--Rodney G. Winkler and Michael Freeling
Seedlings that are low or null for ADH1, in contrast to normal seedlings,
do not germinate under partially anaerobic conditions. The Freeling lab
has utilized this behavior to select revertants of transposon-induced mutants
of Adh1 (C.-H. Chen, M. Freeling, Maydica 31:93-108, 1986). Adh1-3F1124
is a novel mutant that has a Mu3 insertion that duplicates the TATA
box and has been used in a number of selections for revertants (B. Kloeckener-Gruissem,
M. Freeling, MNL 1989). Under the conditions used Adh1-3F1124, which
has 6% of the ADH1 activity of the 3F progenitor, does not germinate,
although revertants with approximately 30% or greater activity do germinate.
Although this selection has yielded a number of revertants, most of the
germinating seedlings show no detectable ADH1 by starch gel analysis. Preliminary
experiments eliminated the possibility that these mutants are simple ADH2
overproducers. In the simplest terms the phenotypes of the 25 mutants being
analyzed are: anaerobic germination positive (unlike the progenitor), less
than 6% ADH1 activity by starch gel analysis in both the dry scutellum
and submerged root, normal levels of ADH1 in pollen (like the progenitor).
In situ analysis of the dry scutellum is also consistent with most of these
derivatives being lower in ADH1 than the progenitor. Several of the derivatives
are still mutable, yielding full ADH1 revertants. Currently these derivatives
are being analyzed at the molecular level to understand their altered pattern
of expression. One of these, Adh1-3F1124d104, has been shown to
delete all but approximately 600bp of the Mu3 insertion.
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