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JXB Advance Access published online on July 7, 2006

Journal of Experimental Botany, doi:10.1093/jxb/erl027
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© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org
Received October 18, 2005
Accepted April 13, 2006

RESEARCH PAPER

The role of arabinogalactan proteins binding to Yariv reagents in the initiation, cell developmental fate, and maintenance of microspore embryogenesis in Brassica napus L. cv. Topas

Xing-Chun Tang 1, Yu-Qing He 1, Ying Wang 1, and Meng-Xiang Sun 1 *

1 Key Laboratory of the MOE for the Development Biology, College of Life Science, Wuhan University, Wuhan 430072 China

* To whom correspondence should be addressed.
Meng-Xiang Sun, E-mail: mxsun{at}whu.edu.cn


   Abstract

Arabinogalactan proteins (AGPs) are extracellular proteoglycans involved in plant growth and development. The addition of {beta}-D-glucosyl Yariv reagent ({beta}GlcY), a synthetic phenylglycoside that specifically reacts with AGPs, to the culture medium notably disturbed microspore embryogenesis in a concentration-dependent manner. The initiation of microspore embryogenesis was clearly inhibited by 30 µM {beta}GlcY and completely inhibited by 50 µM {beta}GlcY. The transfer of microspore-derived embryos at different developmental stages into NLN6 medium containing 50 µM {beta}GlcY prohibited their normal development, as ~21.24, 43.99, and 59.73%, respectively, of the treated globular-, heart-, and torpedo-stage embryos exhibited numerous root hair-like structures. Both heart-stage and torpedo-stage embryos showed a rapid growth of roots with a large number of clustered root hairs. Some root hair-like structures were also observed on the apical portions of embryos. Microscopy of the treated embryos revealed that the basic patterns of cells at both the radial and apical-basal axes were greatly altered, such that the cells lost their ability to carry out programmed embryogenesis. These results show that the {beta}GlcY-AGP interaction modulates the developmental fate of embryonic cells, especially epidermal cells, and thereby strongly affects root generation and development. Immunofluorescence microscopy revealed that both JIM8 and JIM13 binding to AGP co-localize with {beta}GlcY-binding sites. Thus, AGPs binding to {beta}GlcY, co-localized with Jim8- and Jim13-binding protein, appear to play a crucial role in the initiation of Brassica microspore embryogenesis and the maintenance of cell differentiation during embryonic development. In addition, these proteins may also be involved in the regulation of root generation.

Keywords: Arabinogalactan protein; Brassica napus L. cv. Topas; cell fate; development expansion; microspore embryogenesis.
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