Micropropagation and Acclimatization of Gymnocalycium cv. Fancy (Cactaceae): Developmental Responses to Different Explant Types and Hormone Conditions
Abstract
:1. Introduction
2. Results and Discussion
2.1. Callus
2.1.1. Productivity of TDZ Treatments Related to Callus Production
2.1.2. Efficiency of TDZ Treatments in Relation to Callus/Areole Production
2.2. Shoots
2.2.1. Productivity Related to Shoot Production
2.2.2. Efficiency Related to Shoot Production
2.2.3. Shoot Production Based on Treatment Factor Combination
2.3. General Trends Related to the Induction Period
2.4. General Trends Related to the Growth Period
2.5. Descriptions of Morphotypes
2.5.1. Occurrence of Expected Morphotypes
2.5.2. Occurrence of Unexpected Morphotypes
2.6. Root Emergence
2.7. Acclimatization
3. Materials and Methods
3.1. Plant Material and Disinfection
3.2. In Vitro Establishment and Culture Conditions
3.3. Induction and Tissue Culture Conditions
3.4. Experimental Design
3.5. Statistical Analysis
3.6. Acclimatization
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Explant | Total Areoles Average | Initial Size (1) | Areoles Average (2) |
---|---|---|---|
Apices | 15.32 ± 0.14 | L | 15.58 ± 0.20 |
Apices | M | 15.07 ± 0.20 | |
Bases | 12.70 ± 0.23 | L | 12.24 ± 0.32 |
Bases | M | 13.17 ± 0.31 | |
Central Discs | 4.12 ± 0.09 | L | 4.12 ± 0.09 |
Total Number of Explants | Activated Explants | % of Activated Explants | % of Activated Areoles (1) | ||
---|---|---|---|---|---|
APICES | TDZ1 (2) | 16 | 13 | 81.25 | 14.58 c |
TDZ2 | 16 | 11 | 68.75 | 10.12 c | |
TDZ4 | 16 | 12 | 75.00 | 11.30 c | |
BASES | TDZ1 | 16 | 11 | 68.75 | 13.50 c |
TDZ2 | 16 | 8 | 50.00 | 9.66 c | |
TDZ4 | 16 | 9 | 56.25 | 12.54 c | |
CENTRAL DISCS | TDZ1 | 16 | 7 | 43.75 | 47.55 b |
TDZ2 | 16 | 10 | 62.50 | 32.98 a | |
TDZ4 | 16 | 7 | 43.75 | 35.48 a |
Principal Factors | Gl (1) | Induction Period | Development Period | ||||
---|---|---|---|---|---|---|---|
1st | 2nd | 3rd | 4th | 5th | 6th | ||
Hormones (H) | 2 | 0.311 | 0.001 * | 0.007 * | 0.816 | 0.102 | 0.000 * |
Type of Explant (TE) | 2 | 0.228 | 0.000 * | 0.000 * | 0.000 * | 0.000 * | 0.000 * |
Hormone Concentration (HC) | 3 | 0.319 | 0.891 | 0.461 | 0.008 * | 0.006 * | 0.000 * |
Seedling Size (SS) | 1 | 0.436 | 0.238 | 0.497 | 0.907 | 0.317 | 0.855 |
Total Cases | 576 |
Condition (2) | Induction Period in Presence of PGRs (1) | Development Period in Absence of PGRs | ||||
---|---|---|---|---|---|---|
1st Month | 2nd Month | 3rd Month | 4th Month | 5th Month | 6th Month | |
Control_A | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 |
