Skip to main content Accessibility help

Testing mate recognition through reciprocal crosses of two native populations of the whitefly Bemisia tabaci (Gennadius) in Australia

  • W. Wongnikong (a1), S. L. van Brunschot (a1) (a2), J. P. Hereward (a1), P. J. De Barro (a3) and G. H. Walter (a1)...


Bemisia tabaci (Gennadius) represents a relatively large cryptic species complex. Australia has at least two native populations of B. tabaci sensu lato and these were first found on different host plants in different parts of Australia. The species status of these populations has not been resolved, although their mitochondrial sequences differ by 3.82–4.20%. We addressed the question of whether these AUSI and AUSII B. tabaci populations are distinct species. We used reciprocal cross-mating tests to establish whether the insects from these different populations recognize one another as potential mating partners. The results show that the two native Australian populations of B. tabaci have a mating sequence with four phases, each of which is described. Not all pairs in the control crosses mated and the frequency of mating differed across them. Some pairs in the AUSI-M × AUSII-F did mate (15%) and did produce female progeny, but the frequency was extremely low relative to controls. Microsatellite genotyping of the female progeny produced in the crosses showed these matings were successful. None of the AUSII-M × AUSI-F crosses mated although some of the males did search for females. These results demonstrate the critical role of the mate recognition process and the need to assess this directly in cross-mating tests if the species status of different populations is to be tested realistically. In short, AUSI and AUSII B. tabaci populations are distinct species because the individual males and females do not recognize individuals of the alternative population as potential mating partners.


Corresponding author

Author for correspondence: W. Wongnikong, E-mail:


Hide All
Ahman, I and Ekbom, BS (1981) Sexual behaviour of the greenhouse whitefly (Trialeurodes vaporariorum): orientation and courtship. Entomologia Experimentalis et Applicata 29, 330338.
Bellows, TS, Perring, TM, Gill, RJ and Headrick, DH (1994) Description of a species of Bemisia (Homoptera: Aleyrodidae). Annals of the Entomological Society of America 87, 195206.
Brownstein, MJ, Carpten, JD and Smith, JR (1996) Modulation of non-templated nucleotide addition by Taq DNA polymerase: primer modifications that facilitate genotyping. BioTechniques 20, 10041010.
Byrne, DN and Bellows, TS Jr (1991) Whitefly biology. Annual Review of Entomology 36, 431457.
Chapuis, M-P and Estoup, A (2007) Microsatellite null alleles and estimation of population differentiation. Molecular Biology and Evolution 24, 621631.
Darriba, D, Taboada, GL, Doallo, R and Posada, D (2012) Jmodeltest 2: more models, new heuristics and parallel computing. Nature Methods 9, 772772.
De Barro, PJ (2012) The Bemisia tabaci species complex: questions to guide future research. Journal of Integrative Agriculture 11, 187196.
De Barro, PJ and Hart, PJ (2000) Mating interactions between two biotypes of the whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae) in Australia. Bulletin of Entomological Research 90, 103112.
De Barro, PJ, Scott, KD, Graham, GC, Lange, CL and Schutze, MK (2003) Isolation and characterization of microsatellite loci in Bemisia tabaci. Molecular Ecology Notes 3, 4043.
De Barro, PJ, Liu, S-S, Boykin, LM and Dinsdale, AB (2011) Bemisia tabaci: a statement of species status. Annual Review of Entomology 56, 119.
Dempster, AP, Laird, NM and Rubin, DB (1977) Maximum likelihood from incomplete data via the EM algorithm. Journal of the Royal Statistical Society Series B (Methodological) 39, 138.
Dinsdale, A, Cook, L, Riginos, C, Buckley, YM and De Barro, P (2010) Refined global analysis of Bemisia tabaci (Hemiptera: Sternorrhyncha: Aleyrodoidea: Aleyrodidae) mitochondrial cytochrome oxidase 1 to identify species level genetic boundaries. Annals of the Entomological Society of America 103, 196208.
Elbaz, M, Lahav, N and Morin, S (2010) Evidence for pre-zygotic reproductive barrier between the B and Q biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae). Bulletin of Entomological Research 100, 581590.
Falush, D, Stephens, M and Pritchard, JK (2003) Inference of population structure using multilocus genotype data: linked loci and correlated allele frequencies. Genetics 164, 15671587.
Falush, D, Stephens, M and Pritchard, JK (2007) Inference of population structure using multilocus genotype data: dominant markers and null alleles. Molecular Ecology Notes 7, 574578.
Fernando, LCP and Walter, GH (1997) Species status of two host-associated populations of Aphytis lingnanensis (Hymenoptera: Aphelinidae) in citrus. Bulletin of Entomological Research 87, 137144.
Firdaus, S, Vosman, B, Hidayati, N, Jaya Supena, ED, Visser, RGF and van Heusden, AW (2013) The Bemisia tabaci species complex: additions from different parts of the world. Insect Science 20, 723733.
Guindon, S and Gascuel, O (2003) A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Systematic Biology 52, 696704.
Hadjistylli, M, Schwartz, SA, Brown, JK and Roderick, GK (2014) Isolation and characterization of nine microsatellite loci from Bemisia tabaci (Hemiptera: Aleyrodidae) biotype B. Journal of Insect Science 14, 13.
Hsieh, C-H, Ko, C-C, Chung, C-H and Wang, H-Y (2014) Multi locus approach to clarify species status and the divergence history of the Bemisia tabaci (Hemiptera: Aleyrodidae) species complex. Molecular Phylogenetics and Evolution 76, 172180.
Hu, J, De Barro, P, Zhao, H, Wang, J, Nardi, F and Liu, S-S (2011) An extensive field survey combined with a phylogenetic analysis reveals rapid and widespread invasion of two alien whiteflies in China. PLoS ONE 6, e16061.
Hu, J, Chen, Y-D, Jiang, Z-L, Nardi, F, Yang, T-Y, Jin, J and Zhang, Z-K (2015) Global haplotype analysis of the whitefly Bemisia tabaci cryptic species Asia I in Asia. Mitochondrial DNA 26, 232241.
Hubisz, MJ, Falush, D, Stephens, M and Pritchard, JK (2009) Inferring weak population structure with the assistance of sample group information. Molecular Ecology Resources 9, 13221332.
Jombart, T (2008) Adegenet: a R package for the multivariate analysis of genetic markers. Bioinformatics (Oxford, England) 24, 14031405.
