Transcriptome analysis in Ceratitis capitata to unveil genes involved in ageing-maturation process

V. San Andrés, P. Castañera, B. Sabater Muñoz


The sterile insect technique (SIT) is widely used in integrated programmes against the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Information on the age distribution of insects, and more particularly, the knowledge of wild female reproductive status (mature or not) at the time of the sterile male release is one of the key factors for the success of the SIT. In recent years, sequencing analysis has become an important tool in molecular biology. In this work we present a genome-wide expression analysis based on SSH (substractive sequence hybridization) and EST (expressed sequence tag) sequencing and macroarray expression analysis to identify signature genes related to the ageing-maturing process in C. capitata, leading to the successful identification of new putative candidate genes of reproductive status in medfly that would serve as molecular markers for ageing. We have sorted out 94 unigenes from 873 single-pass ESTs, of which 57% have homology with known genes. Ageing-maturing process in C. capitata presents a marked expression pattern accompanied by the increase of transcription level of genes involved in reproduction (vitellogenins, chorion proteins and male-specific serum proteins). Other identified cDNAs (43%) with a differential expression pattern would be also candidates but deserve further studies, as they belong to the unknown function class.


ageing; expression profile; genome analysis; medfly; macroarray; sexual maturation; SSH

Full Text:



Aigaki T, Seong K, Matsuo T, 2002. Longevity determination genes in Drosophila melanogaster. Mech Ageing Develop 123: 1531-1541.

Alberola TM, Garcia-Martinez J, Antunez O, Viladevall L, Barcelo A, Arino J, Perez-Ortin JE, 2004. A new set of DNA macrochips for the yeast Saccharomyces cerevisiae: features and uses. Inter Microbiol 7: 199-206. PMid:15492934

Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipmann DJ, 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucl Acids Res 25(17): 3389-3402. PMid:9254694 PMCid:PMC146917

Bellés X, 2004. Vitellogenesis directed by juvenile hormone. In: Reproductive biology of invertebrates Vol. XII, Part B: Progress in vitellogenesis (Raikhel AS, ed.). John Wiley & Sons Inc, NY, pp: 157-198.

Camin V, Baker P, Carey J, Valenzuela J, Arredondopeter R, 1991. Biochemical age-determination for adult Mediterranean fruit-flies (Diptera, Tephritidae). J Econ Entomol 84: 1283-1288.

Carey JR, 2003. Longevity: the biology and demography of life span. Princeton Univ. Press, Princeton, NJ, USA. 278 pp.

Carey JR, Papadopoulos NT, Kouloussis NA, Katsoyannos BI, Müller HG, Wang JL, Tseng YK, 2006. Age-specific and lifetime behavior patterns in Drosophila melanogaster and the Mediterranean fruit fly, Ceratitis capitata. Exp Geront 41(1): 93-97. PMid:16297586 PMCid:PMC2387245

Carey JR, Papadopoulos NT, Müller H-G, Katsoyannos BI, Kouloussis NA, Wang JL, Wachter K, Yu W, Liedo P, 2008. Age structure changes and extraordinary lifespan in wild medfly populations. Aging Cell 7(3): 426-437. PMid:18363903 PMCid:PMC2398686

Cook PE, Hugo LE, Iturbe-Ormaetxe I, Williams CR, Chenoweth SF, Ritchie SA, Ryan PA, Kay BH, Blows MW, O'Neill SL, 2006. The use of transcriptional profiles to predict adult mosquito age under field conditions. P Natl Acad Sci USA 103: 18060-18065. PMid:17110448 PMCid:PMC1838706

Cook PE, Hugo LE, Iturbe-Ormaetxe I, Williams CR, Chenoweth SF, Ritchie SA, Ryan PA, Kay BH, Blows MW, O'neill SL, 2007. Predicting the age of mosquitoes using transcriptional profiles. Nature Prot 2: 2796-2806. PMid:18007615

Desena ML, Clark JM, Edman JD, Symington SB, Scott TW, Clark GG, Peters TM, 1999. Potential for aging female Aedes aegypti (Diptera: Culicidae) by gas chromatographic analysis of cuticular hydrocarbons, including a field evaluation. J Med Entomol 36: 811-823. PMid:10593085

Diatchenko L, Lau YF, Campbell AP, Chenchik A, Moqadam F, Huang B, Lukyanov S, Lukyanov K, Gurskaya N, Sverdlov ED, Siebert PD, 1996. Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries. P Natl Acad Sci USA 93: 6025-6030. PMid:8650213 PMCid:PMC39182

