Determinants of Insecticide Resistance Evolution: Comparative Analysis Among Heliothines

It is increasingly clear that pest species vary widely in their propensities to develop insecticide resistance. This review uses a comparative approach to analyze the key pest management practices and ecological and biochemical or genetic characteristics of the target that contribute to this variation. We focus on six heliothine species, three of which, Helicoverpa armigera, Heliothis virescens, and Helicoverpa zea, have developed resistances to many pesticide classes. The three others, Helicoverpa punctigera, Helicoverpa assulta, and Helicoverpa gelotopoeon, also significant pests, have developed resistance to very few pesticide classes. We find that host range and movement between alternate hosts are key ecological traits that influence effective selection intensities for resistance. Operational issues are also critical; area-wide, cross-pesticide management practices that account for these ecological factors are key to reducing selection intensity. Without such management, treatment using broad-spectrum chemicals serves to multiply the effects of host plant preference, preadaptive detoxification ability, and high genetic diversity to create a pesticide treadmill for the three high-propensity species.Without rigorous ongoing management, such a treadmill could still develop for newer, more selective chemistries and insecticidal transgenic crops.

Habituation to a Deterrent Plant Alkaloid Develops Faster in the Specialist Herbivore Helicoverpa assulta Than in Its Generalist Congener Helicoverpa armigera and Coincides with Taste Neuron Desensitisation

The two closely related moth species, Helicoverpa armigera and H. assulta differ strongly in their degree of host-plant specialism. In dual-choice leaf disk assays, caterpillars of the two species that had been reared on standard artificial diet were strongly deterred by the plant-derived alkaloid strychnine. However, caterpillars of both species reared on artificial diet containing strychnine from neonate to the 5th instar were insensitive to this compound. Fifth instar caterpillars of H. assulta and 4th or 5th instars of H. armigera not exposed to strychnine before were subjected to strychnine-containing diet for 24 h, 36 h, 48 h, or 72 h. Whereas H. assulta displayed habituation to strychnine after 48 h, it took until 72 h for H. armigera to become habituated. Electrophysiological tests revealed that a deterrent-sensitive neuron in the medial sensillum styloconicum of both species displayed significantly reduced sensitivity to strychnine that correlated with the onset of habituation. We conclude that the specialist H. assulta habituated faster to strychnine than the generalist H. armigera and hypothesis that desensitization of deterrent-sensitive neurons contributed to habituation.

A chemical basis for different oviposition rates of Helicoverpa assulta across two Nicotiana species

The tobacco Nicotiana rustica is widely used as a trap crop in the fields of Nicotiana tabacum in China, by attracting oviposition of Helicoverpa assulta females, thus preventing damage to N. tabacum. The mechanism underlying the differential oviposition rates of H. assulta across these two tobacco species, however, is largely unknown. We investigated the mechanism of host plant acceptance of H. assulta with respect to these two tobaccos by using a two-choice behavioral bioassay and GC–MS. Our results indicate that both the leaves and inflorescences of N. rustica attracted significantly more eggs than the corresponding parts of N. tabacum. Extracts of leaves and inflorescences of N. rustica with two different solvents elicited similar oviposition patterns to the corresponding parts of the plants. Chemical analysis by GC–MS revealed that the volatiles of N. rustica contain larger amounts of nicotine than those of N. tabacum at the flowering stage. In addition, γ-terpinolene and β-elemene are found only in extracts of N. rustica. A two-choice bioassay on the individual compounds showed that γ-terpinolene, which is specific to the vegetative stage of N. rustica, and nicotine attracted oviposition by H. assulta. The volatile β-elemene, which is present only in N. rustica, was also attractive. We conclude that the larger amount of nicotine, and the species-specific γ-terpinolene and β-elemene may mediate the different oviposition rates of H. assulta females across N. rustica and N. tabacum.

A moth odorant receptor highly expressed in the ovipositor is involved in detecting host-plant volatiles

Antennae are often considered to be the nostrils of insects. Here, we sequenced the transcriptome of the pheromone gland-ovipositor complex of Helicoverpa assulta and discovered that an odorant receptor (OR) gene, HassOR31, had much higher expression in the ovipositor than in antennae or other tissues. To determine whether the ovipositor was involved in odorant detection, we co-expressed HassOR31 and its co-receptor, HassORco, in a Xenopus oocyte model system, and demonstrated that the OR was responsive to 12 plant odorants, especially Z-3-hexenyl butyrate. These odorants elicited electrophysiological responses of some sensilla in the ovipositor, and HassOR31 and HassORco were co-expressed within ovipositor sensilla. Two oviposition preference experiments showed that female moths lacking antennae still preferentially selected oviposition sites containing plant volatiles. We suggest that the expression of HassOR31 in the ovipositor of H. assulta helps females to determine precise egg-laying sites in host plants.

Identification of the Differences in Olfactory System Between Male and Female Oriental Tobacco Budworm Helicoverpa assulta

Background: The olfactory system of insects facilitates their search for host and mates, hence it plays an essential role for insect survival and reproduction. Insects recognize odor substances through olfactory neurons and olfactory genes. Previous studies showed that there are significant sex-specific differences in how insects identify odorant substances, especially sex pheromones. However, whether the sex-specific recognition of odorant substances is caused by differences in the expression of olfaction-related genes between males and females remains unclear. Results: In order to clarify this problem, the whole transcriptome sequence of the adult Helicoverpa assulta, an important agricultural pest of tobacco and other Solanaceae plants, was obtained using PacBio sequencing. RNA-seq analysis showed that there were 27 odorant binding proteins (OBPs), 24 chemosensory proteins (CSPs), 4 pheromone-binding proteins (PBPs), 68 odorant receptors (OR) and 2 sensory neuron membrane proteins (SNMPs) genes that were expressed in the antennae of male and female H. assulta. Females had significantly higher expression of GOBP1-like, OBP, OBP3, PBP3 and SNMP1 than males, while males had significantly higher expression of GOBP1, OBP7, OBP13, PBP2 and SNMP2. Conclusions: Our findings improve our understanding of olfactory genes in H. assulta, and can be used to further study pheromone identification, mate search, and sex differences in an insect’s ability to search for hosts. These results improve our understanding of mate search and host differentiation in H. assulta.