Micropropagation and Genetic Fidelity of Fegra Fig (Ficus palmata Forssk.) and Grafting Compatibility of the Regenerated Plants with Ficus carica
Abstract
:1. Introduction
2. Results and Discussion
2.1. Surface Disinfection and Culture Initiation of F. palmata
2.2. Effect of Cytokinin Concentrations on Axillary Shoot Multiplication
2.3. Effect of Light Intensity and Sucrose Concentration on Axillary Shoot Multiplication
2.4. The Effect of Dark Incubation versus Standard Photoperiod Conditions on the Growth and Elongation of Axillary Shoots
2.5. Effect of Auxins Concentrations on In Vitro Rooting of F. palmata Axillary Shoots
2.6. Effect of Initial Dark Incubation versus Standard Photoperiod Conditions on In Vitro Rooting of F. palmata Microshoots
2.7. Acclimatization
2.8. Assessment of Genetic Fidelity Using RAPD, ISSR, and SCoT Molecular Markers
2.9. Grafting Compatibility of Micropropagated F. palmata Plantlets as a Rootstock for F. carica ‘Brown Turkey’
3. Materials and Methods
3.1. Plant Material, Surface Sterilization, and Establishment of Aseptic Culture
3.2. Standard Medium Preparation and Growth Conditions
3.3. The Effect of Cytokinin Concentrations on Axillary Shoot Multiplication
3.4. The Effect of Sucrose Concentrations and Light Intensity on Axillary Shoot Multiplication
3.5. The Effect of Dark Incubation versus Standard Photoperiod Conditions on the Growth and Elongation of Axillary Shoots
3.6. The Effect of Auxins Concentrations on In Vitro Rooting of F. palmata Axillary Shoots
3.7. The Effect of Initial Dark Incubation versus Standard Photoperiod Conditions on In Vitro Rooting of F. palmata Axillary Shoots
3.8. Acclimatization
3.9. The Genetic Fidelity of F. palmata Micropropagated Plants
3.9.1. Plant Materials and Genomic DNA Extraction
3.9.2. Randomly Amplified Polymorphic DNA (RAPD) Analysis
3.9.3. Inter Simple Sequence Repeats (ISSR) Analysis
3.9.4. Start Codon Targeted (SCoT) Analysis
3.10. Grafting
3.11. Experimental Design and Data Analysis
4. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Treatments | Number of Shoots/Explant | Length of the Longest Shoot/Explant (cm) | Fresh Weight/Explant (g) | Dry Weights/Explant (g) | |
---|---|---|---|---|---|
Type of cytokinin | |||||
BAP | 7.4 a | 1.9 a | 2.456 b | 0.302 a | |
TDZ | 8.5 a | 1.1 b | 4.167 a | 0.351 a | |
Zeatin | 3.1 b | 1.3 b | 0.716 c | 0.143 b | |
p-value | <0.0001 *** | <0.0001 *** | <0.0001 *** | <0.0001 *** | |
Concentrations (mg/L) | |||||
Control (C0) | 1.4 d | 1.1 b | 0.142 c | 0.035 c | |
C1 | 4.4 c | 1.3 ab | 1.941 b | 0.215 b | |
C2 | 7.3 a | 1.6 a | 2.621 a | 0.283 a | |
C3 | 7.5 a | 1.6 a | 2.755 a | 0.300 a | |
C4 | 6.0 b | 1.3 ab | 2.470 a | 0.263 ab | |
p-value | <0.0001 *** | 0.027 * | <0.0001 *** | <0.0001 *** | |
Type of cytokinin × concentration (mg/L) | |||||
Control | C0 | 1.4 e | 1.2 def | 0.142 d | 0.035 d |
BAP | C1 | 3.2 de | 1.7 bc | 0.957 d | 0.185 b |
