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BackgroundThe metallic green beetle, Anomala corpulenta (Coleoptera: Scarabaeidae: Rutelinae), is a destructive pest in agriculture and horticulture throughout Asia, including China. Olfaction plays a crucial role in the survival and reproduction of A. corpulenta. As a non-model species, A. corpulenta is poorly understood, and information regarding the molecular mechanisms underlying olfaction in A. corpulenta and other scarab species is scant.Methodology/Principle FindingsWe assembled separate antennal transcriptome for male and female A. corpulenta using Illumina sequencing technology. The relative abundance of transcripts with gene ontology annotations, including those related to olfaction in males and females was highly similar. Transcripts encoding 15 putative odorant binding proteins, five chemosensory proteins, one sensory neuron membrane protein, 43 odorant receptors, eight gustatory receptors, and five ionotropic receptors were identified. The sequences of all of these chemosensory-related transcripts were confirmed using reverse transcription polymerase chain reaction (RT-PCR), and direct DNA sequencing. The expression patterns of 54 putative chemosensory genes were analyzed using quantitative real time RT-PCR (qRT-PCR). Antenna-specific expression was detected for many of these genes, suggesting that they may have important functions in semiochemical detection.ConclusionsThe identification of a large number of chemosensory proteins provides a major resource for the study of the molecular mechanism of odorant detection in A. corpulenta and its chemical ecology. The genes identified, especially those that were expressed at high levels in the antennae may represent novel molecular targets for the development of population control strategies based on the manipulation of chemoreception-driven behaviors.

Holotrichia parallela (Coleoptera: Scarabaeoidea) is a notorious pest of many crops. To improve the effectiveness of its female-produced sex pheromone (L-isoleucine methyl ester:(R)-(-)-linalool = 6:1), 14 plant volatiles, including dodecanoic acid, dodecanal, farnesol, α-farnesene, (Z)-3-hexen-1-ol, (E)-2-hexen-1-ol, (Z)-3-hexenyl acetate, (E)-2-hexenyl acetate, (R)-(+)-limonene, α-phellandrene, α-pinene, ocimene, methyl benzoate, and benzaldehyde, were individually evaluated using electroantennography and olfactometer assays. (E)-2-Hexenyl acetate and (Z)-3-hexenyl acetate were found to elicit the strongest responses in both males and females. Further testing of these two compounds in mixtures with the sex pheromone indicated that (E)-2-hexenyl acetate had a stronger synergistic effect than (Z)-3-hexenyl acetate. Field evaluations showed that mixtures of (E)-2-hexenyl acetate and the sex pheromone resulted in significantly higher catches than the sex pheromone alone. Using a 5:1 mixture of the sex pheromone and (E)-2-hexenyl acetate, the maximum number of females per trap per day was 14, showing a synergistic effect of a factor of four. For males, a 3:1 mixture of the sex pheromone and (E)-2-hexenyl acetate yielded a maximum number of 310 individuals per trap per day, equivalent to a synergistic effect of 175%. These results may provide the basis for the development of efficient pest management systems against H. parallela using plant volatiles and insect sex pheromones.

Neuropeptides are the most abundant and diverse signal molecules in insects. They act as neurohormones and neuromodulators to regulate the physiology and behavior of insects. The majority of neuropeptides initiate downstream signaling pathways through binding to G protein-coupled receptors (GPCRs) on the cell surface. In this study, RNA-seq technology and bioinformatics were used to search for genes encoding neuropeptides and their GPCRs in the cowpea aphid Aphis craccivora. And the expression of these genes at different developmental stages of A. craccivora was analyzed by quantitative real-time PCR (qRT-PCR). A total of 40 candidate genes encoding neuropeptide precursors were identified from the transcriptome data, which is roughly equivalent to the number of neuropeptide genes that have been reported in other insects. On this basis, software analysis combined with homologous prediction estimated that there could be more than 60 mature neuropeptides with biological activity. In addition, 46 neuropeptide GPCRs were obtained, of which 40 belong to rhodopsin-like receptors (A-family GPCRs), including 21 families of neuropeptide receptors and 7 orphan receptors, and 6 belong to secretin-like receptors (B-family GPCRs), including receptors for diuretic hormone 31, diuretic hormone 44 and pigment-dispersing factor (PDF). Compared with holometabolous insects such as Drosophila melanogaster, the coding genes for sulfakinin, corazonin, arginine vasopressin-like peptide (AVLP), and trissin and the corresponding receptors were not found in A. craccivora. It is speculated that A. craccivora likely lacks the above neuropeptide signaling pathways, which is consistent with Acyrthosiphon pisum and that the loss of these pathways may be a common feature of aphids. In addition, expression profiling revealed neuropeptide genes and their GPCR genes that are differentially expressed at different developmental stages and in different wing morphs. This study will help to deepen our understanding of the neuropeptide signaling systems in aphids, thus laying the foundation for the development of new methods for aphid control targeting these signaling systems.

