Skip to main content
×
×
Home

Cathodic amelioration of the adverse effects of oxidative stress accompanying procedures necessary for cryopreservation of embryonic axes of recalcitrant-seeded species

  • Patricia Berjak (a1), Sershen (a1), Boby Varghese (a1) and N.W. Pammenter (a1)
Abstract

Several of the procedures necessary for cryopreservation of excised zygotic embryonic axes are known to be accompanied by emission of damaging levels of reactive oxygen species (ROS). These have been shown to be associated with shoot apical meristem necrosis, curtailing subsequent axis development to root production only, particularly for tropical/subtropical recalcitrant-seeded species. Here we report on the application of the principles of electrochemistry in the generation of strongly reducing, high pH cathodic water, by electrolysis of a solution containing calcium and magnesium chloride. The cathodic water in which Strychnos gerrardii axes were immersed for 30 min following dehydration, and importantly, after dehydration followed by cryopreservation, was shown to have strongly antioxidative properties in counteracting the damaging effects of ROS bursts and promoting shoot development. In a parallel experiment, axes of Boophane disticha exhibited enhanced total antioxidant activity when exposed to cathodic water both immediately following excision, and after flash-drying. For both species, the efficacious effects of cathodic treatment were manifest after the axes had been in culture for 4 h, suggesting that ROS were not quenched at source, but probably counteracted by enhancement of activity of endogenous antioxidants. Cathodic water therefore affords a non-toxic means of amelioration of oxidative, stress-related damage, which, coupled with the strongly fungicidal activity of the acidic, anionic water fraction, offers significant, and apparently non-injurious, advances towards successful cryopreservation of germplasm – and probably generally improved success of in-vitro-based procedures for plant tissues.

Copyright
Corresponding author
*Correspondence Email: berjak@ukzn.ac.za
References
Hide All
Antunes, F. and Cadenas, E. (2000) Estimation of H2O2 gradients across biomembranes. FEBS Letters 475, 121126.
Bailey-Serres, J. and Mittler, R.(Guest and Monitoring Editors) (2006) The roles of reactive oxygen species in plants. Plant Physiology 141, 311.
Bailly, C. (2004) Active oxygen species and antioxidants in seed biology. Seed Science Research 14, 93107.
Benson, E.E. and Bremner, D. (2004) Oxidative stress in the frozen plant: a free radical point of view. pp. 205241in Fuller, B.J.; Lane, N.; Benson, E.E. (Eds) Life in the frozen state. Boca Raton, CRC Press.
Benson, E.E. and Noronha-Dutra, A.A. (1988) Chemiluminescence in cryopreserved plant tissue cultures: the possible role of singlet oxygen in cryo injury. CryoLetters 9, 120131.
Benson, E.E., Lynch, P.T. and Jones, J. (1995) The use of the iron chelating agent desferrioxamine in rice cell cryopreservation: a novel approach for improving recovery. Plant Science 110, 249258.
Benson, E.E., Harding, K. and Johnston, J. (2007) Cryopreservation of shoot-tips and meristems. pp. 163184in Day, J.G.; Stacey, G. (Eds) Methods in molecular biology, Vol. 368, Cryopreservation and freeze drying protocols. Totowa, NJ, Humana Press.
Berjak, P. (1978) Viability extension and improvement of stored seeds. South African Journal of Science 74, 365368.
Berjak, P. and Pammenter, N.W. (2008) From Avicennia to Zizania: seed recalcitrance in perspective. Annals of Botany 101, 213228.
Berjak, P., Farrant, J.M., Mycock, D.J. and Pammenter, N.W. (1990) Recalcitrant (homoiohydrous) seeds: the enigma of their desiccation sensitivity. Seed Science and Technology 18, 297310.
Bhattacharjee, S. (2005) Reactive oxygen species and oxidative burst: roles in stress, senescence and signal transduction in plants. Current Science 89, 11131121.
Bolwell, G.P., Bindschedler, L.V., Blee, K.A., Butt, V.S., Davies, D.R., Gardner, S.L., Gerrish, C. and Minibayeva, F. (2002) The apoplastic oxidative burst in response to biotic stress in plants: a three-component system. Journal of Experimental Botany 53, 13671376.
Cassandra, , Benson, E.E., Berjak, P., Goveia, M. and Pammenter, N.W. (2011) Exploring the use of DMSO and ascorbic acid to promote shoot development by excised embryonic axes of recalcitrant seeds. CryoLetters (in press).
Cheng, H.-Y. and Song, S.-Q. (2008) Possible involvement of reactive oxygen species scavenging enzymes in desiccation sensitivity of Antiaris toxicaria seeds and axes. Journal of Integrative Plant Biology 50, 15491556.
