Abstract
ABSTRACT A system for the production of transgenic plants has been developed for Italian ryegrass( Lolium multiflorum Lam.) via Agrobacterium -mediated transformation of embryogenic callus. Mature seed-derived calli wereinfected and co-cultured with Agrobacterium EHA101 carrying standard binary vector pIG121Hm encoding thehygromycin phosphotransferase(HPT), neomycin phosphotransferase II (NPTII) and intron-containing - βglucuronidase(intron-GUS) genes in the T-DNA region. The effects of several factors on transformation and theexpression of the GUS gene were investigated. Inclusion of 200 M acetosyringone(AS) in inoculation andμco-cultivation media lead to a significant increase in stable transformation efficiency. Increasing Agrobacterium cell density up to 1.0 in OD 600 during infection increased transformation efficiency of embryogenic calli. Thehighest transformation efficiency was obtained when embryogenic calli were incoulated with Agrobacterium inthe presence of 0.1% Tween20 and 200 M AS. Hygromycin resistant calli were developed into complete plantsμvia somatic embryogenesis. GUS histochemical assay and PCR analysis of transgenic plants demonstrated thattransgenes were integrated into the genome of Italian ryegrass.(
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