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KMID : 1025520040460020235
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Journal of Animal Science and Technology
2004 Volume.46 No. 2 p.235 ~ p.242
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Factors Affecting Genetic Transformation of Italian Ryegrass
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Abstract
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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 were infected and co-cultured with Agrobacterium EHA101 carrying standard binary vector pIG121Hm encoding the hygromycin phosphotransferase(HPT), neomycin phosphotransferase Ⅱ (NPTⅡ) and intron-containing β-glucuronidase(intron-GUS) genes in the T-DNA region. The effects of several factors on transformation and the expression of the GUS gene were investigated. Inclusion of 200㎛ acetosyringone(AS) in inoculation and co-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. The highest transformation efficiency was obtained when embryogenic calli were incoulated with Agrobacterium in the presence of 0.1% Tween20 and 200㎛ AS. Hygromycin resistant calli were developed into complete plants via somatic embryogenesis. GUS histochemical assay and PCR analysis of transgenic plants demonstrated that transgenes were integrated into the genome of Italian ryegrass.
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