关键词: 褐飞虱, 膜饲喂法, 噬菌体短肽文库, 噬菌体展示技术, 中肠, 中肠内膜, 短肽

Abstract: 【Aim】 To screen peptides which may bind to the midgut inner membrane in the rice brown planthopper, Nilaparvata lugens, so as to explore the new methods for blocking N. lugens to transmit rice viruses. 【Methods】 Trophic behavior of nymphs of the rice brown planthopper was imitated with improved membrane feeding method. Through the use of this feeding method, the phage peptide library solution was fed to planthopper nymphs to screen peptides which may bind to the midgut inner membrane. Then, the fusion expression of peptide and enhanced green fluorescent protein was conducted, and the purified fusion protein was fed to nymphs of the planthopper. To confirm the binding activity of screened peptide, the green fluorescence of fusion protein in the midgut of the nymphs was observed under the fluorescence microscope. 【Results】 The feeding rates of nymphs of the planthopper detected by improved membrane feeding method at 16 h and 24 h after treatment reached 91% and 95%, respectively. By observing the green fluorescence from planthopper gut, one peptide, named P2S, was determined as the planthopper gut inner membrane binding peptide. 【Conclusion】 Membrane feeding method was well improved for the rice brown planthopper. A peptide P2S which may bind to the midgut inner membrane of the planthopper was effectively screened and its binding activity was confirmed. The results will provide a possibility of blocking interaction between rice viruses and the midgut inner membrane of the rice brown planthopper. Furthermore, the peptide P2S may be used for further study of the membrane insertion toxins binding to proper receptors in the midgut of the rice brown planthopper.

Key words: Nilaparvata lugens, membrane feeding, phage peptide library, phage display, midgut, midgut inner membrane, peptide