Pheromone System Research Articles

Sexy fingers: Pheromones in the glands of male dendrobatid frogs.

Many animals exchange chemicals during courtship and mating. In some amphibians, sexual chemical communication is mediated by pheromones produced in male breeding glands that are transferred to the female's nostrils during mating. This has been mostly studied in salamanders, despite frogs having similar glands and courtship behaviours suggestive of chemical communication. In Neotropical poison frogs (Dendrobatidae and Aromobatidae), males of many species develop breeding glands in their fingers, causing certain fingers to visibly swell. Many also engage in cephalic amplexus, whereby the male's swollen fingers are placed in close contact with the female's nares during courtship. Here, we investigate the possible roles of swollen fingers in pheromone production using whole-transcriptome sequencing (RNAseq). We examined differential gene expression in the swollen versus non-swollen fingers and toes of two dendrobatid species, Leucostethus brachistriatus and Epipedobates anthonyi, both of which have specialised mucous glands in finger IV, the latter of which has cephalic amplexus. The overwhelming pattern of gene expression in both species was strong upregulation of sodefrin precursor-like factors (SPFs) in swollen fingers, a well-known pheromone system in salamanders. The differentially expressed SPF transcripts in each species were very high (>40), suggesting a high abundance of putative protein pheromones in both species. Overall, the high expression of SPFs in the swollen fingers in both species, combined with cephalic amplexus, supports the hypothesis that these traits, widespread across members of the subfamilies Colostethinae and Hyloxalinae (ca. 141 species), are involved in chemical signalling during courtship.

Love Hug-Functional Validation of Nuptial Pad-Secreted Pheromone in Anurans.

Chemical communication is an important mode of communication in the courtship and breeding processes of amphibians. In caudates, multiple components of sexual pheromones have been identified and functionally verified. One of these pheromone systems is plethodontid modulating factor (PMF). In anurans, the pheromone called amplexin was found in nuptial pads of ranids and was considered a member of the PMF system, yet its bio-function has not been tested. In this study, we obtained 18 amplexin transcript sequences from nuptial pads of Nidirana pleuraden (Amphibia, Ranidae) by transcriptome sequencing and found that the proteins translated by these transcripts are diversified, hydrophilic, and relatively stable. We also acquired a N. pleuraden amplexin isoform with the highest expression level in the transcriptome analysis through the prokaryotic expression system. Using two different animal behavioral experimental settings, we have tested the bio-function of the recombinant PMF protein (rPMF) in N. pleuraden's reproduction and found that the rPMF does not attract females but shortens the duration of amplexus significantly. This is the first study to verify the function of the PMF pheromone in Anura, indicating the pervasiveness of chemical communication during breeding in amphibians.

Genetic modules for α-factor pheromone controlled growth regulation of Saccharomyces cerevisiae.

Saccharomyces cerevisiae is a commonly used microorganism in the biotechnological industry. For the industrial heterologous production of compounds, it is of great advantage to work with growth-controllable yeast strains. In our work, we utilized the natural pheromone system of S. cerevisiae and generated a set of different strains possessing an α-pheromone controllable growth behavior. Naturally, the α-factor pheromone is involved in communication between haploid S. cerevisiae cells. Perception of the pheromone initiates several cellular changes, enabling the cells to prepare for an upcoming mating event. We exploited this natural pheromone response system and developed two different plasmid-based modules, in which the target genes, MET15 and FAR1, are under control of the α-factor sensitive FIG1 promoter for a controlled expression in S. cerevisiae. Whereas expression of MET15 led to a growth induction, FAR1 expression inhibited growth. The utilization of low copy number or high copy number plasmids for target gene expression and different concentrations of α-factor allow a finely adjustable control of yeast growth rate.

Presence of sodefrin precursor-like factor pheromone candidates in mental and dorsal tail base glands in the plethodontid salamander, Karsenia koreana.

