Abstract
Panax ginseng C. A. Meyer is a desirable medicinal plant due to its health benefits. With the growing interest in ginseng products, the safe preservation of genetically engineered material with unique attributes is increasingly important. In the present study, an effective protocol for cryopreservation of the ginseng embryogenic callus by vitrification is reported. After 2-wk subculture, the calluses were cold-acclimated at 4°C for 3 wk, then precultured for 2 d with a preculture medium containing 0.3 M sucrose. The precultured embryonic callus was then treated with a loading solution, followed by exposure to plant vitrification solution 2 at 0°C for 90 min. The dehydrated embryogenic calluses were then transferred into fresh 1.5 mL plant vitrification solution 2 and directly immersed in liquid nitrogen. With this protocol, an average of 68.3% regrowth rate for cryopreserved material was obtained, and the number of somatic embryos differentiated from embryogenic callus was significantly increased. Proteomic analysis showed that proteins related to carbohydrate metabolism, the stress response, and oxidative metabolism all significantly up-regulated after cryopreservation. Our results indicate that cold acclimation and preculture may be linked to enhanced cryopreservation tolerance. The up-regulated proteins play an important role in ginseng stress responses and may contribute to cryopreservation tolerance. Carbohydrate metabolism–related protein may be associated with the recovery growth of the cryopreserved embryogenic callus. The increased expression of HSP and 14-3-3-like protein play important roles in the response to stress and could be related with the improvement of somatic embryo production after cryopreservation.
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Funding
This work was supported by Collection of Germplasm Resources and Breeding of High Yield Varieties of Ginseng (2017YFC1702102-02), Investigation, Arrangement and Protection for Ginseng Germplasm Resources and Excellent Genes Mining (20170101011JC), and International Joint Research enter for the Panax ginseng (20200802015GH).
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Lei, X., Wang, Q., Yang, H. et al. Vitrification and proteomic analysis of embryogenic callus of Panax ginseng C. A. Meyer. In Vitro Cell.Dev.Biol.-Plant 57, 118–127 (2021). https://doi.org/10.1007/s11627-020-10117-5
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DOI: https://doi.org/10.1007/s11627-020-10117-5