한국자원식물학회지 (Korean Journal of Plant Resources)

  • 제36권6호
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  • Pages.562-570
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  • 2023
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  • 1226-3591(pISSN)
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  • 2287-8203(eISSN)
한국자원식물학회 (The Plant Resources Society of Korea)
권호 표지
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Cryopreservation of in vitro Grown Shoot Tips of Two Freesia hybrida Cultivars by Droplet-vitrification

  • Jinjoo Bae (National Agrobiodiversity Center, NAS, RDA) ;
  • Jae-Young Song (National Agrobiodiversity Center, NAS, RDA) ;
  • Ji-Won Han (National Agrobiodiversity Center, NAS, RDA) ;
  • Ho Cheol Ko (National Agrobiodiversity Center, NAS, RDA) ;
  • Sung-Hee Nam (National Agrobiodiversity Center, NAS, RDA) ;
  • Jung-Ro Lee (National Agrobiodiversity Center, NAS, RDA) ;
  • Ho-sun Lee (National Agrobiodiversity Center, NAS, RDA)
  • 투고 : 2023.10.09
  • 심사 : 2023.11.25
  • 발행 : 2023.12.01
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초록

The droplet-vitrification technique for cryopreservation has proven successful across a diverse range of germplasm, ensuring safe and effective long term preservation. In this study, we investigate an effective cryopreservation protocol using the droplet-vitrification technique for shoot tips of Freesia hybrida cultivars 'Sunny Gold' and 'Sweet Lemon'. To determine optimal conditions for Freesia cryopreservation, we employed a carefully selected standard procedure along with additional treatments and alternative solutions. For 'Sunny Gold', the highest regrowth rate of 24% was achieved when shoot tips underwent dehydration with PVS3 solution for 120 minutes before direct immersion in liquid nitrogen (LN) for 1 hour, coupled with a standard protocol involving a two-step preculture with 0.3 M - 0.5 M sucrose, loading with C4 for 40 minutes, and unloading with 0.8 M sucrose for 40 minutes. In the case of 'Sweet Lemon,' regrowth of cryopreserved shoot tips was observed with dehydration treatments, including PVS2 (A3) for 60 minutes and PVS3 (B1) for 60 minutes, as well as longer exposure. The results reflect the distinct sensitivity of shoot tips to chemical toxicity and osmotic stress in these two genotypes. This study provides valuable evidence to consistently enhance the effectiveness of cryopreservation methods for the long-term conservation of Freesia germplasm.

키워드

과제정보

This study was conducted with the support of the National Institute of Agricultural Sciences, Rural Development Administration, Republic of Korea, under the project titled "Development and application of cryopreservation protocol for pear and freesia germplasm (Project No. PJ016811)."

