AbstractBACKGROUNDSweet potato whitefly (Bemisia tabaci) is one of the most destructive pests to an extensive range of crops and vegetables. Pesticide‐dependent management programs have led to severe health problems, including pesticide poisoning and cancer in human beings, as well as pesticide resistance in insect pests. Entomopathogenic fungi (EPF) are considered safe and highly effective against many pests. Therefore, identifying the pathogenicity and virulence of EPFs against Bemisia tabaci is a valuable addition to the management of their infestations. In this study, we investigated the efficacy of conidia suspensions of Aschersonia aleyrodis, Isaria fumosorosea, Beauveria bassiana, and Akanthomyces muscarius (= Lecanicillium muscarium) against nymphal stages of Bemisia tabaci in cucumber seedlings under both optimal and suboptimal conditions.RESULTSAll of the EPFs demonstrated significant ovicidal effects, with the highest cumulative mortalities observed in Aschersonia aleyrodis (96.46%) and I. fumosorosea‐treated (94.60%) seedlings against host eggs and crawlers. Similarly, in the L4‐instars experiment, Aschersonia aleyrodis and I. fumosorosea were the most efficient, resulting in cumulative mortalities of 94.82% and 94.75%, respectively. However, Bemisia tabaci cumulative mortalities on seedlings treated with Akanthomyces muscarius (78.36%) and Beauveria bassiana (85.90%) were also significantly different from untreated seedlings (7.10%). Under suboptimal relative humidity (RH) conditions (≤ 45% RH), Aschersonia aleyrodis exhibited greater tolerance to harsh conditions, causing a significantly higher infection rate in L1–L2 nymphs (~92%) compared to the approximately 32% infected young nymphs observed in I. fumosorosea‐treated seedlings.CONCLUSIONAll the selected EPF were more effective against the young nymphal instars. Our results also highlight the efficacy of Aschersonia aleyrodis under suboptimal conditions. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.