Efficient Biolistic Transformation of Immature Citrus Rootstocks Using Phosphomannose-isomerase Selection
Abstract
:1. Introduction
2. Results
2.1. Preliminary Test of Explant Sensitivity to Mannose and Shoot Regeneration
2.2. Biolistic Transformation of Carrizo Citrange
2.3. Biolistic Transformation of Swingle Citrumelo
2.4. Molecular Analyses of the Transgenics
2.5. Chlorophenol Red Assay
3. Discussion
4. Materials and Methods
4.1. Plasmid DNA
4.2. Plant Materials and Tissue Culture
4.3. Preparation of Gold Particles
4.4. Particle Bombardments and Selection on Mannose
4.5. Fluorescence Microscopy and Data Collection
4.6. Micro-Grafting
4.7. Molecular Analyses
4.8. Chlorophenol Red Assay
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Experiment | Paired Shots | Number of Explants | Number of Shoots | Number of Positive Shoots | Number of Shoots Survived/Number Micro-grafted | PCR Positive Shoots |
---|---|---|---|---|---|---|
1 | 1 | 153 | 294 | 2 | 0/1 | 0 |
1 | 2 | 126 | 237 | 3 | 1/2 | 1 |
2 | 3 | 165 | 288 | 4 | 2/2 | 2 |
2 | 4 | 153 | 284 | 2 | 1/1 | 1 |
3 | 5 | 159 | 270 | 1 | 1/1 | 1 |
3 | 6 | 166 | 214 | 1 | 0/1 | 0 |
4 | 7 | 154 | 346 | 1 | 1/1 | 1 |
4 | 8 | 80 | 140 | 0 | 0/0 | 0 |
5 | 9 | 149 | 319 | 3 | 1/1 | 1 |
5 | 10 | 88 | 173 | 2 | 1/1 | 1 |
Total | - | 1393 | 2565 | 19 | 8/11 | 8/8 |
Mean ± SE | - | 139.3 ± 9.9 | 256.5 ± 20.5 | 1.9 ± 0.4 | 72.7% survival | 100% |
Experiment | Paired Shots | Number of Explants | Number of Shoots | Number of Positive Shoots | Number of Shoots Survived/Number Micro-grafted | PCR Positive Shoots |
---|---|---|---|---|---|---|
1 | 1 | 103 | 121 | 0 | 0/0 | 0 |
1 | 2 | 99 | 187 | 2 | 2/2 | 2 |
2 | 3 | 97 | 132 | 1 | 1/1 | 1 |
2 | 4 | 110 | 164 | 2 | 0/1 | 0 |
3 | 5 | 144 | 205 | 3 | 1/2 | 1 |
Total | - | 553 | 809 | 8 | 4/6 | 4/4 |
Mean ± SE | - | 110.6 ± 8.6 | 161.8 ± 15.9 | 1.6 ± 0.5 | 66.7% | 100% |
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Wu, H.; Acanda, Y.; Canton, M.; Zale, J. Efficient Biolistic Transformation of Immature Citrus Rootstocks Using Phosphomannose-isomerase Selection. Plants 2019, 8, 390. https://doi.org/10.3390/plants8100390
Wu H, Acanda Y, Canton M, Zale J. Efficient Biolistic Transformation of Immature Citrus Rootstocks Using Phosphomannose-isomerase Selection. Plants. 2019; 8(10):390. https://doi.org/10.3390/plants8100390
Chicago/Turabian StyleWu, Hao, Yosvanis Acanda, Michel Canton, and Janice Zale. 2019. "Efficient Biolistic Transformation of Immature Citrus Rootstocks Using Phosphomannose-isomerase Selection" Plants 8, no. 10: 390. https://doi.org/10.3390/plants8100390