Isolation, characterization, culturing, and formulation of a new Beauveria bassiana fungus against Diaphorina citri

Abstract The Asian citrus psyllid, Diaphorina citri, is a worldwide destructive citrus pest. It aids in the transmission of a group of phloem-limited bacteria (Candidatus Liberibacter spp.) that leads to Huanglongbing (HLB) disease. In this study, a new Chinese native fungal strain (F-HY006) was isolated from the cadavers of D. citri collected from Ganzhou, Jiangxi in China (114°94′ E, 25°87′ N). The isolate was virulent towards adult female D. citri with corrected mortality of 76% and LT50 of 4.09 d after application of a suspension containing 107 conidia/ml (400 µl/citrus seedling). Morphologic and phylogenetic analyses identified this isolate as Beauveria bassiana. Based on its virulence against D. citri and growth characteristics (2.89 ± 0.28 × 105 conidia/cm2) on the PDA plate, biphasic (liquid-solid) cultivation for conidia production of B. bassiana F-HY006 was optimized using several nutritional and environmental factors. Soybean meal + cottonseed powder (1:1) and continuous fluorescent light resulted in the highest conidial production (5.7 × 109/g dry substrate). Moreover, the oil-based conidial formulation was optimized using sesame oil as an effective carrier, which resulted in a significant enhancement of the conidial shelf life, tolerance to thermal stress, and virulence against D. citri compared to unformulated conidia. Therefore, we have prepared a potential mycopesticide for the biological control of D. citri based on the optimized biphasic culture growth and formulation of an isolated virulent fungus, B. bassiana F-HY006.