Comparative Biochemistry and Physiology Part D: Genomics and Proteomics
Pheromone biosynthetic pathway and chemoreception proteins in sex pheromone gland of Eogystia hippophaecolus
Graphical abstract
Introduction
Pheromone trapping of moths and other insects based on species-specific sex pheromone-elicited sexual communication and reproduction behaviors is important for integrated pest management practices worldwide. In moths, sex pheromones are multicomponent blends of C10–C18 long-chain unsaturated alcohols, aldehydes, and acetate esters produced and released through a modified fatty acid biosynthesis pathway in sex pheromone glands (PGs) which situate in the intersegmental membrane between the 8-9th abdominal segments in females (Roelofs and Rooney, 2003; Cardé and Haynes, 2004; Jurenka, 2004). Adult male moth antennae perceive female sex pheromone components from afar specifically, and this allows them to successfully locate females to initiate mating (Wyatt, 2009). Among Lepidoptera, pheromone components largely differ between species, but some share the same components in different combinations, as demonstrated in Helicoverpa armigera and Helicoverpa assulta that participate in the mutual sex pheromone components Z11-16:Ald and Z9-16:Ald, although the ratio of them is contrary entirely (7:100 in H. assulta and 100:7 in H. armigera) (Wang et al., 2005; Wu et al., 2013; Li et al., 2015). In moths, multiple varying combinations of reactions produce distinct species-specific pheromone mixtures among different species, due to management of relevant sex pheromone enzymes, leading to speciation via changes in mate recognition systems (Li et al., 2015).
Sex pheromone biosynthesis makes use of a revised fatty acid biosynthesis pathway (Jurenka, 2004; Matsumoto, 2010; Tillman et al., 1999) which contains acetylation, desaturation, chain shortening, reduction, and oxidation by gland-specific enzymes processes, either separately or in combination variedly (Jurenka, 2004). Pheromone biosynthesis-activating neuropeptides (PBANs) was released from the subesophageal ganglion and transportation to the PG, where they bind to a pheromone biosynthesis-activating neuropeptide receptor (PBANr), which is the first step of a characteristic moth pheromone biosynthetic pathway (Zhu et al., 1995; Jurenka, 2017). Then in turn activates functional group modification enzymes (Jurenka, 2004; Matsumoto et al., 2007; Matsumoto et al., 2010) or acetyl-coenzyme A (CoA) carboxylase (ACC) (Blomquist et al., 2012). The resulting signal triggers several enzyme pathways began by ACC that stimulates the conversion of acetyl-CoA to malonyl-CoA, leading to synthesize pheromone precursors (mostly 12, 14, 16, or 18 carbon saturated fatty acids) (Volpe and Vagelos, 1973). NADPH and malonyl-CoA are catalyzed by fatty acid synthases (FASs) to generate most saturated palmitic acid (16:0) and stearic acid (18:0) precursors with16 and 18 carbon atoms, respectively, without double bonds (Bjostad and Roelofs, 1984; Tang et al., 1989; Jurenka et al., 1991). Fatty acid transport proteins (FATPs) then transport these precursors into the PG, and double bonds are introduced at peculiar locations (Δ5 (Foster and Roelofs, 1996), Δ6 (Wang et al., 2010), Δ9 (Martinez et al., 1990; Albre et al., 2012), Δ10 (Foster and Roelofs, 1988; Hao et al., 2002), Δ11 (Liénard et al., 2010b; Ding et al., 2016), and Δ14 (Roelofs et al., 2002)) by different desaturases (DESs), many of which were demonstrated by clone and expression in many moth species functionally (Rafaeli, 2005; Blomquist et al., 2012), then β-oxidation shorten the chain (Wang et al., 2005). The carboxyl carbon of formative unsaturated pheromone precursors with a specific chain length is modified by fatty acid reductase (FAR) enzymes to generate alcohols from fatty acyl precursors (Lassance et al., 2013), which is oxidized to aldehydes in some species likely (Teal and Tumlinson, 1987), or transformed to acetate esters (OAcs) by acetyltransferase (ACT) (Roelofs and Wolf, 1988). In addition to FATPs, acyl-CoA-binding proteins (ACBPs) have been demonstrated to work on production of the sex pheromone bombykol in Bombyx mori by in vivo RNA interference (RNAi) (Liénard et al., 2010a; Lassance et al., 2013). Besides DES and FAR B. mori are showed that are responsible for pheromone production by the same way (Moto et al., 2003; Moto et al., 2004; Ohnishi et al., 2006). However, other critical enzymes listed above, such as ACC, PBAN, and FAS, have not been demonstrated at the enzymatic level in insects. What's more, a change in Des3, Des5 and FAR17 in Spodoptera litura could change the ratio of the sex pheromone components, which showed that key genes of the sex pheromone biosynthesis pathway may lead to differential ratios of the sex pheromones in the field (Lin et al., 2018).
