Skip to main content Accessibility help

Development of silverleaf assay, protein and nucleic acid-based diagnostic techniques for the quick and reliable detection and monitoring of biotype B of the whitefly, Bemisia tabaci (Gennadius)

  • K.S. Shankarappa (a1), K.T. Rangaswamy (a1), D.S. Aswatha Narayana (a1), A.R. Rekha (a1), N. Raghavendra (a1), C.N. Lakshminarayana Reddy (a1), T.C.B. Chancellor (a2) and M.N. Maruthi (a2)...


The aim of this study was to develop and optimize silverleaf bioassay, esterase analysis and PCR-based techniques to distinguish quickly and reliably biotype B of the whitefly, Bemisia tabaci (Gennadius), from Indian indigenous biotypes. Zucchini and squash readily develop silverleaf symptoms upon feeding by the B biotype, but they are not readily available in Indian markets. A local pumpkin variety ‘Big’ was, therefore, used in silverleaf assay, which developed symptoms similar to those on zucchini and squash and can be used reliably to detect B biotype. Analysis of non-specific esterases of B and the indigenous biotypes indicated both quantitative and qualitative differences in esterase patterns. Two high molecular weight bands were unique to B biotype and they occurred in abundance. These esterases were used to develop quick and field-based novel detection methods for differentiating B from the indigenous biotypes. Development of these simple and cost-effective protocols has wider application as they can be potentially used to identify other agricultural pests. Mitochondrial cytochrome oxidase I gene sequences and randomly amplified polymorphic DNA (RAPD) polymorphisms, generated using the primer OpB11, were also found useful for detecting B. tabaci biotypes. A B biotype-specific RAPD band of 800 bp was sequenced, which was used to a develop sequence characterized amplified region (SCAR) marker. The SCAR marker involved the development of B biotype-specific primers that amplified 550 bp PCR products only from B biotype genomic DNA. Silverleaf assay, esterases, RAPDs or a SCAR marker were used in combination to analyse whitefly samples collected from selected locations in India, and it was found that any of these techniques can be used singly or in combination to detect B biotype reliably. The B biotype was found in southern parts of India but not in the north in 2004–06.


Corresponding author

*Author for correspondence Fax: +44 (0)1634 883379 E-mail:


Hide All
Banks, G.K., Colvin, J., Chowda Reddy, R.V., Maruthi, M.N., Muniyappa, V., Venkatesh, H.M., Kiran Kumar, M., Padmaja, A.S., Beitia, F.J. & Seal, S.E. (2001) First report of the Bemisia tabaci B biotype in India and an associated tomato leaf curl virus disease epidemic. Plant Disease 85, 231.
Bharathan, N., Narayanan, K.R. & McMillan, R.T. (1992) Characteristics of sweet potato whitefly mediated silverleaf syndrome and associated double stranded RNA in squash. Phytopathology 82, 136141.
Bird, J. (1957) A whitefly transmitted mosaic of Jatropha gossypifolia. Technical Paper, Agricultural Experiment Station, Puerto Rico 22, 135.
Brown, J.K. (2001) Molecular markers for the identification and global tracking of whitefly vector-begomovirus complexes. Virus Research 71, 233260.
Brown, J.K., Costa, H.S. & Laemmlen, F. (1992) First incidence of whitefly-associated squash silverleaf (SSL) of Cucurbita, and of white streaking (WSt) disorder of cole crops in Arizona and California. Plant Disease 76, 426.
Brown, J.K., Frohlich, D.R. & Rosell, R.C. (1995) The sweet potato or silverleaf whiteflies: Biotypes of Bemisia tabaci or a species complex? Annual Review of Entomology 40, 511534.
Brown, J.K., Pering, T.M., Cooper, A.D., Bedford, I.D. & Markham, P.G. (2000) Genetic analysis of Bemisia (Hemiptera: Aleyrodidae) populations by isoelectric focusing electrophoresis. Biochemical Genetics 38, 1325.
Burban, C., Fishpool, L.D.C., Fauquet, C.M., Fargette, D. & Thovenel, J.C. (1992) Host associated biotypes within the West African populations of the whitefly Bemisia tabaci (Genn.). Journal of Applied Entomology 113, 416423.
Byrne, F.J. & Devonshire, A.L. (1991) In vivo inhibition of esterase and acetylcholinesterase activities by profenofos treatments in the tobacco whitefly Bemisia tabaci (Genn): implications for routine biochemical monitoring of these enzymes. Pesticide Biochemistry and Physiology 40, 198204.
Byrne, F.J., Bedford, I.D., Devonshire, A.L. & Markham, P.G. (1995) Esterase variation and squash silverleaf induction in B-type Bemisia tabaci (Homoptera: Aleyrodidae). Bulletin of Entomological Research 85, 175179.
Byrne, F.J., Gorman, K.J., Cahill, M., Denholm, I. & Devonshire, A.L. (2000) The role of B-type esterases in conferring insecticide resistance in the tobacco whitefly, Bemisia tabaci (Genn). Pest Management Science 56, 867874.
Calvert, L.A., Cuervvo, M., Arroyave, J.A., Constantino, L.M., Belloti, A. & Frohlich, D. (2001) Morphological and mitochondrial DNA markers analyses of whiteflies (Homoptera: Aleyrodidae) colonizing cassava and beans in Colombia. Annals of the Entomological Society of America 94, 512517.
Clark, M.A., Baumann, L., Munsom, M.A. & Campbell, B.C. (1992) The eubacterial endosymbionts of whiteflies (Homoptera: Aleyrodidae) constitute a lineage distinct from endosymbionts of aphids and mealy bugs. Current Microbiology 25, 119123.
Costa, H.S. & Brown, J.K. (1991) Variation in biological characteristics and esterase patterns among populations of Bemisia tabaci (Genn.) and the association of one population with silverleaf symptom induction. Entomologia Experimentalis et Applicata 61, 211219.
Costa, H.S. & Russell, L.M. (1975) Failure of Bemisia tabaci to breed on cassava plants in Brazil (Homoptera: Aleyrodidae). Ciencia e Cultura 27, 388390.
Costa, H.S., Brown, J.K., Sivasubramaniam, S. & Bird, J. (1993a) Regional distribution, insecticide resistance and reciprocal crosses between the A and B biotypes of Bemisia tabaci. Insect Science Application 14, 255266.
Costa, H.S., Ullman, D.E., Johnson, M.W. & Tabashnik, B.E. (1993b) Squash silverleaf symptoms induced by immature, but not adult Bemisia tabaci. Phytopathology 83, 763766.
De Barro, P.J. & Driver, F. (1997) Use of RAPD-PCR to distinguish the B biotype from other biotypes of Bemisia tabaci (Genn.) (Hemiptera: Aleyrodidae). Australian Journal of Entomology 36, 149152.
De Barro, P.J., Liebregts, W. & Carver, M. (1998) The distribution and identity of biotypes of Bemisia tabaci (Gennadius) (Hemiptera:Aleyrodidae) in member countries of the Secretariat of the Pacific Community. Australian Journal of Entomology 37, 214218.
De Barro, P.J., Driver, F., Trueman, J.W.H. & Curran, J. (2000) Phylogenetic relationships of world populations of Bemisia tabaci (Genn.) using ribosomal ITS1. Molecular Phylogenetics and Evolution 16, 2936.
