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- Title
Genetic transformation of Rhododendron delavayi for anthocyanin synthesis using Agrobacterium-mediated transformation.
- Authors
Long, Fenfang; Zuo, Weiwei; Li, Huie; Zeng, Liang
- Abstract
Rhododendron delavayi is an evergreen tree species, as a woody plant belonging to Ericaceae characterized by its long lifecycle and high ornamental value. Genetic engineering has made remarkable progress in improving various plant species. However, applying these advancements to Rhododendron remains a complex endeavor, primarily due to challenges associated with genetic transformation technology. In this study, the optimum transformation conditions for Agrobacterium infection of R. delavayi leaf disc were established as follows: the pre-culture time was 0 d, Agrobacterium liquid concentration OD600 = 0.7, infection time was 60 min, Acetosyringone concentration 20 mg/L, and co-culture time was 6 d. After removal of bacteria, first screening culture was 30 days with medium containing Kanamycin (Kan) at 30 mg/L and Cefotaxime (Cef) at 500 mg/L. Subsequent culture was for 60 days with medium supplemented with Kan at 30 mg/L and Cef at 400 mg/Ls. Third screening culture was for 90 days with medium containing Kan at 40 mg/L and Cef at 300 mg/L. Positive transgenic materials were identified after 120 days culture. Gene cloning and transformation proceeded with two MYB genes, RdMYB8 and RdMYB24, which had been isolated from petals of R. delavayi. These genes were individually inserted into overexpression vectors and then successfully transformed into Agrobacterium GV3101. Leaf discs of R. delavayi were inoculated with Agrobacterium carrying the respective genes, resulting in the successful acquisition of positive transgenic materials. The transgenic callus exhibited a red coloration and showed an increase in the total anthocyanin content compared to the control group. The study suggests that RdMYB8 and RdMYB24 are involved in anthocyanin synthesis in R. delavayi, and this Agrobacterium-mediated transformation system holds promise as a fundamental tool for advancing genetic engineering not only in R. delavayi.Key message: Leaf discs from Rhododendron delavayi are the optimal recipient material for Agrobacterium-mediated transformation. The transgenic callus overexpressing RdMYB8 exhibits enhanced anthocyanin biosynthesis.
- Publication
Plant Cell, Tissue & Organ Culture, 2024, Vol 157, Issue 2, p1
- ISSN
0167-6857
- Publication type
Article
- DOI
10.1007/s11240-024-02775-9