Plethodontid salamanders are well known for their distinct courtship rituals and the associated pheromonal signaling. However, little is known about pheromones produced in the lone Asian plethodontid species Karsenia koreana. Here, we examined the localization patterns of proteins of the sodefrin precursor-like factor (SPF) pheromone system in K. koreana. Using an antibody generated against SPF proteins from another plethodontid, Desmognathus ocoee, we tested three types of skin glands in K. koreana males via immunohistochemistry: the mental gland and two types of dorsal tail base glands-caudal courtship glands and dorsal granular glands. SPF immunoreactivity was detected in the known courtship gland, the mental gland, as well as granular glands, but not in caudal courtship glands. Due to immunoreaction specificity, we hypothesize the proteins of the SPF system in K. koreana and D. ocoee are structurally and functionally related and are used as courtship pheromones in K. koreana. Also, we hypothesize that K. koreana males transmit SPF to the female during the tail-straddling walk via dorsal granular glands. Finally, K. koreana male caudal courtship glands may be producing SPF proteins that are not recognized by our SPF antibody or these glands may play a different role in courtship than anticipated.

Evolutionary shifts in pheromone receptors contribute to speciation in four Helicoverpa species.

Male moths utilize their pheromone communication systems to distinguish potential mates from other sympatric species, which contributes to maintaining reproductive isolation and even drives speciation. The molecular mechanisms underlying the evolution of pheromone communication systems are usually studied between closely-related moth species for their similar but divergent traits associated with pheromone production, detection, and/or processing. In this study, we first identified the functional differentiation in two orthologous pheromone receptors, OR14b, and OR16, in four Helicoverpa species, Helicoverpa armigera, H. assulta, H. zea, and H. gelotopoeon. To understand the substrate response specificity of these two PRs, we performed all-atom molecular dynamics simulations of OR14b and OR16 based on AlphaFold2 structural prediction, andmolecular docking, allowing us to predict a few key amino acids involved in substrate binding. These candidate residues were further tested and validated by site-directed mutagenesis and functional analysis. These results together identified two hydrophobic amino acids at positions 164 and 232 are the determinants of the response specificity of HarmOR14b and HzeaOR14b to Z9-14:Ald and Z9-16:Ald by directly interacting with the substrates. Interestingly, in OR16 orthologs, we found that position 66 alone determines the specific binding of Z11-16:OH, likely via allosteric interactions. Overall, we have developed an effective integrated method to identify the critical residues for substrate selectivity of ORs and elucidated the molecular mechanism of the diversification of pheromone recognition systems.

Sex pheromone of the fall armyworm, Spodoptera frugiperda: identification of a trace component that enhances attractiveness and specificity

AbstractThe fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), is a polyphagous pest native to the tropical and subtropical Americas, but it has recently invaded the Eastern Hemisphere and spread rapidly. This migratory moth, which can fly long distances, now poses a global threat as a transboundary pest. To establish an efficient management program against this pest, monitoring of its occurrence is essential. Lures impregnated with four synthetic pheromone components of S. frugiperda – (Z)‐9‐tetradecenyl acetate (Z9‐14:Ac), (Z)‐11‐hexadecenyl acetate, (Z)‐7‐dodecenyl acetate, and (Z)‐9‐dodecenyl acetate – are commercially available, and traps baited with them are used for monitoring. However, in Japan, these pheromone traps frequently capture many non‐target moths, particularly Mythimna loreyi (Duponchel) (Lepidoptera: Noctuidae). We therefore reexamined the sex pheromone of S. frugiperda in an attempt to improve the monitoring lures and discovered trace amounts of (Z,E)‐9,12‐tetradecadienyl acetate (Z9E12‐14:Ac) in female extracts at ca. 0.2% of the primary component (Z9‐14:Ac). We verified its activity in males by field trap bioassays as well as antennal electrophysiology and demonstrated that this fifth component enhances the attractiveness of synthetic pheromones composed of the other four components to S. frugiperda and reduces the bycatch of M. loreyi. As Z9E12‐14:Ac is found in pheromones of most Spodoptera species, it may have been a pheromone component in the common ancestor and played a key role in divergence of pheromone systems in the genus Spodoptera.

