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ENVIRONMENTAL STRESS AND ADAPTATION TO STRESS

Detection and Quantification of Unbound Phytochelatin 2 in Plant Extracts of Brassica napus Grown with Different Levels of Mercury¹

Departament de Biologia Vegetal, Facultat de Biologia (S.I.-T., A.F., C.C., J.L.A., J.B.), and Scientific and Technical Services (O.J.), University of Barcelona, 08028 Barcelona, Spain

The mercury (Hg) accumulation mechanism was studied in rape (Brassica napus) plants grown under a Hg concentration gradient (0 µM–1,000 µM). Hg mainly accumulated in roots. Therefore, the presence of phytochelatins (PCs) was studied in the roots of the plants. The high stability of the PC-Hg multicomplexes (mPC-nHg) seems to be the main reason for the lack of previous Hg-PC characterization studies. We propose a modification of the method to detect and quantify unbound PC of Hg in plant extracts via high-performance liquid chromatography coupled to electrospray tandem mass spectrometry and inductively coupled plasma mass spectrometry in parallel. We separated the PC from the Hg by adding the chelating agent sodium 2,3-dimercaptopropanesulfonate monohydrate. We only detected the presence of PC after the addition of the chelating agent. Some multicomplexes mPC-nHg could be formed but, due to their large sizes, could not be detected. In this study, only PC₂ was observed in plant samples. Hg accumulation was correlated with PC₂ concentration (r² = 0.98).

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Anna Febrero (annafebrero{at}ub.edu).

www.plantphysiol.org/cgi/doi/10.1104/pp.106.085068

^* Corresponding author; e-mail annafebrero{at}ub.edu; fax 34–934112842.

Received June 12, 2006; accepted July 21, 2006; published August 18, 2006.