De Barro, P.J., Trueman, J.W.H. & 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. Bulletin of Entomological Research 95, 111.
De Barro, P.J., Bourne, A., Khan, S.A. & Brancatini, V.A.L. (2006) Host plant and biotype density interactions – their role in the establishment of the invasive B biotype of Bemisia tabaci. Biological Invasions 8, 287294.
Duffus, J.K. (1987) Whitefly transmission of plant viruses. Current Topics in Vector Research 4, 7391.
Frohlich, D.R., Torres-Jerez, I., Bedford, I.D., Markham, P.G. & Brown, J.K. (1999) A phylogeographical analysis of Bemisia tabaci species complex based on mitochondrial DNA markers. Molecular Ecology 8, 16831691.
Guirao, P., Beitia, F. & Cenis, J.L. (1997) Biotype determination of Spanish population of Bemisia tabaci (Homoptera: Aleyrodidae). Bulletin of Entomological Research 87, 587593.
Jones, D.R. (2003) Plant viruses transmitted by whiteflies. European Journal of Plant Pathology 109, 195219.
Khasdan, V., Levin, I., Rosner, A., Morin, S., Kontsedalov, S., Maslenin, L. & Horowitz, A.R. (2005) DNA markers for identifying biotypes B and Q of Bemisia tabaci (Hemiptera, Aleyrodidae) and studying population dynamics. Bulletin of Entomological Research 95, 605613.
Maruthi, M.N., Colvin, J. & Seal, S.E. (2001) Mating compatibility, life history traits and RAPD-PCR variation in Bemisia tabaci associated with the cassava mosaic disease pandemic in East Africa. Entomologia Experimentalis et Applicata 29, 1323.
Maruthi, M.N, Colvin, J., Gibson, G., Thwaites, R.M., Banks, G.K. & Seal, S.E. (2004) Reproductive incompatibility and cytochrome oxidase I gene-sequence variability amongst host-adapted and geographically separate Bemisia tabaci populations. Systematic Entomology 29, 560568.
Perring, T.M. (2001) The Bemisia tabaci species complex. Crop Protection 20, 725737.
Perring, T.M., Cooper, A.D., Rodriguez, R.J., Farrar, C.A. & Bellows, T.S. (1993) Identification of a whitefly species by genomic and behavioural studies. Science 259, 7477.
Polston, J.E. & Anderson, P.K. (1997) The emergence of whitefly transmitted geminiviruses in tomato in the western hemisphere. Plant Disease 81, 13581369.
Prabhaker, N., Coudriet, D.L. & Meyerdick, D.E. (1987) Determination of three whitefly species by electrophoresis of non-specific esterases. Journal of Applied Entomology 103, 447451.
Rekha, A.R., Maruthi, M.N., Muniyappa, V. & Colvin, J. (2005) Occurrence of three genotypic clusters of Bemisia tabaci and the rapid spread of the B biotype in south India. Entomologia Experimentalis et Applicata 117, 221233.
Schuster, D.J., Kring, J.B. & Price, J.F. (1991) Association of sweet potato whitefly with leaf disorder of squash. HortScience 26, 155156.
Secker, A.E., Bedford, I.A., Markham, P.G. & William, M.E.C. (1998) Squash, a reliable field indicator for the presence of B biotype of tobacco whitefly Bemisia tabaci. pp. 837842 in Brighton Crop Protection Conference: Pests and Diseases. British Crop Protection Council, 1998, Farnham, UK.
Simon, B., Cenis, J.L., Beitia, F., Khalid, S., Moreno, I.M., Fraile, A. & Arenal, F.G. (2003) Genetic structure of field populations of begomoviruses and of their vector Bemisia tabaci in Pakistan. Phytopathology 93, 14221429.
Sseruwagi, P., Legg, J.P., Maruthi, M.N., Colvin, J., Rey, M.E.C. & Brown, J.K. (2005) Genetic diversity of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) populations and presence of the B biotype and a non-B biotype that can induce silverleaf symptoms in squash in Uganda. Annals of Applied Biology 147, 253265.
Swofford, D. (2002) Phylogenetic Analysis Using Parsimony (PAUP), version 4.0 10 beta (and other methods). 246 pp. Champaign, IL, USA, University of Illinois.
Varma, A. & Malathi, V.G. (2003) Emerging geminivirus problems: A serious threat to crop production. Annals of Applied Biology 142, 145164.
Yokomi, R.K., Hoelmer, K.A. & Osborne, L.S. (1990) Relationship between the sweet potato whitefly and the squash silverleaf disorder. Phytopathology 80, 895900.
Recommend this journal

Email your librarian or administrator to recommend adding this journal to your organisation's collection.

Bulletin of Entomological Research
  • ISSN: 0007-4853
  • EISSN: 1475-2670
  • URL: /core/journals/bulletin-of-entomological-research
Please enter your name
Please enter a valid email address
Who would you like to send this to? *



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