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ROSEA1-Based Visual Selection Reduces Plant Regeneration and Alters Developmental Regulator Expression
Tao Jiang1,2, Sameena Ejaz Tanwir1,2, Fangchen Liu1,2
1Crop Transformation Center, Department of Horticulture Sciences, IFAS, University of Florida, Gainesville, FL 32611, USA.
Constitutive ROSEA1 expression, used for identifying transgenic plants, can negatively impact plant regeneration. Optimizing hormones and genotypes may help overcome this regeneration penalty in some species.
Area of Science:
- Plant Biotechnology
- Molecular Biology
- Developmental Biology
Background:
- Anthocyanin-based visual reporters offer non-destructive identification of transgenic tissues.
- Concerns exist regarding the physiological impact of anthocyanin pigment production on plant organogenesis and regeneration.
Purpose of the Study:
- To investigate the regeneration performance of constitutive ROSEA1 expression across diverse eudicot species.
- To determine if ROSEA1 expression affects regeneration frequencies and gene expression during organogenesis.
Main Methods:
- Generated and evaluated stable ROSEA1-overexpressing lines in tomato and petunia.
- Performed transformation-stage assays in begonia and marigold.
- Quantified regeneration frequencies, assessed gene expression (anthocyanin biosynthesis, regeneration regulators), and conducted complementation assays.
Main Results:
- ROSEA1 overexpression reduced regeneration frequencies and increased anthocyanin accumulation in tomato and petunia.
- Stronger pigment production correlated with more severe regeneration defects in petunia.
- ROSEA1 modulated expression of key regeneration genes (e.g., PLT5, WUS) and anthocyanin pathway genes.
- Co-expression of PLT5 partially rescued regeneration, suggesting pigment output isn't the sole factor.
- A regeneration penalty was observed across all tested species, varying in severity by genotype and hormone treatment.
Conclusions:
- Constitutive ROSEA1 expression imposes a context-dependent regeneration penalty.
- Mitigation strategies may involve hormone optimization, careful genotype selection, and developmental support.
- Findings highlight the need to consider physiological impacts of visual reporters in plant transformation.
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