Abstract: In order to breeding high tartronic acid content cucumber special varieties, three cucumber varieties with different tartronic acid content were chosen as the test materials. Using the L9(34) orthogonal test design to analyze the effects of four factors(different varieties, different sampling regions , different nitrogen concentration and different physiological periods) on tartronic acid content. The result showed that stems and leaves contain tartornic acid in spite of the fruits and the content is lower than fruits. When the concentration of nitrogen is in the normal level, the tartronic acid content is higher than in the lower and higher level. In addition, the tartronic acid in full bearing age is slightly higher than in smoke trailing period and in fruiting early stage. Furthermore, the combination of the highest cucumber tartronic acid content: cucumber varieties ‘C11-35’, sampling positions: fruit, nitrogen concentration: 0.014 mol/L, cucumber physiological period: full productive age.