참고문헌

  1. Bae, J., S.Y. Lee, J.Y. Song, J.R. Lee, M.Yoon, J.Y. Yi, H.H. Kim and Y.Y. Lee. 2021. Efficient cryopreservation of in vitro grown shoot tips of strawberry (Fragaria × ananassa Duch.) germplasm using droplet-vitrification. Korean J. Plant Res. 34(6):600-607.
  2. Bae, J., Y. Choi, J.Y. Song, J.R. Lee, M. Yoon and Y.Y. Lee. 2022. Genetic stability assessment of six cryopreserved strawberry (Fragaria × ananassa Duch.) accessions by phenotypic and molecular studies. Biology 11:1746.
  3. Baghdadi, S.H., I. Makhadmeh, M. Syouf, A. Arabiat, R.A. Shibli and M.A. Shatnawai. 2011. Cryopreservation by vitrification of embryogenic callus of wild crocus (Crocus Hyemalis and Crocus Moabiticus). Acta Hort. 908:30.
  4. Chmielarz, P., G. March and M.T. de Boucaud. 2005. Cryopreservation of Quercus robur L. embryogenic calli. Cryo Lett. 26:349-356.
  5. Choi, Y.J., H.R. Cho, H.Y. Jeong, D.H. Goo, Y.I. Kang, B.N. Chung, J.Y. Yoon and S.K. Choi. 2016. Virus identification using RT-PCR method from meristem cultured and ribavirin treated plantlet for producing FreMV-free Freesia 'Shiny Gold'. J. Korean Soc. People Plants Environ. 19(1):19-23. https://doi.org/10.11628/ksppe.2016.19.1.019
  6. Chung, J.W., Y. Lee, G.A. Lee and J.Y. Yi. 2014. Development of cryo-banking system of Lilium species. Flower Res. J. 22 (3):185-189. https://doi.org/10.11623/frj.2014.22.3.13
  7. Engelmann, F. 2011. Use of biotechnologies for the conservation of plant biodiversity. In Vitro Cell. Dev. Biol. 47:5-16. https://doi.org/10.1007/s11627-010-9327-2
  8. Folgado, R., B. Panis, K. Sergeant, J. Renaut, R. Swennen and J.F. Hausman. 2015. Unravelling the effect of sucrose and cold pretreatment on cryopreservation of potato through sugar analysis and proteomics. Cryobiology 71:432-441. https://doi.org/10.1016/j.cryobiol.2015.09.006
  9. Gao, X., D. Yang, D. Cao, M. Ao, X. Sui, Q. Wang, J.N. Kimatu and L. Wang. 2010. In vitro micropropagation of Freesia hybrida and the assessment of genetic and epigenetic stability in regenerated plantlets. J. Plant Growth Regul. 29:257-267. https://doi.org/10.1007/s00344-009-9133-4
  10. Kaczmarczyk, A., S.R. Turner, E. Bunn, R.L. Mancera and K.W. Dixon. 2011. Cryopreservation of threatened native Australian species - what have we learned and where to. In Vitro Cell. Dev. Biol. Plant 47:17-25. https://doi.org/10.1007/s11627-010-9318-3
  11. Kang, J.S., K.J. Jeong, Y.J. Choi and J.G. Yun. 2017. Optimal culture conditions for the shoot growth of Freesia 'Shiny Gold' during bioreactor culture. Korean J. Plant Res. 30(6):699-706.
  12. Kim, H.H. and E. Popova. 2023. Unifying principles of cryopreservation protocols for new plant materials based on alternative cryoprotective agents (CPAS) and a systematic approach. Cryo Lett. 44(1):1-12. https://doi.org/10.54680/fr23110110112
  13. Kim, H.H., J.G. Lee, D.J. Shin, H.C. Ko, J.G. Gwag, E.G. Cho and F. Engelmann. 2009. Development of alternative plant vitrification solutions in droplet-vitrification procedures. Cryo Lett. 30:320-334.
  14. Kulus, D. and M. Zalewska. 2014. Cryopreservation as a tool used in long-term storage of ornamental species. a review. Sci. Hort. 168(26):88-107. https://doi.org/10.1016/j.scienta.2014.01.014
  15. Lee, Y.Y., S.Y. Lee, J.Y. Song, M. Yoon, J.Y. Yi, J.R. Lee and H.H. Kim. 2020. Preculture condition for stable recovery of strawberry (Fragaria × ananassa Duch.) shoot tips after cryopreservation using droplet vitrification. Korean J. Plant Res. 33(6):675-681.
  16. Liu, X.X., Y.B. Wen, Z.H. Cheng, and S.W. Moub. 2017. Establishment of a garlic cryopreservation protocol for shoot apices from adventitious buds in vitro. Sci. Hort. 226:10-18. https://doi.org/10.1016/j.scienta.2017.08.016
  17. Maslanka, M. and A. Szewczyk. 2021. Droplet-vitrifcation cryopreservation of Tulipa tarda Stapf. apical meristems. Plant Cell, Tissue Organ Cult. 144:91-95. https://doi.org/10.1007/s11240-020-01910-6
  18. Maslanka, M., B. Panis and A. Bach. 2013. Cryopreservation of Galanthus elwesii Hook. apical meristems by droplet vitrification. Cryo Lett. 34:1-9.
  19. Maslanka, M., B. Panis and M. Malik. 2016. Cryopreservation of Narcissus L. 'Carlton' somatic embryos by droplet vitrification. Propag. Ornam. Plants 16:28-35.
  20. Merzougui, S.E., C. Benelli, R.E. Boullani and M.A. Serghini. 2023. The cryopreservation of medicinal and ornamental geophytes: application and challenges. a review. Plants 12: 2143.
  21. Mubbarakh, S.A., S. Rahmah, Z.A. Rahman, N.N.M. Sah and S. Subramaniam. 2014. Cryopreservation of Brassidium shooting star orchid using the PVS3 method supported with preliminary histological analysis. Appl. Biochem. Biotechnol. 172:1131-1145. https://doi.org/10.1007/s12010-013-0597-0
  22. Murashige, T. and F. Skoog. 1962. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 15:473-497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
  23. Panis, B., B. Piette and R. Swennen. 2005. Droplet-vitrification of apical meristems: a cryopreservation protocol applicable to all Musaceae. Plant Sci. 168:45-55. https://doi.org/10.1016/j.plantsci.2004.07.022
  24. Park, S.U. and H.H. Kim. 2015. Cryopreservation of sweet potato tips using a droplet-vitrification procedure. Cryo Lett. 36:344-352.
  25. Popova, E., I. Kulichenko and H.H. Kim. 2023. Critical role of regrowth conditions in post-cryopreservation of in vitro plant germplasm. Biology 12:542.
  26. Pourkhaloee, A. and M. Khosh-Khui. 2015. Investigation of callogenesis and indirect regeneration of Freesia × hybrida Bailey 'Argenta'. Folia Hort. 27(2):123-134. https://doi.org/10.1515/fhort-2015-0022
  27. Rajasekharan, P.E., T.M. Rao, T. Janakiram, and S. Ganeshan. 1994. Freeze preservation of gladiolus pollen. Euphytica 80: 105-109. https://doi.org/10.1007/BF00039304
  28. Shibli, R.A. 2000. Cryopreservation of black iris (Iris nigricans) somatic embryos by encapsulation-dehydration. Cryo Lett. 21(1):39-46.
  29. Song, J.Y., Y.Y. Lee, J.Y. Yi, J.R. Lee and M. Yoon. 2020. Preconditioning for cryopreservation of in vitro grown bulblets of lily using droplet-vitrification. Korean J. Plant Res. 33(6):689-695.
  30. Vujovic, T., T. Andelic, B. Vasilijevic, D. Jevremovic and F. Engelmann. 2023. Cryopreservation of indigenous plums and monitoring of multiplication and rooting capacity of shoots obtained from cryopreserved specimens. Plants 12: 3108.
  31. Wang, M.R., M. Lambardi, F. Engelmann, R. Pathirana, B. Panis, G.M. Volk and Q.C. Wang. 2021. Advances in cryopreservation of in vitro-derived propagules: technologies and explant sources. a review. Plant Cell, Tissue and Organ Cult. 144:7-20. https://doi.org/10.1007/s11240-020-01770-0
  32. Yi, J.Y., K. Balaraju, H.J. Baek, M.S. Yoon, H.H. Kim and Y.Y. Lee. 2018. A successful regeneration from shoot tips of Chrysanthemum morifolium (Ramat.) following cryopreservation by droplet-vitrification. Korean J. Plant Res. 31(6): 675-683.