Males have developed a subtle antennal olfaction system that can trace exceptionally small amounts of sex pheromone components which delivered from females in long distance (Vogt, 2005). The first step of olfactory recognition process involves in the abundant odorant-binding proteins (OBPs) and chemosensory proteins (CSPs) in the lymph of sensilla and pheromones and plant volatiles were binding, transported and solvated by them (Kaissling, 2001; Ban et al., 2002; Leal et al., 2005; Pelosi et al., 2006). Pheromone-binding proteins (PBPs) are a subfamily of OBPs (Zhou, 2010) that bind to pheromone compounds and participate in the pheromone recognition process. After capturing odors, transport odors and OBP-odor complexes were transported to four membrane receptor types: odorant receptors (ORs), gustatory receptors (GRs), ionotropic receptors (IRs), and sensory neuron membrane proteins (SNMPs), which are situated in the outer dendrites of olfactory receptor neurons. Depending on the different pheromones functional groups, aldehydes, alcohols, or esters, degradation involves specific enzymes in odor degradation pathways, such as multi-functional odorant-degrading enzymes (ODEs), pheromone-degrading enzymes (PDEs), and antennal esterases (AES).
In 2003, the sea buckthorn carpenterworm Eogystia hippophaecolus (Hua et al.) (Lepidoptera: Cossidae) destroy 66,500 ha of sea buckthorn Hippophae rhamnoides L. (Rosales: Elaeagnaceae) in western and northern China that stems soil erosion and desertification (Marchal et al., 2011), besides this pest species can destroy Ulmus pumila L. (Urticales: Ulmaceae) and two or three species in Rosaceae (Zong et al., 2006). Due to E. hippophaecolus with sophisticated ecological and life history traits, such as larvae bore into trunks and roots, and accomplish one generation among 3–4 years, there is no highly effective method for preventing damage to larvae, which is deemed a key danger to sea buckthorn plantations in China (Zong et al., 2005). Sex pheromones of E. hippophaecolus female PGs (E)-3-tetradecenyl acetate (E3-14:Ac) and (Z)-7-tetradecenyl acetate (Z7-14:Ac) have been identified (Fang et al., 2005), which have been developed specific and efficient artificial sex pheromone traps (Shixiang Zong et al., 2010). And through the antennal transcriptome identified 137 olfactory genes including three PBPs, and found antennal biased expressed OBPs (Hu et al., 2016).
In the present study, we examined the female sex PG transcriptome of E. hippophaecolus, identified genes and enzymes related to pheromone biosynthesis, explored pheromone biosynthesis pathways, and identified putative genes related to chemoreception. Analysis of factors referred to the production of specific pheromones may provide insight into the regulation of the pheromone components, and hence the evolution of sexual communication in moth. Understanding interactions between pheromone biosynthesis and chemoreception genes in the sex PG and chemoreception genes in the antennae may reveal connections between pheromone biosynthesis in females and pheromone recognition in males, and provide knowledge pertinent to integrated pest management strategies.
Section snippets
Ethics statement
The seabuckthorn carpenterworm Eogystia hippophaecolus (Lepidoptera: Cossidae) is a common forestry pest in China, which collections were made with the direct permission of Jianping forestry bureau. All operations were performed according to ethical guidelines in order to minimize pain and discomfort to the insects.
Insect and tissue collection
E. hippophaecolus were collected from damaging seabuckthorn forest by light during middle of June to end of July 2015 to 2016 in Jianping, Liaoning, China. Female sexual glands were
Transcriptome sequencing and sequence assembly
In total, we generated 28.53 million clean reads from the cDNA library of the female sex PG of E. hippophaecolus, of which 87.63% had q30 quality scores. After trimming adapters, removing low-quality raw sequences using Trimmomatic (http://www.usadellab.org/cms/index.php?page=trimmomatic), and splicing and assembly (using Trinity), we obtained 84,796 transcripts with an N50 of 2021 bp and an average length of 1068 bp, and 52,219 unigenes with an N50 of 1410 bp (Fig. 2). Raw reads have been
Overview of the PG transcriptome
The PG transcriptome of E. hippophaecolus was sequenced and analyzed. Among the identified 20,363 unigenes, only 50.60% were annotated to one or more GO terms, which indicated that massive E. hippophaecolus genes are either non-coding or share homology with genes not associated with GO terms, similar to the results of transcriptome analysis of the sex PG of Spodoptera litura (Zhang et al., 2015), H. armigera and H. assulta (Li et al., 2015), Plutella xylostella (He et al., 2017), Ephestia
Conclusion
Herein, we investigated the E. hippophaecolus sex PG transcriptome, in which we identified 77 unigenes involved in female pheromone biosynthesis, providing a foundation for further elucidation of the molecular mechanisms of sex pheromone biosynthesis. Additionally, 31 chemoreception proteins were identified, including 13 new genes, along with 11 novel odorant degradation enzymes, indicating that pheromone recognition may occur in the sex pheromone gland. The pheromone biosynthesis pathway and
Abbreviations
- CSPs
chemosensory proteins
- DEET
N,N-diethyl-meta-toluamide
- GO
Gene Ontology
- GPCRs
G protein-coupled receptors
- GRs
gustatory receptors
- IRs
ionotropic receptors
- iGluRs
ionotropic glutamate receptors
- OBPs
odorant binding proteins
- ORFs
open reading frame
- ORs
odorant receptors
- Orco
odorant receptor co-receptor
- ORNs
olfactory receptor neuron
- PR
pheromone receptor
- SNMPs
sensory neuron membrane proteins
- ODEs
odorant degrading enzymes
- PBAN
pheromone biosynthesis activating neuropeptide
- PBANr
pheromone biosynthesis activating
Declaration of competing interest
The authors declare no conflict of interest.
