Abstract: Due to the importance of wheat and barley production in Jordan, prevention of the cereal leafminer, Syringopais temperatella Led. (Lepidoptera: Scythrididae) is of vital importance. The insect is a severe pest and plays an important role in limiting the production of these crops. The use of insecticides is neither economic nor sustainable, so that there is an urgent need to initiate a viable alternative to chemical control. Therefore, this study aimed at investigating the effect of crop rotation on S. temperatella and its preference to different wheat and barley cultivars. The field history experiment was conducted using five different crop rotations. For the preference experiments in the laboratory, two major experimental groups were conducted. The first was set up to measure the consumed leaf area and the second one to record the number of larval attaches. Each group consisted of four different subgroups; 6 wheat cultivars, 6 barley cultivars, two and one cultivar of each crop. The results indicated that the infestation percentage and number of larvae were significantly the lowest in the crop rotation, wheat/chickpea/wheat while the highest were recorded for barley/barley/barley. The preference results showed that wheat cultivar, Horani Nawawi is significantly the most preferred while Horani 27 is the least cultivar. In case of barley, Mutah was the most preferred cultivar and the least preference was recorded for Athroh. Also, wheat was significantly less preferred than barley. The wheat cultivars, Sham, Em-Qees and Acsad 65 had the highest number of attaches and Deer Alla the least. In contrast, the barley cultivar, Acsad 176 had the highest and Athroh and Rum 1 had the least attaches. Barley cultivars had higher attaches than wheat ones. There was a positive relation between the infestation percent and number of larvae, as well as the consumed area and number of attaches in all of the four subgroups. In conclusion, the crop rotation, wheat/chickpea/wheat should be followed and there is a preference variation among the tested wheat and barley cultivars.