Cryopreservation of Bleeding Heart (Lamprocapnos Spectabilis (L.) Fukuhara) Shoot Tips Using Encapsulation-Dehydration

BACKGROUND: Bleeding heart (Lamprocapnos spectabilis (L.) Fukuhara) is a popular vegetatively-propagated ornamental and medicinal plant species with recalcitrant seeds. Efforts have been made to increase the species diversity. However, there has been no research related to cryopreservation of any Fumariaceae species. OBJECTIVE: The aim of this study was to develop a cryopreservation protocol of Lamprocapnos spectabilis 'White Gold', and to evaluate genetic stability of the recovered plantlets. MATERIALS AND METHODS: Shoot tips precultured on Murashige and Skoog medium enriched with 9% sucrose and 10 μM abscisic acid were embedded in 3% calcium alginate, osmotically dehydrated in a sucrose gradient, and then air-desiccated for 0 to 5 h. After storing in liquid nitrogen (LN), the rewarmed shoot tips were placed on recovery media with various plant growth regulators. Morphometric analysis of the recovered shoots was performed. The genetic stability was evaluated using RAPD, ISSR and SCoT markers. RESULTS: The highest recovery of cryopreservation-derived explants (31.3 – 36.4%) was reported after 4-h desiccation and regrowth on cytokinin-supplemented medium. Shoots recovered from explants subjected to this cryoprocedure were longer and of higher fresh weight compared with the control. Cryopreservation also positively affected the further development of leaves. No genetic variation was detected by any of the three marker systems tested. CONCLUSION: Lamprocapnos spectabilis shoot tips have a limited dehydration tolerance. However, stress related to the cryoprocedure positively affects the development of shoots. Even though cryopreservation affects the viability of explants, it does not alter their DNA sequence.