AbstractThe striped flea beetle, Phyllotreta striolata, is a globally notorious pest of cruciferous vegetables. In this study, we sequenced the genome of a Chinese population of P. striolata and aligned it with the reference genome from a Canadian population. Through Illumina sequencing and reference‐based genome polishing, we obtained a high‐quality genome of 131.31 Mb with an N50 size of 8.68 Mb. Genome comparison revealed that the Chinese and Canadian populations of P. striolata presented 175 and 122 population‐specific genes, respectively. Through a genome‐wide screening, six genes were identified as potential RNA interference targets influencing the development or survival of P. striolata. Additionally, we identified 1.16 million high‐confidence variants in the Chinese P. striolata population, including 0.69 million single‐nucleotide polymorphisms and 0.43 million insertions/deletions. Variant effect analysis and functional annotation revealed 7571 variants with high‐impact effects, and the related genes were involved in various basic biological processes in P. striolata. Overall, our findings provide valuable resources for advancing genomic research on P. striolata as well as for those pursuing targeted biopesticides for effective control strategies.