Korean Journal of Plant Resources (한국자원식물학회지)

The Plant Resources Society of Korea (한국자원식물학회)
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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)
  • Received : 2023.10.09
  • Accepted : 2023.11.25
  • Published : 2023.12.01
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Abstract

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.

Keywords

Acknowledgement

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)."

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