Printer Friendly

Prevalence of a new genetic group, MEAM-K, of the whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) in Karnataka, India, as evident from mtCOI sequences.

Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a pest of global importance that causes significant crop loss as a direct pest and as a vector of 120 geminiviruses, especially begomoviruses (Jones 2003). It is highly polyphagous, feeding on an estimated 900 hosts of agricultural, fiber, vegetable, and ornamental crops (Cahill et al. 1996; Jones 2003). It stands as one of the world's 100 invasive species (International Union for Conservation of Nature and Natural Resources [IUCN] list: Emerging new genetic groups of B. tabaci increase the risk of transmission of geminiviruses to many crops worldwide and of development od high levels of resistance of this insect to various insecticides, especially neonicotinoids (Horowitz et al. 2003, 2004; Rauch &Nauen 2003). Bemisia tabaci is a species complex that contains morphologically indistinguishable biotypes or cryptic species or genetic groups (Dinsdale et al. 2010; De Barro et al. 2011; Liu et al. 2012). However, biotypes--as designated based on esterase banding pattern (Costa &Brown 1991)--and genotypes have recently been elevated to putative species of B. tabaci (Dinsdale et al. 2010; De Barro et al. 2011).

The nymphs and adults actively feed on the phloem sap and excrete copious amounts of honeydew, which supports the growth of sooty mold that in turn impedes photosynthesis (Byrne &Bellows 1991). Several studies examined the host-related phenotypic variations in B. tabaci and concluded that variations are evident not only among the populations on different hosts but also among individuals occurring on the same host (Mound 1963; Palaniswami et al. 1996; Lisha et al. 2003). Morphometric analysis of 4th instar nymphs revealed phenotypic variations corresponding to variations in the leaf anatomy of the host plant (Maruthi et al. 2007). Bemisia tabaci is known to be an aggressive colonizer of crops, with varying traits at the morphological (Bellows et al. 1994; Costa et al. 1995; Rosell et al. 1997), biochemical (Costa &Brown 1991; Brown et al. 2000; Perring 2001), and molecular levels (Gawel & Bartlett 1993; De Barro et al. 2005; Boykin et al. 2007).

Different geographical and ecological variations with host plant specialization offer an ideal system for the study of sympatric speciation in B. tabaci, direct damage to crops and vector ability (Chu et al. 2007). De Barro et al. (2011) stated that asymmetrical mating interference provides a clear mechanism that could contribute to formation of global genetic structure. Genetic differentiation of different populations in the species complex was analyzed mainly based on the ribosomal internal transcribed spacer 1 (rITS-1) and mitochondrial cytochrome oxidase I (mtCOI) sequences worldwide (Simon et al. 1994; Hu et al. 2011; Zasada et al. 2014). Dinsdale et al. (2010) identified 3.5% pair-wise genetic divergence as the considered boundary for separating different species; this was further supported with either complete or partial mating isolation between a number of putative B. tabaci "species" (Xu et al. 2010; Wang et al. 2011). Considering all the past taxonomical approaches, Boykin et al. (2012) defined species in the B. tabaci species complex. In the present study, we examined the genetic variation in the mtCOI region of the genome of B. tabaci, collected on 30 different host plants from different locations in Karnataka, India, to assess the presence and prevalence of various genetic groups.

Materials and Methods

Nymphs and adults of B. tabaci were collected with a handheld aspirator from different locations in Karnataka, India, on 30 different hosts (Fig. 1 and Table 1) and preserved in 70% alcohol until further use.


The total DNA was extracted from individual adult B. tabaci specimens using DNAeasy Tissue Kit (Qiagen GmbH, Hilden, Germany) according to the manufacturer's protocol and stored at -80[degrees]C until further use. Polymerase chain reaction (PCR) was performed with mtCOI primers F-C1-J-2195 (5'-TTGATTTTTTGGTCATCCAGAAGT-3') and R-L2N-3014 (5'-TCCAATGCACTAATCTGCCATATTA-3') (Simon et al. 1994; Frohlich et al. 1999). The 25 [micro]L PCR consisted of 50 ng/[micro]L template,10 picomoles of each primer, 0.25 mM dNTP mix, 1.5 mM Mg[Cl.sub.2], 1 U Taq polymerase, and 2.5 [micro]L Taq buffer (Fermentas GmBH, St. Leon-Rot, Germany). The PCR cycling was carried out in a thermal cycler (Applied Biosystems, Veriti 96 wells, USA) with the following parameters: 94[degrees]C for 5 min as initial denaturation followed by 35 cycles of denaturation at 94[degrees]C for 30 s, annealing at 54[degrees]C for 45 s, and extension at 72[degrees]C for 50 s, and 72[degrees]C for 10 min as the final extension. The amplified products were resolved in 1.0% agarose gel, stained with ethidium bromide (10 Hg/mL), and visualized under UV light. Amplified PCR products were eluted using the gel extraction kit Nucleospin[R] Extract II (Macherey-Nagel, Duren, Germany) according to the manufacturer's protocol, and sequencing was performed in M13 forward and reverse directions.


