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Chrysanthemum low-temperature storage and cryopreservation: a review

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Abstract

Chrysanthemum (Dendranthema × grandiflora (Ramat.) Kitamura) is an ornamental plant that responds well to in vitro growth conditions. This receptivity makes it a particularly attractive target for low-temperature storage and cryopreservation studies. This review examines in detail the protocols thus far used to achieve the short- to long-term low-temperature and cryostorage of important chrysanthemum germplasm. Occasionally, medicinal chrysanthemum species have also been cryostored, and these studies are also examined in detail. Since chrysanthemum is sensitive to both osmotic stress and chemical toxicity of vitrification solutions, a generalized protocol for the cryopreservation of chrysanthemum apical or axillary shoot tips is proposed: excision of apical or axillary shoot tips after 4 or 7 weeks, respectively, from final subculture; progressive preculture with 10 % sucrose for 31 h, 17.5 % sucrose for 17 h, then 25 % sucrose for 7 h; osmoprotection with 17.5 % glycerol + 17.5 % sucrose for 40 min; cryoprotection with PVS3 (50 % glycerol + 50 % sucrose) vitrification solution for 60 min (axillary) or 90 min (apical); cooling and warming using aluminium foil strips; unloading with 30 % sucrose for 40 min. When smaller axillary shoot tips are used, cryoprotection of samples with 37.5 % glycerol + 15 % DMSO + 15 % ethylene glycol + 22.5 % sucrose at 0 °C for about 60 min can be applied.

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Abbreviations

EG:

Ethylene glycol

LN:

Liquid nitrogen

LNC:

Liquid nitrogen control (dehydrated control)

LS:

Loading solution

LTS:

Low-temperature storage

PVS2:

Plant vitrification solution 2

PVS3:

Plant vitrification solution 3

SIM:

Shoot induction medium

Synseed:

Synthetic seed

VS:

Vitrification solution

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Acknowledgments

The authors are grateful to Dr. Jinmei Zhang, National Genebank of CAAS, for providing relevant Chinese articles.

Conflict of interest

The authors declare no conflicts of interest, financial or other.

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Authors and Affiliations

  1. P. O. Box 7, Miki-cho post office, Ikenobe 3011-2, Kagawa-ken, 761-0799, Japan

    Jaime A. Teixeira da Silva

  2. Department of Well-being Resources, Sunchon National University, Suncheon, 540-742, Korea

    Haenghoon Kim

  3. IRD, UMR DIADE, 911 Avenue Agropolis, BP 64501, 34394, Montpellier Cedex 5, Montpellier, France

    Florent Engelmann

Authors
  1. Jaime A. Teixeira da Silva
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  2. Haenghoon Kim
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  3. Florent Engelmann
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Corresponding authors

Correspondence to Jaime A. Teixeira da Silva, Haenghoon Kim or Florent Engelmann.

Additional information

In memoriam: We dedicate this review to the legend of Prof. Akira Sakai, who passed away on October 5, 2013. Prof. Sakai advanced our knowledge on cryopreservation for plant tissues, including of chrysanthemum.

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Teixeira da Silva, J.A., Kim, H. & Engelmann, F. Chrysanthemum low-temperature storage and cryopreservation: a review. Plant Cell Tiss Organ Cult 120, 423–440 (2015). https://doi.org/10.1007/s11240-014-0641-y

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