Acknowledgements
We thank Liansheng Zhang and Yunbo Ma for animal collection.
Authors' contributions
Ping Hu methodology, formal analysis, writing original draft preparation. Chenglong Gao and Pengfei Lu software, validation, investigation. Jing Tao and Youqing Luo designed and conceived of the study; Ping Hu and Jing Tao writing original draft preparation, writing review and editing, visualization, and supervision. All authors have read and agreed to the published version of the manuscript.
References (99)
- et al.
Functional architecture of olfactory ionotropic glutamate receptors
Neuron
(2011) - et al.
Pheromone-gland-specific fatty-acyl reductase in the adzuki bean borer, Ostrinia scapulalis (Lepidoptera: Crambidae)
Insect Biochem. Mol. Biol.
(2009) - et al.
Binding properties of a locust’s chemosensory protein
Biochem. Biophys. Res. Commun.
(2002) - et al.
Variant ionotropic glutamate receptors as chemosensory receptors in Drosophila
Cell
(2009) - et al.
Pheromone Production: Biochemistry and Molecular Biology, Insect Endocrinology
(2012) - et al.
Sequence variation determining stereochemistry of a Δ11 desaturase active in moth sex pheromone biosynthesis
Insect Biochem. Mol. Biol.
(2016) - et al.
Transcriptional comparison between pheromone gland-ovipositor and tarsi in the corn earworm moth Helicoverpa zea
Comparative Biochemistry and Physiology Part D Genomics and Proteomics
(2019) - et al.
Discovery of a disused desaturase gene from the pheromone gland of the moth Ascotis selenaria, which secretes an epoxyalkenyl sex pheromone
Biochem. Biophys. Res. Commun.
(2013) - et al.
Acyl-CoA Z9-and Z10-desaturase genes from a New Zealand leafroller moth species, Planotortrix octo
Insect Biochem. Mol. Biol.
(2002) Regulation of pheromone biosynthesis in moths
Current Opinion in Insect Science
(2017)
The pheromone biosynthesis activating neuropeptide (PBAN) receptor of Heliothis virescens: identification, functional expression, and structure–activity relationships of ligand analogs
Peptides
The Drosophila IR20a clade of ionotropic receptors are candidate taste and pheromone receptors
Neuron
Re-evaluation of the PBAN receptor (PBANR) molecule: characterization of PBANR variants expressed in the pheromone glands of moths
Front. Endocrinol.
Elucidation of the sex-pheromone biosynthesis producing 5, 7-dodecadienes in Dendrolimus punctatus (Lepidoptera: Lasiocampidae) reveals Δ11-and Δ9-desaturases with unusual catalytic properties
Insect Biochem. Mol. Biol.
Role of the Halloween genes, Spook and Phantom in ecdysteroidogenesis in the desert locust, Schistocerca gregaria
J. Insect Physiol.
Sex pheromone biosynthetic pathway in Spodoptera littoralis and its activation by a neurohormone
J. Biol. Chem.
Characterization of acyl-CoA-binding protein (ACBP) in the pheromone gland of the silkworm, Bombyx mori
Insect Biochem. Mol. Biol.
Molecular mechanisms underlying sex pheromone production in the silkmoth, Bombyx mori: characterization of the molecular components involved in bombykol biosynthesis
J. Insect Physiol.
Unraveling the Pheromone Biosynthesis Activating Neuropeptide (PBAN) Signal Transduction Cascade That Regulates Sex Pheromone Production in Moths, Vitamins & Hormones
Pheromone biosynthesis: enzymatic studies in Lepidoptera
Cell-specific expression of enhanced green fluorescence protein under the control of neuropeptide gene promoters in the brain of the silkworm, Bombyx mori, using Bombyx mori nucleopolyhedrovirus-derived vectors
Insect Biochem. Mol. Biol.
