Abstract
Lingonberry (Vaccinium vitis-idaea L. ssp. vitis-idaea Britton) cultivars Regal, Splendor, and Erntedank were obtained by conventional softwood cuttings (taken as a control), by in vitro shoot proliferation of node explants, and by adventitious shoot regeneration from excised leaves of micropropagated shoots. In the plants propagated in vitro, the total ascorbate content increased and its pool was more oxidized, the total glutathione content also increased but its pool became more reduced. The leaves of plants obtained from the in vitro culture showed significantly higher antioxidant enzyme activities except for dehydroascorbate reductase which was at a similar level in all plants. Total soluble phenolics, tannins, and flavonoids were enhanced in fruits of in vitro-propagated plants whereas in leaves, the levels of these metabolites (except flavonoids) were higher in ex vitro derived plants. The total radical scavenging capacity was enhanced in berries of the in vitro propagated plants. It is suggested that the active morphogenetic process, characterized by intensive formation and scavenging reactive oxygen species is reflected in the activities of antioxidant enzymes and metabolites. The reduction potential of glutathione is the most important parameter which determines patterns of growth and differentiation in the investigated plants.
Similar content being viewed by others
Abbreviations
- AFR:
-
ascorbate free radical
- APX:
-
ascorbate peroxidase
- AsA:
-
ascorbic acid
- CE:
-
catechin equivalent
- DHA:
-
dehydroascorbate
- DHAR:
-
dehydroascorbate reductase
- DPPH:
-
2,2-diphenyl-1-picrylhydrazyl
- GAE:
-
gallic acid equivalent
- GR:
-
glutathione reductase
- GSH:
-
reduced glutathione
- GSSG:
-
oxidized glutathione
- LC:
-
plants propagated from leaf cultures
- MDA:
-
malondialdehyde
- MDHAR:
-
monodehydroascorbate reductase
- NC:
-
plants propagated from node cultures
- SC:
-
softwood cutting-derived plants
References
Aebi, H. Catalase. — In: Bergmeyer, H.U. (ed.): Methods of Enzymatic Analysis. Vol. 2. Pp. 673–684. Academic Press, NewYork 1974.
Anttonen, J.M., Karajalainen, O.R.: High-performance liquid chromatography analysis of black currant (Ribes nigrum L.) fruit phenolics grown either conventionally or organically. — J. Agr. Food Chem. 54: 7530–7538, 2006.
Arnon, D.I.: Copper enzymes in isolated chloroplasts. Polyphenol oxidase in Beta vulgaris. — Plant Physiol. 24: 1–15, 1949.
Brand-Williams, W., Cuvelier, M.E., Berset, C.: Use of a free radical method to evaluate antioxidant activity. — Lebensm. Wiss. Technol. 28: 25–30, 1995.
Cárdenas, L.: New findings in the mechanisms regulating polar growth in root hair cells. — Plant Signal. Behav. 4: 4–8, 2009.
Carol, R.J., Dolan, L.: The role of reactive oxygen species in cell growth: lessons from root hairs. — J. exp. Bot. 57: 1829–1834, 2006.
Chandrasekara, N., Shahidi, F.: Effect of roasting on phenolic content and antioxidant activities of whole cashew nuts, kernels, and testa. — J. Agr. Food. Chem. 59: 5006–5014, 2011.
Debnath, S.C.: Bioreactors and molecular analysis in berry crop micropropagation. — Can. J. Plant Sci. 91: 147–157, 2011.
Debnath, S.C.: Influence of propagation method and indole-3-butyric acid on growth and development of in vitro- and ex vitro-derived lingonberry plants. — Can. J. Plant Sci. 86: 235–243, 2006.
Debnath, S.C.: Morphological development of lingonberry as affected by in vitro and ex vitro culture methods and source propagule. — HortScience 40: 760–763, 2005a
Debnath, S.C.: Effects of carbon source and concentration on development of lingonberry (Vaccinium vitis-idaea L.) shoots cultivated in vitro from nodal explants. — In Vitro cell. dev. Biol. Plant 41: 145–150, 2005b.
Debnath, S.C.: Micropropagation of lingonberry: Influence of genotype, explant orientation, and overcoming TDZinduced inhibition of shoot elongation using zeatin. — HortScience 40: 185–188, 2005c.
Debnath, S.C., McRae, K.B.: An efficient adventitious shoot regeneration system on excised leaves of micropropagated lingonberry (Vaccinium vitis-idaea L.). — J. hort. Sci. Biotechnol. 77: 744–752, 2002.
Diaz-Vivancos, P., Dong, Y.P., Ziegler, K., Markovic, J., Pallardó, F., Pellny, T.K., Verrier, P., Foyer, C.H.: Recruitment of glutathione into the nucleus during cell proliferation adjusts whole cell redox homeostasis in Arabidopsis thaliana and lowers the oxidative defence shield. — Plant J. 64: 825–838, 2010.
Ek, S., Kartimo, H., Mattila, S., Tolonne, A.: Characterization of phenolic compounds from lingonberry (Vaccinium vitis-idaea). — J. Agr. Food Chem. 54: 9834–9842, 2006.