Gerade BB, Lee SH, Scott TW, Edman JD, Harrington LC, Kitthawee S, Jones JW, Clark JM, 2004. Field validation of Aedes aegypti (Diptera: Culicidae) age estimation by analysis of cuticular hydrocarbons. J Med Entomol 41: 231-238. PMid:15061283

Gomulski LM, Dimopoulos G, Xi Z, Scolari F, Gabrieli P, Siciliano P, Clarke AR, Malacrida AR, Gaspieri G, 2012. Transcriptome profiling of sexual maturation and mating in the Mediterranean fruit fly, Ceratitis capitata. PLOS One 7(1): e30857. PMid:22303464 PMCid:PMC3267753

Hendrichs J, Vreysen MJB, Enkerlin WR, Cayol JP, 2005. Strategic options in using sterile insects for area-wide integrated pest management. In: Sterile insect technique. Principles and practice in area-wide integrated pest management (Dyck VA, Hendrichs J, Robinson AS, eds.). Springer, Dordrecht, The Netherlands. pp: 563-600.

Herrero J, Al-Shahrour F, Díaz-Uriarte R, Mateos A, Vaquerizas JM, Santoyo J, Dopazo J, 2003. GEPAS: A web-based resource for microarray gene expression data analysis. Nucl Acids Res 31: 3461-3467. PMid:12824345 PMCid:PMC168997

Katsoris PG, Mavroidis M, Mintzas AC, 1990. Identification and characterization of male specific serum proteins in the Mediterranean fruit fly Ceratitis capitata. Insect Biochem 20(7): 653-657.

Kim SK, 2008. Genome-wide views of aging gene networks. In: Molecular biology of aging (Guarente LP, Partridge L, Wallace DC (eds.). Cold Spring Harbor Lab. Press, Cold Spring Harbor, USA. 610 pp.

Kirkwood TBL, Austad SN, 2000. Why do we age? Nature 408: 233-238. PMid:11089980

Konsolaki M, Komitopoulou K, Tolias PP, King DL, Swimmer C, Kafatos FC, 1990. The chorion genes of the medfly, Ceratitis capitata, I: Structural and regulatory conservation of the s36 gene relative to two Drosophila species. Nucl Acid Res 18(7): 1731-1737. PMid:1692403 PMCid:PMC330590

Liquido NJ, Shinoda LA, Cunningham RT, 1991. Host plants of the Mediterranean fruit fly (Diptera: Tephritidae): an annotated world review. Misc Pub Entomol Soc Am 77: V-52.

Maga-a C, Hernández-Crespo P, Brun-Barale A, Couso-Ferrer F, Bride M, Casta-era P, Feyereisen R, Ortego F, 2008. Mechanisms of resistance to malathion in the medfly Ceratitis capitata. Insect Biochem Mol Biol 38: 756-762. PMid:18625398

Malacrida AR, Gomulski LM, Bonizzoni M, Bertin S, Gasperi G, Guglielmino CR, 2007. Globalization and fruitfly invasion and expansion: the medfly paradigm. Genetica 131(1): 1-9. PMid:17111234

McCarroll SA, Murphy CT, Zou S, Pletcher SD, Chin CS, Jan YN, Kenyon C, Bargmann CI, Li H, 2004. Comparing genomic expression patterns across species identifies shared transcriptional profile in aging. Nature Genet 36: 197-204. PMid:14730301

Moshitzky P, Miloslavski I, Aizenshtat Z, Applebaum SW, 2003. Methyl palmitate: a novel product of the Medfly (Ceratitis capitata) corpus allatum. Insect Biochem Mol Biol 33(12): 1299-1306. PMid:14599501

Mullens BA, Lehane MJ, 1995. Fluorescence as a tool for age determination in Culicoides variipennis sonorensis (Diptera: Ceratopogonidae). J Med Entomol 32: 569-571. PMid:7650721

Novoseltsev VN, Arking R, Carey JR, Novoseltseva JA, Yashin AI, 2005. Individual fecundity and senescence in Drosophila and medfly. J Geront Series A: Biol Sci Med Sci 60(8): 953-962.

Papadopoulos NT, Carey JR, Katsoyannos BI, Kouloussis NA, Müller HG, Liu X, 2002. Supine behaviour predicts time-to-death in male Mediterranean fruit flies. Proc Royal Soc London: Biol Sci 269: 1633-1637. PMid:12204122 PMCid:PMC1691077

Penilla RP, Rodriguez MH, Lopez AD, Viader-Salvado JM, Sanchez CN, 2002. Pteridine concentrations differ between insectary-reared and field-collected Anopheles albimanus mosquitoes of the same physiological age. Med Veter Entomol 16: 225-234. PMid:12243223

Raikhel AS, Brown MR, Bellés X, 2004. Hormonal control of reproductive processes. In: Comprehensive molecular insect science, Vol. 3 (Gilbert LI, Iatrou K, Gill SS, eds.). Elsevier, Oxford, UK. pp: 433-491.

