An effective plant alkaloid chemical defense requires a variety of transport processes, but few alkaloid transporters have been characterized at the molecular level. Previously, a gene fragment encoding a putative plasma membrane proton symporter was isolated, because it was coordinately regulated with several nicotine biosynthetic genes. Here, we show that this gene fragment corresponds to a Nicotiana tabacum gene encoding a nicotine uptake permease (NUP1). NUP1 belongs to a plant-specific class of purine uptake permease-like transporters that originated after the bryophytes but before or within the lycophytes. NUP1 expressed in yeast cells preferentially transported nicotine relative to other pyridine alkaloids, tropane alkaloids, kinetin, and adenine. NUP1-GFP primarily localized to the plasma membrane of tobacco Bright Yellow-2 protoplasts. WT NUP1 transcripts accumulated to high levels in the roots, particularly in root tips. NUP1-RNAi hairy roots had reduced NUP1 mRNA accumulation levels, reduced total nicotine levels, and increased nicotine accumulation in the hairy root culture media. Regenerated NUP1-RNAi plants showed reduced foliar and root nicotine levels as well as increased seedling root elongation rates. Thus, NUP1 affected nicotine metabolism, localization, and root growth.

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With widespread detection of endocrine disrupting compounds including hormones in wastewater, there is a need to develop cost-effective remediation technologies for their removal from wastewater. Previous research has shown that Fe(3+)-saturated montmorillonite is effective in quickly transforming phenolic organic compounds such as pentachlorophenol, phenolic acids, and triclosan via surface-catalyzed oligomerization. However, little is known about its effectiveness and reaction mechanisms when reacting with hormones. In this study, the reaction kinetics of Fe(3+)-saturated montmorillonite catalyzed 17β-estradiol (βE2) transformation was investigated. The transformation products were identified using liquid chromatography coupled with mass spectrometry, and their structures were further confirmed using computational approach. Rapid βE2 transformation in the presence of Fe(3+)-saturated montmorillonite in an aqueous system was detected. The disappearance of βE2 follows first-order kinetics, while the overall catalytic reaction follows the second-order kinetics with an estimated reaction rate constant of 200 ± 24 (mmol βE2/g mineral)(−1) h(–1). The half-life of βE2 in this system was estimated to be 0.50 ± 0.06 h. βE2 oligomers were found to be the major products of βE2 transformation when exposed to Fe(3+)-saturated montmorillonite. About 98% of βE2 were transformed into βE2 oligomers which are >10(7) times less water-soluble than βE2 and, therefore, are much less bioavailable and mobile then βE2. The formed oligomers quickly settled from the aqueous phase and were not accumulated on the reaction sites of the interlayer surfaces of Fe(3+)-saturated montmorillonite, the major reason for the observed >84% βE2 removal efficiency even after five consecutive usages of the same of Fe(3+)-saturated montmorillonite. The results from this study clearly demonstrated that Fe(3+)-saturated montmorillonite has a great potential to be used as a cost-effective material for efficient removal of phenolic organic compounds from wastewater.

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