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

D. Tjokrokusumo; T. Heinrich; S. Wylie; R. Potter; J. McComb

doi:10.1007/s002990050009

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

Journal article

Peer reviewed

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

D. Tjokrokusumo, T. Heinrich, S. Wylie, R. Potter and J. McComb

Plant Cell Reports, Vol.19(8), pp.792-797

2000

DOI: https://doi.org/10.1007/s002990050009

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Abstract

Genetic transformation of Petunia hybrida with a reporter gene and selectable marker gene (35S-bar) was achieved in similar frequencies by pollinating flowers with pollen vacuum-infiltrated with Agrobacterium tumefaciens or applying a drop of Agrobacterium suspension to the stigma immediately prior to pollination. Nine percent of the T1, and 5% of the T2 progeny germinated in nutrient medium with 3 mgl/l Basta(R). Polymerase chain reaction assays indicated that of the Basta(R)-resistant plants, 66% of the T1 plants, and 61% of the T2 plants harboured the GUS gene. Histochemical assays showed that 10% of the putatively transformed T1 plants and 5% of their progeny expressed GUS in leaf tissue, pistils and young anthers. Southern hybridization confirmed genomic integration of the bar gene in one to three places in selected T1 and T2 progeny.

Details

Title: Vacuum infiltration of Petunia hybrida pollen with Agrobacterium tumefaciens to achieve plant transformation
Authors/Creators: D. Tjokrokusumo (Author/Creator) - Murdoch University
T. Heinrich (Author/Creator) - Murdoch University
S. Wylie (Author/Creator) - Murdoch University
R. Potter (Author/Creator) - Murdoch University
J. McComb (Author/Creator) - Murdoch University
Publication Details: Plant Cell Reports, Vol.19(8), pp.792-797
Publisher: Springer Verlag
Identifiers: 991005544584607891
Murdoch Affiliation: School of Biological Sciences and Biotechnology
Language: English
Resource Type: Journal article

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Citation topics: 3 Agriculture, Environment & Ecology; 3.4 Crop Science; 3.4.119 Micropropagation
Web Of Science research areas: Plant Sciences
ESI research areas: Plant & Animal Science