Vacuum infiltration of Petunia hybrida pollen with Agrobacterium tumefaciens to achieve plant transformation

Citation
D. Tjokrokusumo et al., Vacuum infiltration of Petunia hybrida pollen with Agrobacterium tumefaciens to achieve plant transformation, PL CELL REP, 19(8), 2000, pp. 792-797
Citations number
24
Categorie Soggetti
Plant Sciences","Animal & Plant Sciences
Journal title
PLANT CELL REPORTS
ISSN journal
07217714 → ACNP
Volume
19
Issue
8
Year of publication
2000
Pages
792 - 797
Database
ISI
SICI code
0721-7714(200007)19:8<792:VIOPHP>2.0.ZU;2-J
Abstract
Genetic transformation of Petunia hybrida with a reporter gene and selectab le marker gene (35S-bar) was achieved in similar frequencies by pollinating flowers with pollen vacuum-infiltrated with Agrobacterium tumefaciens or a pplying a drop of Agrobacterium suspension to the stigma immediately prior to pollination. Nine percent of the T-1, and 5% of the T-2 progeny germinat ed in nutrient medium with 3 mgl/l Basta (R). Polymerase chain reaction ass ays indicated that of the Basta (R)-resistant plants, 66% of the T-1 plants , and 61% of the T-2 plants harboured the GUS gene. Histochemical assays sh owed that 10% of the putatively transformed T-1 plants and 5% of their prog eny expressed GUS in leaf tissue, pistils and young anthers. Southern hybri dization confirmed genomic integration of the bar gene in one to three plac es in selected T-1 and T-2 progeny.