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Prevention of hyperhydricity in micropropagated carnation shoots by bottom cooling: implications of oxidative stress

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Abstract

Carnation shoot cultures were micropropagated in two different agar concentrations (0.58 and 0.85%) and placed in a bottom cooling system or control conditions. During the culture period of 28 days, it was observed that relative humidity, hyperhydricity, dry weight, multiplication rate, and the activity of the antioxidant enzymatic system changed in relation to the agar concentration used and the application of bottom cooling. The percentage of hyperhydric shoots also showed a significant decrease under bottom cooling conditions for both agar concentrations. Lipid peroxidation was always lower in shoots cultured with bottom cooling. All the antioxidant enzymatic activities were lower in bottom cooling treatments compared to controls. These results show that the normalization of the environmental conditions in vitro via bottom cooling can prevent the onset of different simultaneous stress reactions concomitant with hyperhydricity. The present work provides for the first time , direct evidence of a reduced H2O2 generation in the tissues cultured in bottom cooling able to reduce oxidative stress.

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Abbreviations

APX :

ascorbate peroxidase

CAT :

catalase

DHAr :

dehydroascorbate reductase

EDTA :

ethylendiaminetetraacetic sodium salt

GPXs :

soluble guaiacol peroxidase

GPXw :

cell-wall bound guaiacol peroxidase

GR :

glutathione reductase

MDA :

malondialdehyde

MDHr :

monodehydroascorbate reductase

NADPH :

nicotinamide adenine dinucleotide phosphate

PVP :

polyvinylpyrrolidone

ROS :

reactive oxygen species

SOD :

superoxide dismutase

TBA :

thiobarbituric acid

TCA :

trichloroacetic acid

References

  • M Aebi (1984) ArticleTitleCatalase in vitro Methods Enzymol. 105 121–126

    Google Scholar 

  • I Böttcher K Zoglauer H Göring (1988) ArticleTitleInduction and reversion of vitrification of plants cultured in vitro Physiol. Plant 72 560–564

    Google Scholar 

  • SX Chen P Schopfer (1999) ArticleTitleHydroxyl-radical production in physiological reactions A novel function of peroxidase. Eur. J. Biochem. 260 726–735

    Google Scholar 

  • DA Dalton L Langeberg NC Treneman (1993) ArticleTitleCorrrelations between the ascorbate-gluthatione pathway and effectiveness in legume root-nodules Physiol. Plant 87 365–370

    Google Scholar 

  • PC Debergh (1983) ArticleTitleEffects of agar brand and concentration on the tissue culture medium Physiol. Plant 59 270–276

    Google Scholar 

  • PC Debergh Y Harbaoui R Lemeur (53) ArticleTitleMass propagation of globe artichoke (Cynara scolymus) : evaluation of different hypotheses to overcome vitrification with special reference to water potential Physiol. Plant 53 181–187

    Google Scholar 

  • J Riek ParticleDe A Piqueras PC Debergh (1997) ArticleTitleSucrose uptake and metabolism in a double layer system for micropropagation of Rosa multiflora Plant Cell Tiss. Org. Cult. 47 269–279

    Google Scholar 

  • LA delRio F Sevilla M Gomez J Yánez J Lopez (1978) ArticleTitleSuperoxide dismutase: an enzyme system for the study of micronutrient interactions in plants Planta 140 221–225

    Google Scholar 

  • RR Dhindsa P Plunb-Dhindsa DM Reid (1982) ArticleTitleLeaf senescence and lipid peroxidation : effecs of some phytohormones and scavengers of free radical and singlet oxygen Physiol. Plant 56 453–457

    Google Scholar 

  • RR Dhindsa P Plunb-Dhindsa TA Thorpe (1981) ArticleTitleLeaf senescence correlated with increased levels of membrane permeability and lipid peroxidation and decreased levels of superoxide dismutase and catalase J. Exp. Bot. 126 93–106

    Google Scholar 

  • ML Droillard D Bureau P Paulin (1989) ArticleTitleIsozymes of superoxide dismutase in mitochondria and peroxisomes isolated from petals of carnation (Dianthus caryophyllus) during senescence Plant Physiol. 94 1187–1192

    Google Scholar 

  • ML Droillard D Bureau P Paulin (1989) ArticleTitleChanges in activities of superoxide dismutase during aging of petals of cut carnations (Dianthus cariophyllus) during senescence Physiol. Plant 76 1149–1154

    Google Scholar 

  • CH Foyer H Lopez-Delgado JF Dat LM Schott (1997) ArticleTitleHydrogen peroxide and gluthatione-associated mechanisms of acclamatory stress tolerance and signalling Physiol. Plant 92 696–717

