Optimisation of introducing foreign genes into egg cells and zygotes of wheat (Triticum aestivum L.) via microinjection

Z. Pónya, P. Finy, A. Fehér, J. Mitykó, D. Dudits, B. Barnabás

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

An expeditious and highly efficient technique of microinjection has been developed with the aim of introducing exogenous DNA into egg cells and zygotes of wheat. Using a mechanical-dissection method and a novel immobilisation approach enabled us to microinject around 15 egg cells of wheat per hour. Exposing the protoplasts to a high-frequency alternating-current field for immobilisation, a significantly higher transient expression rate of the injected genes (46% and 52% for egg cells and zygotes, respectively) could be achieved than reported thus far for plant protoplasts. Whether this high transformation efficiency is due to the high-frequency electrical field applied for immobilising the protoplasts is not known. The transformation rate appeared to be a factor depending upon the time of egg cell isolation. According to the ultrastructural observations this seems to reflect a variation in competence of the egg cells during in situ development. In order to conduct studies directed towards establishing the optimal time-window for DNA delivery into the fertilised egg cell, the time course of DNA dynamics during zygotic development has been quantified via quantitative microspectrofluorometry.

Original languageEnglish
Pages (from-to)163-172
Number of pages10
JournalProtoplasma
Volume208
Issue number1-4
Publication statusPublished - Dec 1 1999

Keywords

  • Alternating-current field
  • Egg cell
  • Green-fluorescent-protein imaging
  • Immobilisation
  • Microinjection
  • Wheat

ASJC Scopus subject areas

  • Plant Science
  • Cell Biology

Fingerprint Dive into the research topics of 'Optimisation of introducing foreign genes into egg cells and zygotes of wheat (Triticum aestivum L.) via microinjection'. Together they form a unique fingerprint.

  • Cite this