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Application of cryobanking for Platycodon grandiflorum in vitro axillary buds using cryo-plate methods

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Abstract

The application of the V cryo-plate method using either PVS2 or PVS3 and the D cryo-plate method to Platycodon grandiflorum in vitro axillary buds was investigated. Precultured buds (1 mm) were attached to cryo-plates using calcium alginate gel. Then, osmoprotection treatment (2 M glycerol and 1 M sucrose) was performed by immersing the cryo-plates for 30 min at 25°C. The procedures for each cryogenic method were as follows: in the V cryo-plate method, the buds on cryo-plates were exposed to PVS2 for 40 min (highest regrowth 71.1%) or PVS3 for 50 min (highest regrowth 82.2%). In the D cryo-plate method, the buds on the cryo-plates were dehydrated under the air current of a laminar air flow cabinet for 60 min (highest regrowth 80.0%) at 25°C. Then, the cryo-plates were plunged directly into liquid nitrogen. After cryopreservation, buds on the cryo-plates were rewarmed and unloaded by immersion in 1 M sucrose solution for 15 min at 25°C for subsequent plant regeneration. The average regrowth seen on the V cryo-plates exposed to PVS3 and in the D cryo-plate method was higher than that of the V cryo-plates exposed to PVS2. The regrowth after cryopreservation in the V cryo-plate method with PVS2 was not stable. Thus, the V cryo-plate method with PVS3 and the D cryo-plate method are considered to be practical cryopreservation methods for P. grandiflorum germplasm.

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Funding

This study was mainly funded by a grant from the Japan Agency for Medical Research and Development (jp20ak0101046). The authors thank the faculty of Life and Environmental Sciences in Shimane University for their partly financial support for the publication of this report.

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Authors and Affiliations

  1. Faculty of Life and Environmental Sciences, Shimane University, Matsue, Shimane, Japan

    Toshikazu Matsumoto

  2. Genetic Resources Center, National Agriculture and Food Research Organization, Tsukuba, Ibaraki, Japan

    Daisuke Tanaka & Shin-ichi Yamamoto

  3. Tsukuba Division, Research Center for Medicinal Plant Resources, National Institutes of Biomedical Innovation, Health and Nutrition, Tsukuba, Ibaraki, Japan

    Kayo Yoshimatsu, Noriaki Kawano & Nobuo Kawahara

  4. Department of Science and Technology Innovation, Nagaoka University of Technology, Nagaoka, Niigata, Japan

    Shinya Maki

  5. Formary University of Tsukuba, Tsukuba, Japan

    Takao Niino

Authors
  1. Toshikazu Matsumoto
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  2. Daisuke Tanaka
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  3. Kayo Yoshimatsu
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  4. Noriaki Kawano
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  5. Nobuo Kawahara
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  6. Shinya Maki
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  7. Shin-ichi Yamamoto
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  8. Takao Niino
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Correspondence to Toshikazu Matsumoto.

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Matsumoto, T., Tanaka, D., Yoshimatsu, K. et al. Application of cryobanking for Platycodon grandiflorum in vitro axillary buds using cryo-plate methods. In Vitro Cell.Dev.Biol.-Plant 57, 15–20 (2021). https://doi.org/10.1007/s11627-020-10119-3

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  • DOI: https://doi.org/10.1007/s11627-020-10119-3

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