03475nam a2200253 a 450000100080000000500110000800800410001910000200006024501550008026001590023550000140039452025990040865000190300765000170302665000210304365300280306465300140309270000180310670000160312470000170314070000210315770000200317870000230319818869162011-04-29       2011    bl uuuu     u00u1 u    #d1 aGOMES, L. M. C.  aCadmium-induced changes in leaf protein profile of Caesalpinia peltophoroides Benth (sibipiruna)bprotein extraction and bidimentional eletrophoresis.  aIn: SIMPÓSIO BRASILEIRO DE GENÉTICA MOLECULAR DE PLANTAS, 3, 2011, Ilhéus. Resumos... [S. l.]: Sociedade Brasileira de Genética, 2011. 1 CD-ROM.c2011  apdf 34634  aThe specie C.peltophoroides is used as bioindicator of environmental pollution, being suitable for revegetation of degraded areas. Works related to cadmium (Cd) tolerance in this species are still being initiated. In this way, analysis of protein profile aims to assist in the identification of stress-induced protein expression. Seedlings growing in nutrient solution were submitted to two treatments: no Cd and high Cd concentration (32 mg L-1) in a completely randomized design. Leaves from the two treatments were collected at different times of exposure to Cd (0, 6, 12, 24, 48, 72 and 96 h) and fixed in liquid N2. All samples were freeze dried, macerated and washed with TCA/acetone, TCA/water and finally acetone. Proteins were quantified using 2-D Quanti kit (GE Healthcare) and analyzed by SDS-PAGE and 2D-PAGE. Preliminary analysis of proteins in SDS-PAGE showed several bands below 25 kDa, when compared with the control (no Cd), in differents exposure times. The two samples from different exposure times to Cd (24 and 72 h) were selected after analysis of SDS-PAGE and were used in two-dimensional electrophoresis. For isoelectric focusing, we used 380 ?g of protein sample diluted in rehydration buffer strips on linear 13 cm (pH 4-7) using equipment IPGphor 3. The second dimension was performed on polyacrylamide gels with 12.5% in the system Hoefer SE 600 Ruby ?. The gels were fixed with 10% acetic acid and 40% ethanol, stained with Coomassie Brilliant Blue and faded washes in Milli-Q water until the appearance of spots. The gel images were captured in 2D Scanner Magic and analyzed with the software ImageMaster 2D Platinum (Amersham Biosciences) and manual detection. In 2D-PAGE spots were visualized 99 spots (41 differentially expressed) in the 24 h and 122 spots (29 differentially expressed) in the 72 h, both with quality. Most intense spots were analyzed in the Expasy Tagident (www.expasy.ch/), based on its mass and isoelectric point. Taxonomic group and keywords were used to search in the protein database. Preliminary results identified proteins such as: aquaporins, heat shock (HSP), cold and drought-regulated protein (CORA), photosystem I assembly, NAD(P)H-quinone oxidoreductase subunit and cytochrome b6-f complex subunit in Populus, Arabidopsis and Pisum sativum. Some spots were not found in the database. Spots will be selected for the protein sequencing by mass spectrometry. This is a pioneering study involving proteomic data in order to support the hypothesis that C. peltophoroides can be considered a promising species in tolerance to Cd stress.  aabiotic stress  aheavy metals  aphytoremediation  aEnvironmental pollution  aProteomic1 aCASTRO, A. V.1 aDIAS, L. O.1 aGOMES, F. P.1 aALMEIDA, A-A. F.1 aPIROVANI, C. P.1 aGESTEIRA, A. da S.