Gender differences in AI chatbot utilization and research knowledge acquisition among final-year science education undergraduates was survey-studied with two research questions and two hypotheses. Out of the 259 final-year science education undergraduates in Benue State University (now Rev. Fr. Moses Orshio Adasu University), Makurdi, Nigeria; a simple random sample of 102 was used for the study. A Likert-type data-collecting instrument “Artificial Intelligence Chatbot and Research Questionnaire (AICRQ)”, which was validated and trial tested on 25 students outside the study’s sample, with a Cronbach’s Alpha reliability coefficient of 0.78 was used to collect data for the study. Data were analysed using mean and standard deviation to answer the research questions; while the hypotheses were tested using Mann-Whitney U-test. Results showed a significant difference in the extent to which male and female final-year science education undergraduates utilize AI chatbots for research project writing [U = 0.000, Z = -8.717, p = 0.000 < (∝ = 0.05)]; and a significant difference in the influence of AI chatbots usage on research knowledge acquisition among them [U = 6.000, Z = -8.688, p = 0.000 < (∝ = 0.05)] favouring the males. The study concludes that gender significantly affected how AI chatbots usage contributed to research knowledge acquisition among final-year science education undergraduates. It was therefore, recommended that university administration should organize gender-responsive training, mentorship, and practical workshops with digital literacy support and access to AI tools, prioritizing female science education undergraduates to balance the gender difference in AI chatbot utilization and research knowledge acquisition.