Shoot Tip Cryopreservation of Lamprocapnos spectabilis (L.) Fukuhara Using Different Approaches and Evaluation of Stability on the Molecular, Biochemical, and Plant Architecture Levels.

Publisher: MDPI Country of Publication: Switzerland NLM ID: 101092791 Publication Model: Electronic Cited Medium: Internet ISSN: 1422-0067 (Electronic) Linking ISSN: 14220067 NLM ISO Abbreviation: Int J Mol Sci Subsets: MEDLINE

Original Publication: Basel, Switzerland : MDPI, [2000-

Cryopreservation/*methods ; Papaveraceae/*physiology ; Plant Shoots/*physiology ; Plants, Medicinal/*physiology; Chlorophyll/analysis ; Chlorophyll A/analysis ; Cryoprotective Agents ; Desiccation ; Genetic Markers ; Genetic Variation ; Glycerol ; Polymerase Chain Reaction ; Random Amplified Polymorphic DNA Technique ; Sucrose ; Vitrification

The aim of this study is to optimize and evaluate the effectiveness of vitrification, droplet-vitrification, and encapsulation-vitrification techniques in the cryopreservation of Lamprocapnos spectabilis (L.) Fukuhara 'Gold Heart', a popular medicinal and ornamental plant species. In vitro-derived shoot tips were used in the experiments. All three techniques were based on explant dehydration with plant vitrification solution 3 (PVS3; 50% glycerol and 50% sucrose) for 0, 30, 60, 90, 120, 150, or 180 min. The recovered microshoots were subjected to morphometric, biochemical, and molecular analyses (RAPD, ISSR, SCoT). The highest recovery level was reported with the encapsulation-vitrification protocol based on 150 min dehydration (73.1%), while the vitrification technique was the least effective (maximum 25.8% recovery). Explants cryopreserved with the encapsulation-vitrification technique produced the highest mean number of shoots (4.9); moreover, this technique was optimal in terms of rooting efficiency. The highest fresh weight of shoots, on the other hand, was found with the vitrification protocol based on a 30-min PVS3 treatment. The concentrations of chlorophyll a and b were lower in all cryopreservation-derived plants, compared to the untreated control. On the other hand, short dehydration and cryopreservation of non-encapsulated explants stimulated the synthesis of anthocyanins. A small genetic variation in 5% of all samples analyzed was detected by RAPD and ISSR marker systems. Only plants recovered from the encapsulation-vitrification protocol had no DNA sequence alternations.
Competing Interests: The author declares no conflict of interest.

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Keywords: ISSR; PCR; PVS3; RAPD; SCoT; biochemical array; genetic stability; long-term storage; medicinal plants; molecular markers

0 (Cryoprotective Agents)
0 (Genetic Markers)
1406-65-1 (Chlorophyll)
57-50-1 (Sucrose)
5712ZB110R (chlorophyll b)
PDC6A3C0OX (Glycerol)
YF5Q9EJC8Y (Chlorophyll A)

Date Created: 20200604 Date Completed: 20210330 Latest Revision: 20240730

20240730

PMC7311993

10.3390/ijms21113901

32486149