All mtCOI sequences corresponding to different genetic groups of B. tabaci were downloaded from the National Center for Biotechnology Information (NCBI) GenBank (Suppl. Table 1, available online at Sequence alignment was performed employing MUSCLE implemented in Seaview (Thompson et al. 1994). Genetic divergence was calculated employing MEGA5 (Tamura et al. 2011). The nucleotide substitution model for the best fits and the model parameters were estimated using Akaike Information Criterion implemented in the program MODELTEST 3.7 (Posada &Crandall 1998) in conjunction with PAUP*. Maximum parsimony and maximum likelihood analyses were performed with PAUP* 4.0b10 (Swofford 1998), using the heuristic search procedure with 1,000 random additions of sequences and 10 trees held at each pseudo-rep licate, and the tree bisection reconnection branch swapping method with all characters was treated as unordered and equally weighted. The chosen model with estimated parameters was used to derive the maximum likelihood tree in RaxML with the heuristic search settings. The same software was used to generate consensus trees using the CONSENSE program (, and the tree was rooted with the outgroup Bemisia afer (Priesner & Hosny) (Hemiptera: Aleyrodidae). Phylogenetic analysis was performed using MrBayes (Huelsenbeck & Ronquist 2001). MrBayes 3.1 was run for 10 million generations by using 8 chains and sampled every 1,000 generations. All runs reached a plateau in the likelihood score, and the same was indicated by standard deviations of split frequencies (0.0023). Our 4 Markov Chain Monte-Carlo chains were converged, indicated by the potential scale reduction factor, which was close to one. The burn-in parameter was estimated employing Tracer version 1.5 (Rambaut & Drummond 2009), and the trees corresponding to the first 20% of generations were discarded. We performed 2 independent MrBayes runs to ensure the analyses were not trapped in the local optima, after which topologies and posterior probabilities (PP) from these 2 runs were compared for congruence purpose. The trees obtained were visualized using FigTree v1.3.1 (Rambaut 2009).


The mtCOI gene was successfully amplified and sequenced from 71 individual adult B. tabaci. We analyzed 131 sequences, including the 71 generated from this study. The sequence analysis revealed that out of 816 bp, 332 nucleotides were conserved, 484 were variable, and 379 were parsimony informative. Absence of stop codons indicated that no pseudogenes were amplified within the sequences, and similar base composition indicated no indels. Bemisia tabaci nucleotide frequencies were 24.46% (A), 43.10% (T/U), 19.23% (C), and 13.22% (G). Rates of different transitional substitutions are shown in bold and those of transversional substitutions are shown in italics in Table 2. The base composition of the mtCOI gene fragment was biased toward adenine (A) and thymine (T) with an overall 67.56%. The transition/transversion rate ratios were [k.sub.1] = 6.255 (purines) and [k.sub.2] = 6.190 (pyrimidines). The overall transition/transversion bias was R = 2.629. All sequences were deposited in the NCBI GenBank with the accession numbers KF790634 to KF790689 and HQ268814, HQ268813, HQ268812, HQ268811, HQ331246, HQ331245, and HQ331244.


Based on the phylogenetic analyses, the Indian B. tabaci samples were clustered into Asia-I, Asia-II-7, Asia-II-8, Middle East Asia Minor-1 (MEAM-1), and MEAM-K groups (Figs. 2a and 2b). The Asia-I genetic group was predominant representing 44 of the 71 (61.97%) samples sequenced. The Asia-I population had 17 different host plant species (Table 1), of which Solanum lycopersicum L., Solanum melongena L. (Solanales: Solanaceae), Gossypium hirsutum L., and Abelmoschus esculentus (L.) Moench (Malvales: Malvaceae) were the most frequent host plants and are widely distributed in Karnataka (Fig. 2b). Few samples collected from Karnataka clustered with AsiaII-7 and were found on 9 host plant species. The Asia-II-8 genetic group contained 5 samples, which had been collected on 5 different host plants. The most important genetic groups MEAM-1 and MEAM-2 formed 2 subclusters representing B and B2 genotypes. Here, MEAM-1 comprised 5 sequences, which had been collected on 3 host plants (mostly cabbage). However, 3 sequences from specimens collected on Phaseolus vulgaris L. (Fabales: Fabaceae) at Kolar, H-Cross, and Malur (Karnataka, India) diverged from all the MEAM-1, MEAM-2, Indian Ocean, and Mediterranean groups. Thus, they formed an independent genetic group, which we named MEAM-K group. The genetic divergence ranged from 4.0 to 24.0% with an average of 15.7% (Table 3 and Fig. 3).


"There is probably no other concept in biology that has remained as consistently controversial as the species concept" (Mayr 1982). This quote is remarkably true with B. tabaci, because this species is difficult to be diagnosed using morphological, cytological, behavioral, molecular, and biochemical methods, and its identification is complicated further by reproductive isolation (Rosell et al. 1997; Maruthi et al. 2007). Molecular studies on various insects of agricultural importance have helped to identify new species (Ball &Armstrong 2006), biotypes (Perring 2001), cryptic species (Hebert et al. 2004), and haplotypes (Toda &Murai 2007), which are difficult to identify through morphology due to phenotypic plasticity and lack of distinguishing morphological features (Russell 1957; Mound 1963; Rosell et al. 1997; Maruthi et al. 2007).

Considering the vector potential of B. tabaci, it is necessary to analyze the molecular diversity of the same species or species complex collected on various host plants. In this regard, the resistant tomato varieties Arka Ananya (against tomato leaf curl virus) and Arka Abhay and Arka Anamika (against Bhendi yellow vein mosaic virus) have become susceptible again, and this breakdown of resistance may be correlated with the existence of various species of vectors or virus, but the exact plant physiological mechanism is still unknown. In 1991, the biotype nomenclature was introduced to the B. tabaci species complex based on esterase banding patterns but was no longer used after the advent of Random Amplified Polymorphic DNA-PCR and mtCOI and ITS sequencing (Perring 2001; Simon et al. 2003; Zang et al. 2006; Boykin 2013). In the discussion on B. tabaci nomenclature, it remains unclear whether to call a new biotype a genetic group or a putative species with novel binomial nomenclature, such as Bemisia argentifolii Bellows & Perring (De Barro et al. 2011; Boykin et al. 2013).

Bemisia tabaci can attack a wide range of host plants globally, and its species complex is composed of at least 34 morphologically indistinguishable species (Boykin et al. 2012, 2013; Boykin 2014). Different cropping patterns and diverse climatic conditions may be responsible for the apparent diversity in B. tabaci, which otherwise is grouped by geographic location (Rekha et al. 2005). The current study revealed the existence of 5 genetic groups within Karnataka State, India, namely Asia-I, Asia-II-7, Asia-II-8, MEAM-1, and MEAM-K, with the latter being a new group identified in this study.

