Differential Proteomics Analysis Of Leaves From Hordum Brevisublatum Under Salt Stress

Proteomics provides a complementary and potentially more comprehensive approach to the analysis of signaling mechanisms. Changes in protein profiles during signaling events can be monitored using two-dimensional (2D) gel electrophoresis or related techniques, detecting alterations in expression levels. Hordeum brevisubulatium(Trin.)Link,an excellent forage with high nutritional value and yield,is of high performance great economic.On the basis of previous researches on physiological and biochemical features of Hordeum brevisubulatium(Trin.)Link,its salt-resistant mutant and the salt-tolerant molecular mechanism in our Lab.Objectives of this study were to identify proteins differentially expressed in leaves from Hordeum brevisubulatium(Trin.)Link under Salt stress using two dimensional electrophoresis combined with mass spectrometry to further our understanding of the action mechanism of Hordum brevisublatum under Salt stress.We construct and modified the two dimensional electrophoresis method according to the material Two-dimensional gel electrophoresis which is important in proteomics was improved.Some important steps,such as sample prepare,isoelectric focusing,balance,dyeing were improved. For sample preparation, the tissue sample was grounded in liquid nitrogen and then homogenized in the lysis buffer (8M urea, 2M thiourea, 50mM DTT, 0.5% carrier amphorlyte and 1mM PMSF), then centrifuged at 15 000 rpm for 1 h at room temperature, was carefully recovered, avoiding the unhomogenized material at the bottom of the centrifuge tube. For the preparative gel staining, we tried the"blue silver"method to improve the resolution of the twodimensional electrophoresis map.Using two dimensional electrophoresis and Peptide Mass Fingerprint (PMF) analysis, the differential expressed protein: ribulose-1,5-bisphosphate carboxylase/ oxygenase small subunit was identified. It implyed that a ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit may be involved in the salt stress of Hordeum brevisubulatium(Trin)Link.