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THE PLANT CELL, Vol 5, Issue 11 1555-1566, Copyright © 1993 by American Society of Plant Biologists
Genetic Isolation, Cloning, and Analysis of a Mutator-Induced, Dominant Antimorph of the Maize amylose extender1 Locus
P. S. Stinard, D. S. Robertson and P. S. Schnable
Department of Agronomy, Iowa State University, Ames, Iowa 50011
We report the genetic identification, molecular cloning, and
characterization of a dominant mutant at the amylose extender1 locus,
Ae1-5180. The identities of our clones are corroborated by their ability to
reveal DNA polymorphisms between seven wild-type revertants from Ae1-5180
relative to the Ae1-5180 mutant allele and between four of five
independently derived, Mutator (Mu)-induced recessive ae1 alleles relative
to their respective wild-type progenitor alleles. The Ae1-5180 mutation is
associated with two Mu1 insertions flanked by complex rearrangements of
ae1-related sequences. One of the Mu1 elements is flanked by inverted
repeats of ae1-related DNA of at least 5.0 kb in length. This Mu1 element
and at least some of this flanking inverted repeat DNA are absent or
hypermethylated in six of seven wild-type revertants of Ae1-5180 that were
analyzed. The second Mu1 element is flanked on one side by the 5.0-kb
ae1-specific repeat and on the other side by a sequence that does not
hybridize to the ae1-related repeat sequence. This second Mu1 element is
present in revertants to the wild type and does not, therefore, appear to
affect ae1 gene function. A 2.7-kb ae1 transcript can be detected in
wild-type and homozygous ae1-Ref endosperms 20 days after pollination. This
transcript is absent in endosperms containing one, two, or three doses of
Ae1-5180. This result is consistent with a suppression model to explain the
dominant gene action of Ae1-5180 and establishes Ae1-5180 as an antimorphic
allele. Homozygous wild-type seedlings produce no detectable transcript,
indicating some degree of tissue specificity for ae1 expression. Sequence
analyses establish that ae1 encodes starch branching enzyme II.
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