Isolation, Identification And Analysis Of The Proteins Iduced By Drought In Lathyrus Sativus L. With Proteomics Technique

Grass pea (Lathyrus sativus L., Leguminosae) is an annual legume crop with a number of advantageous biological and agronomic characters, such as extensive tolerance to drought, water-logging and poor semiarid soils; resistance to insects and pests; high grain-yielding capacity and high content of proteins in its seeds.L. sativus has been used as a promissing experimental materials to study mechanism of strss resistance in plant, because of its special physiological characteristic. We used grass pea as a experimental materials to study its drought resistance proteomics and to provide scientific evidence for eluminating mechanism of stress resistence of grass pea.1 The optimized 2-DE technique has been developed successfully in this study, by comparing three kinds of protein exrarct methodes, prolonging the time of IEF(Isoelectric focusing) under low voltage to desalt in the samples, as well as using compound staining method of silve staining and commassia brilliant blue. Using this 2-DE technique, high-quality of proteins could be extracted and satisfactory 2-DE maps with high reproduction and resolution power of protein were obtained, suggesting that the 2-DE technique developed in this study could absolutely meet the requirement of further proteomics study.2 Total proteins from leaves of L. sativus cultivars grown in the hydroponic condition and artificial drought that was induced by different concentrations of PEG-6000 was isolated, using optimized methods of two-dimensional gel electrophoresis. The maps were analyzed with solftware ImageMaster. There were 1326, 1292 and 1265 spots were detected in 2-DE gel of total proteins from L, sativus cultivars grown in the Hoagland Nutrient solution, 15 % and 20% PEG, respectively. The molecular weight of most proteins were distributed in 14.1-116.0 kDa, and their PI were around pH 5-7, and spots were found differentially expressing in leaves of L sativus. 61 and 34 differentially expressing protein spots were detected in the gel from samples treated with 15% and 20% PEG, respectively, compared to contral.3 Three spots in the gel containing samples treated with 20%PEG were cut off,...