| Summary: | The whitefly (<i>Bemisia tabaci</i>), an important invasive pest that causes severe damage to crops worldwide, has developed resistance to a variety of insecticides. Carboxylesterases (COEs) are important multifunctional enzymes involved in the growth, development, and xenobiotic metabolism of insects. However, systematic studies on the COEs of <i>B. tabaci</i> are scarce. Here, 42 putative COEs in different functional categories were identified in the Mediterranean species of <i>B. tabaci</i> (<i>B. tabaci</i> MED) based on a genome database and neighbor-joining phylogeny. The expression patterns of the COEs were affected by the development of <i>B. tabaci</i>. The expression levels of six COEs were positively correlated with the concentration of imidacloprid to which <i>B. tabaci</i> adults were exposed. The mortality of <i>B. tabaci</i> MED adults fed dsBTbe5 (67.5%) and dsBTjhe2 (58.4%) was significantly higher than the adults fed dsEGFP (41.1%) when treated with imidacloprid. Our results provide a basis for functional research on COEs in <i>B. tabaci</i> and provide new insight into the imidacloprid resistance of <i>B. tabaci</i>.
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