Evolution of chemosensory genes in Colorado potato beetle, Leptinotarsa decemlineata.

Associating with plant hosts is thought to have elevated the diversification of insect herbivores, which comprise the majority of global species diversity. In particular, there is considerable interest in understanding the genetic changes that allow host-plant shifts to occur in pest insects and in determining what aspects of functional genomic diversity impact host-plant breadth. Insect chemoreceptors play a central role in mediating insect-plant interactions, as they directly influence plant detection and sensory stimuli during feeding. Although chemosensory genes evolve rapidly, it is unclear how they evolve in response to host shifts and host specialization. We investigate whether selection at chemosensory genes is linked to host-plant expansion from the buffalo burr, Solanum rostratum, to potato, Solanum tuberosum, in the super-pest Colorado potato beetle (CPB), Leptinotarsa decemlineata (Coleoptera: Chrysomelidae). First, to refine our knowledge of CPB chemosensory genes, we developed novel gene expression data for the antennae and maxillary-labial palps. We then examine patterns of selection at these loci within CPB, as well as compare whether rates of selection vary with respect to 9 closely related, non-pest Leptinotarsa species that vary in diet breadth. We find that rates of positive selection on olfactory receptors are higher in host-plant generalists, and this signal is particularly strong in CPB. These results provide strong candidates for further research on the genetic basis of variation in insect chemosensory performance and novel targets for pest control of a notorious super-pest.

Diversity of the Antimicrobial Peptide Genes in Collembola.

Multidrug-resistant bacteria are a current health crisis threatening the world's population, and scientists are looking for new drugs to combat them. Antimicrobial peptides (AMPs), which are part of the organism's innate immune system, are a promising new drug class as they can disrupt bacterial cell membranes. This study explored antimicrobial peptide genes in collembola, a non-insect hexapod lineage that has survived in microbe-rich habitats for millions of years, and their antimicrobial peptides have not been thoroughly investigated. We used in silico analysis (homology-based gene identification, physicochemical and antimicrobial property prediction) to identify AMP genes from the genomes and transcriptomes of five collembola representing three main suborders: Entomobryomorpha (Orchesella cincta, Sinella curviseta), Poduromorpha (Holacanthella duospinosa, Anurida maritima), and Symphypleona (Sminthurus viridis). We identified 45 genes belonging to five AMP families, including (a) cysteine-rich peptides: diapausin, defensin, and Alo; (b) linear α-helical peptide without cysteine: cecropin; (c) glycine-rich peptide: diptericin. Frequent gene gains and losses were observed in their evolution. Based on the functions of their orthologs in insects, these AMPs potentially have broad activity against bacteria, fungi, and viruses. This study provides candidate collembolan AMPs for further functional analysis that could lead to medicinal use.

Genome-Wide Identification of the Gustatory Receptor Gene Family of the Invasive Pest, Red Palm Weevil, Rhynchophorus ferrugineus (Olivier, 1790).

The red palm weevil (Rhynchophorus ferrugineus) is a highly destructive pest of oil palm, date, and coconut in many parts of Asia, Europe, and Africa. The Food and Agriculture Organization of the United Nations has called for international collaboration to develop a multidisciplinary strategy to control this invasive pest. Previous research focused on the molecular basis of chemoreception in this species, particularly olfaction, to develop biosensors for early detection and more effective bait traps for mass trapping. However, the molecular basis of gustation, which plays an essential role in discriminating food and egg-laying sites and chemical communication in this species, is limited because its complete gustatory receptor gene family still has not been characterized. We manually annotated the gene family from the recently available genome and transcriptome data and reported 50 gustatory receptor genes encoding 65 gustatory receptors, including 7 carbon dioxide, 9 sugar, and 49 bitter receptors. This study provides a platform for future functional analysis and comparative chemosensory study. A better understanding of gustation will improve our understanding of this species' complex chemoreception, which is an important step toward developing more effective control methods.

Identification of Novel Toxin Genes from the Stinging Nettle Caterpillar Parasa lepida (Cramer, 1799): Insights into the Evolution of Lepidoptera Toxins.

