Agrobacterium-mediated transformation of the dwarf pomegranate (Punica granatum L. var. nana)

作者：Terakami Shingo; Matsuta Nagao; Yamamoto Toshiya; Sugaya Sumiko^*; Gemma Hiroshi; Soejima Junichi

来源：Plant Cell Reports, 2007, 26(8): 1243-1251.

DOI：10.1007/s00299-007-0347-2

• Punica granatum
• pomegranate
• Agrobacterium tumefaciens
• transformation

The dwarf pomegranate (Punica granatum L. var. nana) is a dwarf ornamental plant that has the potential to be the model plant of perennial fruit trees because it bears fruits within 1 year of seedling. We established an Agrobacterium-mediated transformation system for the dwarf pomegranate. Adventitious shoots regenerated from leaf segments were inoculated with A. tumefaciens strain EHA105 harboring the binary vector pBin19-sgfp, which contains neomycin phosphotransferase (npt II) and green fluorescent protein (gfp) gene as a selectable and visual marker, respectively. After co-cultivation, the inoculated adventitious shoots were cut into small pieces to induce regeneration, and then selected on MS medium supplemented with 0.5 mu M alpha-naphthaleneacetic acid (NAA), 5 mu M N-6-benzyladenine (BA), 0.3% gellan gum, 50 mg/l kanamycin, and 10 mg/l meropenem. Putative transformed shoots were regenerated after 6-8 months of selection. PCR and PCR-Southern blot analysis revealed the integration of the transgene into the plant genome. Transformants bloomed and bore fruits within 3 months of being potted, and the inheritance of the transgene was confirmed in T-1 generations. The advantage of the transformation of dwarf pomegranate was shown to be the high transformation rate. The establishment of this transformation system is invaluable for investigating fruit-tree-specific phenomena.

• 出版日期2007-8