Advertisement

Protoplasma

, Volume 255, Issue 4, pp 1065–1077 | Cite as

Cold, antioxidant and osmotic pre-treatments maintain the structural integrity of meristematic cells and improve plant regeneration in cryopreserved kiwifruit shoot tips

  • Liya Mathew
  • Andrew McLachlan
  • Rubina Jibran
  • David J. Burritt
  • Ranjith Pathirana
Original Article

Abstract

Cryopreservation is a reliable and cost-effective method for the long-term preservation of clonally propagated species. The number of vegetatively propagated species conserved by cryopreservation is increasing through development of vitrification-based methods; droplet vitrification in particular is becoming the preferred method for many species, as it ensures fast freezing and thawing rates. This research investigated if cold, antioxidant and osmotic pre-treatments could maintain the structural integrity of cells, thence aid in developing a droplet vitrification protocol for kiwifruit using Actinidia chinensis var. chinensis ‘Hort16A’ as a model. Cold acclimation of donor plantlets at 4 °C for 2 weeks followed by sucrose pre-culture of shoot tips and supplementation of ascorbic acid (0.4 mM) in all media throughout the procedure registered 40% regeneration after cryopreservation. Transmission electron microscope imaging of meristematic cells confirmed sucrose and ascorbic acid pre-treatment of shoot tips from cold acclimated plantlets following treatment in vitrification solution exhibited severe plasmolysis and some disruption of membrane and vacuoles. In contrast cells without cold acclimation or sucrose and ascorbic acid pre-treatments exhibited minimal change after exposure to vitrification solution. After cryopreservation and recovery, all cells of untreated shoot tips showed rupture of the plasma membrane, loss of cytoplasmic contents and organelle distortions. By comparison, most pre-treated shoot-tip cells from cold acclimated plantlets retained their structural integrity, showing that only those cells that have been dehydrated and plasmolysed can withstand cryopreservation by vitrification.

Keywords

Actinidia chinensis Ascorbic acid Droplet vitrification Tissue culture Sucrose Transmission electron microscopy Ultrastructure 

Notes

Acknowledgements

The authors thank Andrew Mullan and Belinda Diepenheim for the technical assistance, Tony Corbett for the preparation of figures, and Dr. Jayanthi Nadarajan for the critical reading of the manuscript.

Compliance with ethical standards

Conflicts of interest

The authors declare that they have no conflicts of interests.

Supplementary material

709_2018_1215_MOESM1_ESM.pdf (238 kb)
Online Resource 1 (PDF 237 kb)

