Higher level phylogeny and the first divergence time estimation of Heteroptera (Insecta: Hemiptera) based on multiple genes

Heteroptera, or true bugs, are the largest, morphologically diverse and economically important group of insects with incomplete metamorphosis. However, the phylogenetic relationships within Heteroptera are still in dispute and most of the previous studies were based on morphological characters or with single gene (partial or whole 18S rDNA). Besides, so far, divergence time estimates for Heteroptera totally rely on the fossil record, while no studies have been performed on molecular divergence rates. Here, for the first time, we used maximum parsimony (MP), maximum likelihood (ML) and Bayesian inference (BI) with multiple genes (18S rDNA, 28S rDNA, 16S rDNA and COI) to estimate phylogenetic relationships among the infraorders, and meanwhile, the Penalized Likelihood (r8s) and Bayesian (BEAST) molecular dating methods were employed to estimate divergence time of higher taxa of this suborder. Major results of the present study included: Nepomorpha was placed as the most basal clade in all six trees (MP trees, ML trees and Bayesian trees of nuclear gene data and four-gene combined data, respectively) with full support values. The sister-group relationship of Cimicomorpha and Pentatomomorpha was also strongly supported. Nepomorpha originated in early Triassic and the other six infraorders originated in a very short period of time in middle Triassic. Cimicomorpha and Pentatomomorpha underwent a radiation at family level in Cretaceous, paralleling the proliferation of the flowering plants. Our results indicated that the higher-group radiations within hemimetabolous Heteroptera were simultaneously with those of holometabolous Coleoptera and Diptera which took place in the Triassic. While the aquatic habitat was colonized by Nepomorpha already in the Triassic, the Gerromorpha independently adapted to the semi-aquatic habitat in the Early Jurassic.     

Phylogenetic divergences of the true bugs (Insecta: Hemiptera: Heteroptera), with emphasis on the aquatic lineages: the last piece of the aquatic insect jigsaw originated in the Late Permian/Early Triassic

Heteroptera are among the most diverse hemimetabolous insects. Seven infraorders have been recognized within this suborder of Hemiptera. Apart from the well‐established sister‐group relationship between Cimicomorpha and Pentatomomorpha (= Terheteroptera), the two terminal lineages, the relationships among the other five infraorders are still controversial, of which three (Gerromorpha, Nepomorpha and Leptopodomorpha) are intimately connected to aquatic environments. However, the various and often conflicting available phylogeny hypotheses do not offer a clear background for a connection between diversification and palaeoenvironments. In this study, a molecular data set representing 79 taxa and 10 149 homologous sites is used to infer the phylogenetic relationships within Heteroptera. Bayesian inference, maximum‐likelihood and maximum parsimony analyses were employed. The results of phylogenetic inferences largely confirm the widely accepted phylogenetic context. Estimation of the divergence time based on the phylogenetic results revealed that Gerromorpha, Nepomorpha and Leptopodomorpha originated successively during the period from the Late Permian to Early Triassic (269–246 Ma). This timescale is consistent with the origin and radiation time of various aquatic holometabolans. Our results indicate that the aquatic and semi‐aquatic true bugs evolved under environmental conditions of high air temperature and humidity in an evolutionary scenario similar to that of the aquatic holometabolans.     

Revisiting habitat and lifestyle transitions in Heteroptera (Insecta: Hemiptera): insights from a combined morphological and molecular phylogeny