The genetic divergence ranged from 4.0 to 24.0% with an average of 15.7%. According to Dinsdale et al. (2010), the genetic divergence among 198 mtCOI sequences of B. tabaci and sequences of the outgroup species B. afer, B. atriplex (Froggatt), and B. subdecipiens Martin (Hemiptera: Aleyrodidae) ranged from zero to 34%. However, Lee et al. (2013) analyzed the genetic divergence after excluding the outgroup sequences and concluded that it ranged from zero to 24.0%, which was on par with our study.

Of the reported 5 genetic groups, the major group was Asia-I with members collected on 19 different host plants although most had been collected on eggplant; this group was found previously to be prevalent across Asia (Boykin et al. 2007; Dinsdale et al. 2010; Hu et al. 2011). The eggplant-associated Asia-I B. tabaci can transmit several begomo-viruses (Govindappa 2002; Muniyappa et al. 2003) and was shown to transmit eggplant yellow mosaic virus in Thailand (Green et al. 2003). In this study, we report another 17 host plants for Asia-I including cotton, pumpkin, ridge gourd, okra, capsicum, sunflower, potato, carrot, mustard, and tomato. However, a previous study showed that tomato is not a preferred host plant for Asia-I because it is associated with low fecundity (Chowda-Reddy et al. 2012). Host-associated variations in B. tabaci influence its rate of fecundity, which could be due to premating or post-mating selection against migrants and hybrid progeny (Liou & Price 1994; Brunner et al. 2004). However, the most to least preferred hosts, in terms of oviposition, were eggplant, cotton, pumpkin, tomato, and cassava (Venkatesh 2000). Our study also supported this finding, wherein eggplant and cotton were the most frequent host plants for Asia-I. Apart from the Jatropha genetic group, which occurs in a separate ecological niche, most of the putative species exhibit high levels of polyphagy (Burban et al. 1992; Brown et al. 1995). According to Chowda-Reddy et al. (2012), MEAM-1 has a wide host range and produces higher quantities of honeydew than other genetic groups. This is also true for genetic groups Asia-I, Asia-II-7, and Asia-II-8, of which Asia-I had the greatest number of host plants in our study.

We identified Asia-II-7 from Karnataka occurring on 9 different host plants, mostly on ornamental plants. Asia-II-7 was reported first in 1998 in India (Ramappa et al. 1998) and in China (Qiu et al. 2006). A previous study suggested that this putative species adapts readily to ornamental plants rather than vegetables (Shah et al. 2013). The 3rd genetic group identified in this study was Asia-II-8, which was recently named as B. gossypiperda (Boykin 2014) and was collected on 5 different host plants including cotton (Chowda-Reddy et al. 2012). Significantly, we here report the occurrence of a previously unreported subclade for Middle East genetic groups (MEAM-1 and 2). We named it MEAM-K because the samples were collected on P. vulgaris from Kolar, Karnataka, the native location for MEAM-1. For Kolar and nearby areas where both species have become established, our study raises questions about the relative pest status of MEAM-1and MEAM-K. We do not know to what extent these genetic groups differ with respect to important factors such as their ability in feeding and reproduction, their efficiency as vectors, or their susceptibility to insecticides. These high levels of molecular and ecological resolution will be needed to mitigate quarantine disputes arising from the detection of morphologically indistinguishable members of the B. tabaci genetic groups. Thus, our study may stand as a link to detect and identify B. tabaci genetic groups, determine areas of occurrence, identify areas of invasion, and design management strategies.

In conclusion, the current study revealed the existence of 5 genetic groups of B. tabaci in Karnataka, India, identified as Asia-I, Asia-II-7, Asia-II-8, MEAM-1, and a previously unreported genetic group, MEAMK. Thus, our work will help in rapid and accurate identification of these putative genetic groups of B. tabaci, which in turn will help in further elucidating the epidemiology and management of geminiviruses and be of value in the operation of quarantines.


This paper is part of the doctoral degree work of the senior author. We gratefully acknowledge the financial support received for the "Outreach Programme on Management of Sucking Pests in Horticultural Crops" from the Indian Council for Agricultural Research (ICAR), New Delhi, India. We are thankful to the director at the Indian Institute of Horticultural Research (IIHR) for the support and facilities.

References Cited

Ball SL, Armstrong KF.2006. DNA barcodes for insect pest identification: a test case with tussock moths (Lepidoptera: Lymantriidae). Canadian Journal of Forest Research 36: 337-350.

Bellows TS, Perring TM, Gill RJ, Headrick DH. 1994. Description of a species of Bemisia (Homoptera: Aleyrodidae). Annals of the Entomological Society of America 87: 195-206.

Boykin LM. 2014. Bemisia tabaci nomenclature: lessons learned. Pest Management Science 70: 1454-1459.

Boykin LM, Shatters Jr RG, Rosell RC, McKenzie CL, Bagnall RA, De Barro PJ, Frohlich DR. 2007. Global relationships of Bemisia tabaci (Hemiptera: Aleyrodidae) revealed using Bayesian analysis of mitochondrial COI DNA sequences. Molecular Phylogenetics and Evolution 44: 1306-1319.

Boykin LM, Armstrong KF, Kubatko L, De Barro PJ. 2012. Species delimitation and global biosecurity. Evolutionary Bioinformatic Online 8: 1-37.

Boykin LM, Bell CD, Evans G, Small I, De Barro PJ. 2013. Is agriculture driving the diversification of the Bemisia tabaci species complex (Hemiptera: Sternorrhyncha: Aleyrodidae)? Dating, diversification and biogeographic evidence revealed. BMC Evolutionary Biology 13: 228.

Brown JK, Frohlich DR, Rosell RC. 1995. The sweetpotato or silverleaf whiteflies: biotypes of Bemisia tabaci or a species complex. Annual Review of Entomology 40: 511-534.

Brown JK, Perring TM, Cooper AD, Bedford ID, Markham PG. 2000. Genetic analysis of Bemisia tabaci (Hemiptera: Aleyrodidae) populations by isoelectric focusing electrophoresis. Biochemical Genetics 38: 13-25.

