Transformation of Statice (Limonium sinuatum Mill.) by Agrobacterium tumefaciens-Mediated Gene Transfer.

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  • IGAWA Tomoko
    Laboratory of Plant Cell Technology, Faculty of Horticulture, Chiba University
  • MII Masahiro
    Laboratory of Plant Cell Technology, Faculty of Horticulture, Chiba University

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A successful transformation procedure using Agrobacterium has been established for the most important ornamental statice, Limonium sinuatum Mill. Cell suspension cultures derived from friable calli, which were originated from leaf segments, were infected with Agrobacterium tumefaciens strain EHA105 (pIG121Hm) and co-cultivated for 2 days. The concentration of acetosyringone in co-cultivation medium and the growth stage of host plant cell used for infection were important factors for obtaining high transient GUS expression. After selection on medium with 75mgl-1 kanamycin, approximately 20 putative transgenic callus lines with kanamycin-resistance and GUS expression were obtained from 1g F. W. of cell cultures infected. Adventitious shoots were induced from 5 different putative transgenic callus lines on the regeneration medium containing 75mgl-1 kanamycin, and finally 16 transgenic plants which showed GUS expression in leaves were obtained. Presence of NPT II and GUS genes was confirmed by PCR analysis in the genomic DNAs of transgenic callus. Integration of GUS gene into the genome of transgenic plantlets derived from different transgenic callus lines was detected by Southern blot analysis.

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