The involvement of carboxylesterases (CarEs) in resistance to chlorpyrifos has been confirmed by the synergism analysis in Nilaparvata lugens. However, the function of specific CarE gene in chlorpyrifos resistance and the transcriptional regulatory mechanism are obscure. Herein, the expression patterns of 29 CarE genes in the susceptible and chlorpyrifos-resistant strains were analyzed. Among them, CarE3, CarE17 and CarE19 were overexpressed in the resistant strain, and knockdown of either CarE gene by RNA interference significantly increased the susceptibility to chlorpyrifos. Remarkably, knockdown of CarE17 reduced the enzymatic activity of CarE by 88.63 % and showed a much greater effect on increasing chlorpyrifos toxicity than silencing other two CarE genes. Overexpression of CarE17 in Drosophila melanogaster decreased the toxicity of chlorpyrifos to transgenic fruit flies. Furthermore, the region between - 205 to + 256 of CarE17 promoter sequence showed the highest promoter activity, and 16 transcription factors (TFs) were predicted from this region. Among these TFs, Lim1β and C15 were overexpressed in the resistant strain. Knockdown of either TF resulted in reduced CarE17 expression and a decrease in resistance of N. lugens to chlorpyrifos. These results indicate that the constitutive overexpression of Lim1β and C15 induces CarE17 expression thus conferring chlorpyrifos resistance in N. lugens.

Frankliniella occidentalis is a highly destructive and invasive agricultural pest that has developed resistance to a variety of insecticide classes. Different planting structures and insecticide use frequency can directly affect the resistance development of F. occidentalis. In this study, the susceptibility of three field strains of F. occidentalis, collected over one year (April to November) from three habitat conditions (facility agriculture area, FA; open field crop area, OF; agroforestry intersection area, AI), to spinetoram, spinosad, emamectin benzoate, chlorfenapyr, acetamiprid, and imidacloprid were monitored and compared. At the same time, the detoxification enzyme activity of F. occidentalis in different habitats was determined. The results showed that the susceptibility of the F. occidentalis population in FA was significantly lower than that of populations from OF and AI. Among them, the F. occidentalis population in FA had developed low levels of resistance to spinetoram (RR = 9.18-fold), emamectin benzoate (RR = 5.47-fold), chlorfenapyr (RR = 6.67-fold), and acetamiprid (RR = 7.49-fold), and had developed moderate level resistance to imidacloprid (RR = 11.67-fold), while still being relatively sensitive to spinosad. The population of F. occidentalis from OF had developed low level resistance to spinetoram (RR = 5.24-fold) but was still relatively sensitive to the other five insecticides. The resistance of F. occidentalis from AI to six insecticides was at relatively sensitive levels. The results of the enzyme activities of detoxification enzymes, including carboxylesterase (CarE), glutathione S-transferase (GST), acetylcholinesterase (AChE), and the cytochrome P450 enzyme system (CYP450), revealed that the activities of the FA population of F. occidentalis were significantly higher than those of the other two populations. The change of CarE activity in F. occidentalis was consistent with that of spinetoram resistance, indicating that CarE may be involved in the metabolic resistance of F. occidentalis to spinetoram. Among the three populations, the resistance and detoxification enzyme activities of F. occidentalis of the FA population to six insecticides were higher than those of the other two populations. Our findings, along with other strategies, are expected to help with the resistance management of F. occidentalis in different habitats.

The fall armyworm (FAW), Spodoptera frugiperda, is one of the most important invasive species and causes great damage to various host crops in China. In this study, the diversity and function of gut bacteria in the 5th instar larvae of FAW fed on maize, wheat, potato and tobacco leaves were analyzed through 16S rRNA sequencing. A total of 1324.25 ± 199.73, 1313.5 ± 74.87, 1873.00 ± 190.66 and 1435.25 ± 139.87 operational taxonomic units (OTUs) from the gut of FAW fed on these four different host plants were detected, respectively. Firmicutes, Proteobacteria and Bacteroidetes were the most abundant bacterial phyla. Beta diversity analysis showed that the gut bacterial community structure of larvae fed on different host plants was significantly differentiated. At the genus level, the abundance of Enterococcus in larvae fed on wheat was significantly lower than those fed on the other three host plants. Enterobacter and ZOR0006 were dominant in FAW fed on tobacco leaves, and in low abundance in larvae fed on wheat. Interestingly, when fed on Solanaceae (tobacco and potato) leaves which contained relative higher levels of toxic secondary metabolites than Gramineae (wheat and maize), the genera Enterococcus, Enterobacter and Acinetobacter were significantly enriched. The results indicated that gut bacteria were related to the detoxification and adaptation of toxic secondary metabolites of host plants in FAW. Further analysis showed that replication, repair and nucleotide metabolism functions were enriched in the gut bacteria of larvae fed on tobacco and potato. In conclusion, the gut bacterial diversity and community composition in FAW larvae fed on different host plants showed significant differences, and the insect is likely to regulate their gut bacteria for adaptation to different host plants.