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

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

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.

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E.E. Benson and D. Bremner (2004) Oxidative stress in the frozen plant: a free radical point of view. pp. 205241in B.J. Fuller ; N. Lane ; E.E. Benson (Eds) Life in the frozen state. Boca Raton, CRC Press.

E.E. Benson , K. Harding and J. Johnston (2007) Cryopreservation of shoot-tips and meristems. pp. 163184in J.G. Day ; G. Stacey (Eds) Methods in molecular biology, Vol. 368, Cryopreservation and freeze drying protocols. Totowa, NJ, Humana Press.

J.B. Dickie and H.W. Pritchard (2002) Systematic and evolutionary aspects of desiccation tolerance in seeds. pp. 239259in M. Black ; H.W. Pritchard (Eds) Desiccation and survival in plants. Drying without dying. Wallingford, Oxon, CABI Publishing.

N.W. Pammenter , P. Berjak , M. Goveia , Sershen J.I. Kioko , C. Whitaker and R.P. Beckett (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).

B. Varghese , Sershen , P. Berjak , D. Varghese and N.W. Pammenter (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.

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Seed Science Research
  • ISSN: 0960-2585
  • EISSN: 1475-2735
  • URL: /core/journals/seed-science-research
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