Control_B | 0.00 ± 0.00 | 0.09 ± 0.05 | 0.34 ± 0.12 | 0.64 ± 0.31 | 1.06 ± 0.25 | 1.22 ± 0.27 |
Control_CD | 0.00 ± 0.00 | 0.75 ± 0.20 | 1.25 ± 0.22 | 1.49 ± 0.32 | 2.03 ± 0.29 | 2.19 ± 0.30 |
BAP2_A | 0.00 ± 0.00 | 0.13 ± 0.13 | 0.13 ± 0.13 | 0.13 ± 0.13 | 0.19 ± 0.14 | 0.25 ± 0.19 |
BAP4_A | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.32± 0.16 | 0.44 ± 0.22 | 0.44 ± 0.22 |
BAP8_A | 0.00 ± 0.00 | 0.25 ± 0.14 | 0.38 ± 0.18 | 0.38 ± 0.18 | 0.44 ± 0.20 | 0.44 ± 0.20 |
BAP2_B | 0.00 ± 0.00 | 0.44 ± 0.27 | 0.63 ± 0.34 | 0.86 ± 0.35 | 1.38 ± 0.58 | 1.56 ± 0.56 |
BAP4_B | 0.00 ± 0.00 | 0.50 ± 0.22 | 1.00 ± 0.26 | 1.18 ± 0.36 | 1.38 ± 0.33 | 1.38 ± 0.33 |
BAP8_B | 0.00 ± 0.00 | 0.88 ± 0.31 | 1.38 ± 0.41 | 2.36 ± 0.28 | 2.88 ± 0.82 | 2.88 ± 0.82 |
BAP2_CD | 0.31 ± 0.22 | 1.06 ± 0.32 | 1.19 ± 0.34 | 1.44 ± 0.29 | 1.94 ± 0.35 | 2.13 ± 0.40 |
BAP4_CD | 0.00 ± 0.00 | 0.69 ± 0.18 | 1.25 ± 0.23 | 1.49 ± 0.30 | 2.25 ± 0.27 | 2.25 ± 0.27 |
BAP8_CD | 0.00 ± 0.00 | 0.38 ± 0.15 | 0.69 ± 0.27 | 1.12 ± 0.34 | 1.69 ± 0.44 | 1.69 ± 0.44 |
KIN2_A | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 |
KIN4_A | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 |
KIN8_A | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.06 ± 0.06 | 0.06 ± 0.06 |
KIN2_B | 0.00 ± 0.00 | 0.25 ± 0.14 | 0.69 ± 0.20 | 1.00 ± 0.35 | 1.44 ± 0.34 | 1.69 ± 0.43 |
KIN4_B | 0.00 ± 0.00 | 0.25 ± 0.14 | 0.50 ± 0.18 | 0.74 ± 0.22 | 1.25 ± 0.19 | 1.50 ± 0.26 |
KIN8_B | 0.00 ± 0.00 | 0.19 ± 0.10 | 0.63 ± 0.24 | 0.86 ± 0.28 | 1.25 ± 0.25 | 1.44 ± 0.27 |
KIN2_CD | 0.00 ± 0.00 | 0.69 ± 0.25 | 1.13 ± 0.27 | 1.44 ± 0.29 | 1.88 ± 0.29 | 1.88 ± 0.29 |
KIN4_CD | 0.00 ± 0.00 | 0.63 ± 0.18 | 1.50 ± 0.27 | 1.87 ± 0.25 | 2.38 ± 0.18 | 2.56 ± 0.26 |
KIN8_CD | 0.00 ± 0.00 | 0.40 ± 0.16 | 0.73 ± 0.21 | 1.06 ± 0.24 | 1.73 ± 0.27 | 1.80 ± 0.30 |
TDZ1_A | 0.00 ± 0.00 | 0.13 ± 0.13 | 0.13 ± 0.13 | 0.74 ± 0.23 | 1.94 ± 0.35 | 2.69 ± 0.33 |
TDZ2_A | 0.06 ± 0.06 | 0.38 ± 0.26 | 0.81 ± 0.39 | 1.01 ± 0.38 | 1.31 ± 0.48 | 2.81 ± 0.54 |
TDZ4_A | 0.06 ± 0.06 | 0.38 ± 0.26 | 0.50 ± 0.26 | 0.80 ± 0.41 | 1.06 ± 0.35 | 3.00 ± 0.43 |
TDZ1_B | 0.00 ± 0.00 | 0.19 ± 0.14 | 0.31 ± 0.20 | 0.75 ± 0.34 | 2.00 ± 0.45 | 3.00 ± 0.50 |
TDZ2_B | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.06 ± 0.06 | 0.86 ± 0.34 | 1.81 ± 0.31 | 2.88 ± 0.36 |
TDZ4_B | 0.00 ± 0.00 | 0.06 ± 0.06 | 0.06 ± 0.06 | 0.57 ± 0.27 | 0.81 ± 0.26 | 2.56 ± 0.62 |
TDZ1_CD | 0.00 ± 0.00 | 0.13 ± 0.09 | 1.00 ± 0.32 | 1.44 ± 0.34 | 2.06 ± 0.44 | 2.69 ± 0.39 |
TDZ2_CD | 0.00 ± 0.00 | 0.00 ± 0.00 | 0.56 ± 0.22 | 1.12 ± 0.28 | 2.06 ± 0.35 | 2.69 ± 0.41 |
TDZ4_CD | 0.00 ± 0.00 | 0.19 ± 0.19 | 0.19 ± 0.19 | 0.57 ± 0.25 | 1.25 ± 0.32 | 1.88 ± 0.29 |
