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Calluses initiated from thin mature embryo fragments are suitable targets for wheat transformation as assessed by long-term GUS expression studies

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Abstract

Microprojectile- or Agrobacterium-mediated DNA delivery into calluses initiated from immature embryos has proven to be effective in transforming wheat. Yet, obtaining a large number of high quality immature embryos throughout the year is a laborious and delicate process. To circumvent these limitations, we propose an alternative technique applying the particle bombardment technology to calluses derived from fragmented mature embryos rather than immature tissues. The phosphinothricin acetyl transferase (bar) and β-glucuronidase (gus) genes were used as selectable and screenable marker genes, respectively, to assess and optimise the performance of the proposed technique. Primary requirement for genetic transformation method development, the regeneration capacity of bombarded calluses was established. A preculture duration of 6 days was identified as optimal for DNA uptake and β-glucuronidase (GUS) expression. The highest activity was recorded when calluses were selected. Long-term GUS expression studies (1–7 weeks subsequent to bombardment), showed differentiated behaviours for tissues obtained from mature versus immature embryos. Notably, mature embryos exhibited the greatest number of cells stably expressing the reporter gene, thus providing an excellent source material for developing a stable transformation procedure.

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Abbreviations

2,4-D:

2,4-dichlorophenoxyacetic acid

GUS:

β-glucuronidase

PAT:

phosphinothricin acetyl transferase

PPT:

L-phosphinothricin

psi:

pounds per square inch

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Authors and Affiliations

  1. Department of Biotechnology, Agricultural Research Centre, Chaussée de Charleroi 234, Gembloux, B-5030, Belgium

    Fabienne Delporte & Jean-Marie Jacquemin

  2. Qingdao Institute of Agricultural Science, 126, Jiushui Road Licang, Qingdao, 266100, China

    Shirun Li

Authors
  1. Fabienne Delporte
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  2. Shirun Li
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  3. Jean-Marie Jacquemin
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Correspondence to Fabienne Delporte.

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Delporte, F., Li, S. & Jacquemin, JM. Calluses initiated from thin mature embryo fragments are suitable targets for wheat transformation as assessed by long-term GUS expression studies. Plant Cell Tiss Organ Cult 80, 139–149 (2005). https://doi.org/10.1007/s11240-004-9221-x

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  • DOI: https://doi.org/10.1007/s11240-004-9221-x

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