Transformation of American Chestnut (Castanea dentata (Marsh.) Borkh) Using RITA® Temporary Immersion Bioreactors and We Vitro Containers
Abstract
:1. Introduction
2. Materials and Methods
2.1. Plant Material
Embryos for Transformation
2.2. Plasmid DNA
2.2.1. p35S-OxO for Blight Tolerance
2.2.2. pFHI_Gnk2 for Putative Phytophthora-Tolerance
2.3. Transformation Protocol
2.3.1. Transformation of American Chestnut for Blight Tolerance
2.3.2. Transformation of American Chestnut for Putative Phytophthora-Tolerance
2.4. Multiplication, Rooting, and Acclimatization of Regenerated Shoots Expressing OxO
2.5. PCR Analysis
2.6. Data Analysis
3. Results
3.1. Transformations
3.1.1. Transformation of American Chestnut for Blight Tolerance
3.1.2. Transformation of American Chestnut for Putative Phytophthora-Tolerance
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Trial | Clonal Line | Treatment | Starting Mass (mg) | Positive Events | Escapes |
---|---|---|---|---|---|
1 | AL | RITA | 50 | 3 | 5 |
2 | AL | RITA | 110 | 10 | 0 |
1 | AxW3 | RITA | 37 | 6 | 0 |
2 | AxW3 | RITA | 113 | 1 | 0 |
1 | BM5 | RITA | 98 | 0 | 1 |
2 | ML | RITA | 141 | 4 | 0 |
1 | TG | RITA | 86 | 8 | 6 |
2 | TG | RITA | 133 | 8 | 0 |
1 | AL | Semisolid | 86 | 8 | 11 |
2 | AL | Semisolid | 120 | 6 | 1 |
1 | AxW3 | Semisolid | 108 | 6 | 2 |
2 | AxW3 | Semisolid | 135 | 5 | 1 |
1 | BM5 | Semisolid | 65 | 8 | 9 |
2 | ML | Semisolid | 136 | 6 | 0 |
1 | TG | Semisolid | 59 | 8 | 8 |
2 | TG | Semisolid | 92 | 3 | 0 |
1 | AL | We Vitro | 65 | 3 | 6 |
2 | AL | We Vitro | 121 | 9 | 0 |
1 | AxW3 | We Vitro | 36 | 0 | 0 |
2 | AxW3 | We Vitro | 98 | 6 | 0 |
1 | BM5 | We Vitro | 94 | 6 | 6 |
2 | ML | We Vitro | 157 | 3 | 0 |
1 | TG | We Vitro | 63 | 4 | 0 |
2 | TG | We Vitro | 113 | 2 | 0 |
Clonal Line | Vector | Starting Mass (mg) | Putative Events |
---|---|---|---|
AxW3 | pFHI_Gnk2 | 144 | 5 |
SH4 | pFHI_Gnk2 | 103 | 0 |
SH11 | pFHI_Gnk2 | 113 | 2 |
Ellis #1 | pFHI_Gnk2 | 168 | 2 |
pFHI_Gnk2 | 180 | 5 | |
pFHI_GFP | 168 | 3 |
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McGuigan, L.; Fernandes, P.; Oakes, A.; Stewart, K.; Powell, W. Transformation of American Chestnut (Castanea dentata (Marsh.) Borkh) Using RITA® Temporary Immersion Bioreactors and We Vitro Containers. Forests 2020, 11, 1196. https://doi.org/10.3390/f11111196
McGuigan L, Fernandes P, Oakes A, Stewart K, Powell W. Transformation of American Chestnut (Castanea dentata (Marsh.) Borkh) Using RITA® Temporary Immersion Bioreactors and We Vitro Containers. Forests. 2020; 11(11):1196. https://doi.org/10.3390/f11111196
Chicago/Turabian StyleMcGuigan, Linda, Patrícia Fernandes, Allison Oakes, Kristen Stewart, and William Powell. 2020. "Transformation of American Chestnut (Castanea dentata (Marsh.) Borkh) Using RITA® Temporary Immersion Bioreactors and We Vitro Containers" Forests 11, no. 11: 1196. https://doi.org/10.3390/f11111196