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Doubled Haploid Transgenic Wheat Lines by Microspore Transformation.

Sachin Rustgi1,2, Nii O Ankrah3,4, Rhoda A T Brew-Appiah3

  • 1Department of Plant and Environmental Sciences, Clemson University Pee Dee Research and Education Center, 2200 Pocket Road, Florence, SC, 29506, USA. srustgi@clemson.edu.

Methods in Molecular Biology (Clifton, N.J.)
|September 16, 2017
PubMed
Summary

Microspore transformation in wheat offers a faster route to homozygous transformants. Cocultivation with Agrobacterium tumefaciens is the most effective method, yielding homozygous plants across diverse wheat cultivars.

Keywords:
Agrobacterium tumefaciensElectroporationGenetic transformationMicrospore embryogenesisParticle bombardment

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Area of Science:

  • Plant Biotechnology
  • Genetics
  • Agricultural Science

Background:

  • Microspores are ideal explants for plant genetic transformation due to accelerated homozygous line development.
  • Optimizing gene delivery methods is crucial for efficient transformation in wheat.

Purpose of the Study:

  • To optimize and compare established gene delivery methods for wheat genetic transformation using microspores.
  • To assess the efficiency and genotype independence of different transformation techniques.

Main Methods:

  • Particle bombardment, electroporation, and Agrobacterium tumefaciens cocultivation were optimized on microspore-derived tissues.
  • Specific protocols were developed for each method, utilizing either haploid microspore embryoids or freshly isolated microspores.
  • Methods were validated across multiple wheat cultivars of varying market classes.

Main Results:

  • Cocultivation with Agrobacterium tumefaciens successfully produced homozygous transformants.
  • Particle bombardment and electroporation resulted in chimeric plants.
  • Transformation efficiency demonstrated genotype independence across tested wheat cultivars.
  • Reduced explant donor requirements and a timeframe of 8-12 months for homozygous transformants were observed.

Conclusions:

  • Microspore-based transformation, particularly via Agrobacterium cocultivation, is an efficient and rapid strategy for developing homozygous wheat.
  • The optimized methods are robust and applicable to a wide range of wheat genotypes.
  • This approach significantly shortens the timeline for obtaining genetically modified wheat lines.