Molecular Biology and Evolution, Vol 15, 978-987, Copyright © 1998 by Society for Molecular Biology and Evolution
HY Hirano, M Eiguchi and Y Sano
The rice waxy (wx) locus has two functional alleles, Wxa and Wxb, which are
defined by a large difference in the amount of the gene product, called Wx
protein, that accumulates in mature seeds. To elucidate the molecular
mechanism underlying this difference and to identify the base change
causing the alteration of the regulation of the Wx gene during rice
evolution, we determined the nucleotide sequences of the regulatory region
of Wx alleles and analyzed their function in a transient assay system using
rice protoplasts. All Wxa alleles from Oryza sativa Indica, O. rufipogon,
and O. glaberrima have a normal sequence of GT at the 5' splice junction of
the first intron, representing a high expression level of the Wx
transcripts in the endosperm and a high beta-glucuronidase (GUS) activity
in protoplasts. On the other hand, Wxb alleles from two strains of O.
sativa Japonica have TT at the 5' splice junction, representing a low
expression level of the mature transcripts and a low GUS activity. Northern
blot analysis also indicated that a larger transcript, consisting of the
unspliced first intron, is closely correlated with the function of the Wxb
allele. These results suggest that a single base change at the 5' splice
junction causes inefficient splicing and, as a result, reduces the level of
mature transcript and the GUS activity in the Wxb allele. The Wxb allele in
O. saativa Japonica may have been differentiated from the Wxa allele of O.
rufipogon, its wild progenitor, by this mutation, and, therefore, a single
base change that has altered the regulation of the Wx gene at the
posttranscriptional level probably occurred during the domestication of
rice.
ORIGINAL ARTICLE
A single base change altered the regulation of the Waxy gene at the posttranscriptional level during the domestication of rice
National Institute of Genetics, Mishima, Japan. ahirano@hongo.ecc.u- tokyo.ac.jp
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