BackgroundRNA interference (RNAi) is a powerful technique for functional genomics research in insects. Transgenic plants producing double-stranded RNA (dsRNA) directed against insect genes have been reported for lepidopteran and coleopteran insects, showing potential for field-level control of insect pests, but this has not been reported for other insect orders.Methodology/Principal FindingsThe Hemipteran insect brown planthopper (Nilaparvata lugens Stål) is a typical phloem sap feeder specific to rice (Oryza sativa L.). To analyze the potential of exploiting RNAi-mediated effects in this insect, we identified genes (Nlsid-1 and Nlaub) encoding proteins that might be involved in the RNAi pathway in N. lugens. Both genes are expressed ubiquitously in nymphs and adult insects. Three genes (the hexose transporter gene NlHT1, the carboxypeptidase gene Nlcar and the trypsin-like serine protease gene Nltry) that are highly expressed in the N. lugens midgut were isolated and used to develop dsRNA constructs for transforming rice. RNA blot analysis showed that the dsRNAs were transcribed and some of them were processed to siRNAs in the transgenic lines. When nymphs were fed on rice plants expressing dsRNA, levels of transcripts of the targeted genes in the midgut were reduced; however, lethal phenotypic effects after dsRNA feeding were not observed.ConclusionsOur study shows that genes for the RNAi pathway (Nlsid-1 and Nlaub) are present in N. lugens. When insects were fed on rice plant materials expressing dsRNAs, RNA interference was triggered and the target genes transcript levels were suppressed. The gene knockdown technique described here may prove to be a valuable tool for further investigations in N. lugens. The results demonstrate the potential of dsRNA-mediated RNAi for field-level control of planthoppers, but appropriate target genes must be selected when designing the dsRNA-transgenic plants.

Plant β-1,3-glucanases are involved in plant defense and development. In rice (Oryza sativa), 14 genes encoding putative β-1,3-glucanases have been isolated and sequenced. However, only limited information is available on the function of these β-1,3-glucanase genes. In this study, we report a detailed functional characterization of one of these genes, Osg1. Osg1 encodes a glucanase carrying no C-terminal extension. Osg1 was found to be expressed throughout the plant and highly expressed in florets, leaf sheaths, and leaf blades. Investigations using real-time PCR, immunocytochemical analysis, and a GUS-reporter gene driven by the Osg1 promoter indicated that Osg1 was mainly expressed at the late meiosis, early microspore, and middle microspore stages in the florets. To elucidate the role of Osg1, we suppressed expression of the Osg1 gene by RNA interference in transgenic rice. The silencing of Osg1 resulted in male sterility. The pollen mother cells appeared to be normal in Osg1-RI plants, but callose degradation was disrupted around the microspores in the anther locules of the Osg1-RI plants at the early microspore stage. Consequently, the release of the young microspores into the anther locules was delayed, and the microspores began to degenerate later. These results provide evidence that Osg1 is essential for timely callose degradation in the process of tetrad dissolution.

To avoid possible eco-disadvantages of phosphating and zinc plating used for corrosion protection of ferrous metals, this work was to develop an alternative coating technique, called plasma electrolytic aluminating (PEA) process, which can be environmentally friendly in terms of the process itself, and eco-friendly with respect to the coating materials. The PEA process was to form a metal aluminate coating on the metallic surfaces through plasma discharging in an aluminate-based electrolyte when a high voltage was applied to the metals. The aluminating mechanism was investigated using scanning electron microscopy, X-ray photoelectron spectroscopy, and X-ray diffraction (XRD). The research revealed that the hercynite film formed on the metallic surface was indispensable for initiation of the aluminating process. Only after a continuous hercynite film fully covered the metallic surface could the stable plasma discharges be established to sinter the film into a strong ceramic coating. The XRD analysis indicated that a prolonged PEA treatment would result in a hercynite–alumina composite coating. Hardness tests and electrochemical corrosion tests showed that the composite coating could provide the gray cast iron (as an example of ferrous metals) with excellent wear and corrosion protection. With the benign coating process and safe ceramic coating materials, the plasma electrolytic aluminating approach could be used as an eco-friendly and cost-effective strategy for anticorrosion of ferrous metals.

The brown planthopper, Nilaparvata lugens, is a serious pest threatening rice production across the world. To identify the main features of the gene expression and the key components of the midgut of N. lugens responsible for nutrition, xenobiotic metabolism and the immune response, we used pyrosequencing to sample the transcriptome. More than 190,000 clean sequences were generated, which led to about 30,000 unique sequences. Sequence analysis indicated that genes with abundant transcripts in the midgut of N. lugens were mainly sugar hydrolyases and transporters, proteases and detoxification-related proteins. Based on the sequence information, we cloned the candidate sucrase gene; this enzyme is likely to interact with the perimicrovillar membrane through its highly hydrophobic C-terminal region. Many proteases were identified, which supported the hypothesis that N. lugens uses the proteolysis system for digestion. Scores of detoxification genes were newly identified, including cytochrome P450s, glutathione S-transferases, caroxylesterases. A wealth of new transcripts possibly participating in the immune response were described as well. The gene encoding a peptidoglycan recognition protein was cloned. Unlike in Acyrthosiphon pisum, the immunodeficiency pathway may be present in N. lugens. This is the first global analysis of midgut transcriptome from N. lugens.

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