Brunner PC, Chatzivassiliou EK, Katis NI, Frey JE. 2004. Host-associated genetic differentiation in Thrips tabaci (Insecta; Thysanoptera), as determined from mtDNA sequence data. Heredity 93: 364-370.

Burban C, Fishpool LDC, Fauquet C, Fargette D, Thouvenel JC. 1992. Host-associated biotypes within West African populations of the whitefly Bemisia tabaci (Genn.) (Hom., Aleyrodidae). Journal of Applied Entomology 113: 416-423.

Byrne DN, Bellows Jr TS. 1991. Whitefly biology. Annual Review of Entomology 36: 431-457.

Cahill M, Denholm I, Ross G, Gorman K, Johnston D. 1996. Relationship between bioassay data and the simulated field performance of insecticides against susceptible and resistant adult Bemisia tabaci. Bulletin of Entomological Research 86: 109-116.

Chowda-Reddy RV, Kiran Kumar M, Seal SE, Muniyappa V, Valand GB, Govindappa MR, Colvin J. 2012. Bemisia tabaci phylogenetic groups in India and the relative transmission efficacy of tomato leaf curl Bangalore virus by an indigenous and an exotic population. Journal of Integrative Agriculture 11: 235-248.

Chu D, Jiang T, Liu GX, Jiang DF, Tao YL, Fan ZX, Zhou HX, Bi YP. 2007. Biotype status and distribution of Bemisia tabaci (Hemiptera: Aleyrodidae) in Shandong Province of China based on mitochondrial DNA markers. Environmental Entomology 36: 1290-1295.

Costa HS, Brown JK. 1991. Variation in biological characteristics and in esterase patterns among populations of Bemisia tabaci (Genn.) and the association of one population with silver leaf symptom development. Entomologia Experimentalis et Applicata 61: 211-219.

Costa HS, Westcot DM, Ullman DE, Rosell R, Brown JK, Johnson MW. 1995. Morphological variation in Bemisia endosymbionts. Protoplasma 189: 194-202.

De Barro PJ, Trueman JWH, Frohlich DR. 2005. Bemisia argentifolii is a race of B. tabaci (Hemiptera: Aleyrodidae): the molecular genetics differentiation of B. tabaci populations around the world. Bulletin of Entomological Research 95: 193-203.

De Barro PJ, Liu SS, Boykin LM, Dinsdale A. 2011. Bemisia tabaci: a statement of species status. Annual Review of Entomology 56: 1-19.

Dinsdale A, Cook L, Riginos C, Buckley YM, De Barro PJ. 2010. Refined global analysis of Bemisia tabaci (Hemiptera: Sternorrhyncha: Aleyrodoidea: Aleyrodidae) mitochondrial cytochrome oxidase I to identify species level genetic boundaries. Annals of the Entomological Society of America 103: 196-208.

Frohlich DR, Torres-Jerez I, Bedford ID, Markham PG, Brown JK. 1999. A phylogeographical analysis of the Bemisia tabaci species complex based on mitochondrial DNA markers. Molecular Ecology 8: 1683-1691.

Gawel NJ, Bartlett AC. 1993. Characterization of differences between whiteflies using RAPD-PCR. Insect Molecular Biology 2: 33-38.

Govindappa MR. 2002. Detection and transmission of tomato leaf curl virus and interactions studies with host plants and whitefly Bemisia tabaci Genn. in relation to epidemics. Ph.D. thesis, University of Agricultural Sciences, Bangalore, India.

Green SK, Tsai WS, Shih SL.2003. Molecular characterization of a new begomovirus associated with tomato yellow leaf curl and eggplant yellow mosaic diseases in Thailand. Plant Disease 87: 446.

Hebert PDN, Penton EH, Burns JM, Janzen DH, Hallwachs W. 2004. Ten species in one: DNA barcoding reveals cryptic species in the Neotropical skipper butterfly Astraptes fulgerator. Proceedings of the National Academy of Sciences of the USA 101: 14812-14817.

Horowitz AR, Gorman K, Ross G, Denholm I. 2003. Inheritance of pyriproxyfen resistance in the whitefly, Bemisia tabaci (Q biotype). Archives of Insect Biochemistry and Physiology 54: 177-186.

Horowitz AR, Kontsedalov S, Ishaaya I. 2004. Dynamics of resistance to the neonicotinoids acetamiprid and thiamethoxam in Bemisia tabaci (Homoptera: Aleyrodidae). Journal of Economic Entomology 97: 2051-2056.

Hu J, De Barro PJ, Zhao H, Wang J, Nardi F, Liu SS. 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.

Huelsenbeck JP, Ronquist F. 2001. MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics 17: 754-755.

Jones DR. 2003. Plant viruses transmitted by whiteflies. European Journal of Plant Pathology 109: 195-219.

Lee W, Park J, Lee GS, Lee S, Akimoto SI. 2013. Taxonomic status of the Bemisia tabaci complex (Hemiptera: Aleyrodidae) and reassessment of the number of its constituent species. PLoS One 8: e63817.

Liou LW, Price TD. 1994. Speciation by reinforcement of premating isolation. Evolution 48: 1451-1459.

Lisha VS, Antony B, Palaniswami MS, Henneberry TJ. 2003. Bemisia tabaci (Homoptera: Aleyrodidae) biotypes in India. Journal of Economic Entomology 96: 322-327.

Liu SS, Colvin J, De Barro PJ. 2012. Species concepts as applied to the whitefly Bemisia tabaci systematics: How many species are there? Journal of Integrative Agriculture 11: 176-186.

Maruthi MN, Rekha AR, Sseruwagi P, Hillocks RJ. 2007. Mitochondrial DNA variability and development of a PCR diagnostic test for populations of the whitefly Bemisia afer (Priesner and Hosny). Molecular Biotechnology 35: 31-40.

Mayr E. 1982. The Growth of Biological Thought. Harvard University Press, Cambridge, Massachusetts, USA.

Mound AL. 1963. Host-correlated variations in Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae). Proceedings of the Royal Entomological Society of London, Series A, General Entomology 38: 171-180.