    Google Scholar 

  • T Franck C Kevers T Gaspar (1995) ArticleTitleProtective enzymatic systems against activated oxygen species compared in normal and vitrified shoots of Prunus avium L raised in vitro Plant Growth Regul. 16 253–256

    Google Scholar 

  • T Franck C Kevers C Penel H Greppin JF Hausman T Gaspar (1998) ArticleTitleReducing properties, and markers of lipid peroxidation in normal and hyperhydric shoots of Prunus avium L J. Plant Physiol. 153 339–346

    Google Scholar 

  • T Gaspar T Franck B Bisbis C Kevers L Jouve JF Haussman J Dommes (2002) ArticleTitleConcepts in plant stress physiology Application to plant tissue cultures. Plant Growth Regul. 37 263–285

    Google Scholar 

  • EF George (1996) Plant Proagation by Tissue Culture Part 2: In Practice. Exegetics LTD Edington

    Google Scholar 

  • J Ghashghaie F Brenckmann B Saugier (1992) ArticleTitleWater relations and growth of rose plants cultured in vitro under various relative humidities Plant Cell Tiss. Org. Cult. 30 51–57

    Google Scholar 

  • K Gribble (1999) ArticleTitleThe influence of relative humidity on vitrification, growth and morphology of Gypsophila paniculata L Plant Growth Regul. 27 179–188

    Google Scholar 

  • HM Hassan I Fridovich (1977) ArticleTitleRegulation of the synthesis of superoxide dismutase in Escherichia coli J. Biol. Chem. 252 7667–7672

    Google Scholar 

  • RL Heath L Packer (1968) ArticleTitlePhotoperoxidation in isolated chloroplasts I. Kinetics and stoichiometry of fatty acid peroxidation Arch. Biochem. Biophys. 125 189–198

    Google Scholar 

  • DM Hodges CJ Andrews DA Johnson RI Hamilton (1997) ArticleTitleAntioxidant enzyme responses to chilling stress in differentially sensitive inbred maize lines J. Exp. Bot. 48 1105–1113

    Google Scholar 

  • A Jimenez JA Hernandez Rio LA del F Sevilla (1997) ArticleTitleEvidence for the presence of of the ascorbate-glutathione cycle in mitochondria and peroxisomes of pea leaves Plant Physiol. 114 275–284

    Google Scholar 

  • T Kozai K Fujiwara M Hayashi J Aitken-Christie (1992) The in vitro environment and its control in micropropagation K Kurata T Kozai (Eds) Transplant Production Systems Kluwer Academic Publishers Dordrecht The Netherlands 247–282

    Google Scholar 

  • OH Lowry NJ Rosebrough AL Farr RJ Randall (1951) ArticleTitleProtein measurement with the Folin phenol reagent J. Biol. Chem. 193 265–275 Occurrence Handle1:CAS:528:DyaG38XhsVyrsw%3D%3D Occurrence Handle14907713

    CAS  PubMed  Google Scholar 

  • JP Majada M Fal R Sánchez-Tames (1997) ArticleTitleThe effect of ventilation rate on proliferation and nyperhhydricity of Dianthus cariophyllus L In Vitro Cell. Dev. Biol. Plant 33 62–69

    Google Scholar 

  • JP Majada F Tadeo MA Fal R Sanchez-Tames (2000) ArticleTitleImpact of culture vessel ventilation on the anatomy and morphology of micropropagated carnation Plant Cell Tiss. Org. Cult. 63 207–214

    Google Scholar 

  • F Marga L Vebret H Morvan (1997) ArticleTitleAgar fractions could protect apple shoots cultured in liquid media against hyperhydricity Plant Cell Tiss Org Cult 49 1–5

    Google Scholar 

  • JM McCord I Fridovich (1969) ArticleTitleSuperoxide dismutase An enzymic function for erythrocuprein (hemocuprein) J. Biol. Chem. 244 6049–6055

    Google Scholar 

  • T Murashige F Skoog (1962) ArticleTitleA revised medium for rapid growth and bioassays with tobacco tissue cultures Physiol. Plant 15 437–497

    Google Scholar 

  • E Olmos E Hellin (1998) ArticleTitleUltrastructural differences of hyperhydric and normal leaves from regenerated carnation shoots Sci. Hort. 75 91–101

    Google Scholar 

  • E Olmos A Piqueras JR Martinez-Solano E Hellin (1997) ArticleTitleThe subcellular localization of peroxidase and the implication of oxidative stress in hyperhydrated leaves of regenerated carnation shoots Plant Sci. 130 97–105