Heteroptera, the true bugs, are part of the largest clade of non-holometabolous insects, the Hemiptera, and include > 42 000 described species in about 90 families. Despite progress in resolving phylogenetic relationships between and within infraorders since the first combined morphological and molecular analysis published in 1993 (29 taxa, 669 bp, 31 morphological characters), recent hypotheses have relied entirely on molecular data. Weakly supported nodes along the backbone of Heteroptera made these published phylogenies unsuitable for investigations into the evolution of habitats and lifestyles across true bugs. Here we present the first combined morphological and molecular analyses of Heteroptera since 1993, using 135 taxa in 60 families, 4018 aligned bp of ribosomal DNA and 81 morphological characters, and various analytical approaches. The sister-group relationship of the predominantly aquatic Nepomorpha with all remaining Heteroptera is supported in all analyses, and a clade formed by Enicocephalomorpha, Dipsocoromorpha and Gerromorpha in some. All analyses recover Leptopodomorpha + (Cimicomorpha + Pentatomomorpha), mostly with high support. Parsimony- and likelihood-based ancestral state reconstructions of habitats and lifestyles on the combined likelihood phylogeny provide new insights into the evolution of true bugs. The results indicate that aquatic and semi-aquatic true bugs invaded these habitats three times independently from terrestrial habitats in contrast to a recent hypothesis. They further suggest that the most recent common ancestor of Heteroptera was predacious, and that the two large predominantly phytophagous clades (Trichophora and Miroidea) are likely to have derived independently from predatory ancestors. We conclude that by combining morphological and molecular data and employing various analytical methods our analyses have converged on a relatively well-supported hypothesis of heteropteran infraordinal relationships that now requires further testing using phylogenomic and more extensive morphological datasets.     

The rhabdom structure in the ommatidia of the Heteroptera (Insecta), and its phylogenetic significance

In its plesiomorphic state the insect ommatidium consists of eight retinula cells forming a fused rhabdom. It has long been observed that, in contrast to this pattern, Heteroptera have open rhabdoms. However, there has so far been no comprehensive and comparative study of heteropteran ommatidia. For this reason, we investigated the rhabdom structure in 36 species from all higher groups of Heteroptera, as well as from Coleorrhyncha and Auchenorrhyncha as outgroup representatives. In addition we surveyed the data of earlier authors, which brings the number of examined species to a total of more than 70. All examined Heteroptera do have open rhabdoms, with a system of six peripheral and two central rhabdomeres. Outgroup comparison shows that the open rhabdom is an autapomorphy of the Heteroptera. As for the rhabdom structure within the Heteroptera, we found further autapomorphic patterns in Corixidae (Nepomorpha), Gerromorpha, and Leptopodomorpha. Finally, the Cimicomorpha and Pentatomomorpha share a special pattern of the two central rhabdomeres, which we call V-pattern. This is a new synapomorphy of these two taxa. Accepted: 22 November 1999     

Crustacean Pigmentary-Effector Hormones: Chemistry and Functions of RPCH, PDH, and Related Peptides

Integumental color changes and eye pigment movements in crustaceansare regulated by pigmentary-effector hormones. The identifiedhormones include: an octapeptide RPCH (red pigment-concentratinghormone) and several forms of octadecapeptide PDH (pigment-dispersinghormone: -PDH, ß-PDH). RPCH-related peptides (AKHs,adipokinetic hormones) and PDH-related peptides (PDFs, pigment-dispersingfactors) occur in insects, and are recognized as members ofAKH/RPCH and PDH/PDF peptide families. The domain for maturepeptide is located between the signal peptide and precursor-relatedpeptide in AKH/RPCH precursors, and at the C-terminal end inthe PDH/PDF precursors. The precursor-related (associated) peptidesin RPCH and PDH precursors in Crustacea show little or no similarityto corresponding domains of AKH and PDF precursors in insects.Although the functions of precursor-related peptides are unknown,the mature peptides are shown to serve diverse functions. RPCH'sactions in crustaceans include: pigment concentration in oneor more types of chromatophores, dark-adaptational screeningpigment movement in distal eye pigment cells, increase of retinalsensitivity, and neuromodulation. The related AKHs largely influencemetabolism in insects, although they serve additional functions.PDHs trigger pigment dispersion in chromatophores and inducelight-adaptational screening pigment movements in extraretinulareye pigment cells. The related PDFs appear to serve as a transmitterof circadian signals in the regulation of biological rhythmsin insects. Evolutionary relationships among the PDH/PDF peptidesand directions for future research are discussed.     

