Entomopathogenic Fungus Infection Dynamics

Conidia of WT and BbVRF1 strains were harvested, and the concentrations of their suspensions were adjusted to 2 × 10⁶ conidia mL⁻¹. For the in vitro experiment, H. armigera cells were cultured in six-well culture plates (60,000 cells per well) and infected with 1 μL suspension of WT or BbVRF1. The mock-infected (infected with 1 × phosphate-buffered saline [PBS]), WT-infected (infected with WT strain), and BbVRF1-infected (infected with BbVRF1 strain) cells were collected 72 h later. For the in vivo experiment, 5th-instar H. armigera larvae were injected with 5 μL conidia suspension (2 × 10⁶ conidia mL⁻¹) of WT and BbVRF1. The hemocytes were collected at 0 h, 12 h, 24 h, 48 h and 72 h after infection. RNA was extracted from samples using the RNA Easy Fast tissue/cell kit (Tiangen, China). The RNA was treated using the HiScript III All-in-One RT SuperMix Perfect for RT-qPCR (Vazyme, China) for reverse transcription, and qRT-PCR was performed using the StepOne Plus RT-qPCR system (Applied Biosystems, USA) and Taq Pro Universal SYBR qPCR master mix (Vazyme, China) according to the manufacturers’ manuals, using the primers listed in Table S1. The rps3 gene was used as an internal reference (35 (link)).