Identification and characterization of lipocalin-type prostaglandin D2 synthase homologs in the urine of male rockfish

Black rockfish (Sebastes schlegelii) and its relatives are viviparous marine fish. Males produce urinary proteins during the copulation season; however, the identity of these proteins was unknown. In this study, we focused on high-molecular-weight urinary proteins (HMWups) in male black rockfish. The HMWups were identified by liquid chromatography and tandem mass spectrometry (LC-MS/MS) of urine. In silico analyses of RNA-seq data predicted the tissue distribution of candidate HMWup transcripts and their gene structures. Candidate cDNAs were cloned and a recombinant protein of a major candidate was prepared. Western blotting of urine using an antiserum against the recombinant protein was performed to reconfirm the LC-MS/MS results. Quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry were employed to validate the prediction by RNA-seq and identify the cells producing HMWups, respectively. LC-MS/MS, in conjunction with Western blotting and cDNA cloning, identified the HMWups as lipocalin-type prostaglandin D2 synthase (l-PGDS) homologs. RNA-seq analyses and qRT-PCR revealed that the l-PGDS homolog transcripts were dominantly expressed in the testis and male kidney; Sertoli cells and epithelial cells in the renal tubules were immunoreactive. These results indicated that major protein components in the urine of male black rockfish are l-PGDS homologs, potentially produced by the renal tubules in the kidney. Male rockfish (genus Sebastes) are thought to release unknown pheromone substances during mating behavior. The knowledge and tools obtained in this study empower research into the role(s) of HMWups in pheromone systems underlying rockfish reproduction. No protein-type teleost pheromone has heretofore been discovered.

Analysis of sex pheromone production and field trapping of the Asian corn borer (Ostrinia furnacalis Guenée) in Xinjiang, China

Identifying the sex pheromone systems of local pest populations facilitates their management, especially for moth species that show significant geographic variation in sex pheromone communication. We investigated the pheromone production and behavioral responses of the Asian corn borer (Ostrinia furnacalis Guenée; ACB) in Xinjiang, China. The ACB produces three compounds: (Z)-12-tetradecenyl acetate (Z12-14:Ac) and (E)-12-tetradecenyl acetate (E12-14:Ac) which are two sex pheromone compounds, and n-tetradecyl acetate (14:Ac) which has variable roles in mediating behavioral responses. The ratios of these three compounds produced in female gland are geographically distinct among different populations. Quantitative analysis of pheromone production showed that the proportions of Z12-14:Ac in the E/Z isomers (i.e., Z and E12-14:Ac) and the proportions of 14:Ac in the ternary blend respectively averaged 60.46% (SD=5.26) and 25.00% (SD=7.37), with their probabilities normally or near-normally distributed. Trapping experiments in a cornfield indicated that deploying the E/Z isomers and the three compounds in rubber septa close to their gland ratios yielded the most captured males, while other ratios that deviated from the gland ratios showed reduced field captures. The ternary blend was significantly more attractive to males than the E/Z isomers in the field, indicating a functional role of 14:Ac as the third pheromone component used by the local population. Additionally, the dose-response test demonstrated that the application of the three compounds at dosages between 200 and 350 μg attracted significantly more males compared to other dosages. Therefore, the characterization of this local ACB pheromone system provides additional information about its geographic variation and serves as a basis for optimizing the pheromone-mediated control of this pest in Xinjiang.

Sex Pheromone of the Azalea Mealybug With a Non-Terpene Structure.