References

  1. Bachiri Y, Song GQ, Plessis P, Shoar-Ghaffari A, Rekab T, Morisset C (2001) Routine cryopreservation of kiwifruit (Actinidia spp.) germplasm by encapsulation-dehydration: importance of plant growth regulators. Cryo-Lett 22(1):61–74Google Scholar
  2. Beatson RA, Datson PM, Ferguson AR, Montefiori M (2014) Use of kiwifruit germplasm resources for genetic improvement. Acta Hortic (1048):25–34.  https://doi.org/10.17660/ActaHortic.2014.1048.2
  3. Benelli C, De Carlo A, Engelmann F (2013) Recent advances in the cryopreservation of shoot-derived germplasm of economically important fruit trees of Actinidia, Diospyros, Malus, Olea, Prunus, Pyrus and Vitis. Biotechnol Adv 31(2):175–185.  https://doi.org/10.1016/j.biotechadv.2012.09.004 CrossRefPubMedGoogle Scholar
  4. Benson EE (2008) Cryopreservation of phytodiversity: a critical appraisal of theory and practice. Crit Rev Plant Sci 27(3):141–219.  https://doi.org/10.1080/07352680802202034 CrossRefGoogle Scholar
  5. Bettoni JC, Dalla Costa M, Gardin JPP, Kretzschmar AA, Pathirana R (2016) Cryotherapy: a new technique to obtain grapevine plants free of viruses. Rev Bras Frutic 38(2).  https://doi.org/10.1590/0100-29452016833
  6. Chang YJ, Reed BM (1999) Extended cold acclimation and recovery medium alteration improve regrowth of Rubus shoot tips following cryopreservation. CryoLetters 20(6):371–376Google Scholar
  7. Coelho N, Elena Gonzalez-Benito M, Martin C, Romano A (2014) Cryopreservation of Thymus lotocephalus shoot tips and assessment of genetic stability. CryoLetters 35(2):119–128PubMedGoogle Scholar
  8. Danso KE, Ford-Lloyd BV (2004) Cryopreservation of embryogenic calli of cassava using sucrose cryoprotection and air desiccation. Plant Cell Rep 22(9):623–631.  https://doi.org/10.1007/s00299-003-0727-1 CrossRefPubMedGoogle Scholar
  9. Debenham MC, Seelye JF, Mullan AC (2016) An in vitro repository for clonal kiwifruit. Acta Hortic (1113):93–97.  https://doi.org/10.17660/ActaHortic.2016.1113.13
  10. Escobar RH, Muñoz L, Rios A, Núñez A, Tohme J (2014) Using a droplet-vitrification method to partially overcome the recalcitrance of cassava to cryostorage. Acta Hortic 1039(1039):227–232.  https://doi.org/10.17660/ActaHortic.2014.1039.29 CrossRefGoogle Scholar
  11. Ferguson AR, Huang H (2007) Genetic resources of kiwifruit: domestication and breeding. Hortic Rev 33:1–121Google Scholar
  12. Fki L, Bouaziz N, Chkir O, Benjemaa-Masmoudi R, Rival A, Swennen R, Drira N, Panis B (2013) Cold hardening and sucrose treatment improve cryopreservation of date palm meristems. Biol Plant 57(2):375–379.  https://doi.org/10.1007/s10535-012-0284-y CrossRefGoogle Scholar
  13. Folgado R (2015) Cryopreservation of Agave margaritae shoot tips by droplet-vitrification: the effects of sucrose preculture on plant regeneration. Cryobiology 71(3):562.  https://doi.org/10.1016/j.cryobiol.2015.10.105 CrossRefGoogle Scholar
  14. Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50(1):151–158.  https://doi.org/10.1016/0014-4827(68)90403-5 CrossRefPubMedGoogle Scholar
  15. Hakozaki M, Yoshida Y, Suzuki M (1996) Viability of calli from hypocotyl of kiwifruit seedlings exposed to liquid nitrogen. Environment Control in. Biology 34(2):147–151Google Scholar
  16. Helliot B, Swennen R, Poumay Y, Frison E, Lepoivre P, Panis B (2003) Ultrastructural changes associated with cryopreservation of banana (Musa spp.) highly proliferating meristems. Plant Cell Rep 21(7):690–698.  https://doi.org/10.1007/s00299-002-0537-x PubMedCrossRefGoogle Scholar
