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Abstract Spodoptera frugiperda is a highly polyphagous pest worldwide with a wide host range that causes serious losses to many economically important crops. Recently, insect-microbe associations have become a hot spot in current entomology research, and the midgut microbiome of S. frugiperda has been investigated, while the effects of cruciferous vegetables remain unknown. In this study, the growth of S. frugiperda larvae fed on an artificial diet, Brassica campestris and Brassica oleracea for 7 days was analyzed. Besides, the microbial community and functional prediction analyses of the larval midguts of S. frugiperda fed with different diets were performed by high-throughput sequencing. Our results showed that B. oleracea inhibited the growth of S. frugiperda larvae. The larval midgut microbial community composition and structure were significantly affected by different diets. Linear discriminant analysis effect size (LEfSe) suggested 20 bacterial genera and 2 fungal genera contributed to different gut microbial community structures. The functional classification of the midgut microbiome analyzed by PICRUSt and FUNGuild showed that the most COG function categories of midgut bacterial function were changed by B. oleracea, while the guilds of fungal function were altered by B. campestris significantly. These results showed that the diversity and structure of the S. frugiperda midgut microbial community were affected by cruciferous vegetable feeding. Our study provided a preliminary understanding of the role of midgut microbes in S. frugiperda larvae in response to cruciferous vegetables.