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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

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Plants (Basel, Switzerland)
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PubMed
Summary

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.

Keywords:
ROSEA1anthocyanin biosynthesismetabolic fluxregeneration capacity

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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.