Vitellogenin (Vg) is precursor of vitellin. Here, we identified a Vg (NlVg) and two Vg-likes (NlVg-like1 and NlVg-like2) in the brown planthopper, Nilaparvata lugens. Phylogenetic analyses showed that NlVg-like1 and NlVg-like2 are not clustered with the conventional insect Vgs associated with vitellogenesis. Temporo-spatial expression analyses showed that the NlVg and NlVg-like2 transcript levels increased significantly 24 h after emergence and were primarily expressed in female adults. However, NlVg-like1 was expressed during all stages, and in both genders. Tissue-specific analyses showed that all three genes were most highly expressed in the fat body. The injection of double-stranded RNA targeting NlVg showed that NlVg is essential not only for oocyte development but also for nymph development. The knockdown of NlVg-like1 in female adults resulted in failure to hatch or death before eggshell emergence in 18% of offspring embryos, suggesting that NlVg-like1 plays an important role during late embryogenesis. Approximately 65% of eggs laid by females that were treated with double-stranded RNA targeting NlVg-like2 failed to hatch, indicating that NlVg-like2 plays a role in nutrition absorption during oocyte, or embryonic development. Our results illustrate the structural and functional differences among the Vg and Vg-like genes and provide potential targets for RNA-interference-based insect pest management strategies.

BACKGROUND: The brown planthopper (BPH), Nilaparvata lugens, is the most destructive rice insect pest. To exploit potential target genes for applications in transgenic rice to control this sap-sucking insect pest, three ferritin genes were functionally characterized in this study. RESULTS: In this study, three ferritin genes, that is, ferritin 1 Heavy Chain (NlFer1), ferritin 2 Light Chain (NlFer2) and soma ferritin (Nlsoma-Fer), were identified from BPH. Tissue-specific analyses showed that all three genes were highly expressed in the gut. Although double-stranded RNA injection-mediated RNA inference (RNAi) of Nlsoma-Fer expression resulted in only < 14% mortality in BPH, knockdown of NlFer1 or NlFer2 led to retarded growth and 100% mortality in young nymphs, and downregulation of NlFer1 and NlFer2 in newly emerged female adults caused undeveloped ovaries and severely inhibited oocyte growth, resulting in extremely low fecundity and a zero hatching rate. Knockdown of NlFer1 and NlFer2 caused similar phenotypes in BPH, indicating that they function together, as in many other animals. The results demonstrated that NlFer1 and NlFer2 were essential for BPH development and reproduction. BPHs showed high sensitivity to both dsNlFer1 and dsNlFer2, and injection of only 0.625 ng dsNlFer1 per BPH resulted in 100% mortality. Additionally, the effectiveness of feeding dsNlFer1 and dsNlFer2 to BPH nymphs was further proven. CONCLUSION: NlFer1 and NlFer2 are essential for BPH development and reproduction, and the insect is highly sensitive to their depletion, suggesting that the two gut-highly-expressed genes are promising candidates for application in RNAi-based control of this destructive pest.

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