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Plant and Cell Physiology Advance Access published online on April 13, 2006
Plant and Cell Physiology, doi:10.1093/pcp/pcj049
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Plant and Cell Physiology 2006 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received January 25, 2006
Accepted April 5, 2006

Regular Paper

The Correlation between Expression and Localization of a Foreign Gene Product in Rice Endosperm

Hiroshi Yasuda 1, Yuji Hayashi 2, Takahito Jomori 2, and Fumio Takaiwa 1 *

1 Department of Plant Biotechnology, National institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan
2 Sanwa Kagaku Kenkyusyo Co Ltd., Inabe, Mie 511-0406, Japan

* To whom correspondence should be addressed.
Fumio Takaiwa, E-mail: takaiwa{at}nias.affrc.go.jp


  Abstract

Glucagon-like peptide 1 (GLP-1) is a 30 amino acid peptide hormone involved in insulin stimulation that is dependent upon blood glucose levels. We have previously reported that when this short peptide gene was directly expressed under the control of a glutelin promoter and its signal peptide, it was not accumulated in transgenic rice seed due to gene silencing. However, when the modified GLP-1 (mGLP-1) gene was enlarged to 5xmGLP-1 (mGLPx5) by tandem repeat, no silencing was observed. The mGLPx5 peptide could be accumulated in rice seed and its localization was mainly limited to the endoplasmic reticulum (ER). We also investigated alternate cellular localization sites that would increase accumulation. The relationship between expression level and localization was examined by attaching the chitinase targeting signal peptide to mGLPx5 to direct it into the intercellular space (apoplast), or by expression as a fusion protein with glutelin by insertion into a variable region of the acidic subunit, thus directing the peptide to protein body II (PB II). Attachment of the KDEL ER retention signal to the 6xmGLP-1 (mGLPx6) or its fusion to the C-terminus of the 13-kD prolamin directed the peptide to ER or protein body I (PB I), respectively. Unexpectedly, these results indicated that mGLPx5 without any signal except for the glutelin signal peptide was most highly accumulated in rice endosperm. It can thus be concluded that the ER is a suitable intracellular organelle for accumulation of mGLPx5 peptide.

Keywords: rice; endosperm; subaleurone cell; GLP-1; transgenic plant.
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