The complete mitochondrial genome and novel gene arrangement of the unique-headed bug Stenopirates sp. (Hemiptera: Enicocephalidae)

Many of true bugs are important insect pests to cultivated crops and some are important vectors of human diseases, but few cladistic analyses have addressed relationships among the seven infraorders of Heteroptera. The Enicocephalomorpha and Nepomorpha are consider the basal groups of Heteroptera, but the basal-most lineage remains unresolved. Here we report the mitochondrial genome of the unique-headed bug Stenopirates sp., the first mitochondrial genome sequenced from Enicocephalomorpha. The Stenopirates sp. mitochondrial genome is a typical circular DNA molecule of 15, 384 bp in length, and contains 37 genes and a large non-coding fragment. The gene order differs substantially from other known insect mitochondrial genomes, with rearrangements of both tRNA genes and protein-coding genes. The overall AT content (82.5%) of Stenopirates sp. is the highest among all the known heteropteran mitochondrial genomes. The strand bias is consistent with other true bugs with negative GC-skew and positive AT-skew for the J-strand. The heteropteran mitochondrial atp8 exhibits the highest evolutionary rate, whereas cox1 appears to have the lowest rate. Furthermore, a negative correlation was observed between the variation of nucleotide substitutions and the GC content of each protein-coding gene. A microsatellite was identified in the putative control region. Finally, phylogenetic reconstruction suggests that Enicocephalomorpha is the sister group to all the remaining Heteroptera.     

The adipokinetic hormones of African water bugs of the Heteropteran families Nepidae and Belostomatidae

Four African species of true water bugs (Nepomorpha: Hemiptera: Heteroptera) are studied by mass spectrometry and biological assays to gain information on the presence, structure and function of peptides from the adipokinetic hormone (AKH) family, which are produced in the corpora cardiaca (CC). The water scorpion Laccotrephes fabricii Stål (Nepidae) has the peptide code‐named Peram‐CAH‐I with the sequence pGlu‐Val‐Asn‐Phe‐Ser‐Pro‐Asn‐Trp amide, whereas Appasus grassei Poisson (Belostomatidae) produces Anaim‐AKH, which is a Ser⁷ analogue of Peram‐CAH‐I (pGlu‐Val‐Asn‐Phe‐Ser‐Pro‐Ser‐Trp amide). The giant water bug Hydrocyrius columbiae Spinola (Belostomatidae) has two adipokinetic hormone family members: Anaim‐AKH and Letin‐AKH, which again differ only at position 7 (Ser⁷ versus Tyr⁷). When the sequence data are compared with current molecular phylogenetic analyses of Nepomorpha, they are essentially in agreement with the newest ideas on phylogenetic relationships among the families. Functional investigation of these peptides reveals a mainly lipid‐based energy metabolism in these insects, as demonstrated by a hyperlipaemic response after injecting crude CC extract or the appropriate peptide into the respective species. The carbohydrate concentration in the haemolymph is not affected by such injections, and the carbohydrate level in most cases is lower than that of the circulating lipids. During physical exercise, such as swimming for 1 h, carbohydrates may contribute to some extent to the provision of energy; the substantial increase in the concentration of lipids in the haemolymph, however, is a strong indicator that the peptides are released from the CC and act primarily as true adipokinetic hormones during this period of intense muscular activity.     

The adipokinetic hormone of Mantodea in comparison to other Dictyoptera

Six species of the order Mantodea (praying mantises) are investigated for the presence and sequence of putative adipokinetic hormones (AKHs). The selected species span a wide evolutionary range of various families and subfamilies of the clade Mantodea. The corpora cardiaca of the different species are dissected, methanolic extracts prepared, peptides separated by liquid chromatography, and AKHs detected and sequenced by ion trap mass spectrometry. All six species investigated contain an octapeptide with the primary structure pGlu‐Val‐Asn‐Phe‐Thr‐Pro‐Asn‐Trp amide, which is code‐named Emppe‐AKH and had been found earlier in three other species of Mantodea. Conspecific bioassays with the species Creoboter sp. (family Hymenopodidae) reveal an adipokinetic but not a hypertrehalosemic function of Emppe‐AKH. Comparison with other members of the Dictyoptera (cockroaches, termites) show that Emppe‐AKH is only found in certain termites, which have been recently placed into the Blattaria (cockroaches) as sister group to the family Cryptocercidae. Termites and cockroaches both show biodiversity in the sequence of AKHs, and some cockroach species even contain two AKHs. In contrast, all praying mantises—irrespective of their phylogenetic position—synthesize uniformly only one and the same octapeptide Emppe‐AKH.     

