Please wait a minute...
Journal of Integrative Agriculture  2014, Vol. 13 Issue (9): 1972-1981    DOI: 10.1016/S2095-3119(13)60654-1
Plant Protection Advanced Online Publication | Current Issue | Archive | Adv Search |
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
Download:  PDF in ScienceDirect  
Export:  BibTeX | EndNote (RIS)      
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.
Keywords:  brown planthopper       Krüppel-homolog 1       RNA interference       juvenile hormone signaling pathway  
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, 130,2397-2406

Ayoade 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, 45,93-100

Bao 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, 1087-1094

Beck Y, Pecasse F, Richards G. 2004. Kruppel-homolog isessential for the coordination of regulatory gene hierarchiesin early Drosophila development. Developmental Biology,268, 64-75

Benevolenskaya E V, Frolov M V, Birchler J A. 2000. Kruppelhomolog (Kr h) is a dosage-dependent modifier of geneexpression in Drosophila. Genetics Research, 75, 137-142

Bertuso 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, 221-229

Erezyilmaz 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, 313-323

Fichelson 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, 7893-7898

Fussnecker B, Grozinger C. 2008. Dissecting the role of Kr-h1brain gene expression in foraging behavior in honey bees(Apis mellifera). Insect Molecular Biology, 17, 515-522

Ge 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, 172-181

Jindra M, Palli S R, Riddiford L M. 2013. The juvenilehormone signaling pathway in insect development. AnnualReview of Entomology, 58, 181-204

Konopova 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, 123-133

Liu 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, 54,1495-1502

Liu 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, 62,279-282

Livak K J, Schmittgen T D. 2001. Analysis of relative geneexpression data using real-time quantitative PCR and the2-ΔΔCt method. Methods, 25, 402-408

Lozano 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, 341-350

Minakuchi 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, 125-134

Minakuchi C, Zhou X, Riddiford L M. 2008. Kruppel homolog1 (Kr-h1) mediates juvenile hormone action duringmetamorphosis of Drosophila melanogaster. Mechanismof Development, 125, 91-105

Miranda 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, 26-32

Nakashima 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, 139-146

Nakashima N, Noda H. 1995. Nonpathogenic Nilaparvatalugens reovirus is transmitted to the brown planthopperthrough rice plant. Virology, 207, 303-307

Noda H, Ishikawa K, Hibino H, Omura T. 1991. A reovirusin the brown planthopper, Nilaparvata lugens. Journal ofGeneral Virology, 72, 2425-2430

Oliveira 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, 41-49

Parthasarathy 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, 299-313

Pecasse 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, 53-67

Riddiford L M. 2008. Juvenile hormone action: A 2007perspective. Journal of Insect Physiology, 54, 895-901

Riddiford LM. 2012. How does juvenile hormone controlinsect metamorphosis and reproduction? General andComparative Endocrinology, 179, 477-484

Shi 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, 1614-1626

Xue 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, 180-186

Zera 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, 293-306

Zera 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
[1] SU Qin, LÜ Jun, LI Wan-xue, CHEN Wei-wen, LUO Min-shi, ZHANG Chuan-chuan, ZHANG Wen-qing. The combination of NlMIP and Gαi/q coupled-receptor NlA10 promotes abdominal vibration production in female Nilaparvata lugens (Stål)[J]. >Journal of Integrative Agriculture, 2023, 22(8): 2470-2482.
[2] JIN Ji-su, LIU Yi-ran, ZHOU Zhong-shi, WAN Fang-hao, GUO Jian-ying. Halloween genes AhCYP307A2 and AhCYP314A1 modulate last instar larva–pupa–adult transition, ovarian development and oogenesis in Agasicles hygrophila (Coleoptera: Chrysomelidae)[J]. >Journal of Integrative Agriculture, 2023, 22(3): 812-824.
[3] Jelli VENKATESH, Sung Jin KIM, Muhammad Irfan SIDDIQUE, Ju Hyeon KIM, Si Hyeock LEE, Byoung-Cheorl KANG. CopE and TLR6 RNAi-mediated tomato resistance to western flower thrips[J]. >Journal of Integrative Agriculture, 2023, 22(2): 471-480.
[4] YE Qing-ya, LI Zhi-xing, CHEN Qing-ling, SUN Ming-xu, YIN Ming-liang, LIN Tong. Fatty acid-binding protein gene is indispensable for molting process in Heortia vitessoides (Lepidoptera: Crambidae)[J]. >Journal of Integrative Agriculture, 2023, 22(2): 495-504.
[5] FAN Zi-zhen, MA Qin, MA Si-ya, CAO Feng-qin, YAN Ri-hui, LIN Xian-wu.

