Attachment of Agrobacterium tumefaciens to leaf tissue in response to infiltration conditions

Christopher W. Simmons, Jean S. Vandergheynst, N. Nitin

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Transient expression of recombinant proteins in plant tissues following Agrobacterium-mediated gene transfer is a promising technique for rapid protein production. However, transformation rates and transient expression levels can be sub-optimal depending on process conditions. Attachment of Agrobacterium tumefaciens to plant cells is an early, critical step in the gene transfer pathway. Bacterial attachment levels and patterns may influence transformation and, by extension, transient expression. In this study, attachment of A. tumefaciens to lettuce leaf tissue was investigated in response to varying infiltration conditions, including bacterial density, surfactant concentration, and applied vacuum level. Bacterial density was found to most influence attachment levels for the levels tested (108, 109, and 1010 CFU/mL), with the relationship between bacterial density and attachment levels following a saturation trend. Surfactant levels tested (Break-Thru S240: 1, 10, 100, and 1,000 μL/L) also had a significant positive effect on bacterial attachment while vacuum level (5, 25, and 45 kPa) did not significantly affect attachment in areas exposed to bacteria. In planta transgene transient expression levels were measured following infiltration with 108, 109, and 1010 CFU/mL bacterial suspension. Notably, the highest attachment level tested led to a decrease in transient expression, suggesting a potential link between bacterial attachment levels and downstream phenomena that may induce gene silencing. These results illustrate that attachment can be controlled by adjusting infiltration conditions and that attachment levels can impact transgene transient expression in leaf tissue.

Original languageEnglish (US)
Pages (from-to)1137-1144
Number of pages8
JournalBiotechnology Progress
Volume30
Issue number5
DOIs
StatePublished - Sep 1 2014

Fingerprint

Agrobacterium tumefaciens
Vacuum
Transgenes
Surface-Active Agents
Lettuce
Agrobacterium
Plant Cells
Gene Silencing
Recombinant Proteins
Genes
Suspensions
Bacteria
Proteins

Keywords

  • Agrobacterium tumefaciens
  • Agroinfiltration
  • Bacterial attachment
  • Confocal microscopy
  • Plant-microbe interaction

ASJC Scopus subject areas

  • Biotechnology

Cite this

Attachment of Agrobacterium tumefaciens to leaf tissue in response to infiltration conditions. / Simmons, Christopher W.; Vandergheynst, Jean S.; Nitin, N.

In: Biotechnology Progress, Vol. 30, No. 5, 01.09.2014, p. 1137-1144.

Research output: Contribution to journalArticle

Simmons, Christopher W. ; Vandergheynst, Jean S. ; Nitin, N. / Attachment of Agrobacterium tumefaciens to leaf tissue in response to infiltration conditions. In: Biotechnology Progress. 2014 ; Vol. 30, No. 5. pp. 1137-1144.
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