Analysis of oxidised and reduced phytochelatins in pea and lupin plants using HPLC/MSn

Abstract

Plants exposed to heavy metals activate a detoxification system capable of chelating and transporting these harmful ions to vacuoles. Phytochelatins–low molecular weight oligopeptides containing thiols such as glutathione and cystein–have been reported to play a very important role in this respect. High performance liquid chromatography coupled to the electospray ion trap mass spectrometer (HPLC-ESI-IT-MS) was used for identification of phytochelatins induced by Cd2+ and Pb2+ in roots, stems and leaves of pea (Pisum sativum L.) and yellow lupin (Lupinus luteus L.). This approach enabled unambiguous identification of phytochelatins in plant tissues and detection of phytochelatins and homophytochelatins in reduced as well as in oxidised form. Significant differences were detected in phytochelatin relative amounts and profiles in different parts of plants treated with heavy metals. Roots of both plant species contained mainly reduced phytochelatins, reduced and oxidised forms of these peptides were observed in stems in similar amounts, whereas only the oxidised phytochelatins were present in leaves.