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Plant and Cell Physiology Advance Access originally published online on September 6, 2008
Plant and Cell Physiology 2008 49(10):1589-1599; doi:10.1093/pcp/pcn135
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Co-Expression of Soybean Glycinins A1aB1b and A3B4 Enhances Their Accumulation Levels in Transgenic Rice Seed

Fumio Takaiwa1,*, Chiyoko Sakuta1, Seon-Kang Choi2, Yoshifumi Tada1, Takayasu Motoyama2 and Shigeru Utsumi2

1 Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki, 305-8602 Japan
2 Laboratory of Food Quality Design and Development, Graduate School of Agriculture, Kyoto University, Gokasho, Uji, Kyoto, 611-0011 Japan

*Corresponding author: E-mail, takaiwa{at}nias.affrc.go.jp; Fax, +81-29-838-8373.


  Abstract

The soybean major storage protein glycinin is encoded by five genes, which are divided into two subfamilies. Expression of A3B4 glycinin in transgenic rice seed reached about 1.5% of total seed protein, even if expressed under the control of strong endosperm-specific promoters. In contrast, expression of A1aB1b glycinin reached about 4% of total seed protein. Co-expression of the two proteins doubled accumulation levels of both A1aB1b and A3B4 glycinins. This increase can be largely accounted for by their aggregation with rice glutelins, self-assembly and inter-glycinin interactions, resulting in the enrichment of globulin and glutelin fractions and a concomitant reduction of the prolamin fraction. Immunoelectron microscopy indicated that the synthesized A1aB1b glycinin was predominantly deposited in protein body-II (PB-II) storage vacuoles, whereas A3B4 glycinin is targeted to both PB-II and endoplasmic reticulum (ER)-derived protein body-I (PB-I) storage structures. Co-expression with A1aB1b facilitated targeting of A3B4 glycinin into PB-II by sequestration with A1aB1b, resulting in an increase in the accumulation of A3B4 glycinin.

Keywords: Endosperm - Glycinin - Immunolocalization - Protein body - Seed storage protein - Transgenic rice

Abbreviations: BSA, bovine serum albumin; CBB, Coomassie Brilliant Blue; DAF, days after flowering; DTT, dithiothreitol; ER, endoplasmic reticulum; PAC, precursor-accumulating; PB, protein body; PBS, phosphate-buffered saline; PSV, protein storage vacuole; SDG, sucrose density gradient; UTR, untranslated region.

(Received June 22, 2008; Accepted August 29, 2008)
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