Mpp51Aa1 toxicity to Diaphorina citri nymphs demonstrated using a new, long-term bioassay method.

While pesticidal proteins from Bacillus thuringiensis have provided for effective management of several insect pests of agricultural importance, few with toxicity to hemipteran species have been identified. The Asian citrus psyllid, Diaphorina citri transmits Candidatus Liberibacter asiaticus (CLas), the presumed bacterial causative agent of the devastating disease citrus greening. Despite the critical role of D. citri nymphs in the acquisition and inoculation of CLas, the lack of a long-term feeding method impedes the screening of Bt proteins for toxicity against nymphs, which play a key role in CLas transmission. Here, we developed a long-term nymph bioassay and determined the toxicity of the Bt pesticidal protein Mpp51Aa1. The new bioassay method allows nymphs to survive for up to six days when maintained on treated folded wipes. The standard hemipteran membrane feeding assay was used to assess Mpp51Aa1 toxicity against D. citri adults. Mpp51Aa1 was toxic to D. citri nymphs with a median lethal concentration (LC50) of 56.5 µg/ml in wipe feeding assays, and to D. citri adults with an LC50 of 110.4 µg/ml in membrane feeding assays. These results demonstrate the utility of this long-term nymph bioassay method and suggest that Mpp51Aa1 has potential for sustainable use in D. citri management toward mitigation of citrus greening disease.