TRANSGENIC PLANTS OF LOLIUM-MULTIFLORUM, LOLIUM-PERENNE, FESTUCA-ARUNDINACEA AND AGROSTIS-STOLONIFERA BY SILICON-CARBIDE FIBER-MEDIATED TRANSFORMATION OF CELL-SUSPENSION CULTURES
Sj. Dalton et al., TRANSGENIC PLANTS OF LOLIUM-MULTIFLORUM, LOLIUM-PERENNE, FESTUCA-ARUNDINACEA AND AGROSTIS-STOLONIFERA BY SILICON-CARBIDE FIBER-MEDIATED TRANSFORMATION OF CELL-SUSPENSION CULTURES, PLANT SCI, 132(1), 1998, pp. 31-43
Seven Lolium multiflorum, one Lolium perenne, 12 Festuca arundinacea a
nd six Agrostis stolonifera plants were regenerated following transfor
mation with a hygromycin resistance gene and hygromycin selection, fro
m cell suspension colonies treated with silicon-carbide whiskers. Tran
sformation was confirmed by PCR and Southern blotting; the latter also
showed that six of the L. multiflorum plants were independent transfo
rmants (insufficient molecular evidence was obtained for the seventh),
nine of the 12 F. arundinacea plants were independent transformants,
but that all the A. stolonifera plants were derived from a single tran
sformation event. Most plants tested contained fewer than five integra
ted transgene copies. Transgene expression was confirmed by reverse tr
anscriptase-PCR (RT-PCR). Of the one A. stolonifera and three L. multi
florum transformants regenerated after co-transformation with both the
hygromycin resistance gene and the beta-glucuronidase (gusA) gene, no
ne were found to express GUS activity. L. multiflorum regenerants from
older (14-16 week old) cell suspensions showed loss of female fertili
ty, but analysis of the progeny from three plants showed that the tran
sgenes were being inherited as a single dominant allele with a high fr
equency of transmission of hygromycin resistance. (C) 1998 Elsevier Sc
ience Ireland Ltd. All rights reserved.