Mealybug females release sex pheromones to attract conspecific males for mating. It is critical for mealybug males, which are fragile and short-lived, to respond to the pheromone of their species without time- and energy-consuming cross-attractions to other species. Thus, mealybug pheromone systems are considered to have evolved to be species-specific with unique structures in each species and offer an opportunity to study the diversity of pheromone chemistry that mediates intersexual courtship signals. More than 20 mealybug pheromones are reported to be monoterpenes in general, with only one exception, a hemiterpene alcohol esterified with a medium-chain fatty acid (MCFA), found in the Matsumoto mealybug, Crisicoccus matsumotoi. However, it is unknown whether this is truly exceptional, or if similar compounds are used in other related mealybugs. In this study, we isolated and characterized the pheromone of an allied species, the azalea mealybug C. azaleae. Using gas chromatography-mass spectrometry, nuclear magnetic resonance spectroscopy, and bioassays with synthetics, the pheromone was shown to be composed of isopropyl (E)-7-methyl-4-nonenoate, isopropyl (E)-7-methyl-4-octenoate, and ethyl (E)-7-methyl-4-nonenoate. Surprisingly, the structures of these compounds do not include hemiterpene nor monoterpene motifs but have methyl-branched MCFA parts that are similar to an acid moiety of the C. matsumotoi pheromone. This study implies irregular events for the divergence of pheromone structures in ancestors of the genus Crisicoccus and other mealybugs.

Trapping of Retrachydes thoracicus thoracicus (Olivier) and Other Neotropical Cerambycid Beetles in Pheromone- and Kairomone-Baited Traps.

The subfamily Cerambycinae, one of the most diverse in longhorn beetles, is well known for its remarkable chemical parsimony in male-emitted pheromones. Conserved shared structural motifs have been reported in numerous species, sometimes working in combination with plant volatile kairomones. Among other compounds, the most ubiquitous male pheromone in cerambycine species is 3-hydroxyhexan-2-one. We conducted field trials using intercept traps baited with 3-hydroxyhexan-2-one and observed abundant captures of several Neotropical cerambycine species. These were Retrachydes thoracicus thoracicus (Olivier), Megacyllene acuta (Germar), Compsocerus violaceus (White), and Cotyclytus curvatus (Germar) in high numbers, as well as Chydarteres striatus striatus (Fabricius) and Odontocroton flavicauda (Bates) in smaller numbers. When ethanol was added to the traps, a remarkable increase in the attractiveness of 3-hydroxyhexan-2-one was observed for R. thoracicus thoracicus and M. acuta. Adding ethanol also resulted in the capture of Chrysoprasis aurigena (Germar). Finally, incidental catches in pheromone-baited traps of Trachelissa maculicollis (Audinet-Serville), Neoclytus pusillus (Laporte & Gory), Achryson unicolor (Bruch, 1908) and Achryson surinamum (Linnaeus), Megacyllene mellyi (Chevrolat) and Thelgetra adustus (Burmeister) were also observed. Pheromone chemistry has been reported for C. curvatus, M. acuta and N. pusillus, all three producing 3-hydroxyhexan-2-one, and for C. aurigena and A. surinamum, which produce other compounds. Our findings suggest that the captured species probably produce 3-hydroxyhexan-2-one for their pheromone communication system. Alternatively, they might be "eavesdropping" on the pheromones of other cerambycine species. The probable synergistic effect of ethanol is likely explained from its kairomonal role as a volatile cue for plant stress or ripeness.

Mountain Pine Beetle Epidemic: An Interplay of Terpenoids in Host Defense and Insect Pheromones.

The mountain pine beetle epidemic has highlighted the complex interactions of bark beetles with conifer host defenses. In these interactions, oleoresin terpenoids and volatiles, produced and released by the host tree, can be both harmful and beneficial to the beetle's success in colonizing a tree and completing its life cycle. The insect spends almost its entire life, from egg to adult, within the bark and phloem of a pine host, exposed to large quantities of complex mixtures of oleoresin terpenoids. Conifer oleoresin comprises mostly monoterpenes and diterpene resin acids as well as many different sesquiterpenes. It functions as a major chemical and physical defense system. However, the insect has evolved host colonization behavior and enzymes for terpenoid metabolism and detoxification that allow it to overcome some of the terpenoid defenses and, importantly, to co-opt pine monoterpenes as cues for host search and as a precursor for its own pheromone system. The insect-associated microbiome also plays a role in the metabolism of conifer terpenoids.

Formation and function of dauer ascarosides in the nematodes Caenorhabditis briggsae and Caenorhabditis elegans.

