Tuesday, July 31, 2012
Grand Ballroom
In this project, seedlings of wild tomato (Solanum chilense) were grown to 4-leaf-stage in a hydroponic tank filled with ½ Hoagland solution. Drought treatment was applied by letting plants to air-dehydrate in empty tanks overnight. The control plants were maintained in the solution. The next morning when the leaves started to wilt, roots were harvested and frozen immediately in liquid nitrogen. Protein was extracted using TCA/acetone precipitation method. Proteins in the pellets were solubilized in a buffer consisting of 50 mM TEAB (triethylammonium bicarbonate) and 6 M urea. Proteins (100 ug/ sample) were digested with trypsin A and the digests were labeled with taqs in the iTRAQ reagent kit (8Plex, AB Sciex). Labeled proteins were subjected to MS/MS analysis. Mascot v 2.3 was used to quantify the level of expression of each protein identified via the iTRAQ reporter ions. The MS/MS spectra were used to interrogate the ITAG2.3_proteins database. Proteins identified from the analysis were searched for orthologous proteins from Arabidopsis in STRING database, and a protein interaction map was constructed for the molecular regulation to dehydration stress in wild tomato.