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Expression of CoQ10-producing <Emphasis Type="Italic">ddsA</Emphasis> transgene by efficient <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation in <Emphasis Type="Italic">Panicum meyerianum</Emphasis>
Authors:Mi-Suk Seo  Sakiko Takahashi  Koh-ichi Kadowaki  Makoto Kawamukai  Manabu Takahara  Tadashi Takamizo
Institution:(1) Forage Crop Division, National Institute of Livestock and Grassland Science, 768 Senbonmatsu, Nasushiobara, Tochigi 329-2793, Japan;(2) Transgenic Crop Research and Development Center, National Institute of Agrobiological Science, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan;(3) National Institute of Agrobiological Science, 2-1-2 Kannondai, Tsukuba, Ibaraki 305-8602, Japan;(4) Faculty of Life and Environmental Science, Shimane University, 1060 Nishikawatsu, Matsue, Shimane 690-8504, Japan;(5) Present address: Genomics Division, Department of Agricultural Bio-resources, National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA), 150 Suin-ro, Gwonseon-gu, Suwon, 441-707, Korea;
Abstract:Panicum meyerianum Nees is a wild relative of Panicum maximum Jacq. (guinea grass), which is an important warm-season forage grass and biomass crop. We investigated the conditions that maximized the transformation efficiency of P. meyerianum by Agrobacterium infection by monitoring the expression of the β-glucuronidase (GUS) gene. The highest activities of GUS in calli were achieved by the co-cultivation of plants with Agrobacterium at 28°C for 6 days. We transferred the ddsA gene, which encodes decaprenyl diphosphate synthase and is required for coenzyme Q10 (CoQ10) synthesis, into P. meyerianum by using our optimized co-cultivation procedure for transformation. We confirmed by PCR and DNA gel blot hybridization that all hygromycin-resistant plants retained stable insertion of the hpt and ddsA genes. We also demonstrated strong expression of S14:DdsA protein in the leaves of transgenic P. meyerianum. Furthermore, we showed that transgenic P. meyerianum produced CoQ10 at levels 11–20 times higher than that of non-transformants. By comparison, the CoQ9 level in transgenic plants was dramatically reduced. This is the first report of efficient Agrobacterium-mediated transfer of a foreign gene into the warm-season grass P. meyerianum.
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