The aim of this research is to examine the effects of course activities with learning styles and portfolio applications on the attitudes of pre-service science teachers towards biology lesson. The research was studied with 100 teacher candidates in the 2nd grade of Science Teaching at Ondokuz Mayis University. The learning styles of the prospective teachers were determined according to the Kolb Learning Styles Inventory (KÖSE-3), and lesson plans were created within the scope of the General Biology Laboratory course, and also teaching activities which used portfolio evaluation were developed. In this context, the effect of the taught course on the attitudes of pre-service science teachers towards the biology course was examined. The design of the research is quasi-experimental design with pre-test post-test control group (model with unequalized control group). One of the branches of the pre-service science teachers' class was chosen as Experiment1 group, one as Experiment 2 and one as Control group. At the end of the period, it was found that the applications which was completed in a total of 28 weeks were more effective in the attitudes of pre-service science teachers in both experimental groups towards the biology lesson.