Transient and stable transformation of wheat with DNA preparations delivered by a biolistic method |
| |
Authors: | Marta Dobrzańska Cezary Krysiak El?bieta Kraszewska |
| |
Institution: | (1) Institute of Biochemistry and Biophysics, Polish Academy of Sciences, ul. Pawińskiego 5a, 02-106 Warszawa, Poland |
| |
Abstract: | An optimized procedure for transformation of wheat with the use of a Biolistic Particle Delivery System PDS 1000/He to deliver
foreign DNA is described in detail. The bacterial uidA and bar genes (both driven by plant promoters) were utilized as the reporter and selectable marker genes, respectively. Moderately
high gas pressure appeared to be most important to achieve the highest level of transient GUS expression in target tissues.
There was, however, no apparent correlation between transient and stable GUS expression. The presence of telomeric DNA sequences
in an uidA gene-containing vector did not influence transient GUS expression but, apparently, prevented its stable expression. Mechanical
lesions caused by the bombardment (tungsten particles) seemed to be less severe when embryo- derived calli, instead of freshly
excised immature embryos, were used as the target tissue. The limited ability of callus cells for regeneration, together with
a restricted number of cells that receive the foreign DNA by particle bombardment, result in a low efficiency of wheat stable
transformation. |
| |
Keywords: | gene gun microprojectile bombardment somatic embryogenesis transformation telomere Triticum aestivum |
本文献已被 SpringerLink 等数据库收录! |
|