Testing Method Of LOX Activity And Its Dynamic Variation During Development And Germination Of Seeds In Wheat

4lipoxygenase activity stable wheat varieties (lines) were selected as material to do the wheat LOX activity detection method optimization,analyze the changes of wasoenzymes during kernel development and grain germination;14wheat varieties (lines) which lipoxygenase activities have more differences were selected as material to test the wheat dry seed lipoxygenase isozymes,and study the characterwastics of LOX for enzymology.The results from above experiments showed as follows:1.In the single factor test optimization of wheat LOX activity detection method,the optimum amount of whole wheat flour was about0.2g/ml,optimum substrate concentration of linoleic acid was about0.05%,and the optimum extraction temperature should be5℃,the best ice bath extraction time was about30min,the optimum buffer pH value was about6.0. The optimal composition obtained in the interaction of multiple factors showed that linoleic acid concentration of the substrate was0.05%,the amount of whole wheat flour was0.2g/ml, pH value was7,and extraction time of the ice bath was30min.2. The optimum pH was6.0,the optimum reaction temperature was30℃,Km=5.61mol/L,Vmax=0.61mol/L·min,kinetic equation was v=(0.611·[S])/(5.61+[S])3.The test of14wheat varieties showed that the high-LOX activity varieties had one more isoenzymes band than the low-LOX activity varieties.4.In wheat grain maturation, after flowering5d-15d acidic LOX activity was higher, but in the late of mature alkaline LOX activity was higher. In wheat seed germination, after germinated2d-3d LOX activity was maintained at a stable high value,but with a reduced fat content,LOX activity rapidly reduced;4d-5d acidic LOX activity was higher,6d-7d alkaline LOX activity was higher. No matter maturation or germination period, the trend of isozymes and activity is always high related,and the acidity isozymes are the most important factor which decides the influence of isozyme.