The biosynthetic pathways and functions of ascaroside signaling molecules in the nematode Caenorhabditis elegans have been studied to better understand complex, integrative developmental decision-making. Although it is known that ascarosides play multiple roles in the development and behavior of nematode species other than C. elegans, these parallel pheromone systems have not been well-studied. Here, we show that ascarosides in the nematode Caenorhabditis briggsae are biosynthesized in the same manner as C. elegans and act to induce the alternative developmental pathway that generates the stress-resistant dauer lifestage. We show that ascr#2 is the primary component of crude dauer pheromone in C. briggsae; in contrast, C. elegans dauer pheromone relies on a combination of ascr#2, ascr#3, and several other components. We further demonstrate that Cbr-daf-22, like its C. elegans ortholog Cel-daf-22, is necessary to produce short-chain ascarosides. Moreover, Cbr-daf-22 and Cel-daf-22 mutants produce an ascaroside-independent metabolite that acts antagonistically to crude dauer pheromone and inhibits dauer formation.

Expression Levels of Detoxification Enzyme Genes from Dendroctonus armandi (Coleoptera: Curculionidae) Fed on a Solid Diet Containing Pine Phloem and Terpenoids.

Simple SummaryThe bark beetle is the most well-known pest in coniferous trees worldwide. These insects only leave the host pine bark when they disperse to locate a new host. Determining how Dendroctonus armandi overcome the trees’ terpene-based defense systems has been the key problem in the study of bark beetles. Therefore, the aim of this study was to discover the molecular mechanism of insect detoxification enzymes’ ability to confer resistance to terpenes. For this purpose, the genes of cytochrome P450s, glutathione S-transferases, and carboxylesterases were studied in beetles given diets containing terpenes. The results suggest that beetles express different genes in response to terpenoids, and the responses of multiple detoxifying enzymes indicate these insects’ adaption to their chemical environment.Bark beetles overcome the toxic terpenoids produced by pine trees by both detoxifying and converting them into a pheromone system. Detoxification enzymes such as cytochrome P450s, glutathione S-transferases, and carboxylesterases are involved in the ability of Dendroctonus armandi to adapt to its chemical environment. Ten genes from these three major classes of detoxification enzymes were selected to study how these enzymes help D. armandi to respond to the host defenses. The expression profile of these detoxification enzyme genes was observed in adult beetles after feeding on different types of diet. Significant differences were observed between two types of seminatural diet containing the phloem of pines, and a purely artificial diet containing five monoterpenes ((−)-α-pinene, (−)-β-pinene, (+)-3-carene, (±)-limonene, and turpentine oil) also caused differential transcript levels in the detoxification enzyme genes. The results suggest that monoterpenes enter the beetles through different routes (i.e., respiratory and digestive systems) and cause the expression of different genes in response, which might be involved in pheromone metabolism. In addition, the xenobiotic metabolism in bark beetles should be considered as a system comprising multiple detoxifying enzymes.

Development of methods for application of pheromonic materials for monitoring and managing the number of apple phytophages

Synthetic sex pheromones have found wide application in plant protection as a mean of early detection of pests that allows observation of the phenology of insects to optimise protective measures. Insect sex pheromones can be applied for different purposes in protecting crops such as monitoring, determining the species composition and combating harmful species (by using sex pheromones for disorientation). The present work studies the species composition, dynamics of the number of basic pests of an apple tree, synchronisation of the seasonal and circadian activity of phytophages of an apple tree in the central zone of the Krasnodar Territory. The results of the field assessment of the male complex disorientation method of apple and eastern moths are presented. It was shown that the species-specificity of sex pheromones in the apple orchard depends on the faunistic diversity of Lepidoptera species with similar pheromone systems that develop at a given point in space and time. It was revealed that the behaviour of this complex changes during the season and over years, depending on the climate and natural dynamics of insect populations. The quantitative ratio and species-specificity of pheromones will probably be different in ecosystems with various species composition and different geographic zones. A novelty of this research is the division of the studied Lepidoptera phytophage species into three groups according to a decrease in the absolute species-specificity in the forest biotope in comparison with the garden one. The most widespread and coinciding in terms of summer synchronicity are apple (Cydia pomonella L.), plum (Grapholitha funebrana Tr.), eastern (Grapholitha molesta Tr.) and pomegranate moth (Euzophera bigella Zell.). The disorientation method, applied using a complex system of dispensers with apple and eastern moth pheromones, showed that installing 500 dispensers/ha allowed 99.3 % efficiency to be achieved. Fruit damage amounted to 1.2 % and 2.7 % in the experimental and control groups, respectively. The duration of the disorienting effect of the pheromone formulations lasted for over 4 months.

