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An in planta biolistic method for stable wheat transformation.

Haruyasu Hamada1,2, Qianyan Linghu1, Yozo Nagira2

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A new in planta method simplifies wheat transformation without callus culture. This biolistic DNA delivery targets shoot apical meristems, enabling stable genetic modification in elite cultivars.

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Area of Science:

  • Plant Biotechnology
  • Genetics
  • Agricultural Science

Background:

  • Current wheat transformation methods rely on callus culture and regeneration, limiting applicability to elite cultivars.
  • Developing efficient transformation techniques is crucial for advancing wheat breeding and genetic research.

Purpose of the Study:

  • To establish a simple, reproducible, in planta method for wheat (Triticum aestivum L.) transformation.
  • To bypass the need for callus culture and regeneration in wheat genetic modification.

Main Methods:

  • Biolistic DNA delivery targeting shoot apical meristems (SAMs) of wheat embryos.
  • Optimization of gold particle size (0.6-µm) and pressure (1350-psi) for DNA delivery.
  • Assessment of transient GFP expression and stable transgene integration in T0 plants.

Main Results:

  • Achieved significantly increased transient GFP expression in SAMs and transgene integration in the fifth leaf using optimized parameters.
  • Demonstrated stable transgene integration into germ cells in 62% of transformants, enabling inheritance.
  • Successfully transformed both a model cultivar ('Fielder') and a recalcitrant elite cultivar ('Haruyokoi').

Conclusions:

  • The developed in planta method offers a simplified and efficient approach for wheat transformation.
  • This technique broadens the scope for generating transgenic wheat lines across diverse commercial cultivars.
  • The method holds potential for accelerating genetic improvement in wheat.