JXB Advance Access published online on November 2, 2008
Journal of Experimental Botany, doi:10.1093/jxb/ern265
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© 2008 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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RESEARCH PAPER |
Characterization of a new rice glutelin gene GluD-1 expressed in the starchy endosperm
1Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, 2-1-2 Kan-nondai, Tsukuba, 305-8602, Japan
2QTL Genomics Research Center, National Institute of Agrobiological Sciences, 2-1-2 Kan-nondai, Tsukuba, 305-8602, Japan
To whom correspondence should be addressed: E-mail: takaiwa{at}affrc.go.jp
A new glutelin gene, designated GluD-1, has been discovered by comparing the seed storage proteins from 48 japonica and indica rice cultivars on SDS-PAGE gels. Evidence that GluD-1 is a member of the glutelin family was provided by Western blots using anti-glutelin antiserum and by mapping the gene to the chromosomal glutelin gene cluster. The limited GluD-1 size polymorphism among the rice varieties is due to amino acid substitutions rather than to post-transcriptional modification. GluD-1 is maximally expressed in the starchy endosperm starting at 5 d after flowering (DAF) and increasing through 30 DAF, a major difference from the other glutelins which are primarily expressed in the subaleurone from 10–16 DAF. Only about 0.2 kb of the GluD-1 promoter was sufficient to confer inner starchy endosperm-specific expression. The 0.2 kb truncated GluD-1 promoter contains a bifactorial endosperm box consisting of a truncated GCN4 motif (TGA(G/C)TCA) and AAAG Prolamin box (P box), and ACGT and AACA motifs as cis-regulatory elements. Gel retardation assays and trans-activation experiments indicated that the truncated GCN4 and P box are specifically recognized by RISBZ1 b-ZIP and RPBF Dof activators in vitro, respectively, and are synergistically transactivated, indicating that combinatorial interactions of these motifs are involved in essential endosperm-specific regulation. Furthermore, deviation from the cognate GCN4 motif alters tissue-specific expression in the inner starchy endosperm to include other endosperm tissues.
Key words: bZIP, DOF, endosperm specific expression, glutelin, promoter, rice, seed storage protein, transgenic crop
* Present address: Department of Biology, Faculty of Science, University of Toyama, 3190-Gofuku, Toyama 930-8555, Japan.
Received 4 August 2008; Revised 23 September 2008 Accepted 6 October 2008
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