Medium composition potentially regulates the anthocyanin production from suspension culture of Daucus carota
In the present study, an effort has been made to optimize various culture conditions for enhanced production of anthocyanin. Nutrient content of MS medium (ammonium to potassium nitrate ratio and phosphate concentration) had a profound influence on the cell biomass and anthocyanin accumulation in cell suspension cultures of Daucus carota. Suspension cultures were carried out in shake flasks for 18 days and examined for cell growth, anthocyanin synthesis, anthocyanin yield and development of pigmented cells in relation to the uptake of total sugar, extracellular phosphate, nitrate and ammonia. The addition of NH4NO3 to KNO3 ratio (20.0 mM: 37.6 mM) in the suspension culture media resulted in a 2.85-fold increase in anthocyanin content at day 3. Similarly, a lower concentration of KH2PO4 (0.45 mM) in the MS medium resulted in 1.63-fold increase in anthocyanin content at day 9. The total sugar uptake was closely associated with a significant increase in anthocyanin accumulation. Total sugar and nitrate were consumed until 9–12 days, while ammonia and phosphate were completely consumed within 3 days after inoculation. After 9 days, cell lysis was observed and resulted in the leakage of intracellular substances. These observations suggest that anthocyanin was synthesized only by viable pigmented cells and degraded rapidly after cell death and lysis. This study signifies the utility of D. carota suspension culture for further up-scaling studies of anthocyanin.
KeywordsAmmonium nitrate Potassium nitrate Phosphate Anthocyanin Nutrient Suspension culture
The authors are thankful to the Council of Scientific and Industrial Research, New Delhi and Central Food Technological Research Institute, Mysuru (India) for financial assistance. Encouragement by Director CSIR-CFTRI, Mysuru, is also gratefully acknowledged.
Compliance with ethical standards
Conflict of interest
The authors declare that there is no conflict of interest.
- Bongue-Bartelsman M, Phillips DA (1995) Nitrogen stress regulates gene expression of enzymes in the flavonoid biosynthetic pathway of tomato. Plant Physiol Biochem 33:539–546Google Scholar
- Chen Y-Q, Yi F, Cai M, Luo J-X (2003) Effects of amino acids, nitrate, and ammonium on the growth and taxol production in cell cultures of Taxus yunnanensis. Plant Growth Regul 41:265–268. https://doi.org/10.1023/B:GROW.0000007502.72108.e3 CrossRefGoogle Scholar
- Coste A, Vlase L, Halmagyi A et al (2011) Effects of plant growth regulators and elicitors on production of secondary metabolites in shoot cultures of Hypericum hirsutum and Hypericum maculatum. Plant Cell Tissue Organ Cult 106:279–288. https://doi.org/10.1007/s11240-011-9919-5 CrossRefGoogle Scholar
- Dedaldechamp F, Uhel C, Macheix JJ (1995) Enhancement of anthocyanin synthesis and dihydroflavonol reductase (DFR) activity in response to phosphate deprivation in grape cell suspensions. Phytochemistry 40(5):1357–1360Google Scholar
- Enaksha RM, Arteca RN (1994) Taxus cell suspension cultures: optimizing growth and production of taxol. J Plant Physiol 144(2):183–188Google Scholar
- Ghosh D, Konishi T (2007) Anthocyanins and anthocyanin-rich extracts: role in diabetes and eye function. Asia Pac J Clin Nutr 16(2):200–208Google Scholar
- Morandi D, Le Quere JL (1991) Influence of nitrogen on accumulation of isosojagol (a newly detected coumestan in soybean) and associated isoflavonoids in roots and nodules of mycorrhizal and non-mycorrhizal soybean. New Phytol 117:75–79. https://doi.org/10.1111/j.1469-8137.1991.tb00946.x CrossRefGoogle Scholar
- Mulabagal V, Tsay HS (2004) Plant cell cultures-an alternative and efficient source for the production of biologically important secondary metabolites. Int J Appl Sci Eng 2(1):29–48Google Scholar
- Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15(3):473–497Google Scholar
- Pallavi R, Elakkiya S, Tennety SSR, Devi PS (2012) Anthocyanin analysis and its anticancer property from sugarcane (Saccharum officinarum L.) peel. IJRPC 2:338–345Google Scholar
- Takahashi A, Takeda K, Ohnishi T (1991) Light-induced anthocyanin reduces the extent of damage to DNA in UV-irradiated Centaurea cyanus cells in culture. Plant Cell Physiol 32:541–547. https://doi.org/10.1093/oxfordjournals.pcp.a078113 Google Scholar
- Yamakawa T, Kato S, Ishida K, Kodama T, Minoda Y (1983) Production of anthocyanins by Vitis cells in suspension culture. Agric Biol Chem 47(10):2185–2191Google Scholar
- Yamamoto Y, Kinoshita Y, Watanabe S, Yamada Y (1989) Anthocyanin production in suspension cultures of high-producing cells of Euphorbia millii. Agric Biol Chem 53(2):417–423Google Scholar
- Zhang YH, Zhong JJ, Yu JT (1996) Effect of nitrogen source on cell growth and production of ginseng saponin and polysaccharide in suspension cultures of Panax notoginseng. Biotechnol Prog 12(4):567–571Google Scholar