Maleness-on-the-Y (MoY) orthologue is a key regulator of male sex determination in Zeugodacus cucurbitae (Diptera: Tephritidae) [J]. >Journal of Integrative Agriculture, 2023, 22(2): 505-513.

[6] LIU Li-feng, GAO Le, ZHANG Li-xin, CAI Yu-peng, SONG Wen-wen, CHEN Li, YUAN Shan, WU Ting-ting, JIANG Bing-jun, SUN Shi, WU Cun-xiang, HOU Wen-sheng, HAN Tian-fu. Co-silencing E1 and its homologs in an extremely late-maturing soybean cultivar confers super-early maturity and adaptation to high-latitude short-season regions[J]. >Journal of Integrative Agriculture, 2022, 21(2): 326-335.
[7] LI Tian-pu, ZHANG Li-wen, LI Ya-qing, YOU Min-sheng, ZHAO Qian. Functional analysis of the orphan genes Tssor-3 and Tssor-4 in male Plutella xylostella[J]. >Journal of Integrative Agriculture, 2021, 20(7): 1880-1888.
[8] MENG Miao, YU Qi, WANG Qin, LIU Chun, LIU Zhao-yang, REN Chun-jiu, CUI Wei-zheng, LIU Qing-xin. BmApontic is involved in neurodevelopment in the silkworm Bombyx mori[J]. >Journal of Integrative Agriculture, 2020, 19(6): 1439-1446.
[9] LIU Jiao, ZHANG Xue-yao, WU Hai-hua, MA Wen, ZHU Wen-ya, Kun-Yan ZHU, MA En-bo, ZHANG Jian-zhen . Characteristics and roles of cytochrome b5 in cytochrome P450-mediated oxidative reactions in Locusta migratoria[J]. >Journal of Integrative Agriculture, 2020, 19(6): 1512-1521.
[10] MA Mei-qi, HE Wan-wan, XU Shi-jing, XU Le-tian, ZHANG Jiang.
RNA interference in Colorado potato beetle (Leptinotarsa decemlineata): A potential strategy for pest control
[J]. >Journal of Integrative Agriculture, 2020, 19(2): 428-427.
[11] CHEN Meng-yao, YE Wan-yi, XIAO Hua-mei, LI Mei-zhen, CAO Zheng-hong, YE Xin-hai, ZHAO Xian-xin, HE Kang, LI Fei. LncRNAs are potentially involved in the immune interaction between small brown planthopper and rice stripe virus[J]. >Journal of Integrative Agriculture, 2019, 18(12): 2814-2822.
[12] HU Wen-bin, DENG Xiang-yang, DENG Xiao-xiang, DENG Li-hua, XIAO You-lun, HE Xing-jian, FU Xiqin, XIAO Guo-ying. Characteristic analysis of tetra-resistant genetically modified rice[J]. >Journal of Integrative Agriculture, 2018, 17(03): 493-506.
[13] JI Zhi-juan, Yang Shu-dong, ZENG Yu-xiang, LIANG Yan, YANG Chang-deng, QIAN Qian. Pyramiding blast, bacterial blight and brown planthopper resistance genes in rice restorer lines[J]. >Journal of Integrative Agriculture, 2016, 15(7): 1432-1440.
[14] CHEN Tai-yu, HOU Ji-xiang, LIN Yong-jun. Transcriptome datasets supply basic gene information for RNAi pest management and gene functional studies in Nephotettix cincticeps (Uhler)[J]. >Journal of Integrative Agriculture, 2016, 15(4): 840-847.
[15] ZHANG Jiao, XING Yan-ru, HOU Bo-feng, YUAN Zhu-ting, LI Yao, JIE Wen-cai, SUN Yang, LI Fei. Amplification and function analysis of N6-adenine-specific DNA methyltransferase gene in Nilaparvata lugens[J]. >Journal of Integrative Agriculture, 2016, 15(3): 591-599.
No Suggested Reading articles found!