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Effect of desiccation on physiological and biochemical indicators associated with the germination and vigor of cryopreserved seeds of Nicotiana tabacum L. cv. Sancti Spíritus 96

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Abstract

Cryopreservation is a valuable technique for the long-term conservation of plant germplasm and complementary to traditional seed storage methods. However, critical factors such as seed moisture content should be optimized before using this technique as a safe strategy for storing seeds such as those of Nicotiana spp. This study aimed to determine the effect of desiccation on physiological and biochemical indicators associated with germination and vigor in cryopreserved seeds of Nicotiana tabacum cv. Sancti Spíritus 96 (SS-96). The germination and vigor of seeds with a range of moisture content were assessed using electrolyte leakage and accelerated aging tests. In addition, these physiological indicators were related to the oxidative state of the seeds, in terms of the rate of O2 ·− generation and the H2O2 content, and the activity of enzymatic antioxidants superoxide dismutase and catalase. The cryopreserved seeds of N. tabacum SS-96 with a moisture content of 2.1% exhibited higher vigor probably due to the retention of membrane integrity, reflected by lower levels of lipid peroxidation and electrolyte leakage associated with the absence of oxidative stress. The results suggest 2.1% as the optimal moisture content for the storage of seeds of this cultivar, both at cryogenic temperatures and at 5°C.

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References

  • Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126

    Article  CAS  PubMed  Google Scholar 

  • Bailly C (2004) Active oxygen species and antioxidants in seed biology. Seed Sci Res 14:93–107

    Article  CAS  Google Scholar 

  • Bailly C, Audigier C, Ladonne F, Wagner MH, Coste F, Corbineau F, Côme D (2001) Changes in oligosaccharide content and antioxidant enzyme activities in developing bean seeds as related to acquisition of drying tolerance and seed quality. J Exp Bot 52:701–708

    Article  CAS  PubMed  Google Scholar 

  • Bailly C, El-Maarouf-Bouteau H, Corbineau F (2008) From intracellular signaling networks to cell death: the dual role of reactive oxygen species in seed physiology. C R Biologies 331:806–814

    Article  CAS  PubMed  Google Scholar 

  • Benson EE, Bremner D (2004) Oxidative stress in the frozen plant: a free radical point of view. In: Fuller BJ, Lane N, Benson EE (eds) Life in the frozen state. CRC Press, Boca Raton, Florida, USA, pp 205–242

    Chapter  Google Scholar 

  • Berjak P (2006) Unifying perspectives of some mechanisms basic to desiccation tolerance across life forms. Seed Sci Res 16:1–15

    Article  CAS  Google Scholar 

  • Berjak P, Farrant JM, Pammenter NW (2007) Seed desiccation-tolerance mechanisms. In: Jenks MA, Wood AJ (eds) Plant desiccation tolerance, 1st edn. Blackwell Publishing, Iowa, USA, pp 151–192

    Google Scholar 

  • Buege JA, Aust SD (1978) Microsomal lipid peroxidation. Methods Enzymol 52:302–310

    Article  CAS  PubMed  Google Scholar 

  • Buitink J, Claessens MMAE, Hemminga MA, Hoekstra FA (1998) Influence of water content and temperature on molecular mobility and intracellular glasses in seed and pollen. Plant Physiol 118:531–541

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Buitink J, Hemminga MA, Hoekstra FA (1999) Characterization of molecular mobility in seed tissues: an electron paramagnetic resonance spin probe study. Biophys J 76:3315–3322

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Buitink J, Leprince O (2004) Glass formation in plant anhydrobiotes: survival in the dry state. Cryobiology 48:215–228

    Article  CAS  PubMed  Google Scholar 

  • Buitink J, Leprince O (2008) Intracellular glasses and seed survival in the dry state. C R Biologies 331:788–795

    Article  CAS  PubMed  Google Scholar 

  • Du Z, Bramlage WJ (1992) Modified thiobarbituric acid assay for measuring lipid oxidation in sugar-rich plant tissue extracts. J Agric Food Chem 40:1566–1570

    Article  CAS  Google Scholar 

  • Fernández-Marín B, Kranner I, Sebastián MS, Artetxe U, Laza JM, Vilas JL, Pritchard HW, Nadajaran J, Míguez F, Becerril JM, García-Plazaola JI (2013) Evidence for the absence of enzymatic reactions in the glassy state. A case study of xanthophyll cycle pigments in the desiccation-tolerant moss Syntrichia ruralis. J Exp Bot 64:3033–3043

