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Related Experiment Video

Updated: Apr 22, 2026

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Development of an Efficient Regeneration and Agrobacterium-Mediated Transformation Protocol for Hosta 'Light Star'

Hangkuan Zhou1, Yidi Qin1, Shuang Liu1

  • 1College of Horticulture and Landscape, Northeast Agricultural University, Harbin, 150030, China.

Molecular Biotechnology
|April 20, 2026
PubMed
Summary

This study presents the first efficient in vitro regeneration and Agrobacterium-mediated genetic transformation system for Hosta 'Light Star'. The developed protocol utilizes the RUBY reporter gene for effective screening of transgenic ornamental plants.

Keywords:
35S:RUBYAgrobacterium-mediated transformationHosta ‘Light Star’BetacyaninGenetic transformationIn vitro organ culture

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Area of Science:

  • Plant Biotechnology
  • Horticultural Science
  • Molecular Biology

Background:

  • Hosta 'Light Star' is a valuable ornamental cultivar lacking efficient in vitro regeneration and genetic transformation methods.
  • Developing such systems is crucial for advancing research and breeding in ornamental plants.

Purpose of the Study:

  • To establish a highly efficient in vitro regeneration system for Hosta 'Light Star'.
  • To develop an Agrobacterium-mediated genetic transformation protocol for Hosta 'Light Star' using the RUBY reporter gene.

Main Methods:

  • Optimization of callus induction, adventitious bud proliferation, and rooting media using various plant growth regulators.
  • Development of an Agrobacterium-mediated transformation protocol, including optimizing infection parameters and selection conditions.
  • Utilized the RUBY visual reporter gene for non-invasive screening of transformants.
  • Confirmed gene integration and expression using PCR, RT-PCR, and RT-qPCR.

Main Results:

  • Achieved high efficiency in callus induction (53.33%), adventitious bud proliferation (coefficient 5.87), and rooting (100%).
  • Established optimal Agrobacterium infection parameters (OD600=0.6, 10 min infection, 200 μM acetosyringone) and selection conditions (300 mg/L cefotaxime, 30 mg/L hygromycin).
  • Obtained a stable transformation efficiency of 2.50% with distinct visual markers (purplish-red coloration) in transgenic plants due to betacyanin accumulation.

Conclusions:

  • Successfully established the first efficient in vitro regeneration and Agrobacterium-mediated transformation system for Hosta 'Light Star'.
  • Validated the RUBY reporter gene as an effective visual marker for Hosta transformation.
  • The developed system provides a foundation for functional genomics, gene editing, and breeding of ornamental traits in Hosta.