Molecular Biotechnology

, Volume 35, Issue 1, pp 31–40 | Cite as

Mitochondrial DNA variability and development of a PCR diagnostic test for populations of the whitefly Bemisia afer (priesner and hosny)

  • M. N. Maruthi
  • A. R. Rekha
  • P. Sseruwagi
  • R. J. Hillocks
Research

Abstract

The whitefly, Bemisia afer (Hemiptera; Aleyrodidae), is emerging as a major agricultural pest. The current identification methods based on adult and pupal morphology are laborious and unreliable. A diagnostic polymerase chain reaction (PCR) protocol was developed for the first time in this study to discriminate B. afer from other whitefly species. Primers specific to mitochondrial cytochrome oxidase 1 gene (mtCOI) were designed to amplify a band of approx 650 bp. The PCR products were sequenced from B. afer samples collected from Malawi, Tanzania, Uganda, Zanzibar, and the United Kingdom. Phylogenetic analyses of mtCOI sequences and those of reference B. afer sequences clustered the African B. afer separately from the UK and Chinese populations and from other whitefly species. The African cluster was divided into two clades by parsimony and neighbor-joining methods. This indicates the existence of at least two genotypic clusters of B. afer, which are diverged by 0.8 to 3.2% nucleotide (nt) identities. Analysis of molecular variance indicated that these differences were the result of within population variation but were insufficient to identify discrete populations. Among the whitefly species used in the analysis, B. afer was equally dissimilar to Bemisia tabaci and Bemisia tuberculata (21.3–26.2% nt identities). As is the case for B. tabaci, these data show that mtCOI sequences are informative also for identifying B. afer variants, which lack distinguishing morphological features.

Index Entries

Aleyrodidae cytochromoe oxidase I gene genetic diversity genotypes molecular markers 

