ISSN:
1615-6102
Keywords:
Wheat
;
Egg cell
;
Alternating-current field
;
Immobilisation
;
Microinjection
;
Green-fluorescent-protein imaging
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary 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 highfrequency 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 timewindow for DNA delivery into the fertilised egg cell, the time course of DNA dynamics during zygotic development has been quantified via quantitative microspectrofluorometry.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01279087
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