Distinct protocerebral neuropils associated with attractive and aversive female-produced odorants in the male moth brain.

The pheromone system of heliothine moths is an optimal model for studying principles underlying higher-order olfactory processing. In Helicoverpa armigera, three male-specific glomeruli receive input about three female-produced signals, the primary pheromone component, serving as an attractant, and two minor constituents, serving a dual function, that is, attraction versus inhibition of attraction. From the antennal-lobe glomeruli, the information is conveyed to higher olfactory centers, including the lateral protocerebrum, via three main paths - of which the medial tract is the most prominent. In this study, we traced physiologically identified medial-tract projection neurons from each of the three male-specific glomeruli with the aim of mapping their terminal branches in the lateral protocerebrum. Our data suggest that the neurons' widespread projections are organized according to behavioral significance, including a spatial separation of signals representing attraction versus inhibition - however, with a unique capacity of switching behavioral consequence based on the amount of the minor components.

Bric \xe0 brac controls sex pheromone choice by male European corn borer moths

The sex pheromone system of ~160,000 moth species acts as a powerful form of assortative mating whereby females attract conspecific males with a species-specific blend of volatile compounds. Understanding how female pheromone production and male preference coevolve to produce this diversity requires knowledge of the genes underlying change in both traits. In the European corn borer moth, pheromone blend variation is controlled by two alleles of an autosomal fatty-acyl reductase gene expressed in the female pheromone gland (pgFAR). Here we show that asymmetric male preference is controlled by cis-acting variation in a sex-linked transcription factor expressed in the developing male antenna, bric à brac (bab). A genome-wide association study of preference using pheromone-trapped males implicates variation in the 293 kb bab intron 1, rather than the coding sequence. Linkage disequilibrium between bab intron 1 and pgFAR further validates bab as the preference locus, and demonstrates that the two genes interact to contribute to assortative mating. Thus, lack of physical linkage is not a constraint for coevolutionary divergence of female pheromone production and male behavioral response genes, in contrast to what is often predicted by evolutionary theory.

Monoterpenoid signals and their transcriptional responses to feeding and juvenile hormone regulation in bark beetle Ips hauseri.

Hauser's engraver beetle, Ips hauseri, is a serious pest in spruce forest ecosystems in Central Asia. Its monoterpenoid signal production, transcriptome responses and potential regulatory mechanisms remain poorly understood. The quality and quantity of volatile metabolites in hindgut extracts of I. hauseri were found to differ between males and females and among three groups: beetles that were newly emerged, those with a topical application of juvenile hormone III (JHIII) and those that had been feeding for 24 h. Feeding males definitively dominated monoterpenoid signal production in I. hauseri, which uses (4S)-(-)-ipsenol and (S)-(-)-cis-verbenol to implement reproductive segregation from Ipstypographus and Ipsshangrila. Feeding stimulation induced higher expression of most genes related to the biosynthesis of (4S)-(-)-ipsenol than JHIII induction, and showed a male-specific mode in I. hauseri. JHIII stimulated males to produce large amounts of (-)-verbenone and also upregulated the expression of several CYP6 genes, to a greater extent in males than in females. The expression of genes involved in the metabolism of JHIII in females and males was also found to be upregulated. Our results indicate that a species-specific aggregation pheromone system for I. hauseri, consisting of (4S)-(-)-ipsenol and S-(-)-cis-verbenol, can be used to monitor population dynamics or mass trap killing. Our results also enable a better understanding of the bottom-up role of feeding behaviors in mediating population reproduction/aggregation and interspecific interactions.

