Gissella M. Vásquez

Functional characterization of pheromone receptors in the tobacco budworm Heliothis virescens.

Functional analyses of candidate Heliothis virescens pheromone odorant receptors (HvORs) were conducted using heterologous expression in Xenopus oocytes. HvOR6 was found to be highly tuned to Z9‐14:Ald, while HvOR13, HvOR14 and HvOR16 showed specificity for Z11‐16:Ald, Z11‐16:OAc and Z11‐16:OH, respectively. HvOR15, which had been considered a candidate receptor for Z9‐14:Ald did not respond to any of the pheromone compounds tested, nor to 50 other general odorants. Thus, while HvOR15 is specifically expressed in H. virescens male antennae, its role in pheromone reception remains unknown. Based on our results and previous research we can now assign pheromone receptors in H. virescens males to each of the critical H. virescens agonistic pheromone compounds and two antagonistic compounds produced by heterospecific females.

Intraspecific aggression and colony fusion in the Argentine ant

Unicolonial ants possess an unusual social system characterized by the absence of internest aggression resulting in expansive networks where individuals move freely among distant nests. The Argentine ant, Linepithema humile (Mayr), can form geographically vast and numerically large unicolonial populations, or supercolonies, a trait that has been linked to its ecological success in the introduced range, and is one of the few invasive ants in which native and introduced populations have been examined to elucidate the origins and maintenance of unicoloniality. Supercolony formation may result from mixing of genetically homogenous and nonaggressive colonies, or initially aggressive colonies harbouring the most common recognition alleles. In this study, we examined interactions between mutually aggressive L. humile colonies in the absence of barriers limiting intercolony encounters to determine whether aggressive interactions result in either colony elimination or fusion into new nonaggressive colonies. By pairing experimental laboratory and field colonies displaying varying levels of intraspecific aggression, we determined that pairs that did not fuse had higher numbers of workers fighting and killed than colony pairs that fused and that genetic and cuticular hydrocarbon similarity between colony pairs was correlated with both levels of intraspecific aggression and colony fusion. We suggest that selective fusion of initially aggressive colonies sharing certain recognition cues may be a proximate mechanism shaping L. humile social structure, leading ultimately to extreme unicoloniality in introduced populations when ecological conditions are favourable.

Queen acceptance and the complexity of nestmate discrimination in the Argentine ant

In most social insect species, individuals recognize and behave aggressively towards non-nestmate conspecifics to maintain colony integrity. However, introduced populations of the invasive Argentine ant, Linepithema humile, exhibit pronounced variation in intraspecific aggression denoting diversity in nestmate recognition behavior, which possibly shapes their social structure and the varying levels of unicoloniality observed among these populations. One approach to better understand differential aggression behaviors towards conspecifics and recognition cue perception and response in L. humile is to examine variation in nestmate discrimination capability among genetically distinct colonies under different social contexts. Consequently, we investigated the dynamics of queen and worker recognition in southeastern US L. humile queenless and queenright colonies by measuring rates of non-nestmate worker and queen adoption and intercolony genetic similarity. Aggression levels between colony pairs differed and were associated with non-nestmate worker, but not queen adoption. Adoption of queens and workers was a function of host colony origin, while colony queen number affected adoption of queens, but not workers, with queens more readily accepted by queenless hosts. Fecundity of adopted non-nestmate queens was comparable to that of rejected non-nestmate and host colony queens, suggesting that queen fecundity did not affect adoption decisions. Genetic similarity between colonies ranged from 30 to 77% alleles shared, with more genetically similar colonies showing lower levels of intraspecific aggression. Non-nestmate queens and workers that were more genetically similar to host colony workers were more likely to be adopted. We provide the first evidence for the role of L. humile colony queen number on queen discrimination and suggest an effect of resident queens on worker conspecific acceptance thresholds. Our findings indicate a role for genetically based cues in L. humile nestmate recognition. However, subtle discrimination capability seems to be influenced by the social context, as demonstrated by more frequent recognition errors in queenless colonies.

Geographic variation in sexual attraction of Spodoptera frugiperda corn- and rice-strain males to pheromone lures

The corn- and rice-strains of Spodoptera frugiperda exhibit several genetic and behavioral differences and appear to be undergoing ecological speciation in sympatry. Previous studies reported conflicting results when investigating male attraction to pheromone lures in different regions, but this could have been due to inter-strain and/or geographic differences. Therefore, we investigated whether corn- and rice-strain males differed in their response to different synthetic pheromone blends in different regions in North America, the Caribbean and South America. All trapped males were strain-typed by two strain-specific mitochondrial DNA markers. In the first experiment, we found a nearly similar response of corn- and rice-strain males to two different 4-component blends, resembling the corn- and rice-strain female blend we previously described from females in Florida. This response showed some geographic variation in fields in Canada, North Carolina, Florida, Puerto Rico, and South America (Peru, Argentina). In dose-response experiments with the critical secondary sex pheromone component (Z)-7-dodecenyl acetate (Z7-12:OAc), we found some strain-specific differences in male attraction. While the response to Z7-12:OAc varied geographically in the corn-strain, rice-strain males showed almost no variation. We also found that the minor compound (Z)-11-hexadecenyl acetate (Z11-16:OAc) did not increase attraction of both strains in Florida and of corn-strain males in Peru. In a fourth experiment, where we added the stereo-isomer of the critical sex pheromone component, (E)-7-dodecenyl acetate, to the major pheromone component (Z)-9-tetradecenyl acetate (Z9-14:OAc), we found that this compound was attractive to males in North Carolina, but not to males in Peru. Overall, our results suggest that both strains show rather geographic than strain-specific differences in their response to pheromone lures, and that regional sexual communication differences might cause geographic differentiation between populations.

Colony fusion in Argentine ants is guided by worker and queen cuticular hydrocarbon profile similarity.

Introduced populations of the Argentine ant, Linepithema humile, have experienced moderate to severe losses of genetic diversity, which may have affected nestmate recognition to various degrees. We hypothesized that cuticular hydrocarbons (CHC) serve as nestmate recognition cues, and facilitate colony fusion of unrelated L. humile colonies that share similar CHC profiles. In this study, we paired six southeastern U.S. L. humile colonies in a 6-month laboratory fusion assay, and determined if worker and queen CHC profile similarity between colonies was associated with colony fusion and intercolony genetic similarity. We also compared worker and queen CHC profiles between fused colony pairs and unpaired controls to determine if worker and queen chemical profiles changed after fusion. We found that colony fusion correlated with the CHC similarity of workers and queens, with the frequency of fusion increasing with greater CHC profile similarity between colonies. Worker and queen CHC profile similarity between colonies also was associated with genetic similarity between colonies. Queen CHC profiles in fused colonies appeared to be a mix of the two colony phenotypes. In contrast, when only one of the paired colonies survived, the CHC profile of the surviving queens did not diverge from that of the colony of origin. Similarly, workers in non-fused colonies maintained their colony-specific CHC, whereas in fused colonies the worker CHC profiles were intermediate between those of the two colonies. These results suggest a role for CHC in regulating interactions among mutually aggressive L. humile colonies, and demonstrate that colony fusion correlates with both genetic and CHC similarities. Further, changes in worker and queen chemical profiles in fused colonies suggest that CHC plasticity may sustain the cohesion of unrelated L. humile colonies that had fused.

Differential expression of odorant receptor genes involved in the sexual isolation of two Heliothis moths

Moth sexual communication systems are highly diverse, but the mechanisms underlying their evolutionary diversification remain unclear. Recently, genes coding for odorant receptors (ORs) OR6, OR14, OR15 and OR16 have been genetically associated with species‐specific male response to female pheromone blends in Heliothis virescens (Hv) and Heliothis subflexa (Hs). Quantitative real‐time PCR analysis indicates that expression of HvOR6, HsOR6, HvOR14, HsOR14, HvOR15 and HsOR15 is male biased, which supports the hypothesis that they have a role in mediating female sex pheromone detection. The genes HvOR14, HvOR15 and HvOR16 are expressed at higher levels than their corresponding orthologues HsOR14, HsOR15 and HsOR16 in male antennae, while HvOR6 and HsOR6 transcripts are equally abundant in male antennae. The lack of higher expression of any of the receptor genes in H. subflexa antennae suggests that interspecific sequence differences, rather than gene regulation differences, underly the species‐specific male response to pheromone components.

Cuticular hydrocarbons as queen adoption cues in the invasive Argentine ant

SUMMARY In social insects, individuals typically recognize and behave aggressively towards alien conspecifics, thereby maintaining colony integrity. This is presumably achieved via a nestmate recognition system in which cuticular compounds, usually cuticular hydrocarbons (CHC), of genetic and/or environmental origin serve as recognition cues. Most invasive populations of the Argentine ant, Linepithema humile (Mayr), display minimal nestmate–non-nestmate discrimination, resulting in low levels of intraspecific aggression allowing free movement of workers and queens among nests. However, invasive L. humile in the southeastern United States show relatively high levels of intraspecific aggression, and selectively adopt non-nestmate queens. Using behavioral assays and gas chromatography, we found an association between non-nestmate queen adoption and similarity of the CHC profiles of adopted and host colony queens. Also, nestmate and non-nestmate queen CHC profiles became more similar after adoption by queenless colonies. Furthermore, queens treated with non-nestmate queen CHC had distinct CHC profiles and were generally attacked by nestmate workers. We suggest that in L. humile, CHC are used as queen recognition cues, and that queen recognition errors are more likely to occur when the CHC profiles of non-nestmate and host colony queens are similar. Our findings provide further evidence for the complex and dynamic nature of L. humile nestmate discrimination, which may in part underlie the success of introduced populations of this invasive ant.

Specificity of the Receptor for the Major Sex Pheromone Component in Heliothis virescens

Abstract In a previous study, the Drosophila melanogaster OR67dGAL4;UAS system was used to functionally characterize the receptor for the major component of the sex pheromone in the tobacco budworm, Heliothis virescens Fabricius (Lepidoptera: Noctuidae), HvOR13. Electrophysiological and behavioral assays showed that transgenic flies expressing HvOR13 responded to (Z)-11-hexadecenal (Z11-16:Ald). However, tests were not performed to determine whether these flies would also respond to secondary components of the H. virescens sex pheromone. Thus, in this study the response spectrum of HvOR13 expressed in this system was examined by performing single cell recordings from odor receptor neuron in trichoid T1 sensilla on antennae of two Or67dGAL4 [1]; UAS-HvOR13 lines stimulated with Z11-16:Ald and six H. virescens secondary pheromone components. Fly courtship assays were also performed to examine the behavioral response of the Or67dGAL4[1]; UAS-HvOR13 flies to Z11-16:Ald and the secondary component Z9-14:Ald. Our combined electrophysiological and behavioral studies indicated high specificity and sensitivity of HvOR13 to Z11-16:Ald. Interestingly, a mutation leading to truncation in the HvOR13 C-terminal region affected but did not abolish pheromone receptor response to Z11-16:Ald. The findings are assessed in relationship to other HvOR13 heterologous expression studies, and the role of the C-terminal domain in receptor function is discussed. A third line expressing HvOR15 was also tested but did not respond to any of the seven pheromone components.

Quality Assessment of Selected Commercially Available Whitefly and Aphid Biological Control Agents in the United States

Abstract This study assessed the quality of three commercially available natural enemies used for pest management in greenhouses: the whitefly parasitoid Encarsia formosa Gahan (Hymenoptera: Aphelinidae), the aphid parasitoid Aphidius colemani Viereck (Hymenoptera: Braconidae), and the aphid predatory midge Aphidoletes aphidimyza (Rondani) (Diptera: Cecidomyiidae). Shipment packaging was consistent for all natural enemies. However, there was high variability in delivery punctuality, product cost, and product information provided by each of the six selected companies. Product quantity, percentage of emergence upon arrival, percentage of total emergence, percentage of females, and percentage of flying insects were assessed using International Organization for Biological Control (IOBC) recommended procedures. The parameters with greatest variability between companies were percentage of emergence upon arrival (0.9–10.5%) and percentage of flying insects (35.4–85.0%) for E. formosa; product quantity (623.3–833.8 aphid mummies), percentage of emergence upon arrival (6.1–41.2%) and percentage of females (51.1–54.8%) for A. colemani; and percentage of emergence upon arrival (0.0–7.7%) and percentage of females (54.6–76.2%) for A. aphidimyza. Results are discussed in terms of the value to consumers and compared with IOBC standards.

Fusion between Southeastern United States Argentine Ant Colonies and Its Effect on Colony Size and Productivity

ABSTRACT The ecological success of invasive ants has been linked to their ability to form expansive supercolonies. In the Argentine ant, Linepithema humile (Mayr), increased productivity and competitive ability of introduced supercolonies in several places, e.g., California and southern Europe, has been linked to high population densities that could have been attained via fusion of nonaggressive and genetically similar nests. Recently, we have found that introduced L. humile colonies in the southeastern United States, which have higher levels of intraspecific aggression and genetic diversity than those in California and southern Europe, sometimes also fuse; yet it is unclear what the longer term consequences of such colony fusion might be. In this study, we examined whether fusion of these southeastern United States L. humile colonies results in larger colonies by recording colony size and productivity in pairs that fused and in pairs that did not fuse. After 6 mo, colonies that fused produced 47% more workers and had twice as many queens as colony pairs that did not fuse. Also, fused colonies had an overall per capita colony productivity (number of brood and workers produced per queen and per worker) comparable to that of nonfused pairs and unpaired controls. Furthermore, all queens contributed to worker pupae production in fused colonies. Thus, fusion of initially aggressive southeastern United States L. humile colonies results in colonies with higher worker number without decreasing per capita productivity. Moreover, offspring contribution by all queens in fused colonies may alter colony genotypic composition resulting in reduced intraspecific aggression that in turn promotes further fusion. This process may be relevant to the establishment of incipient colonies in areas where multiple introductions have occurred.