Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 20, Issue 10, Pages -Publisher
MDPI
DOI: 10.3390/ijms20102594
Keywords
Agrobacterium tumefaciens; leaf explants; regeneration transformation; P. alba x P; glandulosa Uyeki
Funding
- Fundamental Research Funds for the Central Universities [2019ZY29]
- State '13.5' Key Research Program of China [2016YFD0600102]
- National Natural Science Foundation of China [31670182, 31761133009, 31530084, 31401149, 3180030099]
- China Postdoctoral Science Foundation [2018M631246]
- Program of Introducing Talents of Discipline to Universities (111 project) [B13007]
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Transgenic technology is a powerful tool for gene functional characterization, and poplar is a model system for genetic transformation of perennial woody plants. However, the poplar genetic transformation system is limited to a number of model genotypes. Herein, we developed a transformation system based on efficient Agrobacterium-mediated transformation for the hybrid poplar Populus Alba x Populus glandulosa Uyeki, which is a fast-growing poplar species that is suitably grown in the northern part of China. Importantly, we optimized many independent factors and showed that the transformation efficiency was improved significantly using juvenile leaf explants. Explants were infected by an Agrobacterium suspension with the OD600 = 0.6 for 15 min and then co-cultured in dark conditions for 3 days. Using the improved transformation system, we obtained the transgenic poplar with overexpression of -glucuronidase (GUS) via direct organogenesis without callus induction. Furthermore, we analyzed the GUS gene in the transgenic poplars using PCR, qRT-PCR, and GUS staining. These analyses revealed that the GUS gene was efficiently transformed, and it exhibited various expression levels. Taken together, these results represent a simple, fast, and efficient transformation system of hybrid poplar plants. Our findings may facilitate future studies of gene functions in perennial woody plants and tree breeding via transgenic technology assisted design.
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