Cross-resistance to two fipronil analogs, butene-fipronil and ethiprole, was detected in fipronil-resistant field populations and a resistant laboratory strain of the planthopper Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), although the two analogs have not been used widely in rice-growing areas in China. The results showed that six field populations with 23.8-43.3-fold resistance to fipronil had reached a higher level of cross-resistance to ethiprole (resistance ratio [RR] = 47.1-100.9-fold) and had a minor level of cross-resistance (RR = 3.4- 8.1-fold) to butene-fipronil. After 10 generations of selection, the RR to fipronil increased from 7.3-fold to 41.3-fold. At the same time, the insect increased cross-RR to ethiprole from 16.3-fold to 65.6-fold, whereas it had only minor increase in cross-resistance to butene-fipronil from 2.8-fold to 4.0-fold. These results confirmed that fipronil-resistant N. lugens could develop a higher level of cross-resistance to ethiprole, although it still maintained a lower level cross-resistance to butene-fipronil. Our data suggest that ethiprole is not a suitable alternative for controlling N. lugens, once the insect has developed a high level resistance to fipronil. Further investigation is necessary to understand the cross-resistance mechanisms in N. lugens.

The vanilloid-type transient receptor potential (TRPV) channel is reported to be the molecular target of the commercial insecticide pymetrozine, which specifically disrupts the feeding of plant sap-sucking insects. However, the functions of TRPV channels in plant sap-sucking insects have not been fully elucidated. In the present study, RNA interference was used to investigate the effects of the knockdown of TRPV genes (Nan and Iav) on the mortality, locomotion, and feeding behavior of an important plant-feeding insect pest in rice, the brown planthopper, Nilaparvata lugens. Injecting dsRNA of Nan and Iav into fourth-instar nymphs significantly knocked down the target genes. The injection of dsNan or dsIav did not affect any morphological phenotype (including leg extension) of N. lugens nymphs and adults. Knockdown of Nan or Iav resulted in significantly decreased climbing activity against top plants but did not influence the leg-griping strength of adults. Knockdown of Nan resulted in a significantly elevated mortality of N. lugens in the observation period of 7 d after injection, whereas no significant difference in survival rates 7 d after injection was found between dsIav-injected and dsGFP-injected insects. Electropenetrographic (EPG) recordings indicated that knockdown of Nan and Iav reduced the ingestion activity in the rice phloem tissues of N. lugens. Knockdown of Nan and Iav significantly reduced the amount of honeydew excreted by N. lugens. Our findings indicated a relationship between TRPV and N. lugens locomotion and feeding behavior, which may help to fully elucidate the functions of TRPV in insects.

Honeydew is a watery fluid excreted by plant sap-feeding insects. It is a waste product for the insect hosts. However, it plays important roles for other organisms, such as serving as a nutritional source for beneficial insects and bacteria, as well as elicitors and effectors modulating plant responses. In this study, shotgun LC–MS/MS analyses were used to identify the proteins in the honeydew from two important rice hemipteran pests, the brown planthopper (Nilaparvata lugens, BPH) and green rice leafhopper (Nephotettix cincticeps, GRH). A total of 277 and 210 proteins annotated to insect proteins were identified in the BPH and GRH honeydews, respectively. These included saliva proteins that may have similar functions as the saliva proteins, such as calcium-binding proteins and apolipophorin, involved in rice plant defenses. Additionally, a total of 52 and 32 Oryza proteins were identified in the BPH and GRH honeydews, respectively, some of which are involved in the plant immune system, such as Pathogen-Related Protein 10, ascorbate peroxidase, thioredoxin and glutaredoxin. Coincidently, 570 and 494 bacteria proteins were identified from the BPH and GRH honeydews, respectively, which included several well-known proteins involved in the plant immune system: elongation factor Tu, flagellin, GroEL and cold-shock proteins. The results of our study indicate that the insect honeydew is a complex fluid cocktail that contains abundant proteins from insects, plants and microbes, which may be involved in the multitrophic interactions of plants–insects–microbes.

The predatory stink bug P. Lewisi shows potential for Integrated Pest Management programs for controlling Lepidoptera pest insects in crops and forests. The importance of this insect for biological control has stimulated several studies into its biology and ecology. However, P. lewisi has little genetic information available. In the present study, PacBio single-molecule real-time (SMRT) sequencing and Illumina RNA-seq sequencing technologies were used to reveal the full-length transcriptome profiling and tissue-specific expression patterns of P. lewisi. A total of 12,997 high-quality transcripts with an average length of 2,292 bp were obtained from different stages of P. lewisi using SMRT sequencing. Among these, 12,101 were successfully annotated in seven public databases. A total of 67 genes of cytochrome P450 monooxygenases, 43 carboxylesterase genes, and 18 glutathione S-transferase genes were identified, most of which were obtained with full-length ORFs. Then, tissue-specific expression patterns of 5th instar nymphs were analyzed using Illumina sequencing. Several candidate genes related to detoxification of insecticides and other xenobiotics as well as the degradation of odors, were identified in the guts and antennae of P. lewisi. The current study offered in-depth knowledge to understand the biology and ecology of this beneficial predator and related species.