Vitrification and proteomic analysis of embryogenic callus of Panax ginseng C. A. Meyer

• Published in: In vitro cellular & developmental biology. Plant 2021-02, Vol.57 (1), p.118-127
• Main Authors: Lei, Xiujuan, Wang, Qi, Yang, He, Qi, Yanran, Hao, Xiaoli, Wang, Yingping
• Format: Article
• Language: English
• Subjects: 14-3-3 protein; Acclimation; Acclimatization; Biomedical and Life Sciences; Callus; Carbohydrate metabolism; Carbohydrates; Cell Biology; Cold; Cold acclimation; Cold tolerance; Cryopreservation; Dehydration; Developmental Biology; Embryos; Genetic engineering; Germplasm; Ginseng; Glycerol; Herbal medicine; Life Sciences; Liquid nitrogen; Medicinal plants; Metabolism; Nitrogen; Oxidative metabolism; Panax ginseng; Plant Breeding/Biotechnology; Plant Genetics and Genomics; Plant Sciences; PLANT TISSUE CULTURE; Protein turnover; Proteins; Regrowth; Somatic embryos; Subculture; Sucrose; Sugar; Vitrification
• ISSN: 1054-5476; 1475-2689
• DOI: 10.1007/s11627-020-10117-5

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 crypreservation 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.