Many animal species can produce venom for defense, predation, and competition. The venom usually contains diverse peptide and protein toxins, including neurotoxins, proteolytic enzymes, protease inhibitors, and allergens. Some drugs for cancer, neurological disorders, and analgesics were developed based on animal toxin structures and functions. Several caterpillar species possess venoms that cause varying effects on humans both locally and systemically. However, toxins from only a few species have been investigated, limiting the full understanding of the Lepidoptera toxin diversity and evolution. We used the RNA-seq technique to identify toxin genes from the stinging nettle caterpillar, Parasa lepida (Cramer, 1799). We constructed a transcriptome from caterpillar urticating hairs and reported 34,968 unique transcripts. Using our toxin gene annotation pipeline, we identified 168 candidate toxin genes, including protease inhibitors, proteolytic enzymes, and allergens. The 21 P. lepida novel Knottin-like peptides, which do not show sequence similarity to any known peptide, have predicted 3D structures similar to tarantula, scorpion, and cone snail neurotoxins. We highlighted the importance of convergent evolution in the Lepidoptera toxin evolution and the possible mechanisms. This study opens a new path to understanding the hidden diversity of Lepidoptera toxins, which could be a fruitful source for developing new drugs.

Molecular characterization and evolution of a chemosensory receptor gene family in three notorious rice planthoppers, Nilaparvata lugens, Sogatella furcifera and Laodelphax striatellus, based on genome and transcriptome analyses.

BACKGROUND: The white-backed planthopper (WBPH) Sogatella furcifera, the brown planthopper (BPH) Nilaparvata lugens, and the small brown planthopper (SBPH) Laodelphax striatellus (Hemiptera: Delphacidae) are rice pests that damage rice plants by sap-sucking and by transmitting viruses. Host-seeking behavior involves chemosensory receptor genes that include odorant receptors (ORs), ionotropic receptors (IRs) and gustatory receptors (GRs). RESULTS: We used genome and transcriptome data to identify 141 ORs, 28 GRs and 25 IRs in BPH; 135 ORs, 18 GRs and 16 IRs in WBPH; and 37 ORs, 14 GRs and 6 IRs in SBPH. A phylogenetic analysis identified several specific OR clades of rice planthoppers, the results indicating that these OR members might be used to respond to specific host volatiles. OR co-receptor (Orco) is the most conserved and essential OR gene among these species and RNA interference (RNAi) can decrease their mRNA expression level to <50%. RNAi knockdown rice planthoppers were anosmia and were unable to seek or locate rice plants in behavioral tests. CONCLUSION: The results demonstrate the importance of the planthopper Orco genes in locating rice plants. This information may aid in the development of RNAi-based transgenic rice and other pest management technologies. © 2018 Society of Chemical Industry.

A model species for agricultural pest genomics: the genome of the Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae).

The Colorado potato beetle is one of the most challenging agricultural pests to manage. It has shown a spectacular ability to adapt to a variety of solanaceaeous plants and variable climates during its global invasion, and, notably, to rapidly evolve insecticide resistance. To examine evidence of rapid evolutionary change, and to understand the genetic basis of herbivory and insecticide resistance, we tested for structural and functional genomic changes relative to other arthropod species using genome sequencing, transcriptomics, and community annotation. Two factors that might facilitate rapid evolutionary change include transposable elements, which comprise at least 17% of the genome and are rapidly evolving compared to other Coleoptera, and high levels of nucleotide diversity in rapidly growing pest populations. Adaptations to plant feeding are evident in gene expansions and differential expression of digestive enzymes in gut tissues, as well as expansions of gustatory receptors for bitter tasting. Surprisingly, the suite of genes involved in insecticide resistance is similar to other beetles. Finally, duplications in the RNAi pathway might explain why Leptinotarsa decemlineata has high sensitivity to dsRNA. The L. decemlineata genome provides opportunities to investigate a broad range of phenotypes and to develop sustainable methods to control this widely successful pest.

Diversification of the ant odorant receptor gene family and positive selection on candidate cuticular hydrocarbon receptors.