Jombart, T and Ahmed, I (2011) Adegenet 1.3-1: new tools for the analysis of genome-wide SNP data. Bioinformatics (Oxford, England) 27, 30703071.
Kanakala, S and Ghanim, M (2019) Global genetic diversity and geographical distribution of Bemisia tabaci and its bacterial endosymbionts. PLoS ONE 14, e0213946.
Kanmiya, K (2006) Mating behaviour and vibratory signals in whiteflies (Hemiptera: Aleyrodidae). In Drosopoulos, S and Claridge, MF (eds), Insect Sounds and Communication: Physiology, Behaviour, Ecology, and Evolution. Boca Raton: Taylor and Francis, pp. 365379.
Kopelman, NM, Mayzel, J, Jakobsson, M, Rosenberg, NA and Mayrose, I (2015) Clumpak: a program for identifying clustering modes and packaging population structure inferences across K. Molecular Ecology Resources 15, 11791191.
Las, A (1980) Male courtship persistence in the greenhouse whitefly, Trialeurodes vaporariorum Westwood (Homoptera: Aleyrodidae). Behaviour 72, 107126.
Lee, W, Park, J, Lee, G-S, Lee, S and Akimoto, S (2013) Taxonomic status of the Bemisia tabaci complex (Hemiptera: Aleyrodidae) and reassessment of the number of its constituent species. PLoS ONE 8, e63817.
Li, T-Y, Vinson, SB and Gerling, D (1989) Courtship and mating behavior of Bemisia tabaci (Homoptera: Aleyrodidae). Environmental Entomology 18, 800806.
Li, X-X, Li, S-J, Xue, X, Ahmed, MZ, Ren, S-X, Cuthbertson, AGS and Qiu, B-L (2012) The effects of space dimension and temperature on the cross mating of three cryptic species of the Bemisia tabaci complex in China. Journal of Integrative Agriculture 11, 197205.
Liu, S-S, Colvin, J and De Barro, PJ (2012) Species concepts as applied to the whitefly Bemisia tabaci systematics: how many species are there? Journal of Integrative Agriculture 11, 176186.
Luan, J-B, Ruan, Y-M, Zhang, L and Liu, S-S (2008) Pre-copulation intervals, copulation frequencies, and initial progeny sex ratios in two biotypes of whitefly, Bemisia tabaci. Entomologia Experimentalis et Applicata 129, 316324.
Luo, R, Liu, B, Xie, Y, Li, Z, Huang, W, Yuan, J, He, G, Chen, Y, Pan, Q, Liu, Y, Tang, J, Wu, G, Zhang, H, Shi, Y, Liu, Y, Yu, C, Wang, B, Lu, Y, Han, C, Cheung, DW, Yiu, S-M, Peng, S, Xiaoqian, Z, Liu, G, Liao, X, Li, Y, Yang, H, Wang, J, Lam, T-W and Wang, J (2012) SOAPdenovo2: an empirically improved memory-efficient short-read de novo assembler. GigaScience 1, 16.
Marková, S, Dufresne, F, Manca, M and Kotlík, P (2013) Mitochondrial capture misleads about ecological speciation in the Daphnia pulex complex. PLoS ONE 8, e69497.
Maruthi, MN, 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. Entomologia Experimentalis et Applicata 99, 1323.
Maruthi, MN, Colvin, J, Thwaites, RM, Banks, GK, Gibson, G and Seal, SE (2004) Reproductive incompatibility and cytochrome oxidase I gene sequence variability amongst host-adapted and geographically separate Bemisia tabaci populations (Hemiptera: Aleyrodidae). Systematic Entomology 29, 560568.
Meglécz, E, Costedoat, C, Dubut, V, Gilles, A, Malausa, T, Pech, N and Martin, JF (2010) QDD: a user-friendly program to select microsatellite markers and design primers from large sequencing projects. Bioinformatics (Oxford, England) 26, 403404.
Meglécz, E, Pech, N, Gilles, A, Dubut, V, Hingamp, P, Trilles, A, Grenier, R and Martin, J-F (2014) QDD version 3.1: a user-friendly computer program for microsatellite selection and primer design revisited: experimental validation of variables determining genotyping success rate. Molecular Ecology Resources 14, 13021313.
Mound, LA and Halsey, SH (1978) Whitefly of the World: A Systematic Catalogue of the Aleyrodidae (Homoptera) with Host Plant and Natural Enemy Data. London: British Museum (Natural History).
Muñiz, M and Nombela, G (2001) Bemisia tabaci: A New Clip-Cage for Biological Studies. Norwich: European Whitefly Studies Network (EWSN).
Omondi, BA, Sseruwagi, P, Obeng-Ofori, D, Danquah, EY and Kyerematen, RA (2005) Mating interactions between okra and cassava biotypes of Bemisia tabaci (Homoptera: Aleyrodidae) on eggplant. International Journal of Tropical Insect Science 25, 159167.
Paterson, HEH (1985) The recognition concept of species. In Vrba, ES (ed.), Species and Speciation. Pretoria: Transvaal Museum, pp. 2129.
Peakall, R and Smouse, PE (2006) Genalex 6: genetic analysis in Excel. Population genetic software for teaching and research. Molecular Ecology Notes 6, 288295.
