T. Koprek et al., FERTILE TRANSGENIC BARLEY OF DIFFERENT CULTIVARS OBTAINED BY ADJUSTMENT OF BOMBARDMENT CONDITIONS TO TISSUE-RESPONSE, PLANT SCI, 119(1-2), 1996, pp. 79-91
Fertile transgenic barley (Hordeum vulgare L.) plants of five cultivar
s were obtained by an improved transformation protocol using different
particle delivery devices. Varieties of barley which are hardly amena
ble for genetic transformation due to low tissue culture response were
stably transformed by optimizing transformation parameters for each g
enotype separately according to their tissue culture response. Scutell
a of immature-zygotic embryos were transformed with the plasmid pAHC25
carrying the selectable marker gene bar and the reporter gene uidA. G
ene delivery was optimized using the uidA-expression as transient mark
er for the transformation efficiency and taking into account the maint
enance of the regeneration capacity after the bombardment. After selec
tion on media with bialaphos as selective agent, 40 independently tran
sformed plants were regenerated and analyzed for functional expression
of the bar and uidA gene. Molecular analysis by Southern hybridizatio
n confirmed the integration of both genes into the nuclear genome of t
he plants. Thirty-five T-0-plants of five barley cultivars reached mat
urity so far. Analysis of the progeny confirmed the inheritance of the
marker genes.