Foley, S.L., Debnath, S.C.: Influence of in vitro and ex vitro propagation on anthocyanin content and antioxidant activity of lingonberries. — J. hort. Sci. Biotechnol. 82: 114–118, 2007.
Fiorani, F., Umbach, A.L., Siedow, J.N.: The alternative oxidase of plant mitochondria is involved in the acclimation of shoot growth at low temperature. A study of Arabidopsis AOX1a transgenic plants. — Plant Physiol. 139: 1795–1805, 2005.
Foreman, J., Demidchik, V., Bothwell, J.H., Mylona, P., Miedema, H., Torres, M.A., Linstead, P., Costa, S., Brownlee, C., Jones, J.D., Davies, J.M., Dolan, L.: Reactive oxygen species produced by NADPH oxidase regulate plant cell growth. — Nature 422: 442–446, 2003.
Frederico, A.M., Campos, M.D., Cardoso, H.G., Imani, J., Arnholdt-Schmitt, B.: Alternative oxidase involvement in Daucus carota somatic embryogenesis. — Physiol. Plant. 137: 498–508, 2009.
Gupta, S.D., Datta, S.: Antioxidant enzyme activities during in vitro morphogenesis of gladiolus and the effect of application of antioxidants on plant regeneration. — Biol. Plant. 47: 179–183, 2003.
Hilal, M., Castagnaro, A., Moreno, H., Massa, E.M.: Specific localization of the respiratory alternative oxidase in meristematic and xylematic tissues from developing soybean roots and hypocotyls. — Plant Physiol. 115: 1499–1503, 1997.
Horemans, N., Foyer, C.H., Asard, H.: Transport and action of ascorbate at the plant plasma membrane. — Trends Plant Sci. 5: 263–267, 2000.
Igamberdiev, A.U., Bykova, N.V., Hill, R.D.: Structural and functional properties of class 1 plant hemoglobins. — IUBMB Life 63: 146–152, 2011.
Igamberdiev, A.U., Stoimenova, M., Seregélyes, C., Hill, R.D.: Class-1 hemoglobin and antioxidant metabolism in alfalfa roots. — Planta 223: 1041–1046, 2006.
Jaakola, L., Tolvanen, A., Laine, K., Hohtola, A.: Effect of N-6-isopentenyladenine concentration on growth initiation in vitro and rooting of bilberry and lingonberry microshoots. — Plant Cell Tissue Organ Cult. 66: 73–77, 2001.
Jana, S., Shekhawat, G.S.: In vitro regeneration of Anethum graveolens, antioxidative enzymes during organogenesis and RAPD analysis for clonal fidelity. — Biol. Plant. 56: 9–14, 2012.
Jiang, K., Feldman, L.: Positioning of the auxin maximum affects the character of cells occupying the root stem cell niche. — Plant Signal. Behav. 5: 1–3, 2010.
Kampfenkel, K., Montagu, M.V., Inzé, D.: Extraction and determination of ascorbate and dehydroascorbate from plant tissue. — Anal. Biochem. 225: 165–167, 1995.
Kranner, I., Birtic, S., Anderson, K.M., Pritchard, H.W.: Glutathione half-cell reduction potential: a universal stress marker and modulator of programmed cell death. — Free Radicals Biol. Med. 40: 2155–2165, 2006.
Lätti, A.K., Riihinen, K.R., Jaakola, L.: Phenolic compounds in berries and flowers of a natural hybrid between bilberry and lingonberry (Vaccinium × intermedium Ruthe). — Phytochemistry 72: 810–815, 2011.
McNulty, A.K., Cummins, W.R., Pellizzari, A.: A field survey of respiration rates in leaves of arctic plants. — Arctic 41: 1–5, 1988.
Mitrovic, A., Janosevic, D., Budimir, S., Pristov, J.B.: Changes in antioxidative enzymes activities during Tacitus bellus direct shoot organogenesis. — Biol. Plant. 56: 357–361, 2012.
Murshed, R., Lopez-Lauri, F., Sallanon, H.: Microplate quantification of enzymes of the plant ascorbate-glutathione cycle. — Anal. Biochem. 383: 320–322, 2008.
Mülle, K., Linkies, A., Vreeburg, R.A., Fry, S.C., Krieger-Liszkay, A., Leubner-Metzger, G.: In vivo cell wall loosening by hydroxyl radicals during cress seed germination and elongation growth. — Plant Physiol. 150: 1855–1865, 2009.
Mullinieaux, P.M., Rausch, T.: Glutathione, photosynthesis and the redox regulation of stress-responsive gene expression. — Photosynth. Res. 86: 459–474, 2005.
Meyer, A.J.: Glutathione homeostasis and redox signalling. — J. Plant Physiol. 165: 1390–1403, 2008.
Noctor, G., Foyer, C.: Ascorbate and glutathione: keeping active oxygen under control. — Annu. Rev. Plant Physiol. Plant mol. Biol. 49: 249–279, 1998.
