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Published on October 21, 2009; 10.1104/pp.109.146159

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Received August 13, 2009
Accepted October 18, 2009

Arabidopsis FAB1 / PIKfyve proteins are essential for development of viable pollen

Paul Whitley , Steven Hinz , and James Doughty *

Department of Biology and Biochemistry, University of Bath, Bath, Somerset, BA2 7AY, UK

* Corresponding author; email: bssjd{at}bath.ac.uk.

Phosphatidylinositol 3,5 bisphosphate (PtdIns(3,5)P2) is a phospholipid that has a role in controlling membrane trafficking events in yeast and animal cells. The function of this lipid in plants is unknown although its synthesis has been shown to be upregulated upon osmotic stress in plant cells. PtdIns(3,5)P2 is synthesised by the PIKfyve/Fab1 family of proteins with two orthologues, FAB1A and FAB1B, being present in Arabidopsis thaliana. In this study we attempt to address the role of this lipid by analysing the phenotypes of plants mutated in FAB1A and FAB1B. It was not possible to generate plants homozygous for mutations in both genes though single mutants were isolated. Both homozygous single mutant plant lines exhibited a leaf curl phenotype which was more marked in FAB1B mutants. Genetic transmission analysis revealed that failure to generate double mutant lines was entirely due to inviability of pollen carrying mutant alleles of both FAB1A and FAB1B. This pollen displayed severe defects in vacuolar reorganisation following the first mitotic division of development. The presence of abnormally large vacuoles in pollen at the tricellular stage resulted in the collapse of the majority of grains carrying both mutant alleles. This demonstrates a crucial role for PtdIns(3,5)P2 in modulating the dynamics of vacuolar rearrangement essential for successful pollen development. Taken together our results are consistent with PtdIns(3,5)P2 production being central to cellular responses to changes in osmotic conditions.






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