Skip to main content Accessibility help

Molecular characterization, expression analysis and RNAi knock-down of elongation factor 1α and 1γ from Nilaparvata lugens and its yeast-like symbiont

  • W.X. Wang (a1), T.H. Zhu (a2), K.L. Li (a1), L.F. Chen (a1) (a2), F.X. Lai (a1) and Q. Fu (a1)...


In the present paper, four cDNAs encoding the alpha and gamma subunits of elongation factor 1 (EF-1) were cloned and sequenced from Nilaparvata lugens, named NlEF-1α, NlEF-1γ, and its yeast-like symbiont (YLS), named YsEF-1α and YsEF-1γ, respectively. Comparisons with sequences from other species indicated a greater conservation for EF-1α than for EF-1γ. NlEF-1α has two identical copies. The deduced amino acid sequence homology of NlEF-1α and NlEF-1γ is 96 and 64%, respectively, compared with Homalodisca vitripennis and Locusta migratoria. The deduced amino acid sequence homology of YsEF-1α and YsEF-1γ is 96 and 74%, respectively, compared with Metarhizium anisopliae and Ophiocordyceps sinensis. Reverse transcription-quantitative polymerase chain reaction (RT–qPCR) analysis revealed that the expression level of NlEF-1α and NlEF-1γ mRNA in hemolymph, ovary, fat body and salivary glands were higher than the midgut and leg tissue. YsEF-1α and YsEF-1γ was highly expressed in fat body. The expression level of NlEF-1α was higher than that of NlEF-1γ. Through RNA interference (RNAi) of the two genes, the mortality of nymph reached 92.2% at the 11th day after treatment and the ovarian development was severely hindered. The RT–qPCR analysis verified the correlation between mortality, sterility and the down-regulation of the target genes. The expression and synthesis of vitellogenin (Vg) protein in insects injected with NlEF-1α and NlEF-1γ double-stranded RNA (dsRNA) was significantly lower than control groups. Attempts to knockdown the YsEF-1 genes in the YLS was unsuccessful. However, the phenotype of N. lugens injected with YsEF-1α dsRNA was the same as that injected with NlEF-1α dsRNA, possibly due to the high similarity (up to 71.9%) in the nucleotide sequences between NlEF-1α and YsEF-1α. We demonstrated that partial silencing of NlEF-1α and NlEF-1γ genes caused lethal and sterility effect on N. lugens. NlEF-1γ shares low identity with that of other insects and therefore it could be a potential target for RNAi-based pest management.