Noctor, G., Mhamdi, A., Chaouch, S., Han, Y., Neukermans, J., Marquez-Garcia, B., Queval, G., Foyer, C.H.: Glutathione in plants: an integrated overview. — Plant Cell Environ. 35: 454–484, 2012.
Saez, P.L., Bravo, L.A., Saez, K.L., Sanchez-Olate, M., Latsague, M.I., Rios, D.G.: Photosynthetic and leaf anatomical characteristics of Castanea sativa: a comparison between in vitro and nursery plants. — Biol. Plant. 56: 15–24, 2012.
Sanchez-Fernandez, R., Fricker, M., Corben, L., White, N.S., Sheard, N., Leaver, C.J., Van Montagu, M., Inze, D., May, M.J.: Cell proliferation and hair tip growth in the Arabidopsis root under mechanistically different forms of redox control. — Proc. nat. Acad. Sci. USA 94: 2745–2750, 1997.
Schafer, F.Q., Buettner, G.R.: Redox environment of the cell as viewed through the redox state of the glutathione disulfide/glutathione couple. — Free Radicals Biol. Med. 30: 1191–1212, 2001.
Shekhawat, G.S., Verma, K., Jana, S., Singh, K., Teotia, P., Prasad, A.: In vitro biochemical evaluation of cadmium tolerance mechanism in callus and seedlings of Brassica juncea. — Protoplasma 239: 31–38, 2010.
Stasolla, C., Belmonte, M.F., Van Zyl, L., Craig, D.L., Liu, W., Yeung, E.C., Sederoff, R.: The effect of reduced glutathione on morphology and gene expression of white spruce (Picea glauca) somatic embryos. — J. exp. Bot. 55: 695–709, 2004.
Stasolla, C., Yeung, E.C.: Exogenous applications of ascorbic acid induce shoot apical meristem growth in germinating white spruce (Picea glauca) somatic embryos. — Int. J. Plant Sci. 167: 429–436, 2006.
Talukdar, D.: Ascorbate deficient semi-dwarf asfL1 mutant of Lathyrus sativus exhibits alterations in antioxidant defense. — Biol. Plant. 56: 675–682, 2012.
Thomas, C.E., McLean, L.R., Parker, R.A., Ohlweiler, D.F.: Ascorbate and phenolic antioxidant interactions in prevention of liposomal oxidation. — Lipids 27: 543–550, 1992.
Uhrig, J.F., Hülskamp, M.: Plant GTPases: regulation of morphogenesis by ROPs and ROS. — Curr Biol. 16: R211–213, 2006.
Vatankhah, E., Niknam, V., Ebrahimzadeh, H.: Activity of antioxidant enzymes during in vitro organogenesis in Crocus sativus. — Biol. Plant. 54: 509–514, 2010.
Verma, K., Shekhawat, G.S., Sharma, A., Mehta, S.K., Sharma, V.: Cadmium induced oxidative stress and changes in soluble and ionically bound cell wall peroxidase activities in roots of seedling and 3–4 leaf stage plants of Brassica juncea (L.) Czern. — Plant Cell Rep. 27: 1261–1269, 2008.
Vernoux, T., Wilson, R.C., Seeley, K.A., Reichheld, J.P., Muroy, S., Brown, S., Maughan, S.C., Cobbett, C.S., Van Montagu, M., Inzé, D., May, M.J., Sung, Z.R.: The root MERISTEMLESS1/CADMIUM SENSITIVE2 gene defines a glutathione-dependent pathway involved in initiation and maintenance of cell division during postembryonic root development. — Plant Cell 12: 97–110, 2000.
Wang, S.Y., Feng, R,. Bowman, L., Penhallegon, R., Ding, M., Lu, Y.: Antioxidant activity in lingonberries (Vaccinium vitis-idaea L.) and its inhibitory effect on activator protein-1, nuclear factor-kappa B, and mitogen-activated protein kinases activation. — J. Agr. Food Chem. 53: 3156–3166, 2005.
Zaharieva, T.B., Abadía, J.: Iron deficiency enhances the levels of ascorbate, glutathione, and related enzymes in sugar beet roots. — Protoplasma 221: 269–275, 2003.
Zechmann, B., Koffler, B.E., Russell, S.D.: Glutathione synthesis is essential for pollen germination in vitro. — BMC Plant Biol. 11: 54, 2011.
Zhishen, J., Mengcheng, T., Jianming, W.: The determination of flavonoid contents in mulberry and their scavenging effect on superoxide radicals. — Food Chem. 64: 555–559, 1999.
Author information
Authors and Affiliations
Corresponding author
Additional information
Acknowledgements: This work was supported by the Natural Sciences and Engineering Research Council of Canada. The authors thank Neel Chandrasekara, Sarah Leonard, Glenn Chubbs, and Darryl Martin for their excellent technical help.
Rights and permissions
About this article
Cite this article
Vyas, P., Debnath, S.C. & Igamberdiev, A.U. Metabolism of glutathione and ascorbate in lingonberry cultivars during in vitro and ex vitro propagation. Biol Plant 57, 603–612 (2013). https://doi.org/10.1007/s10535-013-0339-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10535-013-0339-8