Isolation and expression analysis of cadmium-induced genes from Cd/Mn hyperaccumulator Phytolacca americana inresponse to high Cd exposure.

Abstract

Phytolacca americana is recognised as a hyperaccumulator that accumulates cadmium (Cd) and manganese (Mn). Although most studies have provided abundant physiological evidence, little is known about the molecular mechanisms of Cd accumulation in P.americana. In this study, Cd-induced genes were isolated using suppression subtractive hybridisation (SSH) library construction, and gene expression patterns under Cd stress were quantified using real-time quantitative PCR. The functions of PaGST, PaMT and PaFe-SOD were confirmed in transformant yeast. Reactive oxygen species (ROS) formation and cell death in root tips were detected, and SOD and POD activities in leaf tissue were also analysed. There were about 447 expressed sequence tags (ESTs) identified and confirmed. GO analysis showed those genes were mainly involved in metabolism, cell stress and defence, transcription and translation, signal transduction, transport, energy and ion transport, which formed the basis for a molecular understanding of P.americana Cd tolerance mechanisms. Cd also stimulated ROS formation and modified the antioxidant systems. Taken together, our results indicate that ROS formation and Cd-induced gene expression favour P.americana tolerance by activating the defence system and permitting subsequent adaptation to Cd toxicity.