The Physiological Characteristics Of The Third Generation Mutagenic Wheat And The Study Of Its Proteome On Salt Stress

This research used the chemical mutagenic wheat GAO602(Triticum aestivum L.) as the experimental materials to study the effects on seed germination and seedling growth under the condition of the salt stresses,and then identified the one which have the excellent salt resistance. we measured changes in the proteome of the mutagenic wheat using two-dimensional gel electrophoresis(2-DE)via liquid chromatography-tandem mass spectrometry,illustrating the function of salt stress response proteins in the fine mutagenesis,This result provided the evidences to reveal the salt tolerance mechanisms.The experiment results showed as follows:As the test duration prolonged, there was a gradual increase in the content of soluble protein and MDA of the four varieties under the400mmol·L-1NaCl content stress, so was the antioxidase activity (SOD, POD, CAT). Five physical signs were selected including soluble protein, MDA, SOD, POD and CAT to evaluate the capacity of salt-tolerance by empirical distribution function and fuzzy membership function comprehensively. As a result,the mutagenic wheat GAO8113K reveals a better capacity of salt-tolerance than others,and that400mmol·L-1NaCl concentration should be the critical salt concentration to illustrate the germination of wheat seeds.We have established a suitable and effective protocol of protein extraction for two-dimensional gel electrophoresis(2-DE) analysis in the wheat leaf tissues by three extraction methods(Trichloroacetic acid/acetone method,Urea/thiourea method,Phenol extraction method).Then the results were compared in regard to protein extraction efficiency, sodiumdodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)and2-DE gels.Only protein identifications with score greater than P<0.05were considered to be positive.Statistical analysis of the density of differential spots in the2-DE gels.Twenty-three protein spots were significantly altered by exposure to the salinity environment.These proteins including ribulose-1,5-bisphosphate carboxylase activase,fructose-bisphosphate aldolase,ATP synthase subunit beta,heat shock protein,S-adenosylmethionine synthase2,oxygen evolving enhancer protein and so on. These proteins were associated with a variety of functions,including energy and material metabolism, protein metabolism,photosynthesis.Above all,the chemical mutagenesis improved the salt resistance of GAO602wheat variety and the seed germination and antioxidant enzyme activity of the M3generation wheat increased in the salt conditions,laying a foundation to the in-depth study about the mutagenic wheat salt tolerance mechanisms in the protein level.