Vitrification and proteomic analysis of embryogenic callus of Panax ginseng C. A. Meyer
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PLANT TISSUE CULTURE
Vitrification and proteomic analysis of embryogenic callus of Panax ginseng C. A. Meyer Xiujuan Lei 1
&
Qi Wang 1 & He Yang 1 & Yanran Qi 1 & Xiaoli Hao 1 & Yingping Wang 1
Received: 24 February 2020 / Accepted: 17 August 2020 / Editor: Barbara Reed # The Society for In Vitro Biology 2020
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. Keywords Panax ginseng . Embryogenic callus . Vitrification . Cryopreservation . Proteomic analysis
Introduction Panax ginseng C. A. Meyer is a valued medicinal plant in the Araliaceae family. It has been commonly used for thousands of years in East Asian countries such as China, Korea, and Japan (Jung et al. 2018; Kim et al. 2018). The plant provides abundant ginsenosides to millions of cancer, diabetes, and Parkinson’s disease patients (Lee et al. 2018). However, the ginseng commercial and research sectors are facing serious supply challenges. For instance, wild ginseng is almost Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11627-020-10117-5) contains supplementary material, which is available to authorized users. * Yingping Wang [email protected] 1
State Local Joint Engineering Research Center of Ginseng Breeding and Application, Jilin Agricultural University, Changchun 130118, People’s Republic of China
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