Multidrug and Toxic Compound Extrusion-type Transporters Implicated in Vacuolar Sequestration of Nicotine in Tobacco Roots

Overview

Journal Plant Physiol

Specialty Physiology

Date 2008 Dec 23

PMID 19098091

Citations 88

Authors

Tsubasa Shoji

Koji Inai

Yoshiaki Yazaki

Yasutaka Sato

Hisabumi Takase

Nobukazu Shitan

Kazufumi Yazaki

Yumi Goto

Kiminori Toyooka

Ken Matsuoka

Takashi Hashimoto

Affiliations

Soon will be listed here.

Abstract

Nicotine is a major alkaloid accumulating in the vacuole of tobacco (Nicotiana tabacum), 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 electron microscopy, and expression of a green fluorescent protein 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 nonelicited conditions. Expression of NtMATE1 in yeast (Saccharomyces cerevisiae) 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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References

Marinova K, Pourcel L, Weder B, Schwarz M, Barron D, Routaboul J . The Arabidopsis MATE transporter TT12 acts as a vacuolar flavonoid/H+ -antiporter active in proanthocyanidin-accumulating cells of the seed coat. Plant Cell. 2007; 19(6):2023-38. PMC: 1955721. DOI: 10.1105/tpc.106.046029. View

Debeaujon I, Peeters A, Leon-Kloosterziel K, Koornneef M . The TRANSPARENT TESTA12 gene of Arabidopsis encodes a multidrug secondary transporter-like protein required for flavonoid sequestration in vacuoles of the seed coat endothelium. Plant Cell. 2001; 13(4):853-71. PMC: 135529. DOI: 10.1105/tpc.13.4.853. View

. A simple and general method for transferring genes into plants. Science. 1985; 227(4691):1229-31. DOI: 10.1126/science.227.4691.1229. View

An G . High efficiency transformation of cultured tobacco cells. Plant Physiol. 1985; 79(2):568-70. PMC: 1074928. DOI: 10.1104/pp.79.2.568. View

Li L, He Z, Pandey G, Tsuchiya T, Luan S . Functional cloning and characterization of a plant efflux carrier for multidrug and heavy metal detoxification. J Biol Chem. 2001; 277(7):5360-8. DOI: 10.1074/jbc.M108777200. View

Kajikawa M, Hirai N, Hashimoto T . A PIP-family protein is required for biosynthesis of tobacco alkaloids. Plant Mol Biol. 2008; 69(3):287-98. DOI: 10.1007/s11103-008-9424-3. View

LOCHMANN H, Bazzanella A, Kropsch S, Bachmann K . Determination of tobacco alkaloids in single plant cells by capillary electrophoresis. J Chromatogr A. 2001; 917(1-2):311-7. DOI: 10.1016/s0021-9673(01)00627-6. View

Heim W, Sykes K, Hildreth S, Sun J, Lu R, Jelesko J . Cloning and characterization of a Nicotiana tabacum methylputrescine oxidase transcript. Phytochemistry. 2006; 68(4):454-63. DOI: 10.1016/j.phytochem.2006.11.003. View

Steppuhn A, Gase K, Krock B, Halitschke R, Baldwin I . Nicotine's defensive function in nature. PLoS Biol. 2004; 2(8):E217. PMC: 509292. DOI: 10.1371/journal.pbio.0020217. View

10.

Cane K, Mayer M, Lidgett A, Michael A, Hamill J . Molecular analysis of alkaloid metabolism in AABB v. aabb genotype Nicotiana tabacum in response to wounding of aerial tissues and methyl jasmonate treatment of cultured roots. Funct Plant Biol. 2020; 32(4):305-320. DOI: 10.1071/FP04008. View

11.

Koes R, Verweij W, Quattrocchio F . Flavonoids: a colorful model for the regulation and evolution of biochemical pathways. Trends Plant Sci. 2005; 10(5):236-42. DOI: 10.1016/j.tplants.2005.03.002. View

12.

Facchini P . ALKALOID BIOSYNTHESIS IN PLANTS: Biochemistry, Cell Biology, Molecular Regulation, and Metabolic Engineering Applications. Annu Rev Plant Physiol Plant Mol Biol. 2001; 52:29-66. DOI: 10.1146/annurev.arplant.52.1.29. View

13.

Wesley S, Helliwell C, Smith N, Wang M, Rouse D, Liu Q . Construct design for efficient, effective and high-throughput gene silencing in plants. Plant J. 2001; 27(6):581-90. DOI: 10.1046/j.1365-313x.2001.01105.x. View

14.

Shoji T, Nakajima K, Hashimoto T . Ethylene suppresses jasmonate-induced gene expression in nicotine biosynthesis. Plant Cell Physiol. 2000; 41(9):1072-6. DOI: 10.1093/pcp/pcd027. View

15.

Katoh A, Shoji T, Hashimoto T . Molecular cloning of N-methylputrescine oxidase from tobacco. Plant Cell Physiol. 2007; 48(3):550-4. DOI: 10.1093/pcp/pcm018. View

16.

Shoji T, Winz R, Iwase T, Nakajima K, Yamada Y, Hashimoto T . Expression patterns of two tobacco isoflavone reductase-like genes and their possible roles in secondary metabolism in tobacco. Plant Mol Biol. 2002; 50(3):427-40. DOI: 10.1023/a:1019867732278. View

17.

Fulnecek J, Lim K, Leitch A, Kovarik A, Matyasek R . Evolution and structure of 5S rDNA loci in allotetraploid Nicotiana tabacum and its putative parental species. Heredity (Edinb). 2002; 88(1):19-25. DOI: 10.1038/sj.hdy.6800001. View

18.

Kidd S, Melillo A, Lu R, Reed D, Kuno N, Uchida K . The A and B loci in tobacco regulate a network of stress response genes, few of which are associated with nicotine biosynthesis. Plant Mol Biol. 2006; 60(5):699-716. DOI: 10.1007/s11103-005-5546-z. View

19.

Durrett T, Gassmann W, Rogers E . The FRD3-mediated efflux of citrate into the root vasculature is necessary for efficient iron translocation. Plant Physiol. 2007; 144(1):197-205. PMC: 1913786. DOI: 10.1104/pp.107.097162. View

20.

Baldwin I . An ecologically motivated analysis of plant-herbivore interactions in native tobacco. Plant Physiol. 2001; 127(4):1449-58. PMC: 1540177. View