The adipokinetic hormone of the coleopteran suborder Adephaga: Structure,function, and comparison of distribution in other insects

The aim of the current study is to identify the adipokinetic hormone(s) (AKHs) of a basal suborder of the species‐rich Coleoptera, the Adephaga, and possibly learn more about the ancestral AKH of beetles. Moreover, we wanted to compare the ancestral AKH with AKHs of more advanced beetles, of which a number are pest insects. This would allow us to assess whether AKH mimetics would be suitable as insecticides, that is, be harmful to the pest species but not to the beneficial species. Nine species of the Adephaga were investigated and all synthesize only one octapeptide in the corpus cardiacum, as revealed by Edman degradation sequencing techniques or by mass spectrometry. The amino acid sequence pGlu‐Leu‐Asn‐Phe‐Ser‐Thr‐Gly‐Trp corresponds to Schgr‐AKH‐II that was first identified in the desert locust. It is assumed that Schgr‐AKH‐II—the peptide of a basal beetle clade—is the ancestral AKH for beetles. Some other beetle families, as well as some Hymenoptera (including honey bees) also contain this peptide, whereas most of the pest beetle species have different AKHs. This argues that those peptides and their receptors should be explored for developing mimetics with insecticidal properties. A scenario where Schgr‐AKH‐II (the only AKH of Adephaga) is used as basic molecular structure to derive almost all other known beetle AKHs via single step mutations is very likely, and supports the interpretation that Schgr‐AKH‐II is the ancestral AKH of Coleoptera.     

Distribution of water bugs (Heteroptera: Nepomorpha, Gerromorpha) over latitudinal zones in the northeast of European Russia

Data on the taxonomic composition of water bugs (Heteroptera: Nepomorpha, Gerromorpha) in the northeast of European Russia are given. A total of 34 water-bug species, which belong to 16 genera and 8 families, are recorded. The species diversity of Nepomorpha and Gerromorpha changes northwards: 33 occur in the taiga zone and only 8 species occur in the tundra zone. The bulk of the fauna consists of Eurosiberian (29%) and trans-Eurasian (26%) species; in latitude-related terms, the temperate group is most richly represented (64%). The species composition of the aquatic and semiaquatic heteropteran fauna of the region is especially similar to that of the tundra and taiga zones of Western Siberia.     

The first decapeptide adipokinetic hormone (AKH) in Heteroptera: a novel AKH from a South African saucer bug, Laccocoris spurcus (Naucoridae, Laccocorinae)

A novel peptide of the adipokinetic hormone (AKH)/red pigment-concentrating hormone (RPCH) family has been elucidated by mass spectrometry from the corpora cardiaca of an African saucer bug species, Laccocoris spurcus. It is the first decapeptide member found in the species-rich taxon Heteroptera, has the primary sequence pGlu-Val-Asn-Phe-Ser-Pro-Ser-Trp-Gly-Gly amide and is denoted as Lacsp-AKH. The first eight amino acids are identical to the octapeptide Anaim-AKH of the European saucer bug, Ilyocoris cimicoides. The synthetic peptide Lacsp-AKH elevates lipids upon injection into the hemolymph of L. spurcus at a low dose of 3 pmol. Swimming activity in this saucer bug also causes a significant increase in the lipid concentration in the hemolymph. Thus, both results point to an apparent function of the endogenous new decapeptide Lacsp-AKH in L. spurcus, namely, to regulate lipid mobilization. Isolation of an AKH peptide from the corpora cardiaca of the water bug Aphelocheirus aestivalis (Aphelocheiridae) resulted in the assignment of the octapeptide Anaim-AKH, supporting current phylogenies on the infraorder Nepomorpha.     