Muniyappa V, Maruthi MN, Babitha CR, Colvin J, Briddon RW, Rangaswamy KT. 2003. Characterization of pumpkin yellow vein mosaic virus from India. Annals of Applied Biology 142: 323-331.

Palaniswami MS, Nair RR, Pillai KS, Thankappan M. 1996. Whiteflies on cassava and its role as vector of cassava mosaic disease in India. Journal of Root Crops 22: 1-8.

Perring TM. 2001. The Bemisia tabaci species complex. Crop Protection 20: 725-737.

Posada D, Crandall KA. 1998. MODELTEST: testing the model of DNA substitution. Bioinformatics 14: 817-818.

Qiu BL, Ren SX, Mandour NS, Wen SY 2006. Population differentiation of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) by DNA polymorphism in China. Journal of South China Agricultural University 27: 29-34.

Ramappa HK, Muniyappa V, Colvin J. 1998. The contribution of tomato and alternative host plants to tomato leaf curl virus inoculum pressure in different areas of South India. Annals of Applied Biology 133: 187-198.

Rambaut A. 2009. FigTree Version 1.3.1, figtree/ (last accessed 6 Mar 2015).

Rambaut A, Drummond AJ. 2009. Tracer Version 1.5, Tracer (last accessed 6 Mar 2015).

Rauch N, Nauen R. 2003. Identification of biochemical markers linked to neonicotinoid cross resistance in Bemisia tabaci (Hemiptera: Aleyrodidae). Archives of Insect Biochemistry and Physiology 54: 165-176.

Rekha AR, Maruthi MN, 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.

Rosell RC, Bedford ID, Frohlich DR, Gill RJ, Brown JK, Markham PG. 1997. Analysis of morphological variation in distinct populations of Bemisia tabaci (Homoptera: Aleyrodidae). Annals of the Entomological Society of America 90: 575-589.

Russell LM. 1957. Synonyms of Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae). Bulletin of the Brooklyn Entomological Society 52: 122-123.

Shah SHJ, Malik AH, Qazi J. 2013. Identification of new genetic variant of Bemisia tabaci from Pakistan. International Journal of Entomological Research 1: 16-24.

Simon B, Cenis JL, Demichelis S, Rapisarda C, Caciagli P, Bosco D. 2003. Survey of Bemisia tabaci (Hemiptera: Aleyrodidae) biotypes in Italy with the description of a new biotype (T) from Euphorbia characias. Bulletin of Entomological Research 93: 259-264.

Simon C, Frati F, Beckenbach A, Crespi B, Liu H, 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: 651-701.

Swofford DL. 1998. PAUP* Phylogenetic analysis using parsimony (* and other methods). Version 4. Sinaue Associates, Sunderland, Massachusetts, USA.

Tajima F, Nei M. 1984. Estimation of evolutionary distance between nucleotide sequences. Molecular Biology and Evolution 1: 269-285.

Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. 2011. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution 28: 2731-2739.

Thompson JD, Higgins DG, Gibson JJ. 1994. CLUSTAL W: improving the sensitivity of progressive multiple alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research 11: 4673-4680.

Toda S, Murai T. 2007. Phylogenetic analysis based on mitochondrial COI gene sequences in Thrips tabaci Lindeman (Thysanoptera: Thripidae) in relation to reproductive forms and geographical distribution. Applied Entomology and Zoology 42: 309-316.

Venkatesh HM. 2000. Studies on tomato leaf curl geminivirus and Bemisia tabaci (Gennadius): molecular detection, farmers' perception and sustainable management. Ph.D. thesis, Department of Plant Pathology, University of Agricultural Sciences, Bangalore, India.

Wang P, Sun D, Qiu B, Liu SS. 2011. The presence of six cryptic species of the whitefly Bemisia tabaci complex in China as revealed by crossing experiments. Insect Science 18: 67-77.

Xu J, De Barro PJ, 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: 359-366.

Zang LS, Chen WQ, Liu SS. 2006. Comparison of performance on different host plants between the B biotype and a non-B biotype of Bemisia tabaci from Zhejiang, China. Entomologia Experimentalis et Applicata 121: 221-227.

Zasada IA, Peetz A, Howe DK, Wilhelm LJ, Cheam D, Denver DR, Smythe AB. 2014. Using mitogenomic and nuclear ribosomal sequence data to investigate the phylogeny of the Xiphinema americanum species complex. PLoS One 9: e90035.

H. K. Roopa (1), *, R. Asokan (1), K. B. Rebijith (1), Ranjitha H. Hande (1), Riaz Mahmood (2), and N. K. Krishna Kumar (3)

(1) Division of Biotechnology, Indian Institute of Horticultural Research, Hessaraghatta Lake P.O Karnataka, Bengaluru 560 089, India

(2) Department of Biotechnology and Bioinformatics, Kuvempu University, Jnanasahyadri, Shankaraghatta, Shimoga577 451, India

(3) Division of Horticulture, ICAR, Krishi Anusandhan Bhawan - II, New Delhi 110 012, India

* Corresponding author; E-mail:

Supplementary material for this article in Florida Entomologist 98(4) (Dec 2015) is online at

Caption: Fig. 1. Map indicating different locations of Bemisia tabaci sampling and distribution of genetic groups in Karnataka, India.

Caption: Fig. 2a. Phylogenetic tree showing the relationship of the Bemisia tabacimt mtCOI sequences collected in this study to consensus sequences of Dinsdale et al. (2010). Bayesian analyses were performed employing MrBayes under the best-fit model GTR+I+G of molecular evolution for 20 million generations and 25% discarded as burn-in. Posterior probabilities are shown above the branches and maximum likelihood scores from RaxML indicated below the branches. Asia-I condensed is shown in Fig. 2b.

Caption: Fig. 2b. Phylogenetic tree indicating the Asia-I group, which was the most abundant genetic group in this study.

Caption: Fig. 3. Graph indicating net evolutionary divergence between groups of sequences.