Corresponding author

*Author for correspondence Phone: Fax: +86-571-63370348 E-mail:,


Hide All
Belles, X. (2010) Beyond Drosophila: RNAi in vivo and functional genomics in insects. Annual Review of Entomology 55, 111128.
Billaut-Mulot, O., Fernando-Gomez, R., Loyens, M. & Ouaissi, A. (1996) Trypanosoma cruzi elongation factor 1-alpha, nuclear localization in parasites undergoing apoptosis. Gene 174, 1926.
Chen, C.C., Cheng, L.L., Kuan, C.C. & Hou, R.F. (1981) Studies on the intracellular yeast-like symbiote in the brown planthopper, Nilaparvata-lugens Stal. Histological observations and population-changes of the symbiote. Journal of Applied Entomology 91(1), 321327.
Chen, E., Proestou, G., Bourbeau, D. & Wang, E. (2000) Rapid up-regulation of peptide elongation factor EF-1alpha protein levels is an immediate early event during oxidative stress-induced apoptosis. Experiment of Cell Research 259(1), 140148.
Condeelis, J. (1995) Elongation factor 1a, translation and the cytoskeleton. Trends in Biochemical Sciences 20, 169170.
Danforth, B.N. & Ji, S. (1998) Elongation factor-1 alpha occurs as two copies in bees: implications for phylogenetic analysis of EF-1 alpha sequences in insects. Molecular Biology and Evolution 15, 225235.
Ejiri, S. (2002) Moonlighting functions of polypeptide elongation factor 1: from actin bundling to zinc finger protein R1-associated nuclear localization. Bioscience Biotechnology and Biochemistry 66(1), 121.
Elbashir, S.M., Harborth, J., Lendeckel, W., Yalcin, A., Weber, K. & Tuschl, T. (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411(6836), 494498.
Fan, H.W., Noda, H., Xie, H.Q., Suetsugu, Y., Zhu, Q.H., Zhang, C.X. (2015) Genomic analysis of an ascomycete fungus from the rice planthopper reveals how it adapts to an endosymbiotic lifestyle. Genome Biology and Evolution 7(9), 26232634.
Hongoh, Y., Sasaki, T. & Ishikawa, H. (2000) Cloning sequence analysis and expression in Escherichia coli of the gene encoding a uricase from the yeast-like symbiont of the brown planthopper, Nilaparvata lugens. Insect Biochemistry and Molecular Biology 30, 173182.
Hovemann, B., Richter, S., Walldorf, U. & Cziepluch, C. (1988) Two genes encode related cytoplasmic elongation factors 1 alpha (EF-1 alpha) in Drosophila melanogaster with continuous and stage specific expression. Nucleic Acids Research 16, 31753194.
Janssen, G.M.C. & Möller, W. (1988) Elongation factor 1βγ from Artemia. Purification and properties of its subunits. European Journal of Biochemistry 171, 119129.
Kamiie, K., Taira, H., Ooura, H., Kakuta, A., Matsumoto, S., Ejiri, S. & Katsumata, T. (1993) Nucleotide sequence of the cDNA encoding silk gland elongation factor 1 alpha. Nucleic Acids Research 21, 742.
Kamiie, K., Nomura, Y., Kobayashi, S., Taira, H., Kobayashi, K., Yamashita, T., Kidou, S. & Ejiri, S. (2002) Cloning and expression of Bombyx mori silk gland elongation factor 1γ in Escherichia coli . Bioscience Biotechnology and Biochemistry 66, 558565.
Kamiie, K., Yamashita, T., Taira, H., Kidou, S. & Ejiri, S. (2003) Interaction between Elongation Factors 1β and 1γ from Bombyx mori Silk Gland. Bioscience Biotechnology and Biochemistry 67(7), 15221529.
Kaur, K.J. & Ruben, L. (1994) Protein translation elongation factor-1α from Trypanosoma brucei binds calmodulin. Journal of Biology Chemistry 269, 2304523050.
Kidou, S. & Ejiri, S. (1998) Isolation, characterization and mRNA expression of four cDNAs encoding translation elongation factor 1A from rice (Oryza sativa L.). Plant Molecular Biology 36, 137148.
Kidou, S., Tsukamoto, S., Kobayashi, S. & Ejiri, S. (1998) Isolation and characterization of a rice cDNA encoding the γ-subunit of translation elongation factor 1B (eEF1Bγ). FEBS Letters 434(3), 382386.
Lamberti, A., Sanges, C. & Chambery, A. (2011) Analysis of interaction partners for eukaryotic translation elongation factor 1A M-domain by functional proteomics. Biochemistry 93(10), 17381746.
Laurie, J.D., Linning, R. & Bakkeren, G. (2008) Hallmarks of RNA silencing are found in the smut fungus Ustilago hordei but not in its close relative Ustilago maydis . Current Genetics 53(1), 4958.
Le Sourd, F., Boulben, S., Le Bouffant, R., Cormier, P., Morales, J., Belle, R. & Mulner-Lorillon, O. (2006) eEF1B: at the dawn of the 21st century. Biochimica et Biophysica Acta 1759, 1331.
Liu, S., Ding, Z., Zhang, C., Yang, B. & Liu, Z. (2014) Gene knockdown by intro-thoracic injection of double-stranded RNA in the brown planthopper, Nilaparvata lugens . Insect Biochemisty Molecular Biology 40, 666671.
