in vitro techniques. Phytochem Reviews 6: 459-473.
Piri Kh, Nazarian F (2001). Plant Tissue Culture. Bu Ali Sina Press.
Sato F, Hashimoto T, Hachiya A (2001). Metabolic engineering of plant alkaloid biosynthesis. Proceedings of the National Academy of Sciences of the United States of America. 98: 367-372.
Senoussi MM, Nora B, Joe C (2009). Impact of hypoxia on the growth and alkaloid accumulation in Catharanthus roseus cell suspension. Acta Physiologiae Plantarum 31: 359–362.
Shirin F, Hossain M, Kabir MF, Roy M, Sarker SR (2007). Callus induction and plant regeneration from intermodal and leaf explants of four potato (Solanum tuberosum L.) cultivars. World Journal of Agricultural Sciences 3: 1-6.
Taha HS, El-Bahr MK, Seif-El-Nasr MM (2008). In vitro studies on egyptian Catharanthus roseus (L.) G. Don.: 1- calli Production, direct shootlets Regeneration and alkaloids determination. Journal of Applied Sciences Research 4: 1017-1022.
Valluri JV (2009). Bioreactor production of secondary metabolites from cell cultures of Periwinkle and Sandalwood. Methods in Molecular Biology 547: 325-335.
Verpoorte R, van der Heijden R, Van Gulik W (1991). Plant biotechnology for the production of alkaloids: Present status and prospects. In: Brossi A, (ed(. The Alkaloids. Vol 40. New York: Academic Press.
WaiLeng L, LaiKeng C (2003). Establishment of Orthosiphon stamineus cell suspension culture for cell growth. Plant Cell, Tissue and Organ Culture 78: 101-106.
Waileng LW, LaiKeng C (2004). Plant regeneration from stem nodal segments of Orthosiphon stamineus banth., a medicinal plant with diuretic activity. In vitro Cellular and Developmental Biology - Plant 40: 115-118.
Xu M, Dong J (2005). Nitric oxide stimulates indole alkaloid production in Catharanthus roseus cell suspension cultures through a protein kinase-dependent signal pathway. Enzyme and Microbial Technology 37: 49-53.
Yokoyama M, Inomata S (1998). Catharanthus roseus (periwinkle): in vitro culture and high level production of Arbutine by biotransformation. Biotechnology in Agriculture and Forestry 41: 67-80.
Zhao J, Zhu WH, Hu Q, Guo YQ (2001). Compact callus cluster suspension culture of Catharanthus roseus with enhanced indol alkaloid biosynthesis. In Vitro Cellular and Developmental Biology - Plant 37: 68-72.
Zhao J, Zhu WH, Hu Q (2001). Effects of light and plant growth regulators on the biosynthesis of vindoline and other indole alkaloids in Catharanthus roseus callus cultures. Plant Growth Regulation 33: 43-49.