Binary transformation systems based on 'shooter' mutants of Agrobacterium tumefaciens: A simple, efficient and universal gene transfer technology that permits marker gene elimination

V. Mihálka, E. Balázs, I. Nagy

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

A simple transformation procedure with a positive selection scheme using the expression of the isopentenyl transferase (ipt) gene of transfer DNA (T-DNA) 'shooter' mutants of Agrobacterium tumefaciens was elaborated. After comparing several 'shooter' mutants we found that particular strains frequently produced phenotypically normal shoots after co-culturing with tobacco leaf explants. Shoots selected for normal phenotype showed apical dominance and could be rooted with the same efficiency as non-transformed shoots. When binary vectors were introduced into these strains, stably integrated binary vector T-DNA sequences were found in some regenerants, which were produced under non-selective conditions on growth-regulator-free medium. Such phenotypically normal transformants typically lacked a stably integrated ipt gene. Normal looking shoots could also be produced in tomato, muskmelon and sweet pepper.

Original languageEnglish
Pages (from-to)778-784
Number of pages7
JournalPlant Cell Reports
Volume21
Issue number8
Publication statusPublished - Apr 1 2003

Fingerprint

Agrobacterium radiobacter
gene transfer
transfer DNA
mutants
genetic markers
shoots
transferases
muskmelons
apical dominance
sweet peppers
growth regulators
explants
tobacco
tomatoes
phenotype
nucleotide sequences
leaves
genes

Keywords

  • Isopentenyl transferase
  • Marker-free transformation
  • Shooter mutants
  • Sweet pepper
  • Transgenic plants

ASJC Scopus subject areas

  • Plant Science

Cite this

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