Structural diversity of adipokinetic hormones in the hyperdiverse coleopteran Cucujiformia

Seventeen species of the coleopteran series Cucujiformia are investigated for the presence and sequence of putative adipokinetic hormones (AKHs). Cucujiformia includes species from the major superfamilies, that is, Chrysomeloidea, Curculionoidea, Cucujoidea, and Tenebrionoidea. The clade Phytophaga in which the Chrysomeloidea and Curculionoidea reside, harbor very detrimental species for agriculture and forestry. Thus, this study aims not only to demonstrate the structural biodiversity of AKHs in these beetle species and possible evolutionary trends but also to determine whether the AKHs from harmful pest species can be used as lead substances for a future putative insecticide that is harmless to beneficial insects. Sequence analysis of AKHs is achieved by liquid chromatography coupled to mass spectrometry. Most of the investigated species contain AKH octapeptides in their corpora cardiaca, although previously published work also found a few decapeptides, which we comment on. The signature and sole AKH in cerambycidae Chrysomeloidea and Curculionoidea is Peram‐CAH‐I (pEVNFSPNW amide), which is also found in the majority of chrysomelidae Chrysomeloidea and in the one investigated species of Cucujoidea albeit in a few cases associated with a second AKH which can be either Peram‐CAH‐II (pELTFTPNW amide), Emppe‐AKH (pEVNFTPNW amide), or Micvi‐CC (pEINFTPNW amide). The most often encountered AKH in Tenebrionoidea, family Meloidae as well as family Tenebrionidae, is Tenmo‐HrTH (pELNFSPNW amide) followed by Pyrap‐AKH (pELNFTPNW amide) and a Tenmo‐HrTH extended decapeptide (in Meloidae). Finally, we examine AKH sequences from 43 species of cucujiform beetles, including the superfamily Coccinelloidea for a possible lead compound for producing a cucujiform‐specific pesticide.     

Global diversity of true bugs (Heteroptera; Insecta) in freshwater

The aquatic and semi-aquatic Heteroptera, consisting of the infraorders Leptopodomorpha, Gerromorpha, and Nepomorpha, comprise a significant component of the world’s aquatic insect biota. Within these three infraorders as a whole there are currently 23 families, 343 genera and 4,810 species group taxa considered valid, of which 20 families, 326 genera and 4,656 species inhabit freshwater. In addition, more than 1,100 unequivocally diagnosed species remain to be described. Aquatic Heteroptera occur on all continents except Antarctica, and are most numerous in the tropical regions, although there are many distinctly cold-adapted genera. Overall species richness is highest in the Neotropical and Oriental regions, which harbor 1,289 and 1,103 species, respectively. In comparison to these core tropical regions, species richness is significantly lower in the Afrotropical (799 species), Australasian (654 species), Palearctic (496 species), Nearctic (424 species) and Pacific (37 species) regions. Aquatic Heteroptera are notable for utilizing an exceptionally broad range of habitats, from marine and intertidal to arctic and high alpine, across a global altitudinal range of 0–4,700 m. Species may be found in almost every freshwater biotope, and many exhibit striking morphological adaptations to their aquatic environment, making them excellent subjects for ecological and biogeographic studies. Guest editors: E. V. Balian, C. Lévêque, H. Segers & K. Martens Freshwater Animal Diversity Assessment     

Phylogeny of the true water bugs (Nepomorpha: Hemiptera–Heteroptera) based on 16S and 28S rDNA and morphology

Abstract. Morphological characters and molecular sequence data were for the first time analysed separately and combined for the true water bugs (Hemiptera–Heteroptera, infraorder Nepomorpha). Data from forty species representing all families were included, together with two outgroup species representing the infraorders Gerromorpha and Leptopodomorpha. The morphological data matrix consisted of sixty‐five characters obtained from literature sources. Molecular data included approximately 960 bp from the mitochondrial gene 16S and the nuclear gene 28S for all forty‐two terminal taxa. The morphological dataset was analysed using maximum parsimony and the combined morphological and molecular (16S + 28S rDNA) dataset was analysed using direct optimization. A sensitivity analysis of sixteen different sets of parameters (various combinations of insertion–deletion cost and transversion costs) was undertaken. Character congruence was used as an optimality criterion to choose among competing phylogenetic hypotheses. The final hypothesis was obtained from the analysis of the combined molecular and mor phological dataset with the most congruent parameter set. This hypothesis supports the monophyly of all currently recognized families of Nepomorpha, and of the superfamilies Nepoidea (Nepidae + Belostomatidae), Corixoidea (Corixidae), Ochteroidea Ochteridae + Gelastocoridae), Notonectoidea (Notonectidae), and Pleoidea (Pleidae + Helotrephidae), but not the monophyly of the Naucoroidea (Naucoridae + Aphelocheiridae + Potamocoridae). The close relationship between the Notonectidae and Pleoidea is also supported. Our hypothesis concurs with Mahner in the placement of the Corixidae as a sister group to the remaining nepomorphan superfamilies except the Nepoidea, but differs in the placement of the Ochteroidea as a sister group to the Notonectoidea + Pleoidea. The superfamily Naucoroidea should be limited to only including the family Naucoridae and not the families Aphelocheiridae and Potamocoridae. The present analysis strongly supports a sister group relationship between the families Aphelocheiridae and Potamocoridae, a monophylum for which we propose a new superfamily, Aphelocheiroidea.     

