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Functional analysis of OsPUT1, a rice polyamine uptake transporter

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Abstract

Polyamines are nitrogenous compounds found in all eukaryotic and prokaryotic cells and absolutely essential for cell viability. In plants, they regulate several growth and developmental processes and the levels of polyamines are also correlated with the plant responses to various biotic and abiotic stresses. In plant cells, polyamines are synthesized in plastids and cytosol. This biosynthetic compartmentation indicates that the specific transporters are essential to transport polyamines between the cellular compartments. In the present study, a phylogenetic analysis was used to identify candidate polyamine transporters in rice. A full-length cDNA rice clone AK068055 was heterologously expressed in the Saccharomyces cerevisiae spermidine uptake mutant, agp2∆. Radiological uptake and competitive inhibition studies with putrescine indicated that rice gene encodes a protein that functioned as a spermidine-preferential transporter. In competition experiments with several amino acids at 25-fold higher levels than spermidine, only methionine, asparagine, and glutamine were effective in reducing uptake of spermidine to 60% of control rates. Based on those observations, this rice gene was named polyamine uptake transporter 1 (OsPUT1). Tissue-specific expression of OsPUT1 by semiquantitative RT-PCR showed that the gene was expressed in all tissues except seeds and roots. Transient expression assays in onion epidermal cells and rice protoplasts failed to localize to a cellular compartment. The characterization of the first plant polyamine transporter sets the stage for a systems approach that can be used to build a model to fully define how the biosynthesis, degradation, and transport of polyamines in plants mediate developmental and biotic responses.

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Abbreviations

ADC:

Arginine decarboxylase

GABA:

γ-aminobutyric acid

PUT1:

Polyamine uptake transporter1

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Acknowledgements

This work was supported by a grant from the Ohio Plant Biotechnology Consortium. We thank Dr. John Gray, University of Toledo, for access to Bio-Rad gene gun system, Chan Ho Park and Dr. GL Wang, Ohio State University, for help with isolation of rice protoplasts, and Dr. Fengyu Li, Bowling Green State University, for help in acquiring microscopy images of transiently expressed onion epidermal cells.

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Authors and Affiliations

  1. Department of Biological Sciences, Bowling Green State University, 442, Life Sciences Building, Bowling Green, OH, 43403-09, USA

    Vaishali Mulangi, Vipaporn Phuntumart & Paul Morris

  2. Maisonneuve-Rosemont Hospital, Research Center, 5415, Boul. de l’Assomption, Montreal, QC, H1T 2M4, Canada

    Mustapha Aouida & Dindial Ramotar

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  1. Vaishali Mulangi
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  2. Vipaporn Phuntumart
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  3. Mustapha Aouida
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  4. Dindial Ramotar
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  5. Paul Morris
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Correspondence to Paul Morris.

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Mulangi, V., Phuntumart, V., Aouida, M. et al. Functional analysis of OsPUT1, a rice polyamine uptake transporter. Planta 235, 1–11 (2012). https://doi.org/10.1007/s00425-011-1486-9

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