The Protein Differential Analysis Of Rape Seedling Roots Under Phosphorus Deficency By Two-dimensional Electrophoresis

Rape is a very important oil crop, phosphorus is one of the most important nutrientelements which to maintain their normal growth and development,through the studyfound that the soil can be absorbed by the plant using phosphorus is lack, it is difficultfor rapes to play its potential under the influence of phosphorus deficiency, which tolimit the rape yield and quality, so the rape under phosphorus stress responsemechanism and finding the rape of using phosphorus effectively is becoming a hotspot of research. At present, the main use of genomics study of rape in response tophosphorus stress, and protein is the physiological function and the life of theexecutor of the phenomenon, and the protein is an important part of cells, organs andtissues,proteomics can study of the comprehensive mechanism of rape in response tophosphorus stress from the perspective of the overall.In this work, the Brassica napus “Zhongyouza-9” as the experimental material,optimization of rape seedlings hydroponic method, observe the morphology of rapeseedlings roots under phosphorus deficiency, establish and optimize the extraction ofrape seedling root protein system, using the optimized self-made tube gels suitablesystem of two-dimensional gel electrophoresis technology for rape seedling rootsystem under different phosphorus concentration stress gel maps. Through the studyof rape root analysis the differences in protein maps of the rape seedlings underphosphorus stress related molecular mechanism research lay the foundation. Themain contents includes the following several aspects:1.The morphological characteristics of rape seeding in different phosphorusconcentration stress were obeerved and evaluated. Compared with the conrol, themain roots of rape were longer and slender, and the numbers of lateral root and roothair increased under low phosphorus stressed. The roots showed more red-brown withthe longer culture duration in phosphorus deficiency condition. This indicated that therape improve the absorption and utilization of phosphorus by root morphologicalchanges.2.The method of tube gel two-dimensional electrophoresis was optimized. The methods of protein extraction, loading amount for2-DE were compared andevaluated. The optical protein extraction is TCA-acetone precipitation method. Thelysis buffer for2-DE was7M urea buffer with2M thiourea and4%Chaps, and theoptical loading volume is400μg.3.Using the2-DE system，the expressed proteins of rape seedling root wereanalyzed. The2-DE maps were clear and the protein spots were round. When theconcentration of phosphorus nutrient solution reduced to1/2,1/4and0of sufficientphosphorus concentration afrer4days treated.The correlation coefficient of rape rootprotein differential patterns between1/2-dosage-treatment by phosphorus deficiencyand its control is0.764. There were44proteins showed diffenently expressed underphosphorus deficiency stress, with24up and20down with1/2-dosage-treatment.The correlation coefficient of rape root protein differential patterns between1/4-dosage-treatment by phosphorus deficiency and its control is0.713. There were40proteins showed diffenently expressed under phosphorus deficiency stress, with21up and19down with1/4-dosage-treatment. The correlation coefficient of rape rootprotein differential patterns between phosphorus deficiency and its control is0.673.There were41proteins showed diffenently expressed under phosphorus deficiencystress, with24up and17down with phosphorus deficiency. The up-regulated proteinsis10all in1/2,1/4and0dosage treatments, and the down-regulated proteins is8. Thiswork laid a good basement for the futher analysis of homology.