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Plant and Cell Physiology Advance Access originally published online on September 18, 2008
Plant and Cell Physiology 2008 49(11):1734-1746; doi:10.1093/pcp/pcn145
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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

Plastidial Localization of a Potato ‘Nudix’ Hydrolase of ADP-glucose Linked to Starch Biosynthesis

Francisco José Muñoz1,4, Edurne Baroja-Fernández1,4, Miroslav Ovecka1,2,4, Jun Li1, Toshiaki Mitsui3, María Teresa Sesma1, Manuel Montero1, Abdellatif Bahaji1, Ignacio Ezquer1 and Javier Pozueta-Romero1,*

1 Instituto de Agrobiotecnología, Universidad Pública de Navarra/Consejo Superior de Investigaciones Científicas/Gobierno de Navarra, Mutiloako etorbidea zenbaki gabe, 31192 Mutiloabeti, Nafarroa, Spain
2 Institute of Botany, Slovak Academy of Sciences, Dubravska cesta 14, SK-84523 Bratislava, Slovakia
3 Laboratories of Plant and Microbial Genome Control and Department of Applied Biological Chemistry, Niigata University, 2-8050 Ikarashi, Niigata, 950-2181 Japan

*Corresponding author: E-mail, javier.pozueta{at}unavarra.es; Fax, +34-948232191


  Abstract

Escherichia coli and potato (Solanum tuberosum) ADP-sugar pyrophosphatases (EcASPP and StASPP, respectively) are ‘Nudix’ hydrolases of the bacterial glycogen and starch precursor molecule, ADP-glucose (ADPG). We have previously shown that potato leaves expressing EcASPP either in the cytosol or in the chloroplast exhibited large reductions in the levels of starch, suggesting the occurrence of cytosolic and plastidial pools of ADPG linked to starch biosynthesis. In this work, we produced and characterized potato and Arabidopsis plants expressing EcASPP and StASPP fused with green fluorescent protein (GFP). Confocal fluorescence microscopy analyses of these plants confirmed that EcASPP–GFP has a cytosolic localization, whereas StASPP–GFP occurs in the plastid stroma. Both source leaves and potato tubers from EcASPP–GFP-expressing plants showed a large reduction of the levels of both ADPG and starch. In contrast, StASPP–GFP-expressing leaves and tubers exhibited reduced starch and normal ADPG contents when compared with control plants. With the exception of starch synthase in StASPP–GFP-expressing plants, no pleiotropic changes in maximum catalytic activities of enzymes closely linked to starch metabolism could be detected in EcASPP–GFP- and StASPP–GFP-expressing plants. The overall data (i) show that potato plants possess a plastidial ASPP that has access to ADPG linked to starch biosynthesis and (ii) are consistent with the occurrence of plastidic and cytosolic pools of ADPG linked to starch biosynthesis.

Keywords: Carbohydrate metabolism - Solanum tuberosum - StASPP - Stroma - Stromule

Abbreviations: ADPG, ADP-glucose; AGP, ADPG pyrophosphorylase; ASPP, ADP sugar pyrophosphatase; AtASPP, Arabidopsis thaliana ASPP; EcASPP, Escherichia coli ASPP; GFP, green fluorescent protein; StASPP, Solanum tuberosum ASPP; UDPG, UDP-glucose.

4These authors contributed equally to this work.

(Received August 29, 2008; Accepted September 15, 2008)
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