Characteristics of the sperm structure in heteroptera (Hemiptera,Insecta)

Spermatozoa from eight heteropterans, each representing a different family, have been examined by electron microscopy in order to determine whether there exist characters typical for this insect group. Two such characters were found, namely bridges from the mitochondrial derivatives to the axonemal microtubules nos. 1 and 5, and two or three, rather than one, crystalline bodies within the mitochondrial derivatives. It is suggested that these characters are synapomorphic traits. The heteropteran spermatozoa lack accessory bodies typical of spermatozoa from many related groups of insects. The acrosome of the aquatic or semi-aquatic heteropterans (the infraorders Nepomorpha and Gerromorpha) has a peculiar inner structure consisting of tightly packed tubules. On the common theme of the heteropteran sperm structure, there were many variations, and the spermatozoa of each species examined can be recognized.     

Structure and function of adipokinetic hormones of the large white butterfly Pieris brassicae

The large white butterfly Pieris brassicae L. (also called cabbage white) is very common in Europe, Asia and the northern region of Africa, and has also been found in South Africa during approximately the last 20 years. The species is considered a pest insect, with larvae attacking brassicaceous crops. The adult is a strong migratory flyer and new territory can be infested this way. As a first step to investigate methods for combating this pest species, the present study aims to determine the complement of adipokinetic peptides, here generically referred to as adipokinetic hormones (AKHs), which are required to regulate the mobilization of fuels for insect flight. Biological assays, as well as mass spectrometry, reveal information about the presence, structure and function of AKHs in P. brassicae: a methanolic extract of the corpora cardiaca has hypertrehalosaemic activity in cockroaches, does not cause hyperlipaemia in locusts, and has adipokinetic activity in P. brassicae itself. Liquid‐chromatography electrospray ion trap mass spectrometry reveals three peptides that can be associated with the AKH family: the non‐amidated undecapeptide Vanca‐AKH (pELTFTSSWGGK‐OH), the nonapeptide Manse‐AKH (pELTFTSSWG amide) and the novel octapeptide Piebr‐AKH (pELTFSSGW amide). Sequence confirmation of all three assigned structures is obtained from matching mass spectrometry spectra from synthetic and native peptides. Moreover, the synthetic peptides Manse‐AKH and Piebr‐AKH have significant hyperlipaemic (=adipokinetic) activity when injected into newly‐emerged adult cabbage white butterflies. The non‐amidated Vanca‐AKH is, apparently, incompletely processed Manse‐AKH without hormonal activity. Simulated dispersal flight is able to release AKHs, as indicated by the higher concentration of lipids in the haemolymph of adult P. brassicae after activity and rest periods.     

