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Plant and Cell Physiology Advance Access published online on May 30, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci141
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received March 8, 2005
Accepted May 24, 2005

Regular Paper

Contribution of Gibberellins to the Formation of Arabidopsis Seed Coat through Starch Degradation

Young-Cheon Kim 1, Masatoshi Nakajima 1*, Akira Nakayama 1, and Isomaro Yamaguchi 1

1 Department of Applied Biological Chemistry, The University of Tokyo, 1-1-1, Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan

* To whom correspondence should be addressed.
Masatoshi Nakajima, E-mail: nkjm{at}pgr1.ch.a.u-tokyo.ac.jp


  Abstract

To clarify the role of GAs on the seed development of Arabidopsis, we investigated the sites where GAs are synthesized and induce {alpha}-amylase genes. The spatial and temporal expression of the genes coding GA-biosynthetic enzymes and {alpha}-amylases were examined by RT-PCR and in situ hybridization. The mRNAs of AtGA20ox2, AtGA20ox3, and AtGA3ox4 began to be detectable 5-7 days after pollination. In situ hybridization showed that these genes were expressed almost simultaneously around starch granules in the outer integument, preceding the disappearance of those granules. AtGA20ox2 and AtGA3ox4 but not AtGA20ox3 also showed their signals at the rim of the developing embryo. The {alpha}-amylase gene, Amy3, which responded to GA, was mainly expressed in the developing seed, spatially overlapping with the expression of AtGA20ox2 and AtGA3ox4. These results suggest that GAs function in at least two sites of the seed: the outer integument and part of the embryo. We examined the phenotypes of a T-DNA insertion line of AtGA3ox4 and observed the following: (i) a decrease of {alpha}-amylase genes transcripts in young siliques, (ii) delay of starch degradation in the outer integument, (iii) disarrangement of the seed surface structure, and (iv) abnormal swelling pattern of polysaccharides after imbibition by the mature seed. These characteristics are phenotypes of plants under GA starvation, because the abnormalities could be almost overcome with applied GA, and the GA-treated mutant was indistinguishable with wild-type. These results strongly suggest that GAs in the outer integument would be required for the normal formation of the Arabidopsis seed coat.

Keywords: gibberellin; {alpha}-amylase; developing seed.
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