|
|
|
Molecular Characterization and Functional Analysis of Krüppel-homolog 1 (Kr-h1) in the Brown Planthopper, Nilaparvata lugens (Stål) |
JIN Min-na, XUE Jian, YAO Yun , LIN Xin-da |
1、College of Life Sciences, China Jiliang University, Hangzhou 310018, P.R.China
2、Institute of Insect Science, Zhejiang University, Hangzhou 310058, P.R.China |
|
|
摘要 The brown planthopper, Nilaparvata lugens (Stål), is the most serious insect pest of rice. It has developed high resistance to traditional insecticides because of their intensive use. Juvenile hormone (JH) analogs have been used successfully to control this species and other pest insects. However, the molecular mechanism of JH signaling is not well understood. Krüppel-homolog 1 (Kr-h1) is a transcription factor involved in the JH pathway. In this study, the Kr-h1 cDNA was cloned and characterized from N. lugens by rapid amplification of cDNA ends (RACE) and reverse transcription PCR (RT-PCR). Its spatial and temporal expression profiles were examined by real-time quantitative PCR, and its function was also studied by RNA interference (RNAi). The open reading frame of NlKr-h1 is 1 833 bp encoding for 611 amino acids. The protein contains eight conserved zinc-finger motifs. NlKr-h1 was expressed at all life stages, with the highest mRNA level in the 4-day embryo. NlKr-h1 mRNA levels rose during each nymphal molt after the 2nd instar. In the adults, the mRNA level in males was significantly higher than that in females of either the macropterous or brachypterous type. The highest expression was observed in the female midgut. NlKr-h1 was activated by juvenile hormone III (JH III) in the 3rd-5th instar nymphs. Disruption of Nlkr-h1 expression by RNAi caused stunted wing development and malformations of both male and female external genitalia. Our findings suggest that Kr-h1 may be a useful target for pest insect management.
Abstract The brown planthopper, Nilaparvata lugens (Stål), is the most serious insect pest of rice. It has developed high resistance to traditional insecticides because of their intensive use. Juvenile hormone (JH) analogs have been used successfully to control this species and other pest insects. However, the molecular mechanism of JH signaling is not well understood. Krüppel-homolog 1 (Kr-h1) is a transcription factor involved in the JH pathway. In this study, the Kr-h1 cDNA was cloned and characterized from N. lugens by rapid amplification of cDNA ends (RACE) and reverse transcription PCR (RT-PCR). Its spatial and temporal expression profiles were examined by real-time quantitative PCR, and its function was also studied by RNA interference (RNAi). The open reading frame of NlKr-h1 is 1 833 bp encoding for 611 amino acids. The protein contains eight conserved zinc-finger motifs. NlKr-h1 was expressed at all life stages, with the highest mRNA level in the 4-day embryo. NlKr-h1 mRNA levels rose during each nymphal molt after the 2nd instar. In the adults, the mRNA level in males was significantly higher than that in females of either the macropterous or brachypterous type. The highest expression was observed in the female midgut. NlKr-h1 was activated by juvenile hormone III (JH III) in the 3rd-5th instar nymphs. Disruption of Nlkr-h1 expression by RNAi caused stunted wing development and malformations of both male and female external genitalia. Our findings suggest that Kr-h1 may be a useful target for pest insect management.
|
Received: 10 May 2013
Accepted:
|
Fund: This work was supported by the National 973 Program of China (2010CB126205) and the Zhejiang Provincial Natural Science Foundation of China (LY14C140002). |
Corresponding Authors:
LIN Xinda,Tel: +86-571-86835772, E-mail: 07a0903045@cjlu.edu.cn
E-mail: 07a0903045@cjlu.edu.cn
|
About author: JIN Min-na, Tel: +86-571-86835772, E-mail: 1007101022@cjlu.edu.cn; XUE Jian, Tel: +86-571-88982991, E-mail: alexjxue@gmail.com |
Cite this article:
JIN Min-na, XUE Jian, YAO Yun , LIN Xin-da.
2014.
