Plant Physiol. Drug Metab Dispos
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Published on December 19, 2008; 10.1104/pp.108.132811

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Received November 18, 2008
Accepted December 16, 2008

MATE-type Transporters Implicated in Vacuolar Sequestration of Nicotine in Tobacco Roots

Tsubasa Shoji , Koji Inai , Yoshiaki Yazaki , Yasutaka Sato , Hisabumi Takase , Nobukazu Shitan , Kazufumi Yazaki , Yumi Goto , Kiminori Toyooka , Ken Matsuoka , and Takashi Hashimoto *

Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara 630-0192, Japan; National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan; Research Institute for Sustainable Humanosphere, Kyoto University, Uji 611-0011, Japan; RIKEN Plant Science Center, Yokohama, Kanagawa 230-0045, Japan

* Corresponding author; email: hasimoto{at}bs.naist.jp.

Nicotine is a major alkaloid accumulating in the vacuole of tobacco (Nicotiana tabacum L.), but the transporters involved in the vacuolar sequestration are not known. We here report that tobacco genes (NtMATE1 and NtMATE2) encoding transporters of the Multidrug and Toxic Compound Extrusion (MATE) family are coordinately regulated with structural genes for nicotine biosynthesis in the root, with respect to spatial expression patterns, regulation by NIC regulatory loci, and induction by methyl jasmonate. Subcellular fractionation, immunogold electronmicroscopy, and expression of a GFP fusion protein all suggested that these transporters are localized to the vacuolar membrane. Reduced expression of the transporters rendered tobacco plants more sensitive to the application of nicotine. In contrast, overexpression of NtMATE1 in cultured tobacco cells induced strong acidification of the cytoplasm after jasmonate-elicitation or after the addition of nicotine under non-elicited conditions. Expression of NtMATE1 in yeast cells compromised the accumulation of exogenously supplied nicotine into the yeast cells. The results imply that these MATE-type proteins transport tobacco alkaloids from the cytosol into the vacuole in exchange for protons in alkaloid-synthesizing root cells.




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