BACKGROUND: Chemical communication plays important roles in the social behavior of ants making them one of the most successful groups of animals on earth. However, the molecular evolutionary process responsible for their chemosensory adaptation is still elusive. Recent advances in genomic studies have led to the identification of large odorant receptor (Or) gene repertoires from ant genomes providing fruitful materials for molecular evolution analysis. The aim of this study was to test the hypothesis that diversification of this gene family is involved in olfactory adaptation of each species. RESULTS: We annotated the Or genes from the genome sequences of two leaf-cutter ants, Acromyrmex echinatior and Atta cephalotes (385 and 376 putative functional genes, respectively). These were used, together with Or genes from Camponotus floridanus, Harpegnathos saltator, Pogonomyrmex barbatus, Linepithema humile, Cerapachys biroi, Solenopsis invicta and Apis mellifera, in molecular evolution analysis. Like the Or family in other insects, ant Or genes evolve by the birth-and-death model of gene family evolution. Large gene family expansions involving tandem gene duplications, and gene gains outnumbering losses, are observed. Codon analysis of genes in lineage-specific expansion clades revealed signatures of positive selection on the candidate cuticular hydrocarbon receptor genes (9-exon subfamily) of Cerapachys biroi, Camponotus floridanus, Acromyrmex echinatior and Atta cephalotes. Positively selected amino acid positions are primarily in transmembrane domains 3 and 6, which are hypothesized to contribute to the odor-binding pocket, presumably mediating changing ligand specificity. CONCLUSIONS: This study provides support for the hypothesis that some ant lineage-specific Or genes have evolved under positive selection. Newly duplicated genes particularly in the candidate cuticular hydrocarbon receptor clade that have evolved under positive selection may contribute to the highly sophisticated lineage-specific chemical communication in each ant species.

Molecular evolution of the odorant and gustatory receptor genes in lepidopteran insects: implications for their adaptation and speciation.

Lepidoptera (comprised of butterflies and moths) is one of the largest groups of insects, including more than 160,000 described species. Chemoreception plays important roles in the adaptation of these species to a wide range of niches, e.g., plant hosts, egg-laying sites, and mates. This study investigated the molecular evolution of the lepidopteran odorant (Or) and gustatory receptor (Gr) genes using recently identified genes from Bombyx mori, Danaus plexippus, Heliconius melpomene, Plutella xylostella, Heliothis virescens, Manduca sexta, Cydia pomonella, and Spodoptera littoralis. A limited number of cases of large lineage-specific gene expansion are observed (except in the P. xylostella lineage), possibly due to selection against tandem gene duplication. There has been strong purifying selection during the evolution of both lepidopteran odorant and gustatory genes, as shown by the low ω values estimated through CodeML analysis, ranging from 0.0093 to 0.3926. However, purifying selection has been relaxed on some amino acid sites in these receptors, leading to sequence divergence, which is a precursor of positive selection on these sequences. Signatures of positive selection were detected only in a few loci from the lineage-specific analysis. Estimation of gene gains and losses suggests that the common ancestor of the Lepidoptera had fewer Or genes compared to extant species and an even more reduced number of Gr genes, particularly within the bitter receptor clade. Multiple gene gains and a few gene losses occurred during the evolution of Lepidoptera. Gene family expansion may be associated with the adaptation of lepidopteran species to plant hosts, especially after angiosperm radiation. Phylogenetic analysis of the moth sex pheromone receptor genes suggested that chromosomal translocations have occurred several times. New sex pheromone receptors have arisen through tandem gene duplication. Positive selection was detected at some amino acid sites predicted to be in the extracellular and transmembrane regions of the newly duplicated genes, which might be associated with the evolution of the new pheromone receptors.

The red flour beetle's large nose: an expanded odorant receptor gene family in Tribolium castaneum.

The Tribolium castaneum genome sequence reveals a large number of odorant receptor (Or) genes compared to those found in other insects whose olfactory genomes have been studied-341 Or genes and pseudogenes, encoding 259 intact odorant receptor proteins. An RT-PCR study of larvae and adults revealed that only 145 (64%) of 233 genes with successful genomic DNA amplifications were expressed. No expression of the other 87 genes was detected at any age, suggesting either that these genes are not expressed in this particular strain, or that they are induced only in certain environmental or developmental conditions. TcOR1, the ortholog of the Drosophila Or83b (DmOr83b) gene, which is required for the function of olfactory receptor proteins in Drosophila, was expressed in extracts from adult and larval heads and in extracts from adult bodies. Expression of 41 TcOr genes was detected in extracts from larval head tissue and 111 in extracts from adult head tissue (both figures exclude TcOr1). Twenty-eight TcOrs were detected only in adult bodies. Beetle pupae were injected with TcOr1 dsRNA; unlike sham-injected and control beetles, these knock-down beetles showed no significant response to the Tribolium aggregation pheromone, supporting the hypothesis that TcOr1 plays a similar decisive role in olfaction to DmOr83b. The substantial number of Ors poses the question of why Tribolium has such a large olfactory receptor repertoire, and underlines the need for more studies of the natural history of this species.