C2 | 7.4 c | 2.3 a | 2.226 c | 0.305 a | |
C3 | 11.2 a | 1.9 ab | 3.641 ab | 0.389 a | |
C4 | 7.7 c | 1.7 bc | 3.001 bc | 0.331 a | |
TDZ | C1 | 8.5 bc | 1.4 bcde | 4.601 a | 0.378 a |
C2 | 9.8 ab | 1.0 ef | 4.658 a | 0.364 a | |
C3 | 8.0 bc | 1.0 ef | 3.646 b | 0.328 a | |
C4 | 7.5 c | 1.0 ef | 3.765 ab | 0.334 a | |
Zeatin | C1 | 1.5 e | 0.7 f | 0.265 d | 0.080 cd |
C2 | 4.8 d | 1.5 bcde | 0.978 d | 0.182 b | |
C3 | 3.3 de | 1.8 ab | 0.977 d | 0.183 b | |
C4 | 2.8 e | 1.3 cde | 0.646 d | 0.126 bc | |
p-value | <0.0001 *** | 0.0001 *** | < 0.0001 *** | 0.0001 *** |
Treatments | Number of Shoots/Explant | Length of the Longest Shoot/Explant (cm) | Fresh Weight/Explant (g) | Dry Weights/Explant (g) | |
---|---|---|---|---|---|
Light intensity (PPFD; μmol m−2·s−1) | |||||
15 | 5.1 b | 6.1 a | 3.275 ab | 0.337 a | |
35 | 7.2 a | 6.3 a | 3.791 a | 0.349 a | |
70 | 3.7 b | 5.1 b | 2.376 b | 0.321 a | |
p-value | <0.0001 *** | <0.0001 *** | <0.0001 *** | 0.057 NS | |
Sucrose (%) | |||||
1.5 | 6.6 ab | 6.8 a | 3.752 ab | 0.330 b | |
3.0 | 7.1 a | 6.2 ab | 4.346 a | 0.420 a | |
4.5 | 5.1 b | 5.7 b | 3.139 b | 0.382 ab | |
6.0 | 2.7 c | 4.5 c | 1.352 c | 0.213 c | |
p-value | <0.0001 *** | <0.0001 *** | <0.0001 *** | <0.0001 *** | |
Light intensity × sucrose (%) | |||||
15 | 1.5 | 6.6 bc | 7.0 a | 3.965 b | 0.273 f |
3.0 | 6.0 c | 6.0 bc | 3.669 b | 0.345 d | |
4.5 | 4.5 d | 6.9 ab | 3.525 b | 0.439 b | |
6.0 | 3.6 de | 4.7 ef | 1.944 d | 0.295 ef | |
35 | 1.5 | 7.4 b | 6.7 ab | 3.698 b | 0.340 de |
3.0 | 10.9 a | 7.5 a | 6.659 a | 0.514 a | |
4.5 | 7.5 b | 5.7 cd | 3.562 b | 0.348 d | |
6.0 | 3.0 e | 5.1 de | 1.245 e | 0.197 g | |
70 | 1.5 | 5.8 c | 6.7 ab | 3.593 b | 0.377 cd |
3.0 | 4.3 d | 5.1 de | 2.710 c | 0.401 bc | |
4.5 | 3.2 e | 4.6 ef | 2.332 cd | 0.358 cd | |
6.0 | 1.4 f | 3.9 f | 0.868 e | 0.149 h | |
p-values | <0.0001 *** | 0.001 *** | <0.0001 *** | <0.0001 *** |
Treatments | Rooting (%) | Number of Roots/Explant | Length of Root (cm) | Fresh Weight/Explant (g) | Dry Weight/Explant (g) | |
---|---|---|---|---|---|---|
Type of auxin | ||||||
IBA | 35.0 b | 1.0 a | 0.9 b | 0.852 b | 0.204 b | |
NAA | 30.0 b | 0.8 a | 0.5 b | 1.510 a | 0.263 a | |
IAA | 50.0 a | 1.3 a | 2.3 a | 0.848 b | 0.208 b | |
p-value | 0.011 * | 0.520 NS | 0.002 ** | 0.001 *** | 0.007 ** | |
Concentration (mg/L) | ||||||
Control | 41.6 a | 1.2 ab | 2.8 a | 0.937 a | 0.223 ab | |
0.1 | 41.6 a | 1.8 a | 1.7 abc | 0.928 a | 0.211 b | |
0.5 | 38.8 a | 0.5 b | 0.6 c | 0.964 a | 0.208 b | |
1.0 | 50.0 a | 1.1 ab | 1.8 ab | 1.208 a | 0.257 a | |
2.0 | 19.4 b | 0.6 b | 0.8 bc | 1.179 a | 0.223 ab | |
p-value | 0.013 * | 0.028 * | 0.001 *** | 0.109 NS | 0.168 NS | |
Type of auxin × Concentration (mg/L) | ||||||
Control | 0.0 | 41.7 b | 1.2 abcd | 2.8 ab | 0.937 cde | 0.223 abc |
IBA | 0.1 | 33.3 b | 2.1 ab | 1.6 bcd | 0.754 de | 0.192 bc |