  17. Ibrahim S, Normah M (2013) The survival of in vitro shoot tips of Garcinia mangostana L. after cryopreservation by vitrification. Plant Growth Regul 70(3):237–246.  https://doi.org/10.1007/s10725-013-9795-6 CrossRefGoogle Scholar
  18. Johnston JW, Harding K, Benson EE (2007) Antioxidant status and genotypic tolerance of Ribes in vitro cultures to cryopreservation. Plant Sci 172(3):524–534CrossRefGoogle Scholar
  19. Katkov II, Levine F (2004) Prediction of the glass transition temperature of water solutions: comparison of different models. Cryobiology 49(1):62–82.  https://doi.org/10.1016/j.cryobiol.2004.05.004 CrossRefPubMedGoogle Scholar
  20. Kendall EJ, McKersie BD (1989) Free radical and freezing injury to cell membranes of winter wheat. Physiol Plant 76(1):86–94.  https://doi.org/10.1111/j.1399-3054.1989.tb05457.x CrossRefGoogle Scholar
  21. Kushnarenko SV, Romadanova NV, Reed BM (2009) Cold acclimation improves regrowth of cryopreserved apple shoot tips. CryoLetters 30(1):47–54PubMedGoogle Scholar
  22. Le Bras C, Le Besnerais P-H, Hamama L, Grapin A (2014) Cryopreservation of ex-vitro-grown Rosa chinensis ‘Old Blush’ buds using droplet-vitrification and encapsulation-dehydration. Plant Cell Tissue Organ Cult 116(2):235–242.  https://doi.org/10.1007/s11240-013-0400-5 CrossRefGoogle Scholar
  23. Leunufna S, Keller ERJ (2003) Investigating a new cryopreservation protocol for yams (Dioscorea spp.) Plant Cell Rep 21(12):1159–1166.  https://doi.org/10.1007/s00299-003-0652-3 CrossRefPubMedGoogle Scholar
  24. Li J, Guo Y (1996) Cryopreservation of cultured Actinidia deliciosa calli. J Fruit Sci 13(2):88–91Google Scholar
  25. Linsmaier EM, Skoog F (1965) Organic growth factor requirements of tobacco tissue cultures. Physiol Plant 18(1):100–106.  https://doi.org/10.1111/j.1399-3054.1965.tb06874.x CrossRefGoogle Scholar
  26. Liu YL, Suzuki T, Kasai N, Harada T (1998) Effects of cold acclimation and freezing solution treatment on the survival of frozen lateral buds excised from in vitro-cultured shoots of tara vine (Actinidia arguta). J Jpn Soc Hortic Sci 67(4):562–566.  https://doi.org/10.2503/jjshs.67.562 CrossRefGoogle Scholar
  27. Markovic Z, Preiner D, Stupic D, Andabaka Z, Simon S, Voncina D, Maletic E, Kontic JK, Chatelet P, Engelmann F (2015) Cryopreservation and cryotherapy of grapevine (Vitis vinifera L.) Vitis 54:247–251Google Scholar
  28. Matsumoto T (2017) Cryopreservation of plant genetic resources: conventional and new methods. Reviews in Agricultural Science 5 (13–20).  https://doi.org/10.7831/ras5.13
  29. Muldrew K, McGann LE (1994) The osmotic rupture hypothesis of intracellular freezing injury. Biophys J 66(2 Pt 1):532–541.  https://doi.org/10.1016/S0006-3495(94)80806-9 CrossRefPubMedPubMedCentralGoogle Scholar
  30. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15(3):473–497.  https://doi.org/10.1111/j.1399-3054.1962.tb08052.x CrossRefGoogle Scholar
  31. Ozudogru EA, Kaya E (2012) Cryopreservation of Thymus cariensis and T. vulgaris shoot tips: comparison of three vitrification-based methods. CryoLetters 33(5):363–375PubMedGoogle Scholar
  32. Panis B, Piette B, André E, Van den Houwe I, Swennen R (2011) Droplet vitrification: the first generic cryopreservation protocol for organized plant tissues? Acta Hortic 908:157–163CrossRefGoogle Scholar
  33. Panis B, Piette B, Swennen R (2005) Droplet vitrification of apical meristems: a cryopreservation protocol applicable to all Musaceae. Plant Sci 168:45–55CrossRefGoogle Scholar
