Vigorous Growing of Donor Plantlets by Liquid Overlay in Subcultures Is the Key to Cryopreservation of Endangered Species Pogostemon yatabeanus
Abstract
:1. Introduction
2. Results
2.1. Subculture Medium and Supplements in the Last Subculture Cycle (S-0)
2.1.1. Effect of Subculture Medium and Supplements on In Vitro Growth of Donor Plants
2.1.2. Droplet-Vitrification Procedure
2.2. Revitalization of Donor Plants through the Subsequent Subcultures
2.2.1. Selection of Three Subculture Variants
2.2.2. In Vitro Growth of Donor Plantlets during the Subsequent Subculture Cycles
2.2.3. Droplet-Vitrification Procedure Following the Subsequent Subculture Cycles
3. Discussion
3.1. In Vitro Subculture and Propagation System
3.2. Subculture Conditions and Post-Cryopreservation Regeneration
4. Materials and Methods
4.1. Plant Material, In Vitro Establish and Preparation of Mother Plants
4.2. Effect of Subculture Medium and Conditions in the Last Subculture Cycle (S-0)
4.2.1. In Vitro Growth of Donor Plants
4.2.2. Droplet-Vitrification Procedure
4.3. Revitalization of Donor Plants through the Subsequent Subcultures
4.3.1. In Vitro Growth of Donor Plants during the Subsequent Subculture Cycles, S-1→S-0
4.3.2. Droplet-Vitrification Procedure Following the Subsequent Cycles
4.4. Recovery Assessment and Statistical Analysis
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Treatment Sets | Regrowth Steps (5d-3w-2w) | Height | Dry Weight | ||
---|---|---|---|---|---|
Shoots | Roots | Shoots | Roots | ||
Subculture medium and supplements | RM1-RM2-MSF * | 0.49 | 0.62 | 0.85 | 0.90 |
RM2-RM2-MSF | 0.75 | 0.74 | 0.95 | 0.98 |
Code | Subsequent Subculture Cycle | Mean of Three Subculture Combinations each (S-1→S-0) | ||
---|---|---|---|---|
S-1, 5 Weeks | S-0, 5 Weeks | LN Survival (%) | LN Regeneration (%) | |
1→1, 2, 3 | Node + Gell + Liquid-O | Apical + Gell + Liquid-O Node + Gell + Liquid-O Node + Agar + Liquid-X | 77.3 | 72.4 |
2→1, 2, 3 | Node + Gell + Liquid-X | Apical + Gell + Liquid-O Node + Gell + Liquid-O Node + Agar + Liquid-X | 70.6 | 60.7 |
3→1, 2, 3 | Node + Agar + Liquid-X | Apical + Gell + Liquid-O Node + Gell + Liquid-O Node + Agar + Liquid-X | 42.4 | 37.4 |
1, 2, 3→1 | Node + Gell + Liquid-O Node + Gell + Liquid-X Node + Agar + Liquid-X | Apical+Gell+Liquid-O | 77.9 | 74.1 |
1, 2, 3→2 | Node + Gell + Liquid-O Node + Gell + Liquid-X Node + Agar + Liquid-X | Node+Gell+Liquid-O | 71.3 | 65.1 |
1, 2, 3→3 | Node + Gell + Liquid-O Node + Gell + Liquid-X Node + Agar + Liquid-X | Node+Agar+Liquid-X | 41.1 | 31.4 |
No. | Medium | Gelling Agent | Liquid Overlay | Hormone | AC | Code | Remarks |
---|---|---|---|---|---|---|---|
(g L−1) | (mg L−1) | (g L−1) | |||||
1 | MS | Gellan gum 3 | Liquid-O | - | - | Liquid-O | |
2 | MS | Gellan gum 3 | Liquid-X | - | - | Liquid-X | standard |
3 | MS | Agar 8 | Liquid-X | - | - | Agar8 | |
4 | 1/2 MS | Gellan gum 3 | Liquid-X | - | - | 1/2 MSF | |
5 | MS | Gellan gum 3 | Liquid-X | BA 1 | - | BA1 | |
6 | MS | Gellan gum 3 | Liquid-X | - | AC 1 | AC1 |
Repeated Standard Subcultures for PropAgation (S-2 *, 6w) | Subculture Treatment (Gelling Agent and Liquid Overlay, S-1 ** Cycle, 5w) | Subculture Treatment (Section, Gelling Agent, and Liquid Overlay) for Rejuvenation (S-0 ***, 5w) | Code (S-1→S-0 Cycle) |
---|---|---|---|
Node + Gell + Liquid-O | (1) Node + Gell + Liquid-O | (1) Apical + Gell + Liquid-O (2) Node + Gell + Liquid-O (3) Node + Gell + Liquid-X | 1→1 1→2, standard 1→3 |
(2) Node + Gell + Liquid-X | (1) Apical + Gell + Liquid-O (2) Node + Gell + Liquid-O (3) Node + Gell + Liquid-X | 2→1 2→2 2→3 | |
(3) Node + Agar + Liquid-X | (1) Apical + Gell + Liquid-O (2) Node + Gell + Liquid-O (3) Node + Gell + Liquid-X | 3→1 3→2 3→3 |
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Lee, H.; Kim, H. Vigorous Growing of Donor Plantlets by Liquid Overlay in Subcultures Is the Key to Cryopreservation of Endangered Species Pogostemon yatabeanus. Plants 2022, 11, 3127. https://doi.org/10.3390/plants11223127
Lee H, Kim H. Vigorous Growing of Donor Plantlets by Liquid Overlay in Subcultures Is the Key to Cryopreservation of Endangered Species Pogostemon yatabeanus. Plants. 2022; 11(22):3127. https://doi.org/10.3390/plants11223127
Chicago/Turabian StyleLee, Hyoeun, and Haenghoon Kim. 2022. "Vigorous Growing of Donor Plantlets by Liquid Overlay in Subcultures Is the Key to Cryopreservation of Endangered Species Pogostemon yatabeanus" Plants 11, no. 22: 3127. https://doi.org/10.3390/plants11223127