Chemical Properties and Physiological Actions of Crustacean Chromatophorotropins

The crustacean pigment-translocating hormones, the red pigment-concentratinghormone (RPCH), an octapeptide, and the light-adapting distalretinal pigment hormone (DRPH), an octadecapeptide, are thefirst invertebrate neurohormones to be fully characterized.Studies with both purified and synthetic hormones show that,in certain decapods, RPCH is a general pigment-concentratinghormone (PCH), affecting the pigments of all kinds of chromatophores(erythrophores, xanthophores, leucophores and melanophores);the DRPH seems to serve not only light-adapting function, butalso act as a general chromatophore pigment-dispersing hormone(PDH). The two hormones thus function as antagonists when regulatingthe color-adaptation of the decapod crustaceans. PCH activityis widely distributed within the arthropod endocrine systems.The first characterized insect neurohormones, the locust adipokinetichormones (AKH I and AKH II), show close structural similaritiesto the crustacean hormone, indicating a common evolution ofsome of the arthropod neurohormones. Physiological studies ofthe three hormones (RPCH, AKH I, and AKH II) and their syntheticanalogs show that they crossreact, i.e., they all exhibit pigment-concentratingactivity when tested on decapod crustaceans, adipokinetic activitywhen tested on locusts, and hyperglycemic activity when testedon cockroaches, although each of the hormones is more potentin its own system. Structure-function studies show, however,that quite different binding-site requirements exist for thehormones in activating their receptors on the various targettissues. The physiological specificity in their action thereforeseems to depend on a differential evolution of the hormone receptors.     

A third active AKH is present in the pyrgomorphid grasshoppers Phymateus morbillosus and Dictyophorus spumans

The corpora cardiaca of the African pyrgomorphid grasshoppers Phymateus morbillosus and Dictyophorus spumans contain three adipokinetic hormones (AKHs): besides two already known AKHs, Phm-AKH-I and Scg-AKH-II (G?de et al., 1996 [G?de, G., Kellner, R., Rinehart, K.L., 1996. Pyrgomorphid grasshoppers of the genus Phymateus contain species-specific decapeptides of the AKH/RPCH family regulating lipid-mobilisation during flight. Physiol. Entomol. 21, 193-202]), a new AKH-III, denoted Phm-AKH-III, pGlu-Ile-Asn-Phe-Thr-Pro-Trp-Trp-NH(2), has been characterised. This is only the second AKH-III identified so far, thus, only three insect species - all of them grasshoppers - contain three active AKHs. Phm-AKH-III differs from Lom-AKH-III from the migratory locust, Locusta migratoria, only in position 2: isoleucine is present instead of leucine. The structure of the Phm-AKH-III was confirmed by synthesis, subsequent mass determination and reversed-phase high-performance liquid chromatography. The synthetic peptide also induced hyperlipaemia in D. spumans and L. migratoria.     

The adipokinetic hormone family in Chrysomeloidea: structural and functional considerations

The presented work is a hybrid of an overview and an original research paper on peptides belonging to the adipokinetic hormone (AKH) family that are present in the corpora cardiaca of Chrysomeloidea. First, we introduce the AKH/red pigment-concentrating hormone (RPCH) peptide family. Second, we collate the available primary sequence data on AKH peptides in Cerambycidae and Chrysomelidae, and we present new sequencing data (from previously unstudied species) obtained by liquid-chromatography coupled with ion trap electrospray ionisation mass spectrometry. Our expanded data set encompasses the primary structure of AKHs from seven species of Cerambycidae and three species of Chrysomelidae. All of these species synthesise the octapeptide code-named Peram-CAH-I (pGlu-Val-Asn-Phe-Ser-Pro-Asn-Trp amide). Whereas this is the sole AKH peptide in Cerambycidae, Chrysomelidae demonstrate a probable event of AKH gene duplication, thereby giving rise to an additional AKH. This second AKH peptide may be either Emppe-AKH (pGlu-Val-Asn-Phe-Thr-Pro-Asn-Trp amide) or Peram-CAH-II (pGlu-Leu-Thr-Phe-Thr-Pro-Asn-Trp amide). The peptide distribution and structural data suggest that both families are closely related and that Peram-CAH-I is the ancestral peptide. We hypothesise on the molecular evolution of Emppe-AKH and Peram-CAH-II from the ancestral peptide due to nonsynonymous missense single nucleotide polymorphism in the nucleotide coding sequence of prepro-AKH. Finally, we review the biological significance of the AKH peptides as hyperprolinaemic hormones in Chrysomeloidea, i.e. they cause an increase in the circulating concentration of proline. The mobilisation of proline has been demonstrated during flight in both cerambycid and chrysomelid beetles.