Molecular Characterization and Functional Analysis of Krüppel-homolog 1 (Kr-h1) in the Brown Planthopper, Nilaparvata lugens (Stål). Journal of Integrative Agriculture, 13(9): 1972-1981.
|
Adam G, Perrimon N, Noselli S. 2003. The retinoic-likejuvenile hormone controls the looping of left-rightasymmetric organs in Drosophila. Development, 130,2397-2406Ayoade O, Morooka S, Tojo S. 1999. Enhancement of shortwing formation and ovarian growth in the geneticallydefined macropterous strain of the brown planthopper,Nilaparvata lugens. Journal of Insect Physiology, 45,93-100Bao Y Y, Li B L, Liu Z B, Xue J, Zhu Z R, Cheng J A, ZhangC X. 2010. Triazophos up-regulated gene expression in thefemale brown planthopper, Nilaparvata lugens. Journal ofInsect Physiology, 56, 1087-1094Beck Y, Pecasse F, Richards G. 2004. Kruppel-homolog isessential for the coordination of regulatory gene hierarchiesin early Drosophila development. Developmental Biology,268, 64-75Benevolenskaya E V, Frolov M V, Birchler J A. 2000. Kruppelhomolog (Kr h) is a dosage-dependent modifier of geneexpression in Drosophila. Genetics Research, 75, 137-142Bertuso A G, Morooka S, Tojo S. 2002. Sensitive periods forwing development and precocious metamorphosis afterprecocene treatment of the brown planthopper, Nilaparvatalugens. Journal of Insect Physiology, 48, 221-229Erezyilmaz D F, Riddiford L M, Truman J W. 2004. Juvenilehormone acts at embryonic molts and induces the nymphalcuticle in the direct-developing cricket. DevelopmentGenes and Evolution, 214, 313-323Fichelson P, Brigui A, Pichaud F. 2012. Orthodenticle andKruppel homolog 1 regulate Drosophila photoreceptormaturation. Proceedings of the National Academy ofSciences of the United States of America, 109, 7893-7898Fussnecker B, Grozinger C. 2008. Dissecting the role of Kr-h1brain gene expression in foraging behavior in honey bees(Apis mellifera). Insect Molecular Biology, 17, 515-522Ge L Q, Zhao K F, Huang L J, Wu J C. 2011. The effects oftriazophos on the trehalose content, trehalase activity andtheir gene expression in the brown planthopper Nilaparvatalugens (Stål) (Hemiptera: Delphacidae). PesticideBiochemistry and Physiology, 100, 172-181Jindra M, Palli S R, Riddiford L M. 2013. The juvenilehormone signaling pathway in insect development. AnnualReview of Entomology, 58, 181-204Konopova B, Smykal V, Jindra M. 2011. Common anddistinct roles of juvenile hormone signaling genes inmetamorphosis of holometabolous and hemimetabolousinsects. PLoS ONE, 6, e28728.Koyama T, Obara Y, Iwami M, Sakurai S. 2004. Commencementof pupal commitment in late penultimate instar and itshormonal control in wing imaginal discs of the silkworm,Bombyx mori. Journal of Insect Physiology, 50, 123-133Liu S, Yang B, Gu J, Yao X, Zhang Y, Song F, Liu Z.2008. Molecular cloning and characterization of ajuvenile hormone esterase gene from brown planthopper,Nilaparvata lugens. Journal of Insect Physiology, 54,1495-1502Liu S, Zhang C, Yang B, Gu J, Liu Z. 2010. Cloningand characterization of a putative farnesoic acidO-methyltransferase gene from the brown planthopper,Nilaparvata lugens. Journal of Insect Science, 10, 103.Liu Z, Han Z. 2006. Fitness costs of laboratory-selectedimidacloprid resistance in the brown planthopper,Nilaparvata lugens Stål. Pest Management Science, 62,279-282Livak K J, Schmittgen T D. 2001. Analysis of relative geneexpression data using real-time quantitative PCR and the2-ΔΔCt method. Methods, 25, 402-408Lozano J, Belles X. 2011. Conserved repressive functionof Kruppel homolog 