Côme, D. and Corbineau, F. (1996) Metabolic damage related to desiccation sensitivity. pp. 8397in Ouédraogo, A.-S.; Poulsen, K.; Stubsgaard, F. (Eds) Intermediate/recalcitrant tropical forest tree seeds. Rome, IPGRI.
Dickie, J.B. and Pritchard, H.W. (2002) Systematic and evolutionary aspects of desiccation tolerance in seeds. pp. 239259in Black, M.; Pritchard, H.W. (Eds) Desiccation and survival in plants. Drying without dying. Wallingford, Oxon, CABI Publishing.
Foyer, C.H. and Noctor, G. (2005) Redox homeostasis and antioxidant signalling: a metabolic interface between stress perception and physiological responses. Plant Cell 17, 18661875.
Gill, S.S. and Tuteja, N. (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry 48, 909930.
Goveia, M.J.T. (2007) The effect of developmental status and excision injury on the success of cryopreservation of germplasm from non-orthodox seeds. MSc thesis, University of KwaZulu-Natal (Westville Campus), Durban, South Africa. Available at websitehttp://researchspace.ukzn.ac.za/xmlui/handle/10413/1052 (accessed March 2011).
Goveia, M., Kioko, J.I. and Berjak, P. (2004) Developmental status is a critical factor in the selection of excised recalcitrant axes as explants for cryopreservation. Seed Science Research 14, 241248.
Gutiérrez, R., Alvarado, J.L., Presno, M., Pérez-Veyna, O., Serrano, C.J. and Yahuaca, P. (2010) Oxidative stress modulation by Rosmarinus officinalis in CCl4-induced liver cirrhosis. Phytotherapy Research 24, 595601.
Hajari, E., Berjak, P., Pammenter, N.W. and Watt, M.P. (2009) Micropropagation of Ekebergia capensis Sparrm. using in vitro nodal segments and roots. Journal of Horticultural Science and Biotechnology 84, 8791.
Halliwell, B. (2006) Reactive oxygen species and antioxidants. Redox biology is a fundamental theme of aerobic life. Plant Physiology 141, 312322.
Halliwell, B. and Gutteridge, J.M.C. (1999) Free radicals in biology and medicine (3rd edition). Oxford, UK, Oxford University Press.
Hanaoka, K. (2001) Antioxidant effects of reduced water produced by electrolysis of sodium chloride solutions. Journal of Applied Electrochemistry 31, 13071313.
Hanaoka, K., Sun, D., Lawrence, R., Kamitani, Y. and Fernandes, G. (2004) The mechanism of enhanced antioxidant effects against superoxide anion radicals of reduced water produced by electrolysis. Biophysical Chemistry 107, 7182.
Hazra, B., Sarkar, R., Mandal, S., Biswas, S. and Mandal, N. (2009) Studies on antioxidant and antiradical activities of Dolichus biflorus seed extract. African Journal of Biotechnology 8, 39273933.
Hendry, G.A.F. (1993) Oxygen, free radical processes and seed longevity. Seed Science Research 3, 141153.
Hiraoka, A., Takemoto, M., Suzuki, T., Shinohara, A., Chiba, M., Shirao, M. and Yoshimura, Y. (2004) Studies on the properties and real existence of aqueous solution systems that are assumed to have antioxidant activities by the action of ‘active hydrogen’. Journal of Health Science 50, 456465.
Huang, Y.-R., Hung, Y.-C., Hsu, S.-Y., Huang, Y.-W. and Hwang, D.-F. (2008) Application of electrolyzed water in the food industry. Food Control 19, 329345.
Johnston, J.W., Dussert, S., Gale, S., Nadarajan, J., Harding, K. and Benson, E.E. (2006) Optimisation of the azinobis-3-ethyl-benzothiazoline-6-sulphonic acid radical scavenging assay for physiological studies of total antioxidant activity in woody plant germplasm. Plant Physiology and Biochemistry 44, 193201.
Kioko, J.I., Berjak, P., Pammenter, N.W., Watt, P. and Wesley-Smith, J. (1998) Desiccation and cryopreservation of embryonic axes of Trichilia dregeana Sond. CryoLetters 19, 511.
Kranner, I. and Birtić, S. (2005) A modulating role for antioxidants in desiccation tolerance. Integrative and Comparative Biology 45, 734740.
Kranner, I., Minibayeva, F., Beckett, R.P.andSeal, C.E. (2010a) What is stress? Concepts, definitions and applications in seed science. New Phytologist 188, 655673.
