I have received quite a lot of feedback concerning use of the Tobacco
leaf infiltration protocol for expressing fluorescent protein fusions
transiently in Arabidopsis. The consensus is that it works to a point
but nowhere near as well as in Tobacco. We have used it to express
fluorescent proteins targeted to peroxisomes and Golgi bodies. One
interesting bit of feedback was that the fluorescence is not always
targeted as one would expect it to be when used in Arabidopsis and the
second interesting thing was that this type of protocol might work well
in some ecotypes but not in others. Specifically, I hear that
Wassilewskaija (WS-0) transforms well but that Columbia (Col-0) does
not. That would figure wouldn't it? I am a bit emabarassed to admit
that I don't know the background of the plants that we have
transformed. They looked Columbia-like but...
Clearly, this is a technique that would facilitate many group's research
if we can get it to work better. Most who have used the trick
successfully say that choice of leaf (still growing rosette leaf from
young healthy plant) seems to be critical. Try the technique if you can
and let us know the results.
C. John Runions, Ph.D.
School of Biological and Molecular Sciences
Oxford Brookes University
email: jrunions at brookes.ac.uk
phone: +44 (0) 1865 483 964