Agrobacterium-mediated transformation of the commercially important grapefruit cultivar Rio Red (Citrus paradisi Macf.)

Citation
Zn. Yang et al., Agrobacterium-mediated transformation of the commercially important grapefruit cultivar Rio Red (Citrus paradisi Macf.), PL CELL REP, 19(12), 2000, pp. 1203-1211
Citations number
38
Categorie Soggetti
Plant Sciences","Animal & Plant Sciences
Journal title
PLANT CELL REPORTS
ISSN journal
07217714 → ACNP
Volume
19
Issue
12
Year of publication
2000
Pages
1203 - 1211
Database
ISI
SICI code
0721-7714(200012)19:12<1203:ATOTCI>2.0.ZU;2-P
Abstract
Transgenic plants of grapefruit cv. Rio Red (Citrus paradisi Macf.) have be en obtained by Agrobacterium tumefaciens-mediated gene transfer using seedl ing-derived epicotyl segments as explants and kanamycin as the selective ag ent. The transformation procedure includes a shoot elongation phase with a liquid medium overlay, which provides additional selection against non-tran sgenic shoots. Transformed shoots are invigorated and multiplied on a non-s elective medium prior to grafting, thus assuring that plants can be recover ed from transgenic shoots. We have constructed a binary vector, pBin34SGUS, with an intron-containing P-glucuronidase gene (uidA) under the control of the Figwort mosaic virus 34S promoter. The 34S promoter efficiently drives uidA gene expression both in transient assays and in transgenic Rio Red le af tissue, although at levels five- to sevenfold lower than the Cauliflower mosaic virus 35S promoter. An untranslatable coat protein gene (uncp) of t he Citrus tristeza virus strain SY568 and the Galanthus nivalis agglutinin gene (gna) were inserted into pBin34SGUS and transgenic plants have been ob tained. Stable integration of the uncp and gna genes was confirmed by South ern hybridization and gna gene expression was confirmed by Western blot ana lysis.