0.5 | 33.3 b | 0.7 bcd | 0.2 d | 0.726 e | 0.179 c | |
1.0 | 41.7 b | 0.8 abcd | 1.1 bcd | 0.961 cde | 0.233 abc | |
2.0 | 25.0 bc | 0.4 cd | 0.8 cd | 0.967 cde | 0.212 abc | |
NAA | 0.1 | 41.7 b | 2.3 a | 1.1 bcd | 1.239 bcd | 0.243 abc |
0.5 | 33.3 b | 0.1 d | 0.3 d | 1.352 bc | 0.248 abc | |
1.0 | 33.3 b | 0.8 abcd | 0.5 cd | 1.545 ab | 0.284 a | |
2.0 | 0.0 c | 0.0 d | 0.0 d | 1.903 a | 0.278 a | |
IAA | 0.1 | 50.0 ab | 1.2 abcd | 2.4 abc | 0.791 de | 0.200 bc |
0.5 | 50.0 ab | 0.8 abcd | 1.4 bcd | 0.815 de | 0.198 bc | |
1.0 | 75.0 a | 1.8 abc | 3.9 a | 1.119 bcde | 0.255 ab | |
2.0 | 33.3 b | 1.3 abcd | 1.5 bcd | 0.667 e | 0.178 c | |
p-value | 0.456 NS | 0.529 NS | 0.449 NS | 0.059 NS | 0.753 NS |
Treatments | Rooting (%) | Number of Roots/Explant | Length of Root (cm) | Fresh Weight of Roots/Explant (g) | Dry Weight of Roots/Explant (g) | |
---|---|---|---|---|---|---|
Photoperiod (h) | ||||||
Lightening regime 1 | 62.5 | 3.67 | 4.58 | 0.507 | 0.054 | |
Lightening regime 2 | 100 | 5.75 | 2.93 | 0.101 | 0.019 | |
p-value | 0.006 ** | 0.018 * | 0.021 * | 0.001 *** | 0.008 ** | |
Activated charcoal (mg/L) | ||||||
0.0 | 83.3 | 4.29 | 3.43 | 0.363 | 0.043 | |
1.5 | 79.1 | 5.13 | 4.08 | 0.245 | 0.030 | |
p-value | 0.347 NS | 0.088 NS | 0.124 NS | 0.272 NS | 0.375 NS | |
Photoperiod (h) × Activated charcoal (mg/L) | ||||||
Lightening regime 1 | 0.0 | 67 b | 3.67 b | 3.71 ab | 0.752 a | 0.071 a |
1.5 | 75 b | 3.83 b | 6.21 a | 0.518 a | 0.058 ab | |
Lightening regime 2 | 0.0 | 100 a | 4.67 b | 3.24 b | 0.123 b | 0.026 bc |
1.5 | 100 a | 6.67 a | 3.0 b | 0.087 b | 0.014 c | |
p-value | 0.347 NS | 0.147 NS | 0.063 NS | 0.417 NS | 0.956 NS |
Primers | Primer Sequence 5′-3′ | NO. of Scorble Band per Primer | Range of Amplification (bp) | No. of Monomorphic Bands | No. of Polymorphic Bands | Polymorphism % |
---|---|---|---|---|---|---|
RAPD primers | ||||||
OPA-01 | CAGGCCCTTC | 4 | 500–1000 | 4 | 0 | 0 |
OPA-03 | AGTCAGCCAC | 1 | 1300–1500 | 1 | 0 | 0 |
OPA-07 | GAAACGGGTG | Nil | - | - | - | - |
OPA-10 | GTGATCGCAG | 4 | 400–1000 | 4 | 0 | 0 |
OPB-03 | CATCCCCCTG | 2 | 200–600 | 2 | 0 | 0 |
OPH-05 | AGTCGTCCCC | 4 | 300–800 | 4 | 0 | 0 |
OPA-11 | CAATCGCCGT | 2 | 900–1500 | 2 | 0 | 0 |
OPA-12 | TCGGCGATAG | 3 | 250–1500 | 3 | 0 | 0 |
OPA-13 | CAGCACCCAC | 2 | 300–800 | 2 | 0 | 0 |
OPA-15 | TTCCGAACCC | Nil | - | - | - | - |
Total | 22 | 22 | 0 | |||
ISSR primers | ||||||
ISSR-11 | AGAGAGAGAGAGAGAGT | 8 | 200–1500 | 8 | 0 | 0 |
ISSR-15 | GAGAGAGAGAGAGAGAC | 5 | 250–700 | 5 | 0 | 0 |
UBC815 | CTCTCTCTCTCTCTCTG | 3 | 300–900 | 3 | 0 | 0 |
UBC816 | CACACACACACACACAT | 5 | 400–800 | 5 | 0 | 0 |
UBC823 | TCTCTCTCTCTCTCTCC | 2 | 500–1200 | 2 | 0 | 0 |
UBC861 | ACCACCACCACCACCACC | 5 | 300–1000 | 5 | 0 | 0 |
UBC862 | AGCAGCAGCAGCAGCAGC | 3 | 350–700 | 3 | 0 | 0 |
IMA-5-Z | CACACACACACACACAGT | 8 | 300–1500 | 6 | 2 | 25 |
IMA-5-3 | CACACACACACACACATG | 5 | 250–1000 | 5 | 0 | 0 |