    Article  PubMed  PubMed Central  Google Scholar 

  • Fuchs J, Neuberger T, Rolletschek H, Schiebold S, Nguyen TH, Borisjuk N, Börner A, Melkus G, Jakob P, Borisjuk L (2013) A noninvasive platform for imaging and quantifying oil storage in submillimeter tobacco seed. Plant Physiol 161:583–593

    Article  CAS  PubMed  Google Scholar 

  • Golovina EA, As HV, Hoekstra FA (2010) Membrane chemical stability and seed longevity. Eur Biophys J 39:657–668

    Article  CAS  PubMed  Google Scholar 

  • Golovina EA, Wolkers WF, Hoekstra FA (1997) Long-term stability of protein secondary structure in dry seeds. Comp Biochem Physiol 117:343–348

    Article  Google Scholar 

  • Gonzalez-Arnao M, Martinez-Montero M, Cruz-Cruz C, Engelmann F (2014) Advances in cryogenic techniques for the long-term preservation of plant biodiversity. In: Ahuja M, Ramawat K (eds) Biotechnology and biodiversity, sustainable development and biodiversity. Springer International Publishing, Switzerland, pp 129–170

    Google Scholar 

  • González-Benito ME, Iriondo JM, Pérez-García F (1998) Seed cryopreservation: an alternative method for the conservation of Spanish endemics. Seed Sci Technol 26:257–262

    Google Scholar 

  • Groot SP, de Groot L, Kodde J, van Treuren R (2015) Prolonging the longevity of ex situ conserved seeds by storage under anoxia. Plant Genet Resour 13:18–26

    Article  CAS  Google Scholar 

  • Groot SP, Surki AA, de Vos RCH, Kodde J (2012) Seed storage at elevated partial pressure of oxygen, a fast method for analysing seed ageing under dry conditions. Ann Bot 110:1149–1159

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Guéraud F, Atalay M, Bresgen N, Cipak A, Eckl PM, Huc L, Jouanin I, Siems W, Uchida K (2010) Chemistry and biochemistry of lipid peroxidation products. Free Radic Res 44:1098–1124

    Article  PubMed  Google Scholar 

  • Hor YL, Kim YJ, Ugap A, Chabrillange N, Sinniah UR, Engelmann F, Dussert S (2005) Optimal hydration status for cryopreservation of intermediate oily seeds: citrus as a case study. Ann Bot 95:1153–1161

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • ISTA (2005) International rules for seed testing. International Seed Testing Association, Bassersdorf, Suiza

    Google Scholar 

  • Kumutha D, Ezhilmathi K, Sairam RK, Srivastava GC, Deshmukh PS, Meena RC (2009) Waterlogging induced oxidative stress and antioxidant activity in pigeonpea genotypes. Biol Plant 53:75–84

    Article  CAS  Google Scholar 

  • Lee YP, Baek KH, Lee HS, Kwak SS, Bang JW, Kwon SY (2010) Tobacco seeds simultaneously over-expressing Cu/Zn-superoxide dismutase and ascorbate peroxidase display enhanced seed longevity and germination rates under stress conditions. J Exp Bot 61:2499–2506

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Leprince O, Hoekstra FA (1998) The responses of cytochrome redox state and energy metabolism to dehydration support a role for cytoplasmic viscosity in desiccation tolerance. Plant Physiol 118:1253–1264

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Leprince O, Walters-Vertucci C (1995) A calorimetric study of the glass transition behaviors in axes of Phaseolus vulgaris L. seeds with relevance to storage stability. Plant Physiol 109:1471–1481

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li DZ, Pritchard HW (2009) The science and economics of ex situ plant conservation. Trends Plant Sci 14:614–621

    Article  CAS  PubMed  Google Scholar 

  • Liu K, Li Y, Chen F, Yong F (2016) Lipid oxidation of brown rice stored at different temperatures. Int J Food Sci Technol 52(1):188–195

    Article  Google Scholar 

  • Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. Eur J Biochem 47:469–474

    Article  CAS  PubMed  Google Scholar 

  • Meillng X, Xiuping L (1997) Report of ultra dry preservation technology for tobacco seeds. Seed Industry and AGPD, Cultural Development, Beijlng, China 4

  • Michalak M, Plitta-Mchalak BP, Chmielarz P (2015) A new insight in desiccation tolerance and cryopreservation of mazzard cherry (Prunus avium L.) seeds. Open Life Sci 10:354–364

    Google Scholar 

  • Mira S, Estrelles E, González-Benito ME (2015) Effect of water content and temperature on seed longevity of seven Brassicaceae species after 5 years of storage. Plant Biol 17:153–162

    Article  CAS  PubMed  Google Scholar 

  • Mira S, González ME, Hill LM, Walters C (2010) Characterization of volatile production during storage of lettuce (Lactuca sativa) seed. J Exp Bot 61(14):3915–3924