1 on insect metamorphosis inhemimetabolous and holometabolous species. ScientificReports, 1, 163.Minakuchi C, Namiki T, Shinoda T. 2009. Kruppel homolog1, an early juvenile hormone-response gene downstream ofmethoprene-tolerant, mediates its anti-metamorphic actionin the red flour beetle Tribolium castaneum. DevelopmentalBiology, 325, 341-350Minakuchi C, Tanaka M, Miura K, Tanaka T. 2011.Developmental profile and hormonal regulation of thetranscription factors broad and Kruppel homolog 1 inhemimetabolous thrips. Insect Biochemistry and MolecularBiology, 41, 125-134Minakuchi C, Zhou X, Riddiford L M. 2008. Kruppel homolog1 (Kr-h1) mediates juvenile hormone action duringmetamorphosis of Drosophila melanogaster. Mechanismof Development, 125, 91-105Miranda G J, Azzam O, Shirako Y. 2000. Comparison ofnucleotide sequences between northern and southernphilippine isolates of rice grassy stunt virus indicatesoccurrence of natural genetic reassortment. Virology, 266, 26-32Nakashima N, Koizumi M, Watanabe H, Noda H. 1996.Complete nucleotide sequence of the Nilaparvata lugensreovirus: a putative member of the genus Fijivirus. Journalof General Virology, 77, 139-146Nakashima N, Noda H. 1995. Nonpathogenic Nilaparvatalugens reovirus is transmitted to the brown planthopperthrough rice plant. Virology, 207, 303-307Noda H, Ishikawa K, Hibino H, Omura T. 1991. A reovirusin the brown planthopper, Nilaparvata lugens. Journal ofGeneral Virology, 72, 2425-2430Oliveira Filho A M, Pinchin R, Jurberg J, Rangel E F. 1981.Alterations in the morphology of the external genitaliaof Panstrongylus megistus (Burm., 1835), caused bytreatment with a juvenile hormone analogue (Hemiptera,Reduviidae, Triatominae). Revista Brasileira de Biologia,41, 41-49Parthasarathy R, Tan A, Bai H, Palli S R. 2008. Transcriptionfactor broad suppresses precocious development of adultstructures during larval-pupal metamorphosis in thered flour beetle, Tribolium castaneum. Mechanism ofDevelopment, 125, 299-313Pecasse F, Beck Y, Ruiz C, Richards G. 2000. Kruppel-homolog,a stage-specific modulator of the prepupal ecdysoneresponse, is essential for Drosophila metamorphosis.Developmental Biology, 221, 53-67Riddiford L M. 2008. Juvenile hormone action: A 2007perspective. Journal of Insect Physiology, 54, 895-901Riddiford LM. 2012. How does juvenile hormone controlinsect metamorphosis and reproduction? General andComparative Endocrinology, 179, 477-484Shi L, Lin S, Grinberg Y, Beck Y, Grozinger C M, RobinsonG E, Lee T. 2007. Roles of Drosophila Kruppel-homolog 1in neuronal morphogenesis. Developmental Neurobiology,67, 1614-1626Xue J, Bao Y Y, Li B L, Cheng Y B, Peng Z Y, Liu H,Xu H J, Zhu Z R, Lou Y G, Cheng J A, Zhang C X.2010. Transcriptome analysis of the brown planthopperNilaparvata lugens. PLoS ONE, 5, e14233.Yang C M, Cheng C H. 2001. Spectral characteristics of riceplants infested by brown planthoppers. Proceedings of theNational Science Council, Republic of China, 25, 180-186Zera A J, Cisper G. 2001. Genetic and diurnal variation inthe juvenile hormone titer in a wing-polymorphic cricket:Implications for the evolution of life histories and dispersal.Physiological and Biochemical Zoology, 74, 293-306Zera A J, Zhao Z, Kaliseck K. 2007. Hormones in the field:evolutionary endocrinology of juvenile hormone andecdysteroids in field populations of the wing-dimorphiccricket Gryllus firmus. Physiological and BiochemicalZoology, 80, 592-606 |
No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
|
Shared |
|
|
|
|
|
Discussed |
|
|
|
|