    Google Scholar 

  • O Perez-Tornero J Egea E Olmos L Burgos (2001) ArticleTitleControl of hyperhydrcity in micropropagated apricot cultivars In Vitro Cell. Dev. Biol. Plant 37 250–254

    Google Scholar 

  • A Piqueras M Cortina MD Serna JL Casas (2002) ArticleTitlePolyamines and hyperhydricity in micropropagated carnation shoots Plant Sci. 162 671–678

    Google Scholar 

  • A Piqueras BH Han JM Huylenbroeck ParticleVan PC Debergh (1998) ArticleTitleEffect of different environmental conditions in vitro on sucrose metabolism and antioxidant enzymatic activities in cultured shoots of Nicotiana tabacum L Plant Growth Regul. 25 5–10

    Google Scholar 

  • HD Rabinowitch I Fridovich (1983) ArticleTitleSuperoxide radicals, superoxide dismutases and oxygen toxicity in plants Photochem. Photobiol. 37 679–690

    Google Scholar 

  • S Saher A Piqueras E Hellin E Olmos (2004) ArticleTitleHyperhydricity in micropropagated carnation shoots: the role of oxidative stress Physiol Plant 120 152–161

    Google Scholar 

  • HJ Scholten RLM Pierik (1998) ArticleTitleAgar as a gelling agent: chemical and physical analysis Plant Cell Rep. 17 230–235

    Google Scholar 

  • C Schweikert A Liszkay P Schopfer (2002) ArticleTitlePolysaccharide degradation by fenton reaction- or peroxidase-generated hydroxyl radicals in isolated plant cell walls Phytochemistry 61 31–35

    Google Scholar 

  • I Sylvestre MJ Droillars JM Bureau P Paulin (1989) ArticleTitleEffects of the ethylene rise on peroxidation membrane lipids during the senescence of cut carnation Plant Physiol. Biochem. 29 120–125

    Google Scholar 

  • P Thomas JB Mythili KS Shivashankara (2000) ArticleTitleExplant , medium and vessel aeration affect the incidence of hyperhydricity recovery of normal plants in triploid watermelon J. Hortic. Sci. Biotech. 75 19–25

    Google Scholar 

  • AM Vanderschaeghe PC Debergh (1987) Technical aspects of the control of the relative humidity in tissue culture containers G Ducate M Jacob A Simeon (Eds) Plant Micropropagation in Horticultural Industries (pp 68–76) Presses Universitaires Liege

    Google Scholar 

  • AB Whitehouse TR Marks GA Edwards (2002) ArticleTitleControl of hyperhydricity in Eucalyptus axillary shoot cultures grown in liquid medium Plant Cell Tiss. Org. Cult. 71 245–252

    Google Scholar 

  • D Yue A Gosselin Y Desjardins (1993) ArticleTitleEffects of forced ventilation at different relative humidities on growth, photosynthesis and transpiration of geranium plantlets in vitro Can. J. Plant Sci. 73 249–256

    Google Scholar 

  • M Ziv (1991) Vitrification: morphological and physiological disorders of in vitro shoots. PC Debergh RH Zimmerman (Eds) Micropropagation Kluwer Academic Publishers Dordrecht. 45–69

    Google Scholar 

  • M Ziv G Meir AH Halevy (1983) ArticleTitleFactors influencing the production of hardened glaucous carnation plantlets in vitro Plant Cell Tiss. Org. Cult. 2 55–60

    Google Scholar 

  • SMA Zobayed J Armstrong W Armstrong (2001) ArticleTitleMicropropagation of potato: evaluation of closed, diffusive and forced ventilation on growth and tuberization Ann. Bot. 87 53–59

    Google Scholar 

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Authors and Affiliations

  1. Centro de Edafologia y Biologia Aplicada del Segura, CSIC, P.O. Box 164, 30100, Murcia, Spain

    Sadhy Saher, Abel Piqueras, Eladio Hellin & Enrique Olmos

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  1. Sadhy Saher
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  2. Abel Piqueras
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  3. Eladio Hellin
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  4. Enrique Olmos
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Correspondence to Enrique Olmos.

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Saher, S., Piqueras, A., Hellin, E. et al. Prevention of hyperhydricity in micropropagated carnation shoots by bottom cooling: implications of oxidative stress. Plant Cell Tiss Organ Cult 81, 149–158 (2005). https://doi.org/10.1007/s11240-004-4043-4

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  • DOI: https://doi.org/10.1007/s11240-004-4043-4

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