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Title: Cryopreservation of in vitro-grown shoot tips of Chinese medicinal plant Atractylodes macrocephala Koidz. using a droplet-vitrification method

Author
item ZHANG, JIN-MEI - Chinese Academy Of Agricultural Sciences
item HUANG, BIN - Chinese Academy Of Agricultural Sciences
item LU, XIN-XIONG - Chinese Academy Of Agricultural Sciences
item Volk, Gayle
item XIN, XIA - Chinese Academy Of Agricultural Sciences
item YIN, GUANG-KUN - Chinese Academy Of Agricultural Sciences
item HE, JUAN-JUAN - Chinese Academy Of Agricultural Sciences
item CHEN, XIAO-LING - Chinese Academy Of Agricultural Sciences

Submitted to: CryoLetters
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/1/2015
Publication Date: 6/30/2015
Citation: Zhang, J., Huang, B., Lu, X., Volk, G.M., Xin, X., Yin, G., He, J., Chen, X. 2015. Cryopreservation of in vitro-grown shoot tips of Chinese medicinal plant Atractylodes macrocephala Koidz. using a droplet-vitrification method. CryoLetters. 36(3):195-204.

Interpretive Summary: Atractylodes macrocephala is an important medicinal species from China, equal in significance to Ginseng. Consumed as dried, ground roots, Atractylodes macrocephala has purported antioxidant, hepatoprotective, anti-inflammatory, anti-allergic, antithrombotic, antiviral, and anticarcinogenic properties. This research was performed to identify a cryopreservation method to conserve key clonally propagated cultivars of this species. Shoot tips (2-3 mm) were excised from in vitro plants, precultured, treated with cryoprotectants and then placed into PVS2 cryoprotectant solution prior to plunging into liquid nitrogen. Upon warming, shoot tip survival and regrowth levels were 76% and 62%, respectively, after liquid nitrogen exposure. This method will be used to conserve Atractylodes macrocephala genetic resources in the National Genebank, Institute of Crop Science, Chinese Academy of Agricultural Sciences in Beijing, China.

Technical Abstract: BACKGROUND: Atractylodes macrocephala Koidz. is an important medicinal species from China and has been used for thousands of years because of pharmacological antioxidant, hepatoprotective, anti-inflammatory, anti-allergic, antithrombotic, antiviral, and anticarcinogenic activities. OBJECTIVE: The aim of this research was to develop a droplet-vitrification protocol for A. macrocephala shoot tips which could be used as a strategy for long-term conservation within gene banks. MATERIALS AND METHODS: The durationof preculture, loading, and PVS2 steps, as well as the recovery medium formulation, were optimized to achieve high levels of survival and regrowth for A. macrocephala shoot tips after liquid nitrogen exposure. RESULTS: Survival and regrowth levels after cryopreservation were as high as 76% and 62%, respectively, in cultivar ‘Baizhu’. Thermalanalysis using differential scanning calorimetry suggested that the PVS2 treatment plays a critical role for successful cryopreservation. CONCLUSION: The droplet-vitrification method established in this study could be used to cryopreserve A. macrocephala.