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Expression of CoQ10-producing ddsA transgene by efficient Agrobacterium-mediated transformation in Panicum meyerianum

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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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Abbreviations

GUS:

β-glucuronidase

ddsA :

Decaprenyl diphosphate synthase gene

CoQ:

Coenzyme Q

hpt :

Hygromycin phosphotransferase gene

PCR:

Polymerase chain reaction

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Acknowledgments

We thank Dr. Masumi Ebina for valuable suggestions and Dr. Hiroki Matsuyama for support with the HPLC analysis (National Institute of Livestock and Grassland Science). Also, we thank Taiji Adachi (Osaka, Japan) for critical reading of the manuscript. This work was funded by the Ministry of Agriculture, Forestry and Fisheries (MAFF) research project ‘Development of innovative crops through the molecular analysis of useful genes’.

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Author notes

  1. Mi-Suk Seo

    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

Authors and Affiliations

  1. Forage Crop Division, National Institute of Livestock and Grassland Science, 768 Senbonmatsu, Nasushiobara, Tochigi, 329-2793, Japan

    Mi-Suk Seo, Manabu Takahara & Tadashi Takamizo

  2. Transgenic Crop Research and Development Center, National Institute of Agrobiological Science, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan

    Sakiko Takahashi

  3. National Institute of Agrobiological Science, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan

    Koh-ichi Kadowaki

  4. Faculty of Life and Environmental Science, Shimane University, 1060 Nishikawatsu, Matsue, Shimane, 690-8504, Japan

    Makoto Kawamukai

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  1. Mi-Suk Seo
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Correspondence to Tadashi Takamizo.

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Seo, MS., Takahashi, S., Kadowaki, Ki. et al. Expression of CoQ10-producing ddsA transgene by efficient Agrobacterium-mediated transformation in Panicum meyerianum . Plant Cell Tiss Organ Cult 107, 325–332 (2011). https://doi.org/10.1007/s11240-011-9984-9

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  • DOI: https://doi.org/10.1007/s11240-011-9984-9

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