Functional characterization of the Bombyx mori fatty acid transport protein (BmFATP) within the silkmoth pheromone gland
J. Biol. Chem.
Cloning and functional characterization of a fatty acid transport protein (FATP) from the pheromone gland of a lichen moth, Eilema japonica, which secretes an alkenyl sex pheromone
Insect Biochem. Mol. Biol.
Ionotropic receptors (IRs): chemosensory ionotropic glutamate receptors in Drosophila and beyond
Insect Biochem. Mol. Biol.
Structural analysis and disulfide-bridge pairing of two odorant-binding proteins from Bombyx mori
Biochem. Biophys. Res. Commun.
Drosophila auditory organ genes and genetic hearing defects
Cell
Insect pheromones—an overview of biosynthesis and endocrine regulation
Insect Biochem. Mol. Biol.
Insect SNMP gene family
Insect Biochemistry & Molecular Biology
Comparative study of sex pheromone composition and biosynthesis in Helicoverpa armigera, H. assulta and their hybrid
Insect Biochem. Mol. Biol.
Neofunctionalization in an ancestral insect desaturase lineage led to rare Δ6 pheromone signals in the Chinese tussah silkworm
Insect Biochem. Mol. Biol.
Chapter ten-Odorant-binding proteins in insects
Vitamins & Hormones
Acid sensing by the Drosophila olfactory system
Nature
Sex pheromone evolution is associated with differential regulation of the same desaturase gene in two genera of leafroller moths
PLoS Genet.
Genes involved in sex pheromone biosynthesis of Ephestia cautella, an important food storage pest, are determined by transcriptome sequencing
BMC Genomics
Two fatty acyl reductases involved in moth pheromone biosynthesis
Sci. Rep.
Sex pheromone biosynthesis in Trichoplusia ni: key steps involve delta-11 desaturation and chain-shortening
Science
Biosynthesis of sex pheromone components and glycerolipid precursors from sodium [1-14 C] acetate in redbanded leafroller moth
J. Chem. Ecol.
Structure of the pheromone communication channel in moths
Analysis of the agrotis segetum pheromone gland transcriptome in the light of sex pheromone biosynthesis
BMC Genomics
Degradation of pheromone and plant volatile components by a same odorant-degrading enzyme in the cotton leafworm, Spodoptera littoralis
PLoS One
Sex pheromone components of the sandthorn carpenterworm, Holcocerus hippophaecolus
J. Chem. Ecol.
HMMER web server: interactive sequence similarity searching. Nucleic Acids Res 39: W29-W37
Nucleic Acids Res.
Sex pheromone biosynthesis in the leafroller moth Planotortix excessana by Δ10 desaturation
Arch. Insect Biochem. Physiol.
Sex pheromone biosynthesis in the tortricid moth, Ctenopseustis herana (Felder & Rogenhofer)
Archives of Insect Biochemistry and Physiology
Transcriptome exploration of the sex pheromone gland of Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae)
Parasit. Vectors
High-throughput functional annotation and data mining with the Blast2GO suite
Nucleic Acids Res.
An olfactory receptor for food-derived odours promotes male courtship in Drosophila
Nature
Identification of genes expressed in the sex pheromone gland of the black cutworm Agrotis ipsilon with putative roles in sex pheromone biosynthesis and transport
BMC Genomics
Semi–selective fatty acyl reductases from four heliothine moths influence the specific pheromone composition
PLoS One
Cited by (2)
Genes involved in the Type I pheromone biosynthesis pathway and chemoreception from the sex pheromone gland transcriptome of Dioryctria abietella
2021, Comparative Biochemistry and Physiology - Part D: Genomics and ProteomicsCitation Excerpt :In addition, esterases (especially carboxylesterases (CXEs)) are hydrolases and its hydrolysis occurs during plant volatile and sex pheromone degradation (Prestwich et al., 1986; Luo et al., 2014; He et al., 2015). Recently, more researches on CXEs was discovered in the moth sex glands of Agrotis ipsilon, Ephestia cautella, Chilo suppressalis and Eogystia hippophaecolus (Gu et al., 2013; Antony et al., 2015; Xia et al., 2015; Hu et al., 2020). The sequence similarity between DabiCXE17 and Plodia interpunctella (QEA03452.1) was 93.38% (Table 3).
Identification and Characterization of Aldehyde Oxidase 5 in the Pheromone Gland of the Silkworm (Lepidoptera: Bombycidae)
2021, Journal of Insect Science