Sabater-Mu-oz B, Legeai F, Rispe C, Bonhomme J, Dearden P, Dossat C, Duclert A, Gauthier JP, Ducray DG, Hunter W, et al., 2006. Large-scale gene discovery in the pea aphid Acyrthosiphon pisum (Hemiptera). Genome Biol 7(3): R21. PMid:16542494 PMCid:PMC1557754

Sambrook J, Fritsch EF, Maniatis T, 1989. Molecular cloning. A laboratory manual. Cold Spring Harbor Press, Cold Spring Harbor, USA. 2344 pp.

San Andrés V, Urbaneja A, Sabater-Mu-oz B, Casta-era P, 2007. A novel molecular approach to assess mating success of sterile Medfly males in SIT programs. J Econ Entomol 100(4): 1444-1447.[1444:ANMATA]2.0.CO;2

Satyanarayana K, Bradfield JY, Bhaskaran G, Dahm KH, 1994. Stimulation of vitellogenin production by methoprene in prepupae and pupae of Manduca sexta. Arch Insect Biochem Physiol 25: 21-37.

Scolari F, Gomulski LM, Ribeiro JM, Siciliano P, Meraldi A, Falchetto M, Bonomi A, Manni M, Gabrieli P, Malovini A, Bellazzi R, Aksoy S, Gasperi G, Malacrida AR, 2012. Transcriptional profiles of mating-responsive genes from testes and male accessory glands of the Mediterranean fruit fly, Ceratitis capitata. PLOS One 7(10): e46812. PMid:23071645 PMCid:PMC3469604

Seehuus SC, Norberg K, Gimsa U, Krekling T, Amdam GV, 2006. Reproductive protein protects sterile honey bee workers from oxidative stress. P Natl Acad Sci USA 103: 962-967. PMid:16418279 PMCid:PMC1347965

Staden R, Beal KF, Bonfield JK, 2000. The Staden package, 1998 In: Methods in molecular biology, vol. 132, Bioinformatics: methods and protocols (Misener S, Krawetz S, eds.). Humana Press Inc., Totowa, NJ, USA. pp: 115-130.

Thymianou SP, Chrysanthis G, Petropoulou KA, Mintzas AC, 1995. Developmentally regulated biosynthesis of two male-specific serum protein in the fat body of the Mediterranean fruit fly, Ceratitis capitata. Insect Biochem Mol Biol 25(8): 915-920.

Thymianou SP, Mavroidis M, Konsolaki M, Komitopoulou K, Zacharopoulou A, Mintzas AC, 1998. Cloning and characterization of a cDNA encoding a male-specific serum protein of the Mediterranean fruit fly, Ceratitis capitata, with sequence similarity to odorant-binding proteins. Insect Mol Biol 7(4): 345-353. PMid:9723872

Tolias PP, Konsolaki M, Komitopoulou K, Kafatos FC, 1990. The chorion genes of the medfly, Ceratitis capitata. II. Characterization of three novel cDNA clones obtained by differential screening of an ovarian library. Develop Biol 140: 105-112.

Tyndale-Biscoe M, 1984. Age-grading methods in adult insects a review. Bul Entomol Res 74: 341-377.

Vlachou D, Konsolaki M, Tolias PP, Kafatos FC, Komitopoulou K, 1997. The autosomal chorion locus of the medfly Ceratitis capitata. I. Conserved synteny, amplification and tissue specificity but sequence divergence and altered temporal regulation. Genetics 147: 1829-1842. PMid:9409839 PMCid:PMC1208349

White IM, Elson-Harris MM, 1992. Fruit flies of economic significance: their identification and bionomics. CAB Int. Publ., The Australian Center for International Agricultural Research, Eastbourne, Australia.

Wyatt GR, Davey KG, 1996. Cellular and molecular actions of juvenile hormone. II Roles of juvenile hormone in adult insects. Adv Insect Physiol 26: 1-155.

Yano K, Sakurai MT, Watabe S, Izumi S, Tomino S, 1994. Structure and expression of mRNA for vitellogenin in Bombyx mori. Biochim Biophys Acta 1218 (1): 1-10.

DOI: 10.5424/sjar/2013113-3987