Kranner, I., Roach, T., Beckett, R.P., Whitaker, C. and Minibayeva, F. (2010b) Early production of reactive oxygen species during seed germination and early seedling growth in Pisum sativum. Journal of Plant Physiology 167, 805811.
Lee, M.Y., Kim, Y.K., Ryoo, K.K., Lee, Y.B. and Park, E.J. (2006) Electrolyzed-reduced water protects against oxidative damage to DNA, RNA and protein. Applied Biochemistry and Biotechnology 135, 273289.
Liszkay, A., van der Zalm, E. and Schopfer, P. (2004) Production of reactive oxygen intermediates (O2− , H2O2 and ∙OH) by maize roots and their role in wall loosening and elongation. Plant Physiology 136, 31143123.
Masella, R., Di Benedetto, R., Varì, R., Filesi, C. and Giovannini, C. (2005) Novel mechanisms of natural antioxidant compounds in biological systems: involvement of glutathione and glutathione-related enzymes. Journal of Nutritional Biochemistry 16, 577586.
Minibayeva, F., Kolesnikov, O., Chasov, A., Beckett, R.P., Lüthje, S., Vylegzhanina, N., Buck, F. and Böttger, M. (2009) Wound-induced apoplastic peroxidase activities: their roles in the production and detoxification of reactive oxygen species. Plant, Cell and Environment 32, 497508.
Misra, H.R. and Fridovich, I. (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. Journal of Biological Chemistry 247, 31703175.
Müller, K., Linkies, A., Vreeburg, R.A., Fry, S.C., Krieger-Liszkay, A. and Leubner-Metzger, G. (2009) In vivo wall loosening by hydroxyl radicals during cress seed germination and elongation growth. Plant Physiology 150, 18551865.
Murashige, T. and Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum 15, 473497.
Mycock, D.J. (1999) Addition of calcium and magnesium to a glycerol and sucrose cryoprotectant solution improves the quality of plant embryo recovery from cryostorage. CryoLetters 20, 7782.
Ngobese, N.Z., Berjak, P. and Pammenter, N.W. (2011) The effects of the steps of cryopreservation on the viability of zygotic embryos of selected amaryllid species. 10th Conference of the International Society for Seed Science. Informativo ABRATES 21, 105.
Obert, B., Benson, E.E., Millam, S., Pret'ová, A. and Bremner, D.H. (2005) Moderation of morphogenetic and oxidative stress responses in flax in vitro cultures by hydroxynonenal and desferrioxamine. Journal of Plant Physiology 162, 537547.
Pammenter, N.W., Adamson, J.H. and Berjak, P. (1974) Viability of stored seeds: extension by cathodic protection. Science 186, 11231124.
Pammenter, N.W., Greggains, V., Kioko, J.I., Wesley-Smith, J., Berjak, P. and Finch-Savage, W.E. (1998) The time factor during dehydration of non-orthodox (recalcitrant) seeds: effects of differential drying rates on viability retention in Ekebergia capensis. Seed Science Research 8, 463471.
Pammenter, N.W., Berjak, P., Wesley-Smith, J. and Vander Willigen, C. (2002) Experimental aspects of drying and recovery. pp. 263291in Black, M.; Pritchard, H.W. (Eds) Desiccation and survival in plants. Drying without dying. Wallingford, Oxon, CABI Publishing.
Pammenter, N.W., Berjak, P., Goveia, M., Sershen, Kioko, J.I., Whitaker, C. and Beckett, R.P. (2011) Topography determines the impact of reactive oxygen species on shoot apical meristems of recalcitrant embryos of tropical species during processing for cryopreservation. Acta Horticulturae (in press).
Perán, R., Berjak, P., Pammenter, N.W. and Kioko, J.I. (2006) Cryopreservation, encapsulation and promotion of shoot production of embryonic axes of a recalcitrant species, Ekebergia capensis Sparrm. CryoLetters 27, 112.
Pitzschke, A., Forzani, C. and Hirt, H. (2006) Reactive oxygen species signaling in plants. Antioxidants and Redox Signaling 8, 17571764.
Pukacka, S. and Ratajczak, E. (2006) Antioxidative response of ascorbate–glutathione pathway enzymes and metabolites to desiccation of recalcitrant Acer saccharinum seeds. Journal of Plant Physiology 163, 12591266.
Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M. and Rice-Evans, C. (1999) Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radical Biology and Medicine 26, 12311237.
Roach, T., Ivanova, M., Beckett, R.P., Minibayeva, F.V., Green, I., Pritchard, H.W. and Kranner, I. (2008) An oxidative burst of superoxide in embryonic axes of recalcitrant sweet chestnut seeds as induced by excision and desiccation. Physiologia Plantarum 133, 131139.
