Organogenesis in a Broad Spectrum of Grape Genotypes and Agrobacterium-Mediated Transformation of the Podarok Magaracha Grapevine Cultivar
Abstract
:1. Introduction
2. Results
2.1. Development of a Regeneration Protocol
2.2. Enhancing Efficiencies for the Most Responsible Genotypes
2.3. Development of an Elongation Protocol
2.4. Tissue-Culture Cycle (Direct Organogenesis)
2.5. Agrobacterium-Mediated Transformation of ‘Podarok Magaracha’
3. Discussion
4. Materials and Methods
4.1. Plant Materials
4.2. Development of a Regeneration Protocol
4.3. Development of an Elongation Protocol
4.4. Agrobacterium-Mediated Transformation
4.4.1. Transient Transformation
4.4.2. Stable Transformation
4.5. PCR Analysis
4.6. Southern Blot Analysis
4.7. Statistical Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Genotype | MS | PIV | NN | DKW | WPM | IM |
---|---|---|---|---|---|---|
Kober 5BB | 0 a | 0 a | 9.7 bc | 18.0 c | 1.7 ab | 52.1 d |
Podarok Magaracha | 50.0 a | 43.3 a | 32.7 a | 60.2 a | 64.1 a | 97.6 b |
Duration of Cocultivation of Explants with Agrobacterium (h) | Inoculum Concentrations (OD600) | Duration of Cultivation of Explants on a Selective Medium (Days) | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | 6 | 14 | 21 | 28 | 42 | 56 | 70 | |||
24 | 0.2 | 0.0 | 15.5 | 17.2 | 24.1 | 24.1 | 22.4 | 17.2 | 10.3 | 8.6 | 8.6 | 6.9 | 6.9 | bc |
0.4 | 0.0 | 19.0 | 27.6 | 41.4 | 44.8 | 32.8 | 25.9 | 20.7 | 7.0 | 3.5 | 3.5 | 3.5 | a | |
0.6 | 0.0 | 15.5 | 31.0 | 57.6 | 59.2 | 57.9 | 50.0 | 36.2 | 15.5 | 12.1 | 12.1 | 12.1 | fghij | |
0.8 | 0.0 | 39.7 | 46.6 | 60.3 | 65.5 | 44.8 | 27.6 | 19.0 | 10.3 | 8.6 | 6.9 | 6.9 | bc | |
1.0 | 0.0 | 25.9 | 43.1 | 55.2 | 53.5 | 39.7 | 27.6 | 15.5 | 10.3 | 10.3 | 6.9 | 6.9 | bc | |
48 | 0.2 | - | 1.7 | 36.2 | 75.9 | 67.2 | 58.6 | 41.4 | 39.7 | 31.0 | 24.1 | 8.5 | 8.5 | cdef |
0.4 | - | 10.3 | 58.6 | 82.8 | 79.3 | 74.1 | 48.3 | 34.5 | 19.0 | 10.5 | 8.5 | 8.5 | cdef | |
0.6 | - | 17.2 | 69.0 | 86.2 | 82.8 | 74.1 | 48.3 | 36.2 | 19.0 | 12.1 | 8.6 | 5.1 | ab | |
0.8 | - | 36.2 | 87.9 | 93.1 | 91.4 | 82.8 | 70.7 | 56.9 | 31.0 | 17.2 | 10.3 | 8.5 | cdef | |
1.0 | - | 19.0 | 46.6 | 67.2 | 60.3 | 48.3 | 37.9 | 32.8 | 22.4 | 13.5 | 10.3 | 10.3 | efghi | |
72 | 0.2 | - | - | 32.8 | 74.1 | 67.2 | 50.0 | 36.6 | 49.1 | 22.4 | 19.0 | 17.2 | 17.2 | klmn |
0.4 | - | - | 46.6 | 82.6 | 79.3 | 62.1 | 41.4 | 41.0 | 22.4 | 17.2 | 13.8 | 13.8 | hijkl | |
0.6 | - | - | 51.7 | 94.8 | 86.2 | 72.4 | 65.5 | 50.1 | 34.5 | 24.1 | 20.7 | 19.0 | mn | |
