Quantification of Agrobacterium tumefaciens C58 attachment to Arabidopsis thaliana roots.

Abstract:

:Agrobacterium tumefaciens is the causal agent of crown gall disease and is a vector for DNA transfer in transgenic plants. The transformation process by A. tumefaciens has been widely studied, but the attachment stage has not been well characterized. Most measurements of attachment have used microscopy and colony counting, both of which are labor and time intensive. To reduce the time and effort required to analyze bacteria attaching to plant tissues, we developed a quantitative real-time PCR (qPCR) assay to quantify attached A. tumefaciens using the chvE gene as marker for the presence of the bacteria. The qPCR detection threshold of A. tumefaciens from pure culture was 104 cell equivalents/ml. The A. tumefaciens minimum threshold concentration from root-bound populations was determined to be 105 cell equivalents/ml inoculum to detect attachment above background. The qPCR assay can be used for measuring A. tumefaciens attachment in applications such as testing the effects of mutations on bacterial adhesion molecules or biofilm formation, comparing attachment across various plant species and ecotypes, and detecting mutations in putative attachment receptors expressed in plant roots.

journal_name

FEMS Microbiol Lett

authors

Petrovicheva A,Joyner J,Muth TR

doi

10.1093/femsle/fnx158

subject

Has Abstract

pub_date

2017-10-02 00:00:00

issue

18

eissn

0378-1097

issn

1574-6968

pii

4082725

journal_volume

364

pub_type

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