Transformation of barley microspores by electroporation
DOI:
https://doi.org/10.14720/aas.1999.73.1.15912Abstract
Microspores of the winter barley variety Arda were transformed by three plasmids by electroporation. The pBI121 plasmid contains b-glucuronidase (GUS) reporter gene, NPT II - neomycin phosphotransferase II gene conferring kanamycin resistance, and promoters, NOS (nopaline synthase) and 35S from the cauliflower mosaic virus (CaMV); the pAKC plasmid contains DHDPS (14 kb) - dihydrodipicolinate gene for high lysine content in the seeds, and NPT II resistance gene; and the pAK-CLP plasmid contains a modified DHDPS (7 kb) gene ending protein with leader sequences for chloroplast and AACCl - acetyltransferase gene conferring gentamycin resistance, under the control of the CaMV 35S promoter. Parameters such as composition of media, microspore viability, electric conditions, and electroporation buffer constituents were tested to determine the most suitable conditions for gene transfer. Some of the transgenic microspores divided and formed resistant calli on the selection media.
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Copyright (c) 1999 University of Ljubljana, Biotechnical Faculty

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