Condition (1) | N° of Explants | M1 | M2 | M3 | M4 |
---|---|---|---|---|---|
Control_A | 48 | 0 | 0 | 0 | 0 |
Control_B | 48 | 45 | 0 | 0 | 0 |
Control_CD | 48 | 84 | 0 | 0 | 0 |
BAP2_A | 16 | 1 | 0 | 0 | 2 |
BAP4_A | 16 | 4 | 0 | 0 | 0 |
BAP8_A | 16 | 2 | 0 | 3 | 0 |
BAP2_B | 16 | 13 | 9 | 0 | 0 |
BAP4_B | 16 | 20 | 1 | 0 | 0 |
BAP8_B | 16 | 31 | 14 | 0 | 1 |
BAP2_CD | 16 | 23 | 1 | 1 | 7 |
BAP4_CD | 16 | 34 | 2 | 0 | 0 |
BAP8_CD | 16 | 17 | 8 | 0 | 2 |
KIN2_A | 16 | 0 | 0 | 0 | 0 |
KIN4_A | 16 | 0 | 0 | 0 | 0 |
KIN8_A | 16 | 1 | 0 | 0 | 0 |
KIN2_B | 16 | 20 | 3 | 0 | 0 |
KIN4_B | 16 | 20 | 0 | 0 | 0 |
KIN8_B | 16 | 10 | 9 | 0 | 0 |
KIN2_CD | 16 | 30 | 0 | 0 | 0 |
KIN4_CD | 16 | 36 | 2 | 0 | 0 |
KIN8_CD | 16 | 22 | 4 | 0 | 0 |
TDZ1_A | 16 | 7 | 0 | 18 | 8 |
TDZ2_A | 16 | 11 | 0 | 9 | 1 |
TDZ4_A | 16 | 11 | 0 | 6 | 0 |
TDZ1_B | 16 | 20 | 0 | 8 | 4 |
TDZ2_B | 16 | 18 | 0 | 7 | 3 |
TDZ4_B | 16 | 3 | 0 | 14 | 0 |
TDZ1_CD | 16 | 25 | 1 | 2 | 5 |
TDZ2_CD | 16 | 16 | 0 | 16 | 4 |
TDZ4_CD | 16 | 11 | 0 | 5 | 4 |
Total | 576 | 535 | 54 | 89 | 41 |
Total % | 74.41 | 7.51 | 12.38 | 5.7 |
Expected Morphotypes (1 + 2) | Morphotype 3 | Morphotype 4 | |||||
---|---|---|---|---|---|---|---|
Cases | p-Value (1) | Mean (2) | p-Value | Mean | p-Value | Mean | |
H (3) | 0.01 | 0.00 | 0.00 | ||||
BAP | 192 | 1.20 ± 0.09 b | 0.02 ± 0.03 a | 0.06 ± 0.03 a | |||
KIN | 192 | 1.04 ± 0.09 ab | 0.00 ± 0.03 a | 0.00 ± 0.03 a | |||
TDZ | 192 | 0.82 ± 0.09 a | 0.44 ± 0.03 b | 0.15 ± 0.03 b | |||
TE | 0.00 | 0.45 | 0.14 | ||||
Apices | 192 | 0.19 ± 0.09 a | 0.19 ± 0.03 a | 0.06 ± 0.03 a | |||
Bases | 192 | 1.21 ± 0.09 b | 0.15 ± 0.03 a | 0.04 ± 0.03 a | |||
Central Discs | 192 | 1.66 ± 0.09 c | 0.12 ± 0.03 a | 0.11 ± 0.03 a | |||
HC | 0.06 | 0.00 | 0.00 | ||||
Control | 144 | 0.89 ± 0.11 a | 0.00 ± 0.04 a | 0.00 ± 0.03 a | |||
Low | 144 | 1.06 ± 0.11 a | 0.20 ± 0.04 b | 0.18 ± 0.03 b | |||
Medium | 144 | 1.14 ± 0.11 a | 0.22 ± 0.04 b | 0.06 ± 0.03 a | |||
High | 144 | 0.99 ± 0.11 a | 0.19 ± 0.04 b | 0.05 ± 0.03 a | |||
H × TE | 0.00 | 0.73 | 0.70 | ||||
H × HC | 0.12 | 0.00 | 0.05 | ||||
TE × HC | 0.12 | 0.01 | 0.58 | ||||
H × TE × HC | 0.01 | 0.00 | 0.48 | ||||
Total cases | 576 | ||||||
Total mean | 1.021 ± 0.061 | 0.155 ± 0.023 | 0.071 ± 0.016 |
PGR (1) | Concentration | Type of Explant | Rooting after 4 Weeks (2) | Rooting after 8 Weeks |
---|---|---|---|---|
CONTROL | 0 | A | 84.38 ab | 90.63 a |
0 | CD | - | - | |
0 | B | - | - | |
BAP | 2 | A | 93.75 a | 93.75 a |
2 | CD | 12.50 d | 37.50 b | |
2 | B | - | - | |
4 | A | 56.25 ab | 75.00 a | |
4 | CD | - | - | |
4 | B | - | - | |
8 | A | 50.00 bc | 93.75 a | |
