褐飞虱Yellow基因的克隆及功能

doi:10.3864/j.issn.0578-1752.2015.15.007

Abstract

Abstract: 【Objective】The objectives of this study are to clone NlYellow from the brown planthopper (Nilaparvata lugens), explore the expression profiles during the development in adults and in different tissues, then study its function through RNA interference.【Method】Primers were designed by online version of primer 3 software and NlYellow was cloned, then the nucleotide sequence was translated into amino acid sequence. The alignment of amino acids residues was conducted using N. lugens Yellow and others searched from GenBank. Also a Neighbor-Joining method was used to construct a phylogenetic tree. Quantitative real-time PCR (qRT-PCR) was applied to measure the relative expression levels of N. lugens at different developmental stages (embryo, 1st-5th instar nymphs and adult) and in different tissues (head, thorax, abdomen, leg, wing, midgut, ovary, and testis). NlYellow dsRNA at 0.4 µg was injected into 5th instar nymphs and the adult phenotypes were observed. 【Result】The NlYellow was cloned, and through sequence alignment it was found that the NlYellow was more similar to the ApYellow (98%), but it was less conserved when compared with that of Drosophila melanogaster and Bombyx mori.Phylogenetic analysis showed that it clustered with Acyrthosiphon pisum,too. NlYellow’s expression was variable in the embryo, the expression level in 1st, 3rd, 4th and 5th day were lower than other days. Furthermore, its expression levels were higher in 3rd and 5th instar nymph than others (P＜0.05). Tissue-specific expression analysis elucidated that NlYellow expressed in all tissues including head, thorax, abdomen, leg, wing, midgut and testis in male, but the first four tissues had higher expression level (P＜0.05). On the contrary, NlYellow was only expressed in head, thorax, leg, and wing in female. Furthermore, its expression level in short wing adult was higher than that of long wing adult (P＜0.05). Also in short wing adults, male’s expression level was higher than female (P＜0.05). Using RNA interference to knock down the NlYellow expression, it was found that the color of the whole body turned into yellow, especially in thorax, abdomen and legs.【Conclusion】NlYellow might involved in insect pigmentation and change of the body colour.

Key words: Nilaparvata lugens; Yellow, gene expression profile, RNA interference

WANG Bo, YAO Yun, XU Ze-wei, LIN Xin-da . Molecular Cloning and Function of the Yellow Gene from Nilaparvata lugens[J].Scientia Agricultura Sinica, 2015, 48(15): 2976-2984.

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Molecular Cloning and Function of the Yellow Gene from Nilaparvata lugens

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