Peakall, R and Smouse, PE (2012) Genalex 6.5: genetic analysis in Excel. Population genetic software for teaching and research-an update. Bioinformatics (Oxford, England) 28, 25372539.
Perea, S, Vukić, J, Šanda, R and Doadrio, I (2016) Ancient mitochondrial capture as factor promoting mitonuclear discordance in freshwater fishes: a case study in the genus Squalius (actinopterygii, Cyprinidae) in Greece. PLoS ONE 11, e0166292.
Perring, TM and Symmes, EJ (2006) Courtship behavior of Bemisia argentifolii (Hemiptera: Aleyrodidae) and whitefly mate recognition. Annals of the Entomological Society of America 99, 598606.
Perring, TM, Cooper, AD, Rodriquez, RJ, Farrar, CA and Bellows, TS (1993) Identification of a whitefly species by genomic and behavioral studies. Science 259, 7477.
Pritchard, JK, Stephens, M and Donnelly, P (2000) Inference of population structure using multilocus genotype data. Genetics 155, 945959.
R Core Team (2013) R: A Language and Environment for Statistical Computing. Vienna: R Foundation for Statistical Computing.
Qin, L, Pan, L-L and Liu, S-S (2016) Further insight into reproductive incompatibility between putative cryptic species of the Bemisia tabaci whitefly complex. Insect Science 23, 215224.
Rafter, MA and Walter, GH (2013) Mate recognition in the South African citrus thrips Scirtothrips aurantii (Faure) and cross-mating tests with populations from Australia and South Africa. Journal of Insect Behavior 26, 780795.
Rousset, F (2008) Genepop'007: a complete re-implementation of the genepop software for Windows and Linux. Molecular Ecology Resources 8, 103106.
Ruan, Y-M, Luan, J-B, Zang, L-S and Liu, S-S (2007) Observing and recording copulation events of whiteflies on plants using a video camera. Entomologia Experimentalis et Applicata 124, 229233.
Schuelke, M (2000) An economic method for the fluorescent labeling of PCR fragments. Nature Biotechnology 18, 233234.
Simon, C, Frati, F, Beckenbach, A, Crespi, B, Liu, H and Flook, P (1994) Evolution, weighting, and phylogenetic utility of mitochondrial gene sequences and a compilation of conserved polymerase chain reaction primers. Annals of the Entomological Society of America 87, 651701.
Sun, DB, Xu, J, Luan, JB and Liu, SS (2011) Reproductive incompatibility between the B and Q biotypes of the whitefly Bemisia tabaci in China: genetic and behavioural evidence. Bulletin of Entomological Research 101, 211220.
Toon, A, Daglish, GJ, Ridley, AW, Emery, RN, Holloway, JC and Walter, GH (2016) Random mating between two widely divergent mitochondrial lineages of Cryptolestes ferrugineus (Coleoptera: Laemophloeidae): a test of species limits in a phosphine-resistant stored product pest. Journal of Economic Entomology 109, 22212228.
Untergasser, A, Cutcutache, I, Koressaar, T, Ye, J, Faircloth, BC, Remm, M and Rozen, SG (2012) Primer3 – new capabilities and interfaces. Nucleic Acids Research 40, e115.
Vyskočilová, S, Tay, WT, van Brunschot, S, Seal, S and Colvin, J (2018) An integrative approach to discovering cryptic species within the Bemisia tabaci whitefly species complex. Scientific Reports 8, 10886.
Walter, GH (2003) Insect Pest Management and Ecological Research. Cambridge: Cambridge University Press.
White, JA, Kelly, SE, Perlman, SJ and Hunter, MS (2009) Cytoplasmic incompatibility in the parasitic wasp Encarsia inaron: disentangling the roles of Cardinium and Wolbachia symbionts. Heredity 102, 483489.
Xu, J, De Barro, PJ and Liu, SS (2010) Reproductive incompatibility among genetic groups of Bemisia tabaci supports the proposition that the whitefly is a cryptic species complex. Bulletin of Entomological Research 100, 359366.
Zang, L-S and Liu, S-S (2007) A comparative study on mating behaviour between the B biotype and a non-B biotype of Bemisia tabaci (Hemiptera: Aleyrodidae) from Zhejiang, China. Journal of Insect Behavior 20, 157171.


Type Description Title
Supplementary materials

Wongnikong et al. supplementary material
Figure S1

 Word (390 KB)
390 KB

Testing mate recognition through reciprocal crosses of two native populations of the whitefly Bemisia tabaci (Gennadius) in Australia

  • W. Wongnikong (a1), S. L. van Brunschot (a1) (a2), J. P. Hereward (a1), P. J. De Barro (a3) and G. H. Walter (a1)...


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.