A Strategy for the Production and Molecular Validation of Agrobacterium-Mediated Intragenic Octoploid Strawberry
Abstract
:1. Introduction
2. Results
2.1. Optimization of Shoot Regeneration from Leaf Explants of the Strawberry cv. ‘Shanghai Angel’
2.2. Generation of a Marker-Free Binary Vector with an Intragenic Expression Cassette
2.3. Development of Putative Intragenic Strawberry Plants via Agrobacterium-Mediated Transformation without Selection
2.4. Selection of Transformants Using Two Rounds of PCR
2.5. Determination of the Intragene Copy Number in Transformants Using ddPCR
3. Discussion
4. Materials and Methods
4.1. Plant Materials and Growth Conditions
4.2. Agrobacterium Strain and Binary Vector
4.3. Explant Preparation and Strawberry Transformation
4.4. Direct PCR Analysis of Regenerated Shoots
4.5. DNA Purification and ddPCR Analysis
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Experiment a | No. of Initial Explants | No. of Shoots b | No. of PCR-Positive Shoots | Transformation Efficiency (%) c |
---|---|---|---|---|
I | 30 | 53 | 0 | 0 |
II | 51 | 58 | 1 | 1.96% |
III | 40 | 46 | 1 | 2.50% |
IV | 38 | 84 | 2 | 5.26% |
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Duan, K.; Zhao, Y.-J.; Li, Z.-Y.; Zou, X.-H.; Yang, J.; Guo, C.-L.; Chen, S.-Y.; Yang, X.-R.; Gao, Q.-H. A Strategy for the Production and Molecular Validation of Agrobacterium-Mediated Intragenic Octoploid Strawberry. Plants 2021, 10, 2229. https://doi.org/10.3390/plants10112229
Duan K, Zhao Y-J, Li Z-Y, Zou X-H, Yang J, Guo C-L, Chen S-Y, Yang X-R, Gao Q-H. A Strategy for the Production and Molecular Validation of Agrobacterium-Mediated Intragenic Octoploid Strawberry. Plants. 2021; 10(11):2229. https://doi.org/10.3390/plants10112229
Chicago/Turabian StyleDuan, Ke, Ying-Jie Zhao, Zi-Yi Li, Xiao-Hua Zou, Jing Yang, Cheng-Lin Guo, Si-Yu Chen, Xiu-Rong Yang, and Qing-Hua Gao. 2021. "A Strategy for the Production and Molecular Validation of Agrobacterium-Mediated Intragenic Octoploid Strawberry" Plants 10, no. 11: 2229. https://doi.org/10.3390/plants10112229