Roach, T., Beckett, R.P., Minibayeva, F.V., Colville, L., Whitaker, C., Chen, H., Bailly, C. and Kranner, I. (2010) Extracellular superoxide production, viability and redox poise in response to desiccation in recalcitrant Castanea sativa seeds. Plant, Cell and Environment 33, 5975.
Sacandé, M., Jøker, D., Dulloo, M.E. and Thomsen, K.A. (Eds) (2004) Comparative storage behaviour of tropical tree seeds. Rome, IPGRI.
Samarth, R.M., Panwar, M., Kumar, M., Soni, A., Kumar, M. and Kumar, A. (2008) Evaluation of antioxidant and radical-scavenging activities of certain radio-protective plant extracts. Journal of Food Chemistry 106, 868873.
Scandalios, J.G. (1993) Oxygen stress and superoxide dismutase. Plant Physiology 101, 712.
Schopfer, P. and Liszkay, A. (2006) Plasma membrane-generated reactive oxygen intermediates and their role in cell growth of plants. Biofactors 28, 7381.
Sershen, , Pammenter, N.W., Berjak, P. and Wesley-Smith, J. (2007) Cryopreservation of embryonic axes of selected amaryllid species. CryoLetters 28, 387399.
Shao, H.-B., Chu, L.-Y., Lu, Z.-H. and Kang, C.-M. (2008) Primary antioxidant free radical scavenging and redox signaling pathways in higher plant cells. International Journal of Biological Sciences 4, 814.
Shirahata, S., Kabayama, S., Nakano, M., Miura, T., Kusumoto, K., Gotoh, M., Hayashi, H., Otsubo, K., Morisawa, S. and Katakura, Y. (1997) Electrolyzed-reduced water scavenges active oxygen species and protects DNA from oxidative damage. Biochemical and Biophysical Research Communications 234, 269274.
Sirdaarta, J. and Cock, I.E. (2008) Vitamin E and Trolox™ reduce toxicity of Aloe barbadensis Miller juice in Artemia franciscana nauplii but individually are toxic at high concentrations. The Internet Journal of Toxicology™ 5, ISSN: 1559–3916. Available at http://www.ispub.com/journal/the_internet_journal_of_toxicology/(accessed March 2011).
Smirnoff, N. (1993) The role of active oxygen in the response of plants to water deficit and desiccation. New Phytologist 125, 2758.
Sutherland, J.R., Diekmann, M. and Berjak, P. (2002) Forest tree seed health for germplasm conservation. Rome, IPGRI.
Touchell, D. and Walters, C. (2000) Recovery of embryos of Zizania palustris following exposure to liquid nitrogen. CryoLetters 21, 261270.
Tripathi, R., Mohan, H. and Kamat, J.P. (2007) Modulation of oxidative damage by natural products. Journal of Food Chemistry 100, 8190.
Van Breusegem, F., Bailey-Serres, J. and Mittler, R. (2008) Unraveling the tapestry of networks involving reactive oxygen species in plants. Plant Physiology 147, 978984.
Varghese, B. and Naithani, S.C. (2002) Desiccation-induced changes in lipid peroxidation, superoxide level and antioxidant enzymes activity in neem (Azadirachta indica A. Juss) seeds. Acta Physiologiae Plantarum 24, 7987.
Varghese, B., Sershen, , Berjak, P., Varghese, D. and Pammenter, N.W. (2011) Differential drying rates of recalcitrant Trichilia dregeana embryonic axes: a study of survival and oxidative stress metabolism. Physiologia Plantarum DOI: 10.1111/j.1399-3054.2011.01469.
Walters, C., Pammenter, N.W., Berjak, P. and Crane, J. (2001) Desiccation damage accelerated ageing and respiration in desiccation tolerant and sensitive seeds. Seed Science Research 11, 135148.
Whitaker, C., Beckett, R.P., Minibayeva, F.V. and Kranner, I. (2010) Production of reactive oxygen species in excised and cryopreserved explants of Trichilia dregeana Sond. South African Journal of Botany 76, 112118.
Yin, J.-J., Lao, F., Fu, P.P., Warner, W.G., Zhao, Y., Wang, P.C., Qiu, Y., Sun, B., Xing, G., Dong, J., Liang, X.-J. and Chen, C. (2009) The scavenging of reactive oxygen species and the potential for cell protection by functionalized fullerene materials. Biomaterials 30, 611621.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Seed Science Research
  • ISSN: 0960-2585
  • EISSN: 1475-2735
  • URL: /core/journals/seed-science-research
Please enter your name
Please enter a valid email address
Who would you like to send this to? *
×

Keywords

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed