Association of single-stranded transferred DNA from Agrobacterium tumefaciens with tobacco cells

During the inception of crown gall tumorigenesis, the transferred DNA (T-DNA) is processed from the Ti (tumor inducing) plasmid of Agrobacterium tumefaciens and is transferred to plant cells. T-DNA processing and transfer require the induction of vir (virulence) genes by phenolic compounds secreted...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1994-04, Vol.91 (8), p.2994-2998
Main Authors: Yusibov, V.M, Steck, T.R, Gupta, V, Gelvin, S.B
Format: Article
Language:English
Subjects:
ADN
AGROBACTERIUM TUMEFACIENS
Agrobacterium tumefaciens - genetics
Agronomy. Soil science and plant productions
Bacteria
Base Sequence
Biochemistry
Biological and medical sciences
Biotechnology
Cells, Cultured
Coculture techniques
COMPOSE PHENOLIQUE
COMPUESTOS FENOLICOS
Cytoplasm
Deoxyribonucleic acid
DNA
DNA Primers - chemistry
DNA, Single-Stranded - metabolism
Fundamental and applied biological sciences. Psychology
GENE
Gene Transfer Techniques
GENES
Genes, Bacterial
Genetic engineering
Genetic engineering applications
Genetic technics
Genetics and breeding of economic plants
In vitro gene amplification. Pcr technique
In Vitro Techniques
Methods. Procedures. Technologies
Molecular Sequence Data
Molecules
Nicotiana
NICOTIANA TABACUM
Nucleic acids
Pica
Plant breeding: fundamental aspects and methodology
Plant cells
Plants, Toxic
PLASMIDE
PLASMIDIOS
Plasmids
PODER PATOGENO
Polymerase chain reaction
POUVOIR PATHOGENE
PROTOPLASTE
PROTOPLASTOS
Protoplasts
Tobacco
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Summary:During the inception of crown gall tumorigenesis, the transferred DNA (T-DNA) is processed from the Ti (tumor inducing) plasmid of Agrobacterium tumefaciens and is transferred to plant cells. T-DNA processing and transfer require the induction of vir (virulence) genes by phenolic compounds secreted by wounded plant cells. After vir gene induction, both single-stranded (T-strands) and double-stranded forms of processed T-DNA accumulate in the bacteria. Although current models favor the transfer of T-strands to plants, there has yet been no experimental evidence to show this. In this paper, we show that T-strands disappear from acetosyringone-induced A. tumefaciens within 30 min of bacterial cocultivation with tobacco protoplasts. PCR analysis of T-DNA associated with protoplasts indicates that single-stranded, but not double-stranded, T-DNA can be detected in the plant cells within 30 min of bacterial cocultivation. Control experiments show that this T-DNA does not originate from lysed contaminating bacterial cells. T-DNA transfer depends on a functional bacterial virB operon. Protoplast infections using an A. tumefaciens virE mutant result in a low level of accumulation of T-strands in the plant cells.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.91.8.2994