Pheromone Mediated Sexual Reproduction of Pennate Diatom Cylindrotheca closterium

Benthic diatoms dominate primary production in marine subtidal and intertidal environments. Their extraordinary species diversity and ecological success is thought to be linked with their predominantly heterothallic sexual reproduction. Little is known about pheromone involvement during mating of pennate diatoms. Here we describe pheromone guided mating in the coastal raphid diatom Cylindrotheca closterium. We show that the two mating types (mt+ and mt−) have distinct functions. Similar to other benthic diatoms, mt+ cells are searching for the mt− cells to pair. To enhance mating efficiency mt− exudes an attraction pheromone which we proved by establishing a novel capillary assay. Further, two more pheromones produced by mt− promote the sexual events. One arrests the cell cycle progression of mt+ while the other induces gametogenesis of mt+. We suggest that C. closterium shares a functionally similar pheromone system with other pennate diatoms like Seminavis robusta and Pseudostaurosira trainorii which synchronize sexual events and mate attraction. Remarkably, we found no evidence of mt+ producing pheromones, which differentiates C. closterium from other pennates and suggests a less complex pheromone system in C. closterium.

Pheromonal communication in urodelan amphibians.

Pheromonal communication is an ancient and pervasive sensory modality in urodelan amphibians. One family of salamander pheromones (the sodefrin precursor-like factor (SPF) family) originated 300 million years ago, at the origin of amphibians. Although salamanders are often thought of as relatively simple animals especially when compared to mammals, the pheromonal systems are varied and complex with nuanced effects on behavior. Here, we review the function and evolution of pheromonal signals involved in male-female reproductive interactions. After describing common themes of salamander pheromonal communication, we describe what is known about the rich diversity of pheromonal communication in each salamander family. Several pheromones have been described, ranging from simple, invariant molecules to complex, variable blends of pheromones. While some pheromones elicit overt behavioral responses, others have more nuanced effects. Pheromonal signals have diversified within salamander lineages and have experienced rapid evolution. Once receptors have been matched to pheromonal ligands, rapid advance can be made to better understand the olfactory detection and processing of salamander pheromones. In particular, a large number of salamander species deliver pheromones across the skin of females, perhaps reflecting a novel mode of pheromonal communication. At the end of our review, we list some of the many intriguing unanswered questions. We hope that this review will inspire a new generation of scientists to pursue work in this rewarding field.

A Multiple Pheromone Communication System for Swarm Intelligence

Pheromones are chemical substances essential for communication among social insects. In the application of swarm intelligence to real micro mobile robots, the deployment of a single virtual pheromone has emerged recently as a powerful real-time method for indirect communication. However, these studies usually exploit only one kind of pheromones in their task, neglecting the crucial fact that in the world of real insects, multiple pheromones play important roles in shaping stigmergic behaviors such as foraging or nest building. To explore the multiple pheromones mechanism which enable robots to solve complex collective tasks efficiently, we introduce an artificial multiple pheromone system (ColCOS $\Phi $ ) to support swarm intelligence research by enabling multiple robots to deploy and react to multiple pheromones simultaneously. The proposed system ColCOS $\Phi $ uses optical signals to emulate different evaporating chemical substances i.e. pheromones. These emulated pheromones are represented by trails displayed on a wide LCD display screen positioned horizontally, on which multiple miniature robots can move freely. The color sensors beneath the robots can detect and identify lingering “pheromones” on the screen. Meanwhile, the release of any pheromone from each robot is enabled by monitoring its positional information over time with an overhead camera. No other communication methods apart from virtual pheromones are employed in this system. Two case studies have been carried out which have verified the feasibility and effectiveness of the proposed system in achieving complex swarm tasks as empowered by multiple pheromones. This novel platform is a timely and powerful tool for research into swarm intelligence.