Scientia Agricultura Sinica ›› 2015, Vol. 48 ›› Issue (15): 2976-2984.doi: 10.3864/j.issn.0578-1752.2015.15.007

• PLANT PROTECTION • Previous Articles     Next Articles

Molecular Cloning and Function of the Yellow Gene from Nilaparvata lugens

WANG Bo, YAO Yun, XU Ze-wei, LIN Xin-da   

  1. College of Life Sciences, China Jiliang University, Hangzhou 310018
  • Received:2015-02-16 Online:2015-08-01 Published:2015-08-01

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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

[1]    Geyer P K, Green M M, Corces V G. Tissue-specific transcriptional enhancers may act in trans on the gene located in the homologous chromosome: the molecular basis of transvection in Drosophila. The EMBO Journal, 1990, 9(7): 2247-2256.
[2]    程茂高, 乔卿梅, 原国辉. 昆虫体色分化研究进展. 昆虫知识, 2005, 42(5): 502-505.
Cheng M G, Qiao Q M, Yuan G H. Progress of the research on body-color diversity in insects. Chinese Bulletin of Entomology, 2005, 42(5): 502-505. (in Chinese)
[3]    Tanaka S. Endocrine mechanisms controlling body-color polymorphism in locusts. Archives of Insect Biochemistry and Physiology, 2001, 47(3): 139-149.
[4]    赵惠燕, 张改生, 汪世泽, 王玉梅, 巴桑普赤. 棉蚜体色变化的生态遗传学研究. 昆虫学报, 1993, 36(3): 282-289.
Zhao H Y, Zhang G S, Wang S Z, Wang Y M, Basang P C. An eco-genetical study of body colour in cotton aphids (Aphis gossypii). Acta Entomologica Sinica, 1993, 36(3): 282-289. (in Chinese)
[5]    李超, 谢宝瑜. 光周期与温度的联合作用对棉铃虫种群滞育的影响. 昆虫知识, 1981, 18(2): 58-61.
Li C, Xie B Y. Photoperiod and temperature impact on cotton bollworm populations of diapause. Chinese Bulletin of Entomology, 1981, 18(2): 58-61. (in Chinese)
[6]    Wittkopp P J, Beldade P. Development and evolution of insect pigmentation: genetic mechanisms and the potential consequences of pleiotropy. Seminars in Cell & Developmental Biology, 2009, 20(1): 65-71.
[7]    Valmalette J C, Dombrovsky A, Brat P, Mertz, C, Capovilla M, Robichon A. Light-induced electron transfer and ATP synthesis in a carotene synthesizing insect. Scientific Reports, 2012, 2: 579.
[8]    Wu W J, Wang Y, Huang H J, Bao Y Y, Zhang C X. Ecdysone receptor controls wing morphogenesis and melanization during rice planthopper metamorphosis. Journal of Insect Physiology, 2012, 58(3): 420-426.   
[9]    Nakashima N, Noda H. Nonpathogenic Nilaparvata lugens reovirus is transmitted to the brown planthopper through rice plant. Virology, 1995, 207(1): 303-307.
[10]   Xue J, Zhou X, Zhang C X, Yu L L, Fan H W, Wang Z, Cheng J A. Genomes of the rice pest brown planthopper and its endosymbionts reveal complex complementary contributions for host adaptation. Genome Biology, 2014, 15(12): 521.
[11]   陈慧卿. 褐飞虱体色分化的初步研究[D]. 广州: 中山大学, 2005.
Chen H Q. Primary study of body colour differentiation in Nilaparvata lugens[D]. Guangzhou: Sun Yat-sen University, 2005. (in Chinese)
[12]   Gompel N, Carroll S B. Genetic mechanisms and constraints governing the evolution of correlated traits in drosophilid flies. Nature, 2003, 424(6951): 931-935.
[13]   Wittkopp P J, Vaccaro K, Carroll S B. Evolution of yellow gene regulation and pigmentation in Drosophila. Current Biology, 2002, 12(18): 1547-1556.
[14]   Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 2011, 28(10): 2731-2739.
[15]   Yuan M, Lu Y, Zhu X, Wan H, Shakeel M, Zhan S, Li J. Selection and evaluation of potential reference genes for gene expression analysis in the brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae) using reverse-transcription quantitative PCR. PloS One, 2014, 9(1): e86503.
[16]   Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods, 2001, 25(4): 402-408.
[17]   Nappi A J, Christensen B M. Melanogenesis and associated cytotoxic reactions: applications to insect innate immunity. Insect Biochemistry and Molecular Biology, 2005, 35: 443-459.
[18]   Drapeau M D. A novel hypothesis on the biochemical role of the Drosophila Yellow protein. Biochemical and Biophysical Research Communications, 2003, 311(1): 1-3.
[19]   Busca R, Ballotti R. Cyclic AMP a key messenger in the regulation of skin pigmentation. Pigment Cell Research, 2000, 13(2): 60-69.
[20]   Ferguson L C, Green J, Surridge A, Jiggins C D. Evolution of the insect yellow gene family. Molecular Biology and Evolution, 2011, 28(1): 257-272.
[21]   Wittkopp P J, True J R, Carroll S B. Reciprocal functions of the Drosophila yellow and ebony proteins in the development and evolution of pigment patterns. Development, 2002, 129(8): 1849-1858.
[22]   Riedel F, Vorkel D, Eaton S. Megalin-dependent yellow endocytosis restricts melanization in the Drosophila cuticle. Development, 2011, 138(1): 149-158.
[23]   Ayoade O, Morooka S, Tojo S. Enhancement of short wing formation and ovarian growth in the genetically defined macropterous strain of the brown planthopper, Nilaparvata lugens. Journal of Insect Physiology, 1999, 45(1): 93-100.
[24]   Drapeau M D, Radovic A, Wittkopp P J, Long A D. A gene necessary for normal male courtship, yellow, acts downstream of fruitless in the Drosophila melanogaster larval brain. Journal of Neurobiology, 2003, 55(1): 53-72.
[25]   Drapeau M D, Cyran S A, Viering M M, Geyer P K, Long A D. A cis-regulatory sequence within the yellow locus of Drosophila melanogaster required for normal male mating success. Genetics, 2006, 172(2): 1009-1030.
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