0.8 | - | - | 81.0 | 96.6 | 94.8 | 87.9 | 82.8 | 56.7 | 37.9 | 27.6 | 25.7 | 24.3 | pq | |
1.0 | - | - | 70.7 | 86.2 | 84.5 | 75.9 | 67.2 | 56.7 | 43.1 | 36.2 | 29.3 | 27.6 | qr | |
96 | 0.2 | - | - | - | 68.4 | 79.3 | 77.6 | 56.7 | 34.5 | 25.9 | 22.4 | 19.0 | 17.2 | mn |
0.4 | - | - | - | 82.8 | 82.8 | 77.6 | 65.5 | 46.6 | 37.9 | 19.0 | 17.2 | 13.8 | ghijkl | |
0.6 | - | - | - | 87.9 | 89.3 | 80.7 | 68.4 | 39.2 | 35.7 | 20.7 | 17.2 | 15.5 | jklmn | |
0.8 | - | - | - | 89.7 | 94.8 | 94.8 | 74.1 | 37.9 | 37.9 | 31.0 | 25.5 | 24.1 | opq | |
1.0 | - | - | - | 64.0 | 67.2 | 60.3 | 51.7 | 46.6 | 41.4 | 37.9 | 29.3 | 29.3 | r | |
120 | 0.2 | - | - | - | - | 69.0 | 91.4 | 62.1 | 44.8 | 24.1 | 19.0 | 12.1 | 10.3 | defghi |
0.4 | - | - | - | - | 84.1 | 91.4 | 65.5 | 41.4 | 19.0 | 13.8 | 8.5 | 8.5 | cdef | |
0.6 | - | - | - | - | 91.4 | 93.1 | 79.3 | 58.6 | 33.1 | 25.9 | 19.0 | 19.0 | n | |
0.8 | - | - | - | - | 94.8 | 93.1 | 75.9 | 50.0 | 39.1 | 31.0 | 10.3 | 10.3 | fghi | |
1.0 | - | - | - | - | 50.0 | 51.7 | 43.7 | 41.4 | 34.5 | 31.0 | 17.2 | 13.8 | ijkl |
BA Concentration, mg L−1 | Number of GFP-Expressing Regenerated Plants (pcs per 100 Explants) | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
IAA Concentration (mg L−1) | IBA Concentration (mg L−1) | |||||||||
0.1 | 0.2 | 0.3 | 0.4 | 0.5 | 0.1 | 0.2 | 0.3 | 0.4 | 0.5 | |
1.5 | 0 a | 0 a | 0 a | 1 b | 0 a | 0 a | 0 a | 0 a | 0 a | 0 a |
2.0 | 0 a | 0 a | 0 a | 1 b | 0 a | 0 a | 0 a | 0 a | 0 a | 0 a |
2.5 | 0 a | 0 a | 1 b | 4 c | 0 a | 0 a | 0 a | 0 a | 0 a | 0 a |
3.0 | 0 a | 0 a | 0 a | 2 b | 0 a | 0 a | 0 a | 0 a | 0 a | 0 a |
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Maletich, G.; Pushin, A.; Rybalkin, E.; Plugatar, Y.; Dolgov, S.; Khvatkov, P. Organogenesis in a Broad Spectrum of Grape Genotypes and Agrobacterium-Mediated Transformation of the Podarok Magaracha Grapevine Cultivar. Plants 2024, 13, 2779. https://doi.org/10.3390/plants13192779
Maletich G, Pushin A, Rybalkin E, Plugatar Y, Dolgov S, Khvatkov P. Organogenesis in a Broad Spectrum of Grape Genotypes and Agrobacterium-Mediated Transformation of the Podarok Magaracha Grapevine Cultivar. Plants. 2024; 13(19):2779. https://doi.org/10.3390/plants13192779
Chicago/Turabian StyleMaletich, Galina, Alexander Pushin, Evgeniy Rybalkin, Yuri Plugatar, Sergey Dolgov, and Pavel Khvatkov. 2024. "Organogenesis in a Broad Spectrum of Grape Genotypes and Agrobacterium-Mediated Transformation of the Podarok Magaracha Grapevine Cultivar" Plants 13, no. 19: 2779. https://doi.org/10.3390/plants13192779