Liu, S., Liang, Q.M., Zhou, W.W., Jiang, Y.-D., Zhu, Q.Z., Yu, H., Zhang, C.X., Gurr, G.M. & Zhu, Z.R. (2015) RNA interference of NADPH-cytochrome P450 reductase of the rice brown planthopper, Nilaparvata lugens, increases susceptibility to insecticides. Pest Management Science 71, 3239.
Livak, K.J. & Schmittgen, T.D. (2011) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25, 402408.
Lu, F., Qi, G.J., Qin, R.R., Hu, G., Wang, Z., Zhang, X.X., Cheng, X.N. & Zhai, B.P. (2011) The processes of morphological change and grading criteria for ovarian development in the brown planthopper. Chinese Journal of Applied Entomology 48(5), 13941400.
Nakayashiki, H., Kadotani, N. & Mayama, S. (2006) Evolution and diversification of RNA silencing proteins in fungi. Journal of Molecular Evolution 63(1), 127135.
Negrutskii, B.S. & El'skaya, A.V. (1998) Eukaryotic translation elongation factor 1 alpha: structure, expression, functions, and possible role in aminoacyl-tRNA channeling. Progress in Nucleic Acid Research and Molecular Biology 60, 4778.
Noda, H. & Koizumi, Y. (2003) Sterol biosynthesis by symbiotes: cytochrome P450 sterol C-22 desaturase genes from yeastlike symbiotes of rice planthoppers and anobiid beetles. Insect Biochemistry and Molecular Biology 33, 649658.
Rual, J.F., Klitgord, N. & Achaz, G. (2007) Novel insights into RNAi off-target effects using C. elegans paralogs. BMC Genomics 8, 106.
Shepherd, J.C., Walldorf, U., Hug, P. & Gehring, W.J. (1989) Fruit flies with additional expression of the elongation factor EF-1 alpha live longer. Proceedings of the National Academy of Sciences of the United States of America 86, 75207521.
Slobin, L. (1980) The role of eukaryotic elongation factor Tu in protein synthesis. The measurement of the elongation factor Tu content of rabbit reticulocytes and other mammalian cells by a sensitive radioimmunoassay. European Journal of Biochemistry 110, 555563.
Sogawa, K. (1982) The rice brown planthopper-feeding physiology and host plant interactions. Annual Review of Entomology 27, 4973.
Suh, S.O., Noda, H. & Blackwell, M. (2001) Insect symbiosis: derivation of yeast-like endosymbionts within an entomopathogenic filamentous lineage. Molecular Biology and Evolution 18, 9951000.
Taira, H., Kamiie, K., Kakuta, A., Ooura, H., Matsumoto, S., Ejiri, S. & Katsumata, T. (1992) Nucleotide sequence of the cDNA encoding silk gland elongation factor 1 beta’. Nucleic Acids Research 20, 6734.
Vinkenoog, R., Sperança, M.A., van Breemen, O., Ramesar, J., Williamson, D.H., Ross-MacDonald, P.B., Thomas, A.W., Janse, C.J., del Portillo, H.A. & Waters, A.P. (1998) Malaria parasites contain two identical copies of an elongation factor 1 alpha gene. Molecular and Biochemical Parasitology 94(1), 112.
Wang, W.X., Li, K.L., Chen, Y., Lai, F.X. & Fu, Q. (2015) Identification and function analysis of enolase gene NlEno1 from Nilaparvata lugens (Stål) (Hemiptera: Delphacidae). Journal of Insect Science 15(1), 66.
Webster, G.C. & Webster, S.L. (1983) Decline in synthesis of elongation factor one (EF-1) precedes the decreased synthesis of total protein in aging Drosophila melanogaster . Mechanisms of Ageing and Development 22, 121128.
Xue, J., Zhou, X., Zhang, C.X., Yu, L.L. & Cheng, J.A. (2014) Genomes of the rice pest brown planthopper and its endosymbionts reveal complex complementary contributions for host adaptation. Genome Biology 15, 521.
Zhao, L.N., Qin, Z., Wei, P., Guo, H.S., Dang, X.L., Wang, S.G. & Tang, B. (2012) Elongation Factor 1 beta ’ Gene from Spodoptera exigua: characterization and function identification through RNA Interference. International Journal of Molecular Sciences 13(7), 81268141.
Zhou, S., Zhang, J., Fam, M.D., Wyatt, G.R. & Walker, V.K. (2002) Sequences of elongation factors-1α and -1γ and stimulation by juvenile hormone in Locusta migratoria . Insect Biochemistry and Molecular Biology 32, 15671576.


Type Description Title
Supplementary materials

Wang supplementary material
Fig. S1

 Unknown (4.7 MB)
4.7 MB
Supplementary materials

Wang supplementary material
Fig. S2

 Unknown (4.0 MB)
4.0 MB
Supplementary materials

Wang supplementary material
Fig. S3

 Unknown (2.1 MB)
2.1 MB
Supplementary materials

Wang supplementary material
Fig. S4

 Unknown (25 KB)
25 KB
Supplementary materials

Wang supplementary material
Table S1

 Word (86 KB)
86 KB


Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed