Assessment of cross-resistance potential to neonicotinoid insecticides in Bemisia tabaci (Hemiptera: Aleyrodidae)

Laboratory bioassays were carried out with four neonicotinoid insecticides on multiple strains of Bemisia tabaci (Gennadius) to evaluate resistance and cross-resistance patterns. Three imidacloprid-resistant strains and field populations from three different locations in the southwestern USA were compared in systemic uptake bioassays with acetamiprid, dinotefuran, imidacloprid and thiamethoxam. An imidacloprid-resistant strain (IM-R) with 120-fold resistance originally collected from Imperial Valley, California, did not show cross-resistance to acetamiprid, dinotefuran or thiamethoxam. The Guatemala-resistant strain (GU-R) that was also highly resistant to imidacloprid (RR=109-fold) showed low levels of cross-resistance when bioassayed with acetamiprid and thiamethoxam. However, dinotefuran was more toxic than either imidacloprid or thiamethoxam to both IM-R and GU-R strains as indicated by low LC50s. By contrast, a Q-biotype Spanish-resistant strain (SQ-R) of B. tabaci highly resistant to imidacloprid demonstrated high cross-resistance to the two related neonicotinoids. Field populations from Imperial Valley (California), Maricopa and Yuma (Arizona), showed variable susceptibility to imidacloprid (LC50s ranging from 3.39 to 115 microg ml(-1)) but did not exhibit cross-resistance to the three neonicotinoids suggesting that all three compounds would be effective in managing whiteflies. Yuma populations were the most susceptible to imidacloprid. Dinotefuran was the most toxic of the four neonicotinoids against field populations. Although differences in binding at the target site and metabolic pathways may influence the variability in cross-resistance patterns among whitefly populations, comparison of whitefly responses from various geographic regions to the four neonicotinoids indicates the importance of ecological and operational factors on development of cross-resistance to the neonicotinoids.     

Dynamics of resistance to the neonicotinoids acetamiprid and thiamethoxam in Bemisia tabaci (Homoptera: Aleyrodidae)

The dynamics of resistance in the sweetpotato whitefly, Bemisia tabaci (Gennadius), to the neonicotinoids acetamiprid and thiamethoxam was studied extensively in cotton fields in Israel during the cotton-growing seasons 1999-2003. Whitefly strains were collected in early and late seasons mainly in three locations in northern, central, and southern Israel. The whiteflies were assayed under laboratory conditions for susceptibility to neonicotinoids, as part of the Israeli cotton insecticide resistance management strategy. Selections to both acetamiprid and thiamethoxam and cross-resistance between them also were conducted in the laboratory. Although no appreciable resistance to acetamiprid was observed up to 2001, a slight increase of approximately five-fold resistance was detected during 2002 and 2003. However, from 2001 to 2003 thiamethoxam resistance increased >100-fold in the Ayalon Valley and Carmel Coast cotton fields. In cross-resistance assays with both neonicotinoids, the strain that had been selected with thiamethoxam for 12 generations demonstrated almost no cross-resistance to acetamiprid, whereas the acetamiprid-selected strain exhibited high cross-resistance of >500-fold to thiamethoxam.     

Resistance of insect pests to neonicotinoid insecticides: current status and future prospects

The first neonicotinoid insecticide introduced to the market was imidacloprid in 1991 followed by several others belonging to the same chemical class and with the same mode of action. The development of neonicotinoid insecticides has provided growers with invaluable new tools for managing some of the world's most destructive crop pests, primarily those of the order Hemiptera (aphids, whiteflies, and planthoppers) and Coleoptera (beetles), including species with a long history of resistance to earlier-used products. To date, neonicotinoids have proved relatively resilient to the development of resistance, especially when considering aphids such as Myzus persicae and Phorodon humuli. Although the susceptibility of M. persicae may vary up to 20-fold between populations, this does not appear to compromise the field performance of neonicotinoids. Stronger resistance has been confirmed in some populations of the whitefly, Bemisia tabaci, and the Colorado potato beetle, Leptinotarsa decemlineata. Resistance in B- and Q-type B. tabaci appears to be linked to enhanced oxidative detoxification of neonicotinoids due to overexpression of monooxygenases. No evidence for target-site resistance has been found in whiteflies, whereas the possibility of target-site resistance in L. decemlineata is being investigated further. Strategies to combat neonicotinoid resistance must take account of the cross-resistance characteristics of these mechanisms, the ecology of target pests on different host plants, and the implications of increasing diversification of the neonicotinoid market due to a continuing introduction of new molecules.     

Biorational insecticides: mechanism and cross-resistance

Potency and cross-resistance of various biorational insecticides, exemplified by the whitefly Bemisia tabaci, have been studied. Bemisia tabaci were exposed to the juvenile hormone mimic pyriproxyfen for the past 12 years resulting in an over 2,000-fold resistance, but there was no appreciable cross-resistance with the benzoylphenyl urea novaluron. Similarly, no cross-resistance was found between pyriproxyfen and the two neonicotinoids, acetamiprid and imidacloprid. On the other hand, a slight cross-resistance of 5-13-fold was observed with another neonicotinoid thiamethoxam. Among the neonicotinoids, a resistant strain of B. tabaci to thiamethoxam (approximately 100-fold) showed no appreciable cross-resistance to either acetamiprid or imidacloprid, while another strain 500-fold resistant to thiamethoxam resulted in a mild of 4-6-fold resistance to acetamiprid and imidacloprid. In other assays, B. tabaci strain resistant to thiamethoxam (approximately 100-fold) had no cross-resistance to pyriproxyfen. Our findings indicate that no appreciable cross-resistance was observed between the benzoylphenyl urea novaluron, the juvenile hormone mimic pyriproxyfen, and the neonicotinoids acetamiprid and imidacloprid. Hence, these compounds could be used as components in insecticide resistance management programs.     

Can insecticide mixtures be considered to surmount neonicotinoid resistance in Bemisia tabaci?

Cotton whitefly, Bemisia tabaci is an important polyphagous pest worldwide. It is exposed to various chemical insecticides throughout the year, resulting in the rapid development of insecticide resistance. Mixtures of insecticides with distinct modes of action could enhance the toxicity of chemicals more effectively than sequences or rotations in resistant pest populations. Bioassays were conducted to study the efficacy of mixtures of neonicotinoid and ketoenol insecticides at different ratios against a laboratory susceptible (Lab-WB) and a neonicotinoid resistant (TMX-SEL) strain of B. tabaci Asia I. The results showed that mixtures of imidacloprid, acetamiprid, thiamethoxam or dinotefuran with spiromesifen at 1:1, 1:10 and 1:20 ratios and of imidacloprid, thiamethoxam or dinotefuran with spirotetramat at 1:1 ratio significantly increased (p < 0.05) toxicity to neonicotinoids in TMX-SEL strain. The combination indices of each tested neonicotinoids + ketoenols at 1:1 ratio and of acetamiprid + spiromesifen, and imidacloprid or dinotefuran + spirotetramat at 1:10 ratio for TMX-SEL strain were significantly below 1, suggesting synergistic interactions. The inhibitors PBO and DEF largely overcame resistance to the tested neonicotinoids, while none of the synergists significantly restored the susceptibility of B. tabaci to ketoenols. Increased activities of P450 monooxygenase and esterase were observed in TMX-SEL strain with an elevated 2.76 and 1.32-fold, respectively. Mixtures of neonicotinoids with spiromesifen or spirotetramat at a 1:1 ratio could be used to restore the neonicotinoid susceptibility in B. tabaci.     

Novaluron (Rimon), a novel IGR: potency and cross-resistance

The potency of novaluron on laboratory susceptible and field strains of S. littoralis resembles that of chlorfluazuron and both compounds are about 20-fold more potent than teflubenzuron. No appreciable resistance to novaluron or chlorfluazuron was observed in a field strain of Spodoptera littoralis collected from cucumber field in the central part of Israel. On the other hand, the field strain showed a mild resistance of about 4-fold to teflubenzuron as compared to the laboratory susceptible strain. A very resistant colony of Bemisia tabaci to pyriproxyfen (1,200- to 2,000-fold) showed no appreciable cross-resistance to novaluron. Two field colonies of B. tabaci pressurized with acetamiprid or thiamethoxam for 22 generations, resulting in a 30- to 50-fold resistance to acetamiprid and thiamethoxam, has no cross-resistance to novaluron. The above results are of special interest, indicating a possible alternation between novaluron, pyriproxyfen, and neonicotinoids in insecticide-resistance management programs aiming at preventing resistance development to these novel groups of insecticides against important pests such as whitefly and lepidopteran species.     

Testing for non-target effects of spiromesifen on <Emphasis Type="Italic">Eretmocerus mundus</Emphasis> and <Emphasis Type="Italic">Orius laevigatus</Emphasis> under greenhouse conditions

Spiromesifen is a novel insecticide/acaricide belonging to the new chemical class of spirocyclic phenyl-substituted tetronic acids, and it is especially active against whiteflies and tetranychid spider mite species. In the biologically based integrated pest management (IPM) programs in vegetable crops in southeastern Spain, the key natural enemies include the parasitoid Eretmocerus mundus Mercet (Hymenoptera: Aphelinidae) for sweetpotato whitefly control, and the minute pirate bug, Orius laevigatus (Fieber) (Hemiptera: Anthocoridae) for western flower thrips control. Side effects of spiromesifen on E. mundus and O. laevigatus, were evaluated by laboratory studies and field trials in commercial greenhouses under IPM programs. Results indicate that spiromesifen had favourable selectivity to O. laevigatus and E. mundus and would complement biological control of Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) by E. mundus. Handling editor: Patrick De Clercq     

Biotypes B and Q of Bemisia tabaci and their relevance to neonicotinoid and pyriproxyfen resistance

Resistance monitoring for Bemisia tabaci field populations to the juvenile hormone mimic, pyriproxyfen, was conducted from 1996 to 2003 in commercial cotton fields in two areas of Israel: the Ayalon Valley (central Israel) and the Carmel Coast (northwestern Israel). Although the use of pyriproxyfen ceased in these areas in 1996-1997 (because of the resistance), resistance levels to pyriproxyfen declined to some extent in the fields but remained quite stable, and the susceptibility has not been totally restored. Two strains of B. tabaci collected from the Ayalon Valley in the late 1999 and 2002 cotton seasons (AV99L, AV02L) were assayed for their susceptibility to pyriproxyfen at F1, and subsequently a line of each strain was kept under controlled conditions without exposure to insecticides. After maintenance of more than 20 generations under laboratory conditions, the resistance to pyriproxyfen in the untreated strains substantially declined. This decline was concurrent with a replacement of Q biotype by B-type under non-insecticidal regimes; apparently B biotype was more competitive than the pyriproxyfen-resistant Q-type. Selection under controlled conditions with neonicotinoids on these B. tabaci strains resulted in continued pyriproxyfen resistance, predominantly of Q biotype. Based on our data, applications of either pyriproxyfen or neonicotinoids may select for biotype Q, which would survive to a greater degree where these insecticides are applied.     

Toxicity of sucrose octanoate to egg, nymphal, and adult Bemisia tabaci (Hemiptera: Aleyrodidae) using a novel plant-based bioassay

The sweetpotato whitefly, Bemisia tabaci (Gennadius), B biotype, presents a unique problem for vegetable growers by serving as a vector of plant viruses and by inducing physiological disorders of leaves and fruit. An action threshold of a single whitefly is necessary because of the threat of disease in many areas and growers rely heavily on a single class of insecticides (neonicotinoids) for whitefly control. Additional control methods are needed to manage this pest in commercial vegetables. Extracts of wild tobacco contain natural sugar esters that have previously been shown effective in controlling many soft-bodied insects. We developed a novel tomato leaf bioassay system to assess a synthetic sugar ester derivative, sucrose octanoate, for insecticidal activity against the eggs, nymphs, and adults of B. tabaci. The LC50 values for sucrose octanoate against adults, second instars, and fourth instars of the whitefly were 880, 686, and 1,571 ppm, respectively. The LC50 against whitefly eggs was higher (11,446 ppm) but indicated that some egg mortality occurred at the recommended application rate of 0.8-1.2% (3,200-4,800 ppm [Al]). Toxicity of sugar esters to whitefly eggs has not been reported previously. The tomato leaf bioassay produced reliable and repeatable results for whitefly toxicity studies and predicted that effective nymph and adult whitefly control can be achieved with sucrose octanoate at application rates < or = 1% (4,000 ppm [AI]). Field efficacy studies are warranted to determine whether this biorational pesticide has application in commercial tomato production.     

Host plant influence on susceptibility of Bemisia tabaci (Hemiptera: Aleyrodidae) to insecticides

A resistance monitoring program conducted for the polyphagous whitefly, Bemisia tabaci (Gennadius), in Imperial Valley, CA, USA generated a large set of LC50s for adults collected from broccoli, cantaloupe and cotton crops over a four-year period. A vial bioassay and, subsequently, a yellow-sticky card bioassay produced similar temporal profiles of relative susceptibilities to the pyrethroid insecticide bifenthrin. Both bioassays revealed that whiteflies collected from broccoli were significantly less susceptible to bifenthrin compared to the other two crops. A similar finding was observed for endosulfan and the mixture of bifenthrin+endosulfan in the yellow-sticky card bioassay. The possibility that seasonal differences contributed to the observed differences in susceptibility provided the impetus to conduct a reciprocal transfer experiment using broccoli (or kale) and cantaloupe grown simultaneously in the field and greenhouse. Whitefly adults collected from an organic farm over three consecutive weeks had significantly higher LC50s on kale than those collected the same day on cantaloupe. After culturing in the greenhouse on broccoli or cantaloupe and testing again, LC50s remained significantly higher on broccoli after one week and again at the F1 generation. In contrast, whiteflies originating on kale in the field and transferred to cantaloupes in the greenhouse had significantly reduced LC50s at the F1 generation. When tested against the bifenthrin+endosulfan mixture, significantly higher LC50s were generated for whiteflies reared on broccoli in the greenhouse at one week and the F1 compared to the field source from cantaloupes. The consistently higher LC50s for whiteflies on broccoli and other Brassica spp. crops, compared to cantaloupes or cotton, point to statistically significant host-plant influences that are expressed in both field-collected and greenhouse-reared populations of whiteflies.     

Whitefly population dynamics and evaluation of whitefly-transmitted tomato yellow leaf curl virus (TYLCV)-resistant tomato genotypes as whitefly and TYLCV reservoirs

Sweetpotato whitefly, Bemisia tabaci (Gennadius), and whitefly-transmitted tomato yellow leaf curl virus (TYLCV) are major threats to tomato production in the southeastern United States. TYLCV was introduced to Florida from the Caribbean islands and has spread to other southern states of the United States. In Georgia, in recent years, the incidence of TYLCV has been steadily increasing. Studies were conducted to monitor population dynamics of whiteflies in the vegetable production belt of Georgia, to evaluate TYLCV-resistant genotypes against whiteflies and TYLCV, and to assess the potential role of resistant genotypes in TYLCV epidemiology. Monitoring studies indicated that the peak incidence of whiteflies varied seasonally from year to year. In general, whitefly populations were not uniformly distributed. Tomato genotypes exhibited minor differences in their ability to support whitefly populations. TYLCV symptoms were visually undetectable in all but one resistant genotype. The infection rates (visually) in susceptible genotypes ranged from 40 to 87%. Greenhouse inoculations with viruliferous whiteflies followed by polymerase chain reaction (PCR) indicated that up to 100% of plants of resistant genotypes were infected, although predominantly symptomless. TYLCV acquisition by whiteflies from TYLCV-infected genotypes was tested by PCR; TYLCV acquisition rates from resistant genotypes were less than from susceptible genotypes. Nevertheless, this difference did not influence TYLCV transmission rates from resistant to susceptible genotypes. Results emphasize that resistant genotypes can serve as TYLCV and whitefly reservoirs and potentially influence TYLCV epidemics.     

Host plant effects on resistance to bifenthrin in silverleaf whitefly (Homoptera: Aleyrodidae)

Effects of host plants on resistance to bifenthrin in the silverleaf whitefly, Bemisia argentifolii Bellows & Perring, were determined by LC50 bioassay. In addition, inheritance of resistance to bifenthrin was investigated beginning with a single source of a bifenthrin-susceptible population. Overall, the resistance ratio between the bifenthrin-susceptible population and the selected bifenthrin-resistant population from the same source population was 915-fold after 1 yr in the greenhouse. Responses to bifenthrin among the susceptible and the resistant populations were changed when whiteflies were reared on three different host plants, i.e., cotton, cabbage, and squash. In the resistant populations, the LC50 value of whitefly fed on squash was increased as much as 7.5-fold, while the LC50 value of whitefly fed on cabbage was similar to cotton that served as the control plant. The host plant on which whiteflies feed appears to be an important factor in selection for resistance to bifenthrin, but these effects are crop specific. Based on an analysis using LC50 values of the reciprocal F1 cross on cotton, resistance of whitefly from a single-source whitefly population was inherited as an incompletely dominant factor. A model used to estimate loci numbers showed that resistance of whitefly to bifenthrin is probably controlled primarily by a few or a single locus. In addition, the difference in the ratio of LC50 values between males from unmated mother and males from mated mother was approximately fivefold, suggesting that insecticide resistance in whitefly males is in some way affected by mating.     

Inter-regional differences in baseline toxicity of Bemisia argentifolii (Homoptera: Aleyrodidae) to the two insect growth regulators, buprofezin and pyriproxyfen.

A survey of 53 Bemisia argentifolii Bellows & Perring populations from different agricultural regions in California and Arizona was conducted from 1997 to 1999 to establish baseline toxicological responses to buprofezin and pyriproxyfen. Although both compounds proved to be highly toxic even in minute quantities to specific stages, geographical and temporal differences in responses were detected using a leaf spray bioassay technique. Monitoring for three years revealed that six to seven populations had higher LC50 values but not greater survival when exposed to these two insecticides. A significant difference in relative susceptibility to buprofezin was first observed in late season 1997 in San Joaquin Valley populations with LC50s ranging from 16 to 22 microg (AI)/liter(-1) compared with IC50s of 1 to 3 mg (AI)/liter(-1) in Imperial, Palo Verde Valley and Yuma populations. Whiteflies collected in subsequent years from these and other locations showed an increase in susceptibility to buprofezin. Regional differences in susceptibilities to pyriproxyfen were minimal within the same years. Three years of sampling revealed consistently higher LC50s to pyriproxyfen in populations from Palo Verde Valley, CA, compared with whiteflies from Imperial, San Joaquin Valley or Yuma. As was the case with buprofezin, a decline in LC50s to pyriproxyfen was observed in whiteflies from all locations sampled in 1999. However, no correlation was observed between buprofezin and pyriproxyfen toxicity in any of the strains. The variable toxicities observed to both compounds over a period of 3 yr may be due principally to inherent differences among geographical populations or due to past chemical use which may confer positive or negative cross-resistance to buprofezin or pyriproxyfen.     

Variation in susceptibility of Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) (Lepidoptera: Noctuidae) in Australia to two Bacillus thuringiensis toxins

Intra-specific variation in susceptibility of Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) in Australia to the Cry1Ac and Cry2Ab delta-endotoxins from Bacillus thuringiensis (Berliner) (Bt) was determined to establish a baseline for monitoring changes that might occur with the use of Bt cotton. Strains of H. armigera and H. punctigera were established from populations collected primarily from commercial farms throughout the Australian cotton belts. Strains were evaluated for susceptibility using two bioassay methods (surface treatment and diet incorporation) by measuring the dose response for mortality (LC50) and growth inhibition (IC50). The variation in LC50 among H. armigera (n=17 strains) and H. punctigera (n=12 strains) in response to Cry1Ac was 4.6- and 3.2-fold, respectively. The variation in LC50 among H. armigera (n=19 strains) and H. punctigera (n=12 strains) to Cry2Ab was 6.6- and 3.5-fold, respectively. The range of Cry1Ac induced growth inhibition from the 3rd to 4th instar in H. armigera (n=15 strains) was 3.6-fold and in H. punctigera (n=13 strains) was 2.6-fold, while the range of Cry2Ab induced growth inhibition from neonate to 3rd instar in H. armigera (n=13 strains) was 4.3-fold and in H. punctigera (n=12 strains) was 6.1-fold. Variation in susceptibility was also evaluated for two age classes (neonates and 3rd instars) in laboratory strains of H. armigera and H. punctigera. Neonates of H. punctigera had the same or higher sensitivity to Bt than 3rd instars. Neonates of H. armigera were more sensitive to Cry2Ab than 3rd instars, while being less sensitive to Cry1Ac than 3rd instars. Differences in the two methods of bioassay used affected relative sensitivity of species to Bt toxins, highlighting the need to standardize bioassay protocols.     

Molecular characterization of Bemisia tabaci populations in Tunisia: genetic structure and evidence for multiple acquisition of secondary symbionts

A survey was conducted during 2009-2010 seasons to identify the distribution of Bemisia tabaci (Gennadius) biotypes in Tunisia. The genetic affiliation of collected populations was determined by polymerase chain reaction (PCR)-restriction fragment-length polymorphism (TaqI) of the mitochondrial cytochrom oxidase I (mtCOI) gene. Results, validated by sequencing and phylogenetic analysis, allowed the clustering of sampled sweetpotato whiteflies into B and Q biotypes. As B. tabaci harbors the obligatory bacterium Portiera aleyrodidarum, and a diverse array of secondary symbionts including Rickettsia, Hamiltonella, Wolbachia, Cardinium, Arsenophonus, and Fritschea, we report here the infectious status of Tunisian populations by secondary symbionts to find out a correlation between bacterial composition to biotype. The genetic variability and structure of B. tabaci populations in Tunisia was driven by analysis of molecular variance (AMOVA) and the hypothesis of isolation by distance was explored. Selective neutrality and genetic haplotype network tests suggested that Tunisian sweetpotato whiteflies have been undergoing a potential expansion followed by gene flow restriction.     

Baseline susceptibility of Planococcus ficus (Hemiptera: Pseudococcidae) from California to select insecticides

Between 2006 and 2008, 20 populations of Planococcus ficus (Signoret), from Coachella and San Joaquin Valleys of California were measured in the laboratory for susceptibility to buprofezin, chlorpyrifos, dimethoate, methomyl, and imidacloprid. Toxicity was assessed using a petri dish bioassay technique for contact insecticides and by a systemic uptake technique for imidacloprid. Mixed life stages were tested for susceptibility to all insecticides except for buprofezin, which was measured against early and late instars (first, second, and third). Dose-response regression lines from the mortality data established LC50 and LC99 values by both techniques. Responses of populations from the two geographical locations to all five insecticides varied, in some cases significantly. Variations in susceptibility to each insecticide among sample sites showed a sevenfold difference for buprofezin, 11-fold to chlorpyrifos, ninefold to dimethoate, 24-fold to methomyl, and 8.5-fold to imidacloprid. In spite of susceptibility differences between populations, baseline toxicity data revealed that all five insecticides were quite effective based on low LC50s. Chlorpyrifos was the most toxic compound to Planococcus ficus populations as shown by lowest LC50s. Buprofezin was toxic to all immature stages but was more potent to first instars. The highest LC99 estimated by probit analysis of the bioassay data of all 20 populations for each compound was selected as a candidate discriminating dose for use in future resistance monitoring efforts. Establishment of baseline data and development of resistance monitoring tools such as bioassay methods and discriminating doses are essential elements of a sustainable management program for Planococcus ficus.     

Modeling evolution of resistance to pyriproxyfen by the sweetpotato whitefly (Homoptera: Aleyrodidae)

We used computer simulations to examine evolution of resistance to the insect growth regulator (IGR) pyriproxyfen by the sweetpotato whitefly, Bemisia tabaci (Gennadius), biotype B [=Bemisia argentifolii (Bellows & Perring)]. Consistent with trends seen in cotton (Gossipyium spp.) fields in Arizona and Israel, results suggest that evolution of resistance to pyriproxyfen may occur rapidly in this haplodiploid insect. Similar to results from models of diploid insects, resistance evolved faster with increases in toxin concentration, dominance of resistance in females, the initial frequency of the resistance allele, and the proportion of the region treated with pyriproxyfen. Resistance was delayed by fitness costs associated with resistance. Movement between treated and untreated cotton fields had little effect, probably because untreated cotton leaves provided internal refuges in treated fields and whiteflies were controlled with other insecticides in external refuges. Resistance evolved faster when susceptibility to pyriproxyfen was greater in susceptible males than susceptible females. In contrast, resistance evolved slower when susceptibility to pyriproxyfen was greater in resistant males than resistant females. Results suggest that growers may be able to prolong the usefulness of pyriproxyfen by applying lower toxin concentrations and promoting susceptible populations in refuges.     

Over-expression of cytochrome P450 CYP6CM1 is associated with high resistance to imidacloprid in the B and Q biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae)

The two most damaging biotypes of Bemisia tabaci, B and Q, have both evolved strong resistance to the neonicotinoid insecticide imidacloprid. The major mechanism in all samples investigated so far appeared to be enhanced detoxification by cytochrome P450s monooxygenases (P450s). In this study, a polymerase chain reaction (PCR) technology using degenerate primers based on conserved P450 helix I and heme-binding regions was employed to identify P450 cDNA sequences in B. tabaci that might be involved in imidacloprid resistance. Eleven distinct P450 cDNA sequences were isolated and classified as members of the CYP4 or CYP6 families. The mRNA expression levels of all 11 genes were compared by real-time quantitative RT-PCR across nine B and Q field-derived strains of B. tabaci showing strong resistance, moderate resistance or susceptibility to imidacloprid. We found that constitutive over-expression (up to approximately 17-fold) of a single P450 gene, CYP6CM1, was tightly related to imidacloprid resistance in both the B and Q biotypes. Next, we identified three single-nucleotide polymorphic (SNP) markers in the intron region of CYP6CM1 that discriminate between the resistant and susceptible Q-biotype CYP6CM1 alleles (r-Q and s-Q, respectively), and used a heterogeneous strain to test for association between r-Q and resistance. While survivors of a low imidacloprid dose carried both the r-Q and s-Q alleles, approximately 95% of the survivors of a high imidacloprid dose carried only the r-Q allele. Together with previous evidence, the results reported here identify enhanced activity of P450s as the major mechanism of imidacloprid resistance in B. tabaci, and the CYP6CM1 gene as a leading target for DNA-based screening for resistance to imidacloprid and possibly other neonicotinoids in field populations.     

Yellow sticky trap catches of parasitoids of Bemisia tabaci (Hemiptera: Aleyrodidae) in vegetable crops and their relationship to in-field populations

We examined the relationship of yellow sticky trap captures of Bemisia tabaci (Gennadius) biotype B parasitoids to the local population of parasitoids as measured by leaf samples of parasitized whiteflies and mass release of parasitoids. Traps were placed in experimental collard and cowpea field plots in Charleston, SC, and in commercial organic fields of spring cantaloupe and watermelon in the Imperial Valley, CA. The exotic parasitoid Eretmocerus emiratus Zolnerowich and Rose was released in Imperial Valley fields to ensure parasitoid populations would be present. Bemisia adults were trapped in the greatest numbers on the upper surface of horizontally oriented sticky traps in melon fields. In contrast, the lower trap surfaces consistently captured more Eretmocerus than upper surfaces. Female parasitoids were trapped in greater numbers than males, especially on the lower trap surfaces. Progeny of released exotic Eretmocerus greatly outnumbered native E. eremicus Rose and Zolnerowich and Encarsia spp. on traps. Throughout the season, the trend of increasing numbers of Eretmocerus on traps parallelled the increase in numbers of whiteflies. Over the season, 23-84% of all B. tabaci fourth instars were visibly parasitized by Eretmocerus. The numbers of Eretmocerus caught by traps in cantaloupe were similar in trend to numbers on leaf samples in melons, but not with those in watermelon, where whitefly populations were lower. Parasitoid numbers were low in collard and cowpea samples, and no trend was observed in numbers of parasitoids captured on traps and numbers on leaves for these two crops. Overall, there were no significant correlations between sticky trap catches of parasitoids and numbers of parasitized whiteflies on leaf samples in any test fields. Nevertheless, sticky traps placed within crops may be useful for observing trends in whitefly parasitoid populations at a particular site and for detecting parasitoids at specific locations.     

Fitness costs and morphological change of laboratory‐selected thiamethoxam resistance in the B‐type Bemisia tabaci (Hemiptera: Aleyrodidae)

Thiamethoxam has been used as a key insecticide to control the whitefly, B‐type Bemisia tabaci, for several years in China with no known cases of resistance in field populations. To evaluate the risk of resistance, a field population was collected and resistant strains were developed by exposure to thiamethoxam in the laboratory. After selection for 36 generation, a strain with 60‐fold resistance was successfully identified. Fitness analysis by constructing life tables, demonstrated that resistant B‐type whiteflies had obvious fitness disadvantages in their development and reproduction. The fitness of resistant B‐type whiteflies decreased dramatically, to only one‐half that of the susceptible strain. Some changes in the morphological characteristics of the resistant strain were observed. The lengths of first, second and third instars of the resistant strain were significantly smaller than those of the susceptible strain, and the width of the first and the fourth instars were also significantly smaller than in the susceptible strain. Our results suggest that the B‐type B. tabaci has the potential to develop high resistance to thiamethoxam, and that the resistance changed the morphology of the insects. The slow development of resistance and the lower fitness of resistant B. tabaci strains may result in a quick recovery of sensitivity when the population is no longer in contact with thiamethoxam in the field.