  34. Panta A, Panis B, Ynouye C, Swennen R, Roca W (2014) Development of a PVS2 droplet vitrification method for potato cryopreservation. CryoLetters 35(3):255–266PubMedGoogle Scholar
  35. Park S-U, Kim H-H (2015) Cryopreservation of sweet potato shoot tips using a droplet-vitrification procedure. CryoLetters 36(5):344–352PubMedGoogle Scholar
  36. Pathirana R, McLachlan A, Hedderley D, Carra A, Carimi F, Panis B (2015) Removal of leafroll viruses from infected grapevine plants by droplet vitrification. Acta Hortic (1083):491–498.  https://doi.org/10.17660/ActaHortic.2015.1083.64
  37. Pathirana R, Deroles S, Hoeata K, Montefiori M, Tyson J, Wang T, Datson PM, Hellens RP (2016a) Fast-tracking kiwifruit breeding through mutagenesis. Acta Hortic (1127):217–222.  https://doi.org/10.17660/ActaHortic.2016.1127.34
  38. Pathirana R, Mc Lachlan A, Hedderley D, Panis B, Carimi F (2016b) Pre-treatment with salicylic acid improves plant regeneration after cryopreservation of grapevine (Vitis spp.) by droplet vitrification. Acta Physiol Plant 38(1):1–11.  https://doi.org/10.1007/s11738-015-2026-1 CrossRefGoogle Scholar
  39. Peredo EL, Arroyo-Garcia R, Reed BM, Angeles Revilla M (2008) Genetic and epigenetic stability of cryopreserved and cold-stored hops (Humulus lupulus L.) Cryobiology 57(3):234–241.  https://doi.org/10.1016/j.cryobiol.2008.09.002 CrossRefPubMedGoogle Scholar
  40. Reed BM (2001) Implementing cryogenic storage of clonally propagated plants. CryoLetters 22(2):97–104PubMedGoogle Scholar
  41. Ristic Z, Ashworth E (1993) Changes in leaf ultrastructure and carbohydrates in Arabidopsis thaliana L. (Heyn) cv. Columbia during rapid cold acclimation. Protoplasma 172(2–4):111–123.  https://doi.org/10.1007/BF01379368 CrossRefGoogle Scholar
  42. Sakai A, Engelmann F (2007) Vitrification, encapsulation-vitrification and droplet-vitrification: a review. CryoLetters 28(3):151–172PubMedGoogle Scholar
  43. Sakai A, Kobayashi S, Oiyama I (1990) Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb. var. brasiliensis Tanaka) by vitrification. Plant Cell Rep 9(1):30–33.  https://doi.org/10.1007/BF00232130 CrossRefPubMedGoogle Scholar
  44. Suzuki M, Ishikawa M, Okuda H, Noda K, Kishimoto T, Nakamura T, Ogiwara I, Shimura I, Akihama T (2006) Physiological changes in Gentian axillary buds during two-step preculturing with sucrose that conferred high levels of tolerance to desiccation and cryopreservation. Ann Bot 97(6):1073–1081CrossRefPubMedPubMedCentralGoogle Scholar
  45. Suzuki M, Miura N, Akihama T (1997) Effect of abscisic acid and proline on viability of shoot tips of kiwifruit seedlings during cryopreservation. Bull Faculty of Agric Meiji Univ 113:29–34Google Scholar
  46. Suzuki M, Niino T, Miura N, Akihama T (1996) Viability of cryopreserved shoot tips from kiwifruit seedlings at different growth stages in vitro. Bull Faculty Agric Meiji Univ 107:27–35Google Scholar
  47. Taglienti A, Tiberini A, Barba M (2013) Cryotherapy: a new tool for the elimination of artichoke viruses. J Plant Pathol 95(3):597–602Google Scholar
  48. Tavazza R, Lucioli A, Benelli C, Giorgi D, D'Aloisio E, Papacchioli V (2013) Cryopreservation in artichoke: towards a phytosanitary qualified germplasm collection. Ann Appl Biol 163(2):231–241.  https://doi.org/10.1111/aab.12049 CrossRefGoogle Scholar
  49. Uchendu EE, Leonard SW, Traber MG, Reed BM (2010a) Vitamins C and E improve regrowth and reduce lipid peroxidation of blackberry shoot tips following cryopreservation. Plant Cell Rep 29(1):25–35.  https://doi.org/10.1007/s00299-009-0795-y CrossRefPubMedGoogle Scholar