    Article  CAS  PubMed  Google Scholar 

  • Morscher F, Kranner I, Arc E, Bailly C, Roach T (2015) Glutathione redox state, tocochromanols, fatty acids, antioxidant enzymes and protein carbonylation in sunflower seed embryos associated with after-ripening and ageing. Ann Bot 116:669–678

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nagel M, Kodde J, Pistrick S, Mascher M, Börner A, Groot SPC (2016) Barley seed aging: genetics behind the dry elevated pressure of oxygen aging and moist controlled deterioration. Frontiers in Plant Sci 7:1–11

    Article  Google Scholar 

  • Navarro M, Febles G, Herrera RS (2015) Vigor: essential element for seed quality. Cuban J Agri Sci 49:447–458

    Google Scholar 

  • Perry DA (1984) Manual de métodos de ensayos de vigor. Instituto Nacional de Semillas y Plantas de Vivero. Ministerio de Agricultura, Pesca y Alimentación, Madrid, España

  • Pritchard HW (1995) Cryopreservation of seeds. In: Day JG, McLellan MR (eds) Methods in molecular biology, vol 38. Humana Press Inc., Totowa, NJ, pp 133–144

    Google Scholar 

  • Pritchard HW (2007) Cryopreservation of desiccation-tolerant seeds. In: Day JG, Stacey GN (eds) Methods in molecular biology: cryopreservation and freeze-drying protocols, 2nd edn. Humana Press Inc., Totowa, NJ, pp 185–201

    Chapter  Google Scholar 

  • Probert RJ (2003) Seed viability under ambient conditions, and the importance of drying. In: Smith RD, Dickie JB, Linington SH, Pritchard HW, Probert RJ (eds) Seed conservation: turning science into practice. Kew, UK, Royal Botanic Gardens, pp 725–743

    Google Scholar 

  • Rao NK, Hanson J, Dulloo ME, Ghosh K, Novell D, Larinde M (2007) Manual para el manejo de semillas en bancos de germoplasma. Manuales para Bancos de Germoplasma No. 8. Bioversity International, Roma, Italia

  • Sano N, Rajjou L, North HM, Debeaujon I, Marion-Poll A, Seo M (2016) Staying alive: molecular aspects of seed longevity. Plant Cell Physiol 57:660–674

    Article  CAS  PubMed  Google Scholar 

  • Singh N, Singh AK, Dhillon BS (2003) Effect of ultra-drying on ex situ seed conservation. In: Smith RD, Dickie JB, Linington SH, Pritchard HW, Probert RJ (eds) Seed conservation: turning science into practice. Royal Botanic Gardens, Kew, Reino Unido, pp 797–805

    Google Scholar 

  • Stanwood PC, Bass LN (1981) Seed germplasm preservation using liquid nitrogen. Seed Sci Technol 9:423–437

    Google Scholar 

  • Tappell AL (1966) Effects of low temperatures and freezing on enzymes and enzyme systems. In: Meryman HT (ed) Cryobiology. Academic Press, New York, pp 163–177

    Google Scholar 

  • Thanos CA, Doussi MA (1995) Ecophysiology of seed germination in endemic labiates of Crete. Israel J Plant Sci 43:227–237

    Article  Google Scholar 

  • Touchell DH, Dixon KW (1994) Cryopreservation for seedbanking of Australian species. Ann Bot 74:541–546

    Article  Google Scholar 

  • Varghese B, Naithani SC (2008) Oxidative metabolism-related changes in cryogenically stored neem (Azadirachta indica A. Juss) seeds. J Plant Physiol 165:755–765

    Article  CAS  PubMed  Google Scholar 

  • Veiga-Barbosa L, Mira S, González-Benito ME, Souza MM, Meletti LMM, Pérez-García F (2013) Seed germination, desiccation tolerance and cryopreservation of Passiflora species. Seed Sci Technol 41:89–97

    Article  Google Scholar 

  • Verdier J, Lalanne D, Pelletier S, Torres-Jerez I, Righetti K, Bandyopadhyay K, Leprince O, Chatelain E, Vu BL, Gouzy J, Gamas P, Udvardi MK, Buitink J (2013) A regulatory network-based approach dissects late maturation processes related to the acquisition of desiccation tolerance and longevity of Medicago truncatula seeds. Plant Physiol 163:757–774

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Vertucci CW, Roos EE, Crane J (1994) Theoretical basis of protocols for seed storage III. Optimum moisture contents for pea seeds stored at different temperatures. Ann Bot 74:531–540