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References

  1. 1.
    Anderson, P. K., Martin, J. H., Hernandez, P., and Lagnaout, A. (2001) Bemisia afer sens. lat. (Homoptera: Aleyrodidae) outbreak in the Americas. Fla. Entomol. 84, 316–317.CrossRefGoogle Scholar
  2. 2.
    Malumphy, C. (2003) The status of Bemisia afer (Priesner and Hosny) in Britain (Homoptera: Aleyrodidae). Entomol. Gazette 54, 191–196.Google Scholar
  3. 3.
    Mound, L. A. and Halsey, S. H. (1978) Whitefly of the World. John Wiley and Sons, Chichester.Google Scholar
  4. 4.
    Martin, J. H. (1987) An identification to common whitefly pest species of the world (Homoptera, Aleyrodidae). Trop. Pest Manage. 33, 298–322.CrossRefGoogle Scholar
  5. 5.
    Munthali, D. C. (1992) Effect of cassava variety on the biology of Bemisia afer (Priesner and Hosny) (Hemiptera: Aleyrodidae). Insect Sci. Applic. 13, 459–465.Google Scholar
  6. 6.
    Maruthi, M. N., Navaneethan, S., Colvin, J., and Hillocks, R. J. (2004) Bionomics, morphometrics and molecular characterisation of a cassava Bemisic afer (Priesner and Hosny) population. Int. J. Trop. Insect. Sci. 24, 323–329.CrossRefGoogle Scholar
  7. 7.
    Mound, L. A. (1963) Host correlated variation in Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae). Proc. R. Entomol. Soc. London (A) 38, 171–180.Google Scholar
  8. 8.
    Mohanty, A. K. and Basu, A. N. (1986) Effect of host plant and seasonal factors on intraspecific variations on pupal morphology of the whitefly vector, Bemisia tabaci (Genn.) (Homoptera: Aleyrodidae). J. Entomol. Res. 10, 19–26.Google Scholar
  9. 9.
    Russell, L. M. (1957) Synonyms of Bemisia tabaci (Gennadius) (Homoptera, Aleyrodidae). Bull. Brok. Entomol. Soc. 52, 122–123.Google Scholar
  10. 10.
    Priesner, H. and Hosny, M. (1934) Contribution to a knowledge of the whiteflies (Aleurodidae) of Egypt (III). Bull. Min. Agric. Egypt. Technical and Scientific Service 145, 1–11.Google Scholar
  11. 11.
    Habib, A. and Fang, F. A. (1970) Studies on nine common aleurodids of Egypt. Bull. Soc. Entomol. Egypt 54, 1–41.Google Scholar
  12. 12.
    Corbett, G. H. (1936) New Aleurodidae (Hem.). Proc. R. Entomol. Soc. London (B) 5, 18–22.Google Scholar
  13. 13.
    Bink-Moenen, R. M. (1983) Revision of the African Whiteflies (Alcyrodidae) Mainly Based on a Collection From Tchad. Monografieu van de Nederlanse Entomologisch Vereniging No. 10, 211 pp. Amsterdam.Google Scholar
  14. 14.
    Mound, L. A. (1965) An introduction to the Aleyrodidae of Western Africa (Homoptera). Bull. Br. Mus. (Natural History) 17, 115–160.Google Scholar
  15. 15.
    Frohlich, D. R., Torres-Jerez, I., Bedford, I. D., Markham, P. G. and Brown, J. K. (1999) A phylogeographical analysis of Bemisia tabaci species complex based on mitochondrial DNA markers. Mol. Ecol. 8, 1683–1691.PubMedCrossRefGoogle Scholar
  16. 16.
    Brown, J. K. (2001) Molecular markers for the identification and global tracking of whitefly vectorbegomovirus complexes. Virus Res. 71, 233–260.CrossRefGoogle Scholar
  17. 17.
    Berry, S. D., Fondong, V. N., Rey, C., Rogan, D., Fauquet, C. M. and Brown, J. K. (2004) Molecular evidence for five distinct Bemisia tabaci (Homoptera: Aleyrodidae) geographic haplotypes associated with cassava plants in sub-Saharan Africa. Ann. Entomol. Soc. Am. 97, 852–859.CrossRefGoogle Scholar
  18. 18.
    Maruthi, M. N., Colvin, J., Thwaites, R. M., Banks, G. K., Gibson, G. and Seal, S. E. (2004) Reproductive incompatibility and cytochrome oxidase I gene sequence variability amongst host-adapted and geographically separate Bemisia tabaci populations (Hemiptera: Aleyrodidae). Syst. Entomol. 29, 560–568.CrossRefGoogle Scholar
  19. 19.
    Sseruwagi, P., Maruthi M. N., Colvin, J., Rey, M. E. C., Brown, J. K. and Legg, J. P. (2006) Colonization of non-cassva plant species by cassava whiteflies (Bemisia tabaci) in Uganda. Entomol. Expt. Appl. 119, 145–153.CrossRefGoogle Scholar
  20. 20.
    De Barro, P. J., Driver, F., Trueman, J. W. H., and Curran, J. (2000) Phylogenetic relationship of world populations of Bemisia tabaci (Gennadius) using ribosomal ITS1. Mol. Phylogenet. Evol. 16, 29–36.PubMedCrossRefGoogle Scholar
  21. 21.
    De Barro, P. J., Trueman, J. W. H. and Frohlich, D. R. (2005) Bemisia argentifolii is a race of B. tabaci (Hemiptera: Aleyrodidae): the molecular genetic differentiation of B. tabaci populations around the world. Bull. Entomol. Res. 95, 193–203.PubMedCrossRefGoogle Scholar
  22. 22.
    Abdullahi, I., Winter, S., Atiri, G. I., and Thottappilly, G. (2003) Molecular characterisation of whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae) populations infesting cassava. Bull. Entomol. Res. 93, 97–106.PubMedCrossRefGoogle Scholar
  23. 23.
    Maruthi, M. N., Colvin, J. and Seal, S. (2001) Mating compatibility, life-history traits and RAPD-PCR variation in Bemisia tabaci associated with the cassava mosaic disease pandemic in East Africa. Entomol. Expt. Appl. 99, 13–23.CrossRefGoogle Scholar
  24. 24.
    De Barro, P. J. and Driver, F. (1997) Use of RAPD PCR to distinguish the B biotype from other biotypes of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). Aust. J. Entomol. 36, 149–152.CrossRefGoogle Scholar
  25. 25.
    Zhang, D.-X. and Hewitt, G. M. (1996) Assessment of the universality and utility of a set of conserved mitochondrial COI primers in insects. Insect Mol. Biol. 6, 143–150.CrossRefGoogle Scholar
  26. 26.
    Higgins, D. G., Thompson, J. D. and Gibson, T. J. (1996) Using CLUSTAL for multiple sequence alignments. Method Enzymol 266, 383–402.CrossRefGoogle Scholar
  27. 27.
    Swofford, D. L. (2002) PAUP4 Ob10 Phylogenetic Analysis Using Parsimony (and other methods), Version 4.0 Beta. Sinauer Associates, Inc. Sunderland, MA.Google Scholar
  28. 28.
    Excoffier, L., Smouse, P., and Quattro, J. (1992) Analysis of molecular variance inferred from metric distances among DNA haplotypes: application to human mitochondrial DNA restriction data. Genetics 131, 479–491.PubMedGoogle Scholar
  29. 29.
    Schneider, S., Roessli, D., and Excoffier, L. (2000) Arlequin ver. 2,000: A Software for Population Genetics Data Analysis, Genetics and Biometry Laboratory, University of Geneva, Switzerland.Google Scholar
  30. 30.
    Legg, J. P., French, R., Rogan, D., Okao-Okuja, G., and Brown, J. K. (2002) A distinct Bemisia tabaci (Gennadius) (Hemiptera: Sternorrhyncha: Alcyrodidae) genotype cluster is associated with the epidemic of severe cassava mosaic virus disease in Uganda. Mol. Ecol. 11, 1219–1229.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc 2006

Authors and Affiliations

  • M. N. Maruthi
    • 1
  • A. R. Rekha
    • 1
  • P. Sseruwagi
    • 1
  • R. J. Hillocks
    • 1
  1. 1.Natural Resources InstituteUniversity of GreenwichChatham MaritimeUK

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