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THE PLANT CELL, Vol 7, Issue 1 75-84, Copyright © 1995 by American Society of Plant Biologists
Cloning and Characterization of the Maize An1 Gene
R. J. Bensen, G. S. Johal, V. C. Crane, J. T. Tossberg, P. S. Schnable, R. B. Meeley and S. P. Briggs
Pioneer Hi-Bred International, Inc., P.O. Box 1004, Johnston, Iowa 50131
The Anther ear1 (An1) gene product is involved in the synthesis of
ent-kaurene, the first tetracyclic intermediate in the gibberellin (GA)
biosynthetic pathway. Mutations causing the loss of An1 function result in
a GA-responsive phenotype that includes reduced plant height, delayed
maturity, and development of perfect flowers on normally pistillate ears.
The an1::Mu2-891339 allele was recovered from a Mutator (Mu) F2 family.
Using Mu elements as molecular probes, an An1-containing restriction
fragment was identified and cloned. The identity of the cloned gene as An1
was confirmed by using a reverse genetics screen for maize families that
contain a Mu element inserted into the cloned gene and then by
demonstrating that the insertion causes an an1 phenotype. The predicted
amino acid sequence of the An1 cDNA shares homology with plant cyclases and
contains a basic N-terminal sequence that may target the An1 gene product
to the chloroplast. The sequence is consistent with the predicted
subcellular localization of AN1 in the chloroplast and with its biochemical
role in the cyclization of geranylgeranyl pyrophosphate, a 20-carbon
isoprenoid, to ent-kaurene. The semidwarfed stature of an1 mutants is in
contrast with the more severely dwarfed stature of GA-responsive mutants at
other loci in maize and may be caused by redundancy in this step of the GA
biosynthetic pathway. DNA gel blot analysis indicated that An1 is a
single-copy gene that lies entirely within the deletion of the an1-bz2-6923
mutant. However, homozygous deletion mutants accumulated ent-kaurene to 20%
of the wild-type level, suggesting that the function of An1 is supplemented
by an additional activity.
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