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JXB Advance Access published online on September 12, 2005

Journal of Experimental Botany, doi:10.1093/jxb/eri276
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org
Received February 21, 2005
Accepted August 2, 2005

RESEARCH PAPER

The autophagy-associated Atg8 gene family operates both under favourable growth conditions and under starvation stresses in Arabidopsis plants

Silvia Sláviková 1 *, Galia Shy 2 *, Youli Yao 2 *, Rina Glozman 3, Hanna Levanony 2, Shmuel Pietrokovski 4, Zvulun Elazar 5*, and Gad Galili 2

1 Department of Plant Sciences, The Weizmann Institute of Science, Rehovot 76100, Israel; Institute of Cell Biology, Comenius University, Bratislava 81107, Slovakia
2 Department of Plant Sciences, The Weizmann Institute of Science, Rehovot 76100, Israel
3 Department of Plant Sciences, The Weizmann Institute of Science, Rehovot 76100, Israel; Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel
4 Department of Molecular Genetics, The Weizmann Institute of Science, Rehovot 76100, Israel
5 Department of Biological Chemistry, The Weizmann Institute of Science, Rehovot 76100, Israel

* To whom correspondence should be addressed.
Zvulun Elazar, E-mail: bmzevi{at}wicc.weizmann.ac.il


   Abstract

Arabidopsis plants possess a family of nine AtAtg8 gene homologues of the yeast autophagy-associated Apg8/Aut7 gene. To gain insight into how these genes function in plants, first, the expression patterns of five AtAtg8 homologues were analysed in young Arabidopsis plants grown under favourable growth conditions or following exposure to prolonged darkness or sugar starvation. Promoters, plus the entire coding regions (exons and introns) of the AtAtg8 genes, were fused to the {beta}-glucuronidase reporter gene and transformed into Arabidopsis plants. In all plants, grown under favourable growth conditions, {beta}-glucuronidase staining was much more significant in roots than in shoots. Different genes showed distinct spatial and temporal expression patterns in roots. In some transgenic plants, {beta}-glucuronidase staining in leaves was induced by prolonged darkness or sugar starvation. Next, Arabidopsis plants were transformed with chimeric gene-encoding Atg8f protein fused to N-terminal green fluorescent protein and C-terminal haemagglutinin epitope tags. Analysis of these plants showed that, under favourable growth conditions, the Atg8f protein is efficiently processed and is localized to autophagosome-resembling structures, both in the cytosol and in the central vacuole, in a similar manner to its processing and localization under starvation stresses. Moreover, treatment with a cocktail of proteasome inhibitors did not prevent the turnover of this protein, implying that its turnover takes place in the vacuoles, as occurs in yeasts. The results suggest that, in plants, the cellular processes involving the Atg8 genes function efficiently in young, non-senescing tissues, both under favourable growth conditions and under starvation stresses.

Keywords: Apg8; Arabidopsis; autophagy; germination; starvation; stress.

*These authors contributed equally to this work.


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