GUS staining in transgenic plants

Sat Oct 17 16:32:45 EST 1992

 I would appreciate any information you might have on factors that
affect the activity of the 35S promoter or the activity of the GUS
protein in plant tissues, specifically roots.  I'm seeing variable
staining of the subapical (0.2-2 cm) zone of tomato lateral roots
expressing GUS from the 35S promoter.  I know the problem is not
tissue specificity per se, since a significant percentage of my
roots stain in this zone.  Roots from a single original transformant
are variable, as are roots from different transformants, so I don't
think the problem is related to an insertion site effect.  I have
performed fluorometric assays on roots where the tip (meristematic
zone) and body of the root were heavily stained and the subapical
zone was not, and found that the level of GUS activity was at least
10X lower in the subapical zone.  This rules out problems with
X-Gluc penetration or product oxidation.

I would appreciate any ideas or comments, and I'd also like to hear
from people who have worked with 35S-GUS in roots and either have
seen the same problem or are sure they don't.

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