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Efficient Cryopreservation of in vitro Grown Shoot Tips of Pear (Pyrus spp.) by Droplet-vitrification

이호선|한지원|남성희|윤병현|고호철|신일섭|김금선|송재영|원경호|이정로|배진주
한국자원식물학회 (The Plant Resources Society Of Korea)|2023.12|한국자원식물학회지 (Korean Journal of Plant Reources.)|제36권 제6호|pp. 571-579

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키워드

CryopreservationPyrusRegrowthDroplet-vitrification

초록

In this study, cryopreservation by droplet-vitrification was applied to pear (Pyrus spp.) germplasm. We focused on the development and assessment of various strategies for the selection of suitable tissue, osmoprotection, and dehydration. We also evaluated post-thaw recovery of cryopreserved explants by droplet-vitrification. Preferentially, we tested the effects of preculture and loading treatments to determine which tissues were more suitable, either the apical shoot tips or the axillary buds. Apical shoot tips showed the better regrowth rate than in vitro axillary buds. The most effective techniques for cryopreservation were as follows. Shoots from in vitro seedlings which had been cultured for about 5-6 weeks were coldhardened at 4℃ for one week, excised shoot tips were precultured on liquid MS medium including 0.3 M sucrose for 31 hours and 0.7 M sucrose for 17 hours, osmoprotected in loading solution (LS) for 40 min, and then cryoprotected in dehydration solution (PVS3) for 90 min. In addition, we found that regrowth rates of explants on regrowth medium after exposure to liquid nitrogen (LN) were higher than those on MS medium. Results indicated that the highest regrowth percentage was 95.6% for ‘Bartlett’ cultivar and 68.9% for ‘BaeYun No.3’ cultivar. Consequently, apical shoot tips of two pear cultivars, ‘Bartlett’ (P. communis) and ‘BaeYun No.3’ (P. pyrifolia), were successfully cryopreserved by droplet-vitrification. Results of this study show that the enhanced droplet-vitrification method described in the present study could be used as an effective means for long-term storage of pear genetic resources.

Abstract

In this study, cryopreservation by droplet-vitrification was applied to pear (Pyrus spp.) germplasm. We focused on the development and assessment of various strategies for the selection of suitable tissue, osmoprotection, and dehydration. We also evaluated post-thaw recovery of cryopreserved explants by droplet-vitrification. Preferentially, we tested the effects of preculture and loading treatments to determine which tissues were more suitable, either the apical shoot tips or the axillary buds. Apical shoot tips showed the better regrowth rate than in vitro axillary buds. The most effective techniques for cryopreservation were as follows. Shoots from in vitro seedlings which had been cultured for about 5-6 weeks were coldhardened at 4℃ for one week, excised shoot tips were precultured on liquid MS medium including 0.3 M sucrose for 31 hours and 0.7 M sucrose for 17 hours, osmoprotected in loading solution (LS) for 40 min, and then cryoprotected in dehydration solution (PVS3) for 90 min. In addition, we found that regrowth rates of explants on regrowth medium after exposure to liquid nitrogen (LN) were higher than those on MS medium. Results indicated that the highest regrowth percentage was 95.6% for ‘Bartlett’ cultivar and 68.9% for ‘BaeYun No.3’ cultivar. Consequently, apical shoot tips of two pear cultivars, ‘Bartlett’ (P. communis) and ‘BaeYun No.3’ (P. pyrifolia), were successfully cryopreserved by droplet-vitrification. Results of this study show that the enhanced droplet-vitrification method described in the present study could be used as an effective means for long-term storage of pear genetic resources.

참고문헌 (43)