    Article  Google Scholar 

  • Veselovsky VA, Veselova TV (2012) Lipid peroxidation, carbohydrate hydrolysis, and Amadori–Maillard reaction at early stages of dry seed aging. Russ J Plant Physiol 59:763–770

    Article  Google Scholar 

  • Visscher AM, Seal CE, Newton RJ, Frances AL, Pritchard HW (2016) Dry seeds and environmental extremes: consequences for seed lifespan and germination. Funct Plant Biol 43:656–668

    Article  Google Scholar 

  • Volk GM, Crane J, Caspersen AM, Hill LM, Gardner C, Walters C (2006) Massive cellular disruption occurs during early imbibition of Cuphea seeds containing crystallized triacylglycerols. Planta 224:1415–1426

    Article  CAS  PubMed  Google Scholar 

  • Walters C, Ballesteros D, Vertucci VA (2010) Structural mechanics of seed deterioration: standing the test of time. Plant Sci 179:565–573

    Article  CAS  Google Scholar 

  • Walters C, Hill LM, Wheeler LJ (2005) Dying while dry: kinetics and mechanisms of deterioration in desiccated organisms. Integr Comp Biol 45:751–758

    Article  PubMed  Google Scholar 

  • Walters C, Wheeler L, Stanwood PC (2004) Longevity of cryogenically stored seeds. Cryobiology 48:229–244

    Article  PubMed  Google Scholar 

  • Xin X, Jing X-M, Liu Y, Song S-Q (2010) Viability loss pattern under rapid dehydration of Antiaris toxicaria axes and its relation to oxidative damage. J Integr Plant Biol 52(5):434–441

    CAS  PubMed  Google Scholar 

  • Zagorchev L, Seal CE, Kranner I, Odjakova M (2013) A central role for thiols in plant tolerance to abiotic stress. Int J Mol Sci 14:7405–7432

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhou B, Wang J, Guo Z, Tan H, Zhu X (2006) A simple colorimetric method for determination of hydrogen peroxide in plant tissues. Plant Growth Regul 49:113–118

    Article  Google Scholar 

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Acknowledgements

We thank Magister Gilberto Torrecilla Guerra and Magister Otilio Ruiz Padrón for advising and editing at initial steps of the work and the manuscript. We thank also the participation of Dr. Sershen at the end to help edit the manuscript for language and improved aspects related to data presentation, interpretation, and discussion. This study was part of JLPR’s Magister Thesis from University of Ciego de Avila (Cuba).

Funding

This research was funded by the Cuban Ministry for Agriculture.

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Magister Juan Luis Pérez-Rodríguez (JLPR) was heavily involved in sourcing funding to support the project. Moreover, JLPR was involved in the design of all experiments, acquisition, analysis, and interpretation (statistical assessment) for the study. JLPR was also the primary person responsible for the initial drafting of the work (including the writing of manuscript and construction of figures). JLPR was also a major contributor providing detailed comments, additions, and major text modifications for drafts as the manuscript developed. This study was part of JLPR’s Magister Thesis from the University of Ciego de Avila (Cuba). Magister René Carlos Rodríguez Escriba (RCRE) and Magister Gustavo Lorente González (GLG) were heavily involved in the planning, design, analysis, and interpretation of the accelerated aging test experiment. Moreover, RCRE and GLG contributed to the determination of generation rate of O2 ·−, H2O2 content and enzymatic activities of superoxide dismutase (SOD) and catalase (CAT) of Nicotiana tabacum L. cv. Sancti Spíritus 96 seeds collected. RCRE and GLG were also involved in the interpretation of data and contributed for the initial drafting of the manuscript. Dr. Justo González-Olmedo (JGO) was involved in the design of the all experiments and the acquisition of data and data management to align with the original concept of the work. Dr. JGO was also involved in the writing of manuscript. Dr. Marcos Martinez-Montero (MMM) was heavily involved in sourcing funding to support the project and the conception and design of all experiments. MMM was also heavily involved in the acquisition of the data sets and the interpretation of data and its presentation in the manuscript. MMM was also a major contributor providing detailed comments, additions, and text modifications for different drafts of the manuscript. Finally, all authors approve the submission of this manuscript to be published and agree to be accountable for all aspects of the work.

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Correspondence to Juan Luis Pérez-Rodríguez.

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Pérez-Rodríguez, J.L., Rodríguez Escriba, R.C., Lorente González, G.Y. et al. Effect of desiccation on physiological and biochemical indicators associated with the germination and vigor of cryopreserved seeds of Nicotiana tabacum L. cv. Sancti Spíritus 96. In Vitro Cell.Dev.Biol.-Plant 53, 440–448 (2017). https://doi.org/10.1007/s11627-017-9857-y

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