With the increased demand for high quality maize seeds, the market absorbs innovations that add speed and reliability in the tests that enable differentiation of potential performance of seed lots. This study aims to evaluate the combination of the physiological tests with the expression of enzymes of respiratory route for differences in vigor levels in lots of hybrid seed corn. The percentage of germination, germination first count, emergence on tray, index of emergency speed, accelerated aging and respiration rate were evaluated by Pettenkoffer and Titulation method. Furthermore, the expression of alpha amylase, alcohol dehydrogenase, malate dehydrogenase and pyruvate decarboxylase was analyzed. With the results, it is determined that the vigor tests analyzed, together with the tests of enzymatic expressions of respiratory route were efficient to detect different levels of force in hybrid seed corn. Lot 1 had higher physiological quality while lot 2 had the lowest vigor compared to the other lots. With the vigor tests and expression of enzymes, it was possible to differentiate the quality of the seeds and therefore it can be recommended as reliable tests for defining the quality of seed lots.