Table 1. Details of Bemisia tabaci mtCOI sequences generated
for this study along with place of collection, accession number,
voucher specimen number, respective genetic group, and host plants.

SI no.   Host                            Common name

1        Ocimum sanctum                  Tulsi
2        Parthenium hysterophorus        Aster (white top weed)
3        Raphanus sativus                Radish
4        Unknown Ornamental              Ornamental
5        Manihot esculenta               Tapioca
6        Hibiscus cultivar               Hibiscus
7        Gossypium hirsutum              Cotton
8        Cucumis sativus                 Cucumber
9        Solanum melongena               Eggplant
10       Brassica oleraceo               Cabbage
11       Musa acuminata                  Banana
12       Phaseolus vulgaris              Common beans
13       Brassica oleraceo               Cabbage
14       Brassica oleraceo               Cabbage
15       Lablab purpureus                Hyacintha beans
16       Abelmoschus esculentum          Okra
17       Citrullus la not us             Watermelon
18       Lycopersicon seculentum         Tomato
19       Gossypium hirsutum              Cotton
20       Lagenaria sice rana             Bottle gourd
21       Lycopersicon esculentum         Tomato
22       Musa acuminta                   Banana
23       Luffa spp.                      Ridge gourd
24       Abelmoschus esculentus          Okra
25       Helia nth us annuus             Sunflower
26       Capsicum annuum                 Capsicum
27       Cucurbita moschata              Pumpkin
28       Vigna unguiculata               Cowpea
29       Citrullus lanatus               Watermelon
30       Daucus carota                   Carrot
31       Brassica juncea                 Mustard
32       Crossandra infundibuliformis    Crossandra
33       Cucumis sativus                 Cucumber
34       Gossypium hirsutum              Cotton
35       Solanum melongena               Eggplant
36       Ricinus communis                Castor bean
37       Cucumis melo                    Muskmelon
38       Gossypium hirsutum              Cotton
39       Solanum tuberosum               Potato
40       Brassica rapa                   Turnip
41       Solanum melongena               Eggplant
42       Solanum melongena               Eggplant
43       Solanum melongena               Eggplant
44       Solanum melongena               Eggplant
45       Solanum melongena               Eggplant
46       Abelmoschus esculentum          Bhendi
47       Solanum melongena               Eggplant
48       Solanum melongena               Eggplant
49       Solanum melongena               Eggplant
50       Solanum melongena               Eggplant
51       Solanum tuberosum               Potato
52       Cucumis sativus                 Cucumber
53       Abelmoschus esculentum          Bhendi
54       Lycopersicon esculentum         Tomato
55       Gossypium hirsutum              Cotton
56       Gossypium hirsutum              Cotton
57       Solanum melongena               Eggplant
58       Gossypium hirsutum              Cotton
59       Solanum melongena               Eggplant
60       Gossypium hirsutum              Cotton
62       Brassica oleraceo               Cabbage
63       Gossypium hirsutum              Cotton
64       Solanum melongena               Eggplant
65       Gossypium hirsutum              Cotton
66       Solanum melongena               Eggplant
67       Lycopersicon esculentum         Tomato
68       Gossypium hirsutum              Cotton
69       Solanum melongena               Eggplant
70       Phaseolus vulgaris              Beans
71       Phaseolus vulgaris              Beans
72       Phaseolus vulgaris              Beans

              Bangalore,         Accession
SI no.     Karnataka, India        number     Latitude   Longitude

1        Malleshwarum             KF790634     12.98       77.57
2        Tarabanahalli            KF790635     13.11       77.47
3        IIHR                     KF790636     13.13       77.48
4        Malleshwarum             KF790637     12.98       77.57
5        Malleshwarum             KF790638     12.98       77.57
6        Malleshwarum             KF790639     12.98       77.57
7        Hessaraghatta            KF790653     13.13       77.48
8        Andaman                  KF790667     11.68       92.77
9        Chamarajanagar           KF790678     11.56       77.00
10       Rajankunte               KF790641     12.97       77.59
11       IIHR                     KF790663     13.13       77.48
12       Rajarajeshwarinagar      KF790664     12.80       77.51
13       Kolar                    HQ268814     14.10       77.27
14       Kolar                    HQ268811     14.10       77.27
15       Banashankari             KF790659     12.93       77.55
16       Chikkajala               KF790661     13.16       77.63
17       Chamarajanagar           KF790679     11.56       77.00
18       Kolar                    HQ331244     14.10       77.27
19       Bija pur                 KJ523177     16.83       75.71
20       Bidadi                   KF790640     12.80       77.40
21       Rajarajeshwarinagar      KF790642     12.90       77.51
22       IIHR                     KF790643     13.13       77.48
23       Bidadi                   KF790644     12.80       77.40
24       Chikkajala               KF790645     13.16       77.63
25       IIHR                     KF790646     13.13       77.48
26       IIHR                     KF790647     13.13       77.48
27       IIHR                     KF790648     13.13       77.48
28       Golahalli                KF790649     13.13       77.59
29       IIHR                     KF790650     13.13       77.48
30       Dommasandra              KF790651     12.88       77.75
31       Ranjantunte              KF790652     12.97       77.59
32       IIHR                     KF790654     13.13       77.48
33       IIHR                     KF790655     13.13       77.48
34       Bangalore                KF790666     13.04       77.59
35       Bidadi                   KF790656     12.80       77.40
36       Kadugedi                 KF790657     12.99       77.76
37       Banashankari             KF790658     12.93       77.55
38       Rajajinagar              KF790660     12.97       77.57
39       Anekal                   KF790662     12.70       77.70
40       Makali                   KF790665     13.09       77.39
41       Andaman                  KF790668     11.68       92.77
42       Gujarat                  KF790669     23.21       72.68
43       Kodagu                   KF790670     12.42       75.73
44       Maddur                   KF790671     12.59       77.05
45       Hunasuru                 KF790672     12.31       76.29
46       Mysore                   KF790673     12.18       76.42
47       Mysore (*Periyapatna)    KF790677     12.18       76.42
48       Harangi                  KF790675     12.49       75.90
49       Basavanahalli            KF790676     13.11       76.91
50       Channapattana            KF790674     12.65       77.20
51       Chamarajanagar           KF790680     11.56       77.00
52       Chamarajanagar           KF790681     11.56       77.00
53       Chamarajanagar           KF790682     11.56       77.00
54       Tumkur                   KF790683     13.20       77.08
55       Rannibennur              KF790684     14.61       75.62
56       Davanagere               KF790685     14.31       75.58
57       Davanagere               KF790686     14.31       75.58
58       Chitradurga              KF790687     14.14       76.26
59       Sirsi                    KF790688     14.61       74.82
60       Haveri                   KF790689     14.80       75.39
62       Kolar                    HQ268812     13.01       78.33
63       Kolar                    HQ331246     13.73       75.33
64       Kolar                    HQ331245     13.09       78.11
65       Gulbarga                 KJ523182     17.33       76.83
66       Raichur                  KJ523180     16.20       77.37
67       Chikmangaluru            KJ523179     13.32       75.77
68       Dharward                 KJ523181     15.45       75.00
69       Belgaum                  KJ523178     15.85       74.50
70       Kolar (H Cross)          KJ787661     13.24       77.93
71       Kolar (Malur)            KJ787662     13.02       77.93
72       Kolar                    HQ268813     13.13       78.13

