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Published on May 5, 2006; 10.1104/pp.106.078063

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Received February 2, 2006
Returned for revision March 28, 2006
Accepted May 1, 2006

pho2, a phosphate overaccumulator, is caused by a nonsense mutation in a miR399 target gene

Kyaw Aung , Shu-I Lin , Chia-Chune Wu , Yu-Ting Huang , Chun-lin Su , and Tzyy-Jen Chiou *

Institute of BioAgricultural Sciences, Academia Sinica, Taipei 115, Taiwan R.O.C.; Molecular and Biological Agricultural Sciences Program, Taiwan International Graduate Program, Academia Sinica, Taipei 115, Taiwan R.O.C.; Department of Life Science, National Chung-Hsing University, Taichung 402, Taiwan R.O.C
Institute of BioAgricultural Sciences, Academia Sinica, Taipei 115, Taiwan R.O.C.; Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 114, Taiwan, R.O.C.
Institute of BioAgricultural Sciences, Academia Sinica, Taipei 115, Taiwan R.O.C.
Institute of BioAgricultural Sciences, Academia Sinica, Taipei 115, Taiwan R.O.C.; Molecular and Biological Agricultural Sciences Program, Taiwan International Graduate Program, Academia Sinica, Taipei 115, Taiwan R.O.C.; Department of Life Science, National Chung-Hsing University, Taichung 402, Taiwan R.O.C.; Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 114, Taiwan, R.O.C.

* Corresponding author; email: tjchiou{at}gate.sinica.edu.tw.

We recently demonstrated that miR399 controls phosphate (Pi) homeostasis by regulating the expression of UBC24 encoding a ubiquitin-conjugating E2 enzyme in Arabidopsis (Arabidopsis thaliana). Transgenic plants overexpressing miR399 accumulated excessive Pi in the shoots and displayed Pi toxic symptoms. In this study, we revealed that a previously identified Pi overaccumulator, pho2, is caused by a single nucleotide mutation resulting in early termination within the UBC24 gene. The level of full-length UBC24 mRNA was reduced and no UBC24 protein was detected in the pho2 mutant, whereas the upregulation of miR399 by Pi deficiency was not affected. Several characteristics of Pi toxicity in the pho2 mutant were similar to those in the miR399-overexpressing and UBC24 T-DNA knockout plants: both Pi uptake and translocation of Pi from roots to shoots increased and Pi remobilization within leaves was impaired. These phenotypes of pho2 mutation could be rescued by introduction of a wild-type copy of UBC24. Kinetic analyses revealed that greater Pi uptake in the pho2 and miR399-overexpressing plants is due to increased Vmax. The transcript level of most PHT1 Pi transporter genes was not significantly altered except PHT1;8 whose expression was enhanced in Pi-sufficient roots of pho2 and miR399-overexpressing plants compared to wild-type plants. In addition, changes in the expression of several organelle-specific Pi transporters were noticed, which may be associated with the redistribution of intracellular Pi under excess Pi. Furthermore, miR399 and UBC24 were co-localized in the vascular cylinder. This observation not only provides important insights into the interaction between miR399 and UBC24 mRNA but also supports their systemic function in Pi translocation and remobilization.




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