The whitefly, Bemisia tabaci (Genn.) (Hemiptera: Aleyrodidae), is one of the most damaging pests in field crops as well as in greenhouses. The present study aimed to evaluate the in vitro pathogenicity of 2 fungus strains of Beauveria bassiana (BB-72 and BB-252) and one strain of Lecanicillium lecanii (V-2) against the whitefly (B. tabaci) at 4 different temperature degrees 20, 24, 28, and 32 °C, using a spray method. Three different bioassays were carried out comprised of conidial concentrations and filtrate of fungal strains, BB-72, BB-252 and V-2, and their binary combinations. The 1.5 ml of fungal filtrate was used in filtrate bioassay for each strain of fungus. Three different concentrations (1 × 106, 1 × 107, and 1 × 108 conidia ml−1) were used in conidial bioassay for each strain of fungus, whereas in bioassay for binary combination (1 ml conidia + 1 ml filtrate) of BB-72 × BB-72, BB-252 × BB-252, and V2 × V2 were used for these strains of fungus. According to the outcomes, the maximum mortality against whiteflies was observed on 12th day post-treatment. In conidial bioassay, the maximum mortality of B. tabaci was observed in BB-72 isolate (84%), BB-252 isolate (77%), and V-2 isolate (67%) at the highest concentration (1 × 108 conidia ml−1) at 24 °C, and minimum mortality was recorded in BB-72 isolate (33%), BB-252 isolate (29%), and V-2 isolate (19%) at the lowest concentration 1 × 106 conidia ml−1 at 32 °C on 12th day post-treatment. Infiltrate bioassay, BB-72 isolate exhibited maximum mortality (92%) at 24 °C, and V-2 isolate showed minimum mortality (34%) at 32 °C on 12th day post-treatment. Furthermore, in binary combination bioassay, the highest whitefly mortality was recorded in BB-72 × BB-72 isolate (87%), BB-252 × BB-252 isolate (73%), and V2 × V2 isolate (65%) at 24 °C and the lowest mortality in BB-72 × BB-72 isolate (57%), BB-252 × BB-252 isolate (50%), and V2 × V2 isolate (39%) at 32 °C on 12th day post-treatment. In all bioassays, the BB-72 isolate was the utmost virulent, and application of its filtrate was found to be the most impressive against B. tabaci.