                         Voucher     Year of
SI no.   Genetic group   specimen   collection

1          Asia-II-7      HKR01        2012
2          Asia-II-7      HKR02        2012
3          Asia-II-7      HKR03        2012
4          Asia-II-7      HKR04        2012
5          Asia-II-7      HKR05        2012
6          Asia-II-7      HKR06        2012
7          Asia-II-7      HKR20        2013
8          Asia-II-7      HKR34        2012
9          Asia-II-7      HKR45        2011
10         MEAM-1         HKR08        2012
11         MEAM-1         HKR30        2013
12         MEAM-1         HKR31        2013
13         MEAM-1         NA           2010
14         MEAM-1         NA           2010
15         Asia-II-8      HKR26        2013
16         Asia-II-8      HKR28        2013
17         Asia-II-8      HKR46        2011
18         Asia-II-8      NA           2010
19         Asia-II-8      HKR57        2013
20         Asia-1         HKR07        2012
21         Asia-1         HKR09        2012
22         Asia-1         HKR10        2012
23         Asia-1         HKR11        2012
24         Asia-1         HKR12        2012
25         Asia-1         HKR13        2012
26         Asia-1         HKR14        2012
27         Asia-1         HKR15        2012
28         Asia-1         HKR16        2013
29         Asia-1         HKR17        2013
30         Asia-1         HKR18        2013
31         Asia-1         HKR19        2013
32         Asia-1         HKR21        2013
33         Asia-1         HKR22        2013
34         Asia-1         HKR33        2013
35         Asia-1         HKR23        2013
36         Asia-1         HKR24        2013
37         Asia-1         HKR25        2013
38         Asia-1         HKR27        2013
39         Asia-1         HKR29        2013
40         Asia-1         HKR32        2013
41         Asia-1         HKR35        2012
42         Asia-1         HKR36        2011
43         Asia-1         HKR37        2011
44         Asia-1         HKR38        2011
45         Asia-1         HKR39        2011
46         Asia-1         HKR40        2011
47         Asia-1         HKR44        2011
48         Asia-1         HKR42        2011
49         Asia-1         HKR43        2011
50         Asia-1         HKR41        2011
51         Asia-1         HKR47        2011
52         Asia-1         HKR48        2011
53         Asia-1         HKR49        2011
54         Asia-1         HKR50        2011
55         Asia-1         HKR51        2011
56         Asia-1         HKR52        2011
57         Asia-1         HKR53        2011
58         Asia-1         HKR54        2011
59         Asia-1         HKR55        2011
60         Asia-1         HKR56        2011
62         Asia-1         NA           2010
63         Asia-1         NA           2010
64         Asia-1         NA           2010
65         Asia-1         HKR62        2013
66         Asia-1         HKR60        2013
67         Asia-1         HKR59        2013
68         Asia-1         HKR61        2013
69         Asia-1         HKR58        2013
70         MEAM-K         HKR63        2014
71         MEAM-K         HKR64        2014
72         MEAM-K         NA           2010

Table 2. Maximum composite likelihood estimate of the pattern
of nucleotide substitution from Bemisia tabaci populations
collected on various host plants in Karnataka, India.

       A        T        C        G

A      --     5.24##   1.61##   14.63#
T    2.98##     --     9.96#    2.34##
C    2.98##   32.46#     --     2.34##
G    18.62#   5.24##   1.61##     --

Each entry shows the probability of substitution
from one base (row) to another base (column) instantaneously.
Only entries within a row should be compared. Rates of different
transitional substitutions are shown in bold and those of
transversional substitutions are shown in italics.

Note: Rates of different transitional substitutions shown
are indicated with # and those of transversional substitutions
are shown in ##.

Table 3. Estimates of net evolutionary divergence between
groups of sequences showing pair-wise genetic distances
for Bemisia tabaci populations collected on various host
plants in Karnataka, India.