8 | CD | - | - | |
8 | B | - | - | |
KIN | 2 | A | 75.00 ab | 93.75 a |
2 | CD | - | - | |
2 | B | - | - | |
4 | A | 87.50 ab | 93.75 a | |
4 | CD | 18.75 cd | 25.00 b | |
4 | B | - | - | |
8 | A | 81.25 ab | 87.50 a | |
8 | CD | 18.75 cd | 31.25 b | |
8 | B | - | - | |
TDZ | 1 | A | - | - |
1 | CD | - | - | |
1 | B | - | - | |
2 | A | - | - | |
2 | CD | - | - | |
2 | B | - | - | |
4 | A | - | - | |
4 | CD | - | - | |
4 | B | - | - |
PGRs (1) | Concentration (µM) | Type of Explant | Initial Size (2) | N° of Explants |
---|---|---|---|---|
BAP | 2 | Apex | M | 8 |
Apex | L | 8 | ||
Base | M | 8 | ||
Base | L | 8 | ||
Central Disc | L | 16 | ||
BAP | 4 | Apex | M | 8 |
Apex | L | 8 | ||
Base | M | 8 | ||
Base | L | 8 | ||
Central Disc | L | 16 | ||
BAP | 8 | Apex | M | 8 |
Apex | L | 8 | ||
Base | M | 8 | ||
Base | L | 8 | ||
Central Disc | L | 16 | ||
KIN | 2 | Apex | M | 8 |
Apex | L | 8 | ||
Base | M | 8 | ||
Base | L | 8 | ||
Central Disc | L | 16 | ||
KIN | 4 | Apex | M | 8 |
Apex | L | 8 | ||
Base | M | 8 | ||
Base | L | 8 | ||
Central Disc | L | 16 | ||
KIN | 8 | Apex | M | 8 |
Apex | L | 8 | ||
Base | M | 8 | ||
Base | L | 8 | ||
Central Disc | L | 16 | ||
TDZ | 1 | Apex | M | 8 |
Apex | L | 8 | ||
Base | M | 8 | ||
Base | L | 8 | ||
Central Disc | L | 16 | ||
TDZ | 2 | Apex | M | 8 |
Apex | L | 8 | ||
Base | M | 8 | ||
Base | L | 8 | ||
Central Disc | L | 16 | ||
TDZ | 4 | Apex | M | 8 |
Apex | L | 8 | ||
Base | M | 8 | ||
Base | L | 8 | ||
Central Disc | L | 16 | ||
CONTROL | Apex | M | 16 | |
Apex | L | 16 | ||
Base | M | 16 | ||
Base | L | 16 | ||
Central Disc | L | 32 | ||
Total | 528 |
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Cortés-Olmos, C.; Guerra-Sandoval, V.M.; Blanca-Giménez, V.; Rodríguez-Burruezo, A. Micropropagation and Acclimatization of Gymnocalycium cv. Fancy (Cactaceae): Developmental Responses to Different Explant Types and Hormone Conditions. Plants 2023, 12, 3932. https://doi.org/10.3390/plants12233932
Cortés-Olmos C, Guerra-Sandoval VM, Blanca-Giménez V, Rodríguez-Burruezo A. Micropropagation and Acclimatization of Gymnocalycium cv. Fancy (Cactaceae): Developmental Responses to Different Explant Types and Hormone Conditions. Plants. 2023; 12(23):3932. https://doi.org/10.3390/plants12233932
Chicago/Turabian StyleCortés-Olmos, Carles, Vladimir Marín Guerra-Sandoval, Vicente Blanca-Giménez, and Adrián Rodríguez-Burruezo. 2023. "Micropropagation and Acclimatization of Gymnocalycium cv. Fancy (Cactaceae): Developmental Responses to Different Explant Types and Hormone Conditions" Plants 12, no. 23: 3932. https://doi.org/10.3390/plants12233932