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Plant and Cell Physiology Advance Access published online on March 6, 2008

Plant and Cell Physiology, doi:10.1093/pcp/pcn034
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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 e-mail: journals.permissions@oxfordjournals.org

Comparative mutant analysis of Arabidopsis ABCC-type ABC transporters: AtMRP2 Contributes to Detoxification, Vacuolar Organic Anion Transport and Chlorophyll Degradation.

Annie Frelet-Barrand1,5, H. Üner Kolukisaoglu2,6, Sonia Plaza3, Maika Rüffer2, Louis Azevedo1, Stefan Hörtensteiner1, Krasimira Marinova1,7, Barbara Weder1, Burkhard Schulz4 and Markus Klein1,8

1 Zurich Basel Plant Science Center, University of Zurich, Plant Biology, Zollikerstrasse 107, CH-8008 Zürich, Switzerland
2 University of Rostock, Institute of Biosciences, Albert-Einstein-Str. 3, D-18059 Rostock, Germany
3 University of Fribourg, Plant Biology, 3, Albert-Gockel, CH-1700 Fribourg, Switzerland
4 Purdue University, Horticulture and Landscape Architecture, 625 Agriculture Mall Dr., West Lafayette, IN, 47907-2010, USA

8Corresponding author: Dr. Markus Klein, University of Zurich, Plant Biology, Zollikerstrasse 107, CH-8008 Zurich, Switzerland. Phone: +41-1-634-8282, Fax: +41-1-634-8204, e-mail: markus.klein{at}botinst.uzh.ch


   Abstract

The enormous metabolic plasticity of plants allows to detoxify many harmful compounds that are generated during biosynthetic processes or are present as biotic or abiotic toxins in their environment. Derivatives of toxic compounds such as glutathione conjugates are moved into the central vacuole via ATP binding cassette (ABC)-type transporters of the Multidrug Resistance-associated Protein (MRP) subfamily. The Arabidopsis genome contains 15 AtMRP isogenes, four of which (AtMRP1, 2, 11 and 12) cluster together in one of two major phylogenetic clades. We isolated T-DNA knockout alleles in all four highly homologous AtMRP genes of this clade and subjected them to physiological analysis to assess the function of each AtMRP of this group. None of the single atmrp mutants displayed visible phenotypes under control conditions. In spite of the fact that AtMRP1 and AtMRP2 had been described as efficient ATP-dependent organic anion transporters in heterologous expression experiments, the contribution of three of the AtMRPs (1, 11, 12) to detoxification is marginal. Only knockouts in AtMRP2 exhibited a reduced sensitivity towards 1-chloro-2,4-dinitrobenzene, but not towards other herbicides. AtMRP2 but not AtMRP1, 11 and 12 is involved in chlorophyll degradation since ethylene-treated rosettes of atmrp2 showed reduced senescence and AtMRP2 expression is induced during senescence. This suggests that AtMRP2 is involved in vacuolar transport of chlorophyll catabolites. Vacuolar uptake studies demonstrated that transport of typical MRP substrates was reduced in atmrp2. We conclude that within clade I only AtMRP2 contributes significantly to overall organic anion pump activity in vivo.

Keywords: Arabidopsis thaliana - gene family - herbicide resistance - organic anion transport - senescence - T-DNA knockout


5 present address: Laboratoire de Physiologie Cellulaire Végétale, iRTSV, CNRS (UMR 5168) / UJF / INRA / CEA, F-38054 GRENOBLE cedex 9, France.

6 present address: University of Rostock, Center for Life Science Automation (celisca), Fiedrich-Barnewitz-Str. 8, D-18119 Rostock, Germany

7 present address: University Hospital Zurich, Clinical Pharmacology and Toxicology, Department of Internal Medicine, Ramistrasse 100/SHO2, CH-8091 Zurich, Switzerland

(Received July 11, 2007; Accepted February 16, 2008)
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