  1. Bell, R. L, "Genetic Resources of Temperate Fruit and Nut Crops", pp.657~697 (1991)
  2. Benson, E. E., "Developmental competence and ploidy stability in plants regenerated from cryopreserved potato shoot tips", Cryo Lett, 17, pp.119~128 (1996)
  3. Bi, W. -L., "Droplet-vitrification cryopreservation of in vitrogrown shoot tips of grapevine (Vitis spp.)", Vitr. Cell. Dev.Biol. Anim, 54, pp.590~599 (2018)
  4. Chang, Y., "Extended alternating-temperature cold acclimation and culture duration improve pear shoot cryopreservation", Cryobiology, 40, pp.311~322 (2000)
  5. Chang, Y., "Preculture conditions influence cold hardiness and regrowth of Pyrus cordata shoot tips after cryopreservation", Hort. Science, 36, pp.1329~1333 (2001)
  6. Chen, X. L., "Cryopreservation of in vitro-grown apical meristems of Lilium by droplet-vitrification", S. African J. Bot, 77, pp.397~403 (2011)
  7. Condello, E., "Cryopreservation of apple in vitro axillary buds using droplet-vitrification", Cryo Lett, 32, pp.175~185 (2011)
  8. Condello, E., "Genetic stability of wild pear (Pyrus pyraster B.) after cryopreservation by encapsulation-dehydration", Agric. Food Sci, 18, pp.136~143 (2009)
  9. Guyader, A., "Development of the droplet-vitrification cryopreservation technique as a viable alternative to the dormant bud technique for recalcitrant pears", Acta Hortic, 1234, pp.219~224 (2019)
  10. Hao, Y., "Stable maintenance and expression of a foreign gene in transgenic pear shoots retrieved from in vitro conservation", J. Plant Physiol, 162, pp.237~243 (2005)
  11. Hofer, M., "Cryopreservation of Malus and Pyrus wild species in the ‘Fruit Genebank’ in Dresden-Pillnitz, Germany", Biology, 12, pp.200~ (2023)
  12. Hong, S. -Y., "A review of pears (Pyrus spp.), ancient functional food for modern times", BMC Complement. Med. Ther, 21, pp.219~ (2021)
  13. Kim, H. H., "Cryobanking of Korean allium germplasm collections: Results from a 10 year experiences", Cryo Lett, 33, pp.45~57 (2012)
  14. Kim, H. H., "Development of alternative plant vitrification solutions in droplet-vitrification procedures", Cryo Lett, 30, pp.320~334 (2009)
  15. Kim, H. H., "Cryopreservation of garlic germplasm collections using the droplet-vitrification technique", Cryo Lett, 28, pp.471~482 (2007)
  16. Kopnick, C., "Changes of soluble sugars and ATP content during DMSO droplet freezing and PVS3 droplet vitrification of potato shoot tips", Cryobiology, 85, pp.79~86 (2018)
  17. Lambardi, M., "Micropropagation of Woody Trees and Fruits", pp.815~840 (2003), Kluwer Academy Publishers
  18. Leunufna, S., "Investigating a new cryopreservation protocol for yams (Dioscorea spp.)", Plant Cell Rep, 21, pp.1159~1166 (2003)
  19. Li, B. -Q., "Recovery patterns, histological observations and genetic integrity in Malus shoot tips cryopreserved using droplet-vitrification and encapsulation-dehydration procedures", J. Biotechnol, 214, pp.182~191 (2015)
  20. Li, J., "Pear genetics: Recent advances, new prospects, and a roadmap for the future", Hort. Res, 9, pp.uhab040~ (2022)
  21. Murashige, T., "A revised medium for rapid growth and bioassays with tobacco tissue cultures", Physiol. Plant, 15, pp.473~497 (1962)
  22. Niino, T., "Cryopreservation of in vitro-grown shoot tips of apple and pear by vitrification", Plant Cell, Tissue Organ Cult, 28, pp.261~266 (1992)
  23. Pathirana, R., "Pre-treatment with salicylic acid improves plant regeneration after cryopreservation of grapevine (Vitis spp.) by droplet vitrification", Acta Physiol. Plant, 38, pp.12~ (2016)
  24. Potter, D., "Phylogeny and classification of Rosaceae", Plant Syst. Evol, 266, pp.5~43 (2007)
  25. Quinet, M, "The Pear Genome. Compendium of Plant Genomes" (2019), Springer
  26. Rall, W. F., "Cryopreservation of mouse embryo at -196℃ by vitrification", Nature, 313, pp.573~575 (1985)
  27. Reed, B, "Cryoperservation and long-term storage of pear germplasm", Vitr. Cell. Dev. Biol. Plant, 34, pp.256~260 (1998)
  28. Reed, B. M, "Survival of in vitro-grown apical meristems of Pyrus following cryopreservation", Hort. Sci, 25, pp.111~113 (1990)
  29. Reed, B. M, "Implementing cryogenic storage of clonally propagated plants", Cryo Lett, 22, pp.97~104 (2001)
  30. Roque-Borda, C. A., "Cryopreservation of agronomic plant germplasm using vitrification-based methods : an overview of selected case studies", Int. J. Mol. Sci, 22, pp.6157~ (2021)
  31. Ruzic, D., "Cryopreservation of cherry rootstock Gisela 5(Prunus cerasus x Prunus canescens)shoot tips by droplet-vitrification technique", J. Hortic. Res, 21, pp.79~85 (2013)
  32. Scottez, C., "Cryopreservation of cold-acclimated shoot tips of pear in vitro cultures after encapsulationdehydration", Cryobiology, 29, pp.691~700 (1992)
  33. Silva, G. J., "Origin, domestication, and dispersing of pear (Pyrus spp.)", Adv. Agr, 2014, pp.1~8 (2014)
  34. 송재영, "In vitro Root Induction from Shoot Explants of Pear (Pyrus spp.)", 한국자원식물학회지, 35(6), pp.770~777 (2022), 한국자원식물학회
  35. Tahtamouni, R. W., "Preservation at low temperature and cryopreservation in wild pear(Pyrus syriaca)", Adv. Hort. Sci, 13, pp.156~160 (1999)
  36. Towill, L. E, "Genetic considerations for germplasm preservation of clonal materials", Hort. Sci, 23, pp.91~95 (1988)
  37. Volk, G. M., "Citrus cryopreservation: viability of diverse taxa and histological observations", Plant Cell, Tissue Organ Cult, 128, pp.327~334 (2016)
  38. Vujovic, T. I., "Cryopreservation of Serbian autochthonous Prunus spp. by dropletvitrification", Biologia, 70, pp.1359~1365 (2015)
  39. Wang, L. Y., "An efficient droplet-vitrification cryopreservation for valuable blueberry germplasm", Sci. Hort, 219, pp.60~69 (2017)
  40. Yamamoto, S., "Cryopreservation of in vitro-grown shoot tips of strawberry by the vitrification method using aluminum cryo-plates", Plant Gen. Res, 10, pp.14~19 (2012)
  41. 이정윤, "Efficient Micropropagation of Pear Germplasm Using Soot Tips and Nodal Explants", 한국자원식물학회지, 28(6), pp.690~696 (2015), 한국자원식물학회[DOI]
  42. 이정윤, "Regeneration of Cryopreserved Pear Shoot Tips Grown in Vitro by Encapsulation-Dehydration", 한국자원식물학회지, 30(6), pp.612~617 (2017), 한국자원식물학회[DOI]
  43. Zamecnik, J., "Cryoprotocols used for cryopreservation of vegetatively propagated plants in the Czech cryobank", Adv. Hort. Sci, 21, pp.247~250 (2007)

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