The dark black chafer, Holotrichia parallela, is an economically important pest in China and worldwide. Traps based on chemical communication are being developed as an alternative control measure to pesticides for this pest, and studies to reveal chemical communication mechanisms in this pest are highly desirable. To systematically analyze genes potentially involved in chemical communication in this pest, we generated a comprehensive transcriptome with combined samples derived from multiple tissues and developmental stages. A total of 43,967 nonredundant sequences (unigenes) with average length of 806 bp were obtained. These unigenes were annotated into different pathways using gene ontology analysis and cluster analysis of orthologous groups of proteins, and kyoto encyclopedia of genes and genomes. In total, 25 transcripts encoding odorant-binding proteins (OBPs) and 16 transcripts encoding chemosensory proteins (CSPs) were identified based on homology searches. Tissue-specific expression profile indicates that OBP17 and CSP7 are likely responsible for male sex pheromone recognition, whereas OBP1-4, OBP9, OBP13-14, OBP17-18, OBP20, OBP22, OBP25, CSP1-7, CSP11, and CSP12-15 are likely responsible for chemical communication between the beetle and environments. Our data shall provide a foundation for further research on the molecular aspects of chemical communication of this insect, and for comparative genomic studies with other species.

Weed resistance to herbicide can be conferred by gene mutations, and some mutations can cause pleiotropic effects in some cases. We investigated the pleiotropic effects associated with five specific ACCase mutations (Ile1781Leu, Trp2027Cys, Ile2041Asn, Asp2078Gly, and Gly2096Ala) on the plant growth, seed production, and resource competitiveness in American sloughgrass. Resistant plants (M/M) homozygous for specific ACCase mutation and susceptible wild‐type plants (W/W) were derived from single heterozygous mother plant (M/W) by genotyping. Plant growth assay and neighborhood experiments were performed to quantify variation between M/M plants and W/W plants. The Ile1781Leu mutation resulted in slight increases in plant growth in pure stands and improved resource competitiveness under low‐competition conditions in pot experiments, but no clear variation was observed under high competitive pressure or field conditions. During competition with wheat plants under field conditions, American sloughgrass plants containing Ile2041Asn ACCase exhibited a significantly lower (12.5%) aboveground biomass but no distinct differences in seed production or resource competitiveness. No significant detrimental pleiotropic effects associated with Gly2096Ala were detected in American sloughgrass. The Trp2027Cys mutation distinctly reduced seed production, especially under high competitive pressure, but did not significantly alter plant growth. The Asp2078Gly mutation consistently reduced not only plant growth and seed production but also resource competitiveness. Synthesis . The Trp2027Cys and Asp2078Gly mutations led to significant fitness costs, which may reduce the frequency of resistance alleles and reduce the propagation speed of resistant weeds in the absence of ACCase inhibitor herbicides. The Ile1781Leu, Ile2041Asn, and Gly2096Ala mutations displayed no obvious fitness costs or displayed very small fitness penalties, which would likely have no effect on the establishment of resistant weeds in the field.

The dark black chafer, Holotrichia parallela Motschulsky, is an economically important pest worldwide. Odorant-based lures and traps are being developed as a key kind of alternative control measures for this pest, and studies to reveal the mechanisms for chemotaxis in this pest are necessary. Two full-length cDNAs encoding different odorant-binding proteins (OBPs) were cloned. The predicted proteins were found to have the functional domains characteristic of typical OBPs and share a high degree of sequence similarity with OBP1 and OBP2 from other insects and were therefore designated as H. parallela OBP-1 and H. parallela OBP-2 (HparOBP-1 and HparOBP-2, respectively). These two OBPs were specifically expressed in antennae. The binding affinity of two purified proteins indicated that HparOBP-1 and HparOBP-2 could selectively interact with various volatiles emitted from host plants and pheromone components. Among the 10 chemicals tested, HparOBP-1 could bind to six of the tested compounds with a dissociation concentration (Ki) less than 20, and HparOBP-2 could bind to three of the compounds. The two OBPs are probably involved in chemotaxis of the dark black chafer. This discovery should accelerate research on chemical communications of this pest, which could potentially lead to the improvement of control measures based on lures and traps.

The dark black chafer (DBC), Holotrichia parallela, is an important pest of multiple crops. Insect host-searching behaviors are regulated by host plant volatiles. Therefore, a better understanding of the mechanism linking the chemosensory system to plant volatiles at the molecular level will benefit DBC control strategies. Based on antenna transcriptome data, two highly expressed antenna-specific odorant-binding proteins (HparOBP20 and 49) were selected to identify novel DBC attractants using reverse chemical ecology methods. We expressed these proteins, mapped their binding specificity, and tested the activity of the plant volatiles in the field. The ligands used in the binding specificity assays included 31 host-plant-associated volatiles and two sex pheromone components. The results showed that (1) HparOBP20 and 49 are involved in odor recognition; (2) these proteins bind attractive plant volatiles strongly and can therefore be employed to develop environmentally friendly DBC management strategies; and (3) the green-leaf volatile (Z)-3-hexenyl acetate shows a high binding affinity to HparOBP20 (Ki = 18.51 μM) and HparOBP49 (Ki = 39.65 μM) and is highly attractive to DBC adults, especially females. In the field test, a (Z)-3-hexenyl acetate trap caught an average of 13 ± 1.202 females per day, which was significantly greater than the corresponding male catch (F2,6 = 74.18, P < 0.0001). (Z)-3-Hexenyl acetate may represent a useful supplement to the known sex pheromone for DBC attraction. In the present study, the binding characteristics of two HparOBPs with host plant volatiles were screened, providing behaviourally active compounds that might be useful for DBC control, based on reverse chemical ecology.