HB15 | GTGGTGGTGGC | 6 | 400–1250 | 6 | 0 | 0 |
Total | 50 | 48 | 2 | 4 | ||
SCoT primers | ||||||
S1 | CAACAATGGCTACCACCA | 4 | 250–800 | 4 | 0 | 0 |
S2 | CAACAATGGTACCACCC | 1 | 1500 | 1 | 0 | 0 |
S3 | AACAATGGCTACCACCG | 5 | 300–1400 | 5 | 0 | 0 |
S4 | CAACAATGGCTACCACCT | 4 | 350–800 | 4 | 0 | 0 |
S5 | CAACAATGGCTACCACGA | Nil | - | - | - | - |
S6 | CAACAATGGCTACCACGC | Nil | - | - | - | - |
S7 | AGCAGCAGCAGCAGCAGC | 6 | 300–900 | 4 | 2 | 33.3 |
S8 | ACGACATGGCGACCAACG | 4 | 300–1400 | 4 | 0 | 0 |
S9 | ACCATGGCTACCACCGAC | 2 | 800–1000 | 2 | 0 | 0 |
S10 | CCATGGCTACCACCGCAG | 4 | 400–1400 | 4 | 0 | 0 |
Total | 30 | 28 | 2 | 6.6 |
Primer No. | Primer Name | Primer Sequence | Annealing Temperature |
---|---|---|---|
RAPD primers | |||
1 | OPA-01 | CAGGCCCTTC | 37 |
2 | OPA-03 | AGTCAGCCAC | 37 |
3 | OPA-07 | GAAACGGGTG | 37 |
4 | OPA-10 | GTGATCGCAG | 35 |
5 | OPB-03 | CATCCCCCTG | 37 |
6 | OPH-05 | AGTCGTCCCC | 37 |
7 | OPA-11 | CAATCGCCGT | 37 |
8 | OPA-12 | TCGGCGATAG | 37 |
9 | OPA-13 | CAGCACCCAC | 37 |
10 | OPA-15 | TTCCGAACCC | 37 |
ISSR primers | |||
1 | ISSR-11 | AGAGAGAGAGAGAGAGT | 50 |
2 | ISSR-15 | GAGAGAGAGAGAGAGAC | 50 |
3 | UBC815 | CTCTCTCTCTCTCTCTG | 50 |
4 | UBC816 | CACACACACACACACAT | 50 |
5 | UBC823 | TCTCTCTCTCTCTCTCC | 50 |
6 | UBC861 | ACCACCACCACCACCACC | 55 |
7 | UBC862 | AGCAGCAGCAGCAGCAGC | 55 |
8 | IMA-5-Z | CACACACACACACACAGT | 50 |
9 | IMA-5-3 | CACACACACACACACATG | 50 |
10 | HB15 | GTGGTGGTGGC | 40 |
SCoT primers | |||
1 | S1 | CAACAATGGCTACCACCA | 55 |
2 | S2 | CAACAATGGTACCACCC | 55 |
3 | S3 | AACAATGGCTACCACCG | 56 |
4 | S4 | CAACAATGGCTACCACCT | 56 |
5 | S5 | CAACAATGGCTACCACGA | 55 |
6 | S6 | CAACAATGGCTACCACGC | 55 |
7 | S7 | AGCAGCAGCAGCAGCAGC | 57 |
8 | S8 | ACGACATGGCGACCAACG | 55 |
9 | S9 | ACCATGGCTACCACCGAC | 55 |
10 | S10 | CCATGGCTACCACCGCAG | 60 |
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© 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
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Al-Aizari, A.A.; Dewir, Y.H.; Ghazy, A.-H.; Al-Doss, A.; Al-Obeed, R.S. Micropropagation and Genetic Fidelity of Fegra Fig (Ficus palmata Forssk.) and Grafting Compatibility of the Regenerated Plants with Ficus carica. Plants 2024, 13, 1278. https://doi.org/10.3390/plants13091278
Al-Aizari AA, Dewir YH, Ghazy A-H, Al-Doss A, Al-Obeed RS. Micropropagation and Genetic Fidelity of Fegra Fig (Ficus palmata Forssk.) and Grafting Compatibility of the Regenerated Plants with Ficus carica. Plants. 2024; 13(9):1278. https://doi.org/10.3390/plants13091278
Chicago/Turabian StyleAl-Aizari, Ahmed Ali, Yaser Hassan Dewir, Abdel-Halim Ghazy, Abdullah Al-Doss, and Rashid Sultan Al-Obeed. 2024. "Micropropagation and Genetic Fidelity of Fegra Fig (Ficus palmata Forssk.) and Grafting Compatibility of the Regenerated Plants with Ficus carica" Plants 13, no. 9: 1278. https://doi.org/10.3390/plants13091278