  50. Uchendu EE, Muminova M, Gupta S, Reed BM (2010b) Antioxidant and anti-stress compounds improve regrowth of cryopreserved Rubus shoot tips. In Vitro Cell Dev Biol-Plant 46(4):386–393.  https://doi.org/10.1007/s11627-010-9292-9 CrossRefGoogle Scholar
  51. Uchendu EE, Shukla M, Saxena PK, Keller JER (2016) Cryopreservation of potato microtubers: the critical roles of sucrose and desiccation. Plant Cell Tissue Org Cult 124(3):649–656.  https://doi.org/10.1007/s11240-015-0916-y CrossRefGoogle Scholar
  52. Vieira RL, da Silva AL, Zaffari GR, Steinmacher DA, de Freitas Fraga HP, Guerra MP (2015) Efficient elimination of virus complex from garlic (Allium sativum L.) by cryotherapy of shoot tips. Acta Physiol Plant 37(1):1733.  https://doi.org/10.1007/s11738-014-1733-3 CrossRefGoogle Scholar
  53. Volk GM, Bonnart R, Krueger R, Lee R (2012) Cryopreservation of Citrus shoot tips using micrografting for recovery. CryoLetters 33(6):418–426PubMedGoogle Scholar
  54. Volk GM, Walters C (2003) Preservation of genetic resources in the national plant germplasm clonal collections. Plant Breeding Rev 23:291–344Google Scholar
  55. Wang B, Wang R-R, Li J-W, Ma Y-L, Sheng W-M, Li M-F, Wang Q-C (2013) Development of three-vitrification-based cryopreservation procedures for shoot tips of China’s potato. CryoLetters 34(4):369–380PubMedGoogle Scholar
  56. Wang L-Y, Li Y-D, Sun H-Y, Liu H-G, Tang X-D, Wang Q-C, Zhang Z-D (2017) An efficient droplet-vitrification cryopreservation for valuable blueberry germplasm. Sci Hortic 219:60–69.  https://doi.org/10.1016/j.scienta.2017.03.007 CrossRefGoogle Scholar
  57. Wang QC, Panis B, Engelmann F, Lambardi M, Valkonen JPT (2009) Cryotherapy of shoot tips: a technique for pathogen eradication to produce healthy planting materials and prepare healthy plant genetic resources for cryopreservation. Ann Appl Biol 154(3):351–363.  https://doi.org/10.1111/j.1744-7348.2008.00308.x CrossRefGoogle Scholar
  58. Wang QC, Valkonen JPT (2009) Improved recovery of cryotherapy-treated shoot tips following thermotherapy of in vitro grown stock shoots of raspberry (Rubus idaeus L.) CryoLetters 30(3):171–182Google Scholar
  59. Wu Y, Zhao Y, Engelmann F, Zhou M (2000) Cryopreservation of kiwi shoot tips. Cryo letters 22(5):277–284Google Scholar
  60. Xu X, Cai Z, Gu Q, Zhang Q (2006) Cell ultrastructure of kiwifruit (Actinidia chinensis) shoot tips during cryopreservation. Agric Sci China 5(8):587–590.  https://doi.org/10.1016/S1671-2927(06)60096-5 CrossRefGoogle Scholar
  61. Yamamoto S, Rafique T, Priyantha WS, Fukui K, Matsumoto T, Niino T (2011) Development of a cryopreservation procedure using aluminium cryo-plates. CryoLetters 32(3):256–265PubMedGoogle Scholar
  62. Zhai ZY, Wu YJ, Engelmann F, Chen RZ, Zhao YH (2003) Genetic stability assessments of plantlets regenerated from cryopreserved in vitro cultured grape and kiwi shoot-tips using RAPD. CryoLetters 24(5):315–322PubMedGoogle Scholar
  63. Zhang JH, Huang WD, Liu YP, Pan QH (2005) Effects of temperature acclimation pretreatment on the ultrastructure of mesophyll cells in young grape plants (Vitis vinifera L. cv. Jingxiu) under cross-temperature stresses. J Integr Plant Biol 47(8):959–970.  https://doi.org/10.1111/j.1744-7909.2005.00109.x CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.The New Zealand Institute for Plant and Food Research LimitedPalmerston NorthNew Zealand
  2. 2.Department of BotanyUniversity of OtagoDunedinNew Zealand

Personalised recommendations