ABSTRACT
In this work we analysed the protein variations which occurred in two Thlaspi caerulescens populations when subjected to 0 and 10 microM nickel (Ni) treatments: the Ni hyperaccumulator T. caerulescensfrom a metalliferous soil in Italy and T. caerulescens from Czech Republic, adapted to grow on a non-metalliferous soil. Ni accumulation in roots and shoots and the effect on growth and morphology were examined. Leaves proteins profiles of Ni treated and untreated samples were analysed by two dimensional liquid chromatography technique. From the comparison of more than 500 proteins, few differences were observed between treated and untreated plants of the same population. Differences were found between the two Thlaspi populations, instead. Proteins involved in transport, metal chelation, and signal transduction increased in abundance in the 10 microM Ni treated samples while, in condition of absence of Ni, proteins involved in sulphur metabolism, protection against reactive oxygen species and stress response showed to increase in abundance in the two populations. These proteins can be used as biomarkers both for monitoring biodiversity in indigenous plants and for selection of Ni phytoremediation plants.
Subject(s)
Plant Proteins/genetics , Soil/analysis , Thlaspi/genetics , Czech Republic , Enzymes/genetics , Gene Expression Regulation, Plant , Genetic Variation , Italy , Metals/analysis , Nickel/analysis , Nickel/pharmacology , Plant Leaves/anatomy & histology , Plant Leaves/enzymology , Plant Leaves/metabolism , Plant Roots/enzymology , Plant Roots/growth & development , Thioredoxins/genetics , Thlaspi/drug effects , Thlaspi/enzymology , Thlaspi/growth & developmentABSTRACT
Two-dimensional liquid chromatography based on a high-performance chromatofocusing in the first dimension followed by high-resolution reversed-phase chromatography in the second dimension can be used as a complementary approach to protein separation with two-dimensional gel electrophoresis. In this work, Arabidopsis thaliana proteins obtained from different tissue extracts were resolved by using a new automated system, ProteomeLab PF 2D commercialized by Beckman Coulter (Fullerton, CA, USA). In particular, protein patterns obtained after two different extraction procedures (MgSO4 and urea buffer) were compared. Reproducibility of the protein patterns was also confirmed in different injections of the same sample and in the comparative analyses of some proteins by MALDI-TOF/MS. Computer analysis of the chromatograms revealed that with this two-dimensional liquid phase technique, hundreds of "virtual bands" can be identified and compared in crude plant protein lysates.