Asia-I                 0.139
MEAM-1                 0.173    0.161
Asia-II-8              0.119    0.152    0.161
MEAM-l-Karnataka       0.153    0.164    0.074    0.157
Asia-III               0.135    0.078    0.174    0.163    0.175
Asia-II-10             0.109    0.137    0.172    0.120    0.155
Asia-II-1              0.092    0.157    0.172    0.133    0.160
Asia-II-2              0.111    0.144    0.129    0.120    0.130
Asia-II-3              0.122    0.176    0.174    0.118    0.165
Asia-II-4              0.129    0.169    0.160    0.142    0.166
Asia-II-5              0.109    0.173    0.172    0.117    0.095
Asia-II-6              0.096    0.159    0.171    0.136    0.124
Asia-II-9              0.108    0.169    0.169    0.118    0.155
Australia              0.156    0.117    0.163    0.155    0.153
Australia-Indonesia    0.178    0.146    0.174    0.176    0.166
China-I                0.145    0.128    0.161    0.148    0.168
Indian_Ocean           0.165    0.152    0.076    0.169    0.126
Italy                  0.124    0.145    0.153    0.128    0.155
Mediterranean          0.170    0.171    0.052    0.164    0.110
New-World-1            0.159    0.134    0.139    0.163    0.166
Subgroup-Af-1          0.189    0.196    0.164    0.184    0.195
Subgroup-Af-2          0.205    0.186    0.165    0.187    0.198
Subgroup-Af-3          0.199    0.172    0.172    0.176    0.197
Subgroup-Af-4          0.195    0.183    0.175    0.189    0.201
Uganda                 0.229    0.228    0.202    0.207    0.209
MEAM-2                 0.162    0.160    0.041    0.150    0.098
China-II               0.135    0.127    0.153    0.154    0.144
China-III              0.151    0.150    0.156    0.135    0.154


Asia-II-10             0.139
Asia-II-1              0.158    0.131
Asia-II-2              0.145    0.108    0.067
Asia-II-3              0.166    0.107    0.133    0.075
Asia-II-4              0.158    0.119    0.125    0.051    0.048
Asia-II-5              0.168    0.139    0.106    0.107    0.136
Asia-II-6              0.147    0.116    0.107    0.118    0.136
Asia-II-9              0.160    0.098    0.121    0.091    0.050
Australia              0.124    0.154    0.148    0.130    0.152
Australia-Indonesia    0.139    0.170    0.179    0.151    0.176
China-I                0.122    0.134    0.156    0.135    0.156
Indian_Ocean           0.171    0.168    0.167    0.153    0.176
Italy                  0.131    0.131    0.140    0.134    0.134
Mediterranean          0.185    0.177    0.179    0.150    0.182
New-World-1            0.152    0.147    0.165    0.148    0.157
Subgroup-Af-1          0.187    0.180    0.193    0.182    0.184
Subgroup-Af-2          0.191    0.184    0.205    0.183    0.175
Subgroup-Af-3          0.180    0.178    0.190    0.180    0.188
Subgroup-Af-4          0.185    0.172    0.202    0.188    0.193
Uganda                 0.220    0.198    0.212    0.211    0.237
MEAM-2                 0.166    0.167    0.163    0.137    0.169
China-II               0.130    0.134    0.157    0.139    0.158
China-III              0.142    0.138    0.161    0.140    0.162


Asia-II-5              0.133
Asia-II-6              0.141    0.067
Asia-II-9              0.080    0.120    0.126
Australia              0.142    0.133    0.150    0.147
Australia-Indonesia    0.180    0.172    0.174    0.163    0.136
China-I                0.154    0.147    0.141    0.148    0.147
Indian_Ocean           0.183    0.177    0.175    0.163    0.180
Italy                  0.148    0.142    0.129    0.123    0.131
Mediterranean          0.186    0.187    0.171    0.165    0.191
New-World-1            0.159    0.162    0.173    0.145    0.164
Subgroup-Af-1          0.198    0.202    0.199    0.178    0.182
Subgroup-Af-2          0.191    0.205    0.190    0.164    0.175
Subgroup-Af-3          0.197    0.216    0.204    0.180    0.189
Subgroup-Af-4          0.203    0.208    0.206    0.187    0.183
Uganda                 0.239    0.218    0.225    0.212    0.233
MEAM-2                 0.171    0.172    0.157    0.162    0.174
China-II               0.152    0.142    0.145    0.144    0.146
China-III              0.168    0.165    0.160    0.140    0.150


China-I                0.138
Indian_Ocean           0.180    0.165
Italy                  0.122    0.123    0.153
Mediterranean          0.180    0.168    0.070    0.161
New-World-1            0.179    0.150    0.156    0.128    0.148
Subgroup-Af-1          0.180    0.186    0.187    0.155    0.181
Subgroup-Af-2          0.186    0.198    0.176    0.149    0.176
Subgroup-Af-3          0.183    0.189    0.183    0.165    0.185
Subgroup-Af-4          0.181    0.199    0.182    0.156    0.180
Uganda                 0.207    0.206    0.213    0.209    0.207
MEAM-2                 0.177    0.154    0.069    0.148    0.040
China-II               0.142    0.044    0.160    0.121    0.173
China-III              0.115    0.123    0.191    0.142    0.179


Subgroup-Af-1          0.149
Subgroup-Af-2          0.147    0.076
Subgroup-Af-3          0.151    0.072    0.061
Subgroup-Af-4          0.150    0.078    0.072    0.071
Uganda                 0.194    0.190    0.196    0.194
MEAM-2                 0.144    0.167    0.163    0.175
China-II               0.144    0.190    0.195    0.189
China-III              0.174    0.176    0.195    0.191


Uganda                 0.202
MEAM-2                 0.170    0.195
China-II               0.204    0.209    0.159
China-III              0.196    0.205    0.157    0.120

Analyses were conducted using the maximum composite likelihood model
(Tajima & Nei 1984). The analysis involved 135 nucleotide sequences.
All positions containing gaps and missing data were eliminated.
There were 620 positions in the final dataset. Evolutionary analyses
were conducted in MEGA5 (Tamura et al. 2011).


Please note: Some tables or figures were omitted from this article.
COPYRIGHT 2015 Florida Entomological Society
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2015 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Title Annotation:Research Papers
Author:Roopa, H.K.; Asokan, R.; Rebijith, K.B.; Hande, Ranjitha H.; Mahmood, Riaz; Kumar, N.K. Krishna
Publication:Florida Entomologist
Article Type:Report
Date:Dec 1, 2015
Previous Article:Heteroptera attracted to butterfly traps baited with fish or shrimp carrion.
Next Article:First record of Conotrachelus perseae (Coleoptera: Curculionidae) in Comitan, Chiapas, Mexico.

Terms of use | Privacy policy | Copyright © 2020 Farlex, Inc. | Feedback | For webmasters