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Published on November 16, 2007; 10.1104/pp.107.110817

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Received October 15, 2007
Accepted November 11, 2007

ECA3, a Golgi-localised P2A-type-ATPase, plays a crucial role in manganese nutrition in Arabidopsis

Rebecca F. Mills , Melissa Louise Doherty , Rosa L. Lopez-Marques , Thilo Weimar , Paul Dupree , Michael G. Palmgren , Jon K. Pittman , and Lorraine E. Williams *

School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton, SO16 7PX, UK; Centre for Membrane Pumps in Cells and Disease - PUMPKIN, Danish National Research Foundation, Department of Plant Biology, University of Copenhagen, Thorvaldsensvej 40, DK-1871 Frederiksberg, Denmark; Department of Biochemistry, Building O, Downing Site, Cambridge CB2 1QW, UK; Faculty of Life Sciences, University of Manchester, 3.614 Stopford Building, Oxford Road, Manchester M13 9PT, UK

* Corresponding author; email: l.e.williams{at}soton.ac.uk.

Calcium and manganese are essential nutrients required for normal plant growth and development, and transport processes play a key role in regulating their cellular levels. Arabidopsis contains four P2A-type ATPase genes, AtECA1-4, which are expressed in all major organs of Arabidopsis. To elucidate the physiological role of AtECA2 and AtECA3 in Arabidopsis, several independent T-DNA insertion mutant alleles were isolated. When grown on medium lacking Mn, eca3 mutants, but not eca2 mutants, displayed a striking difference from wild-type plants. After approximately 8-9 d on this medium, the eca3 mutants became chlorotic and root and shoot growth were strongly inhibited compared to wild-type plants. These severe deficiency symptoms were suppressed by low levels of Mn indicating a crucial role for ECA3 in Mn nutrition in Arabidopsis. eca3 mutants were also more sensitive than wild-type plants and eca2 mutants on media lacking Ca, however the differences were not so striking as in this case all plants were severely affected. ECA3 partially restored the growth defect on high Mn of the S. cerevisiae pmr1 mutant, which is defective in a Golgi Ca/Mn pump (PMR1), and the S. cerevisiae K616 mutant (pmc1 pmr1 cnb1), defective in Golgi and vacuolar Ca/Mn pumps. ECA3 also rescued the growth defect of K616 on low Ca. Promoter:GUS studies show that ECA3 is expressed in a range of tissues and cells including primary root tips, root vascular tissue, hydathodes, and guard cells. When transiently expressed in tobacco, an ECA3-YFP fusion protein showed overlapping expression with the Golgi protein, GONST1. We propose that ECA3 is important for Mn and Ca homeostasis, possibly functioning in the transport of these ions into the Golgi. ECA3 is the first P-type ATPase to be identified in plants that is required under Mn-deficient conditions.




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