Regeneration and transformation of Egyptian maize inbred lines via immature embryo culture and a biolistic particle delivery system |
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Authors: | Email author" target="_blank">Hanaiya?A?El-ItribyEmail author Shireen?K?Assem Ebtissam?H?A?Hussein Fathy?M?Abdel-Calil Magdy?A?Madkour |
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Institution: | (1) Agricultural Genetic Engineering Research Institute (AGERI), ARC, Giza, Egypt;(2) Department of Genetics, Faculty of Agriculture, Cairo University, Giza, Egypt;(3) Department of Biochemistry, Faculty of Science, Cairo University, Egypt |
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Abstract: | Summary A regeneration system was developed for elite Egyptain maize inbred lines using immature embryos as explants. This system
proved to be highly genotype-dependent. Line Gz 643 was identified as the best line, revealing the highest regeneration frequency
(42.2%). Addition of l-proline and silver nitrate to culture media greatly enhanced the formation of embryogenic type II callus and the regenerability
of some of the tested lines. Transformation of the scutellar tissue of immature embryos from inbred line Gz643 was performed
with the particle delivery system using a single plasmid carrying both the GUS and Bar genes (pAB-6) or by co-transformation with two plasmids, pAct1-F (GUS) and pTW-a(Bar). Different transformation parameters were evaluated, i.e. ostomic treatment, acceleration pressure, and number of shots.
Osmotic treatment (0.25 M sorbitol + 0.25 M mannitol) along with the use of either acceleration pressure 1300 psi and one shot per plate (for co-transformation with
pAB-6) or 1100 psi and two shots per plate (for transformation with pAct1-F and pTW-a) gave the best results, as expressed
by the number of blue spots in the β-glucuronidase (GUS) assay. Stable transformation was confirmed in Ro transformed plants
by means of histochemical GUS assay and herbicide application. PCR and Southern blot analysis proved the integration of the
full-length genes in some of the transgenics. |
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Keywords: | maize immature embryos embryogenic callus plant regeneration tranformation |
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