Substrate specificity of insect trypsins and the role of their subsites in catalysis

Trypsins have high sequence similarity, although the responses of insect trypsins to chemical and natural inhibitors suggest they differ in specificities. Purified digestive trypsins from insects of four different orders were assayed with internally quenched fluorescent oligopeptides with two different amino acids at P1 (Arg/Lys) and 15 amino acid replacements in positions P1', P2', P2, and P3. The binding energy (deltaG(s), calculated from Km values) and the activation energy (deltaG(T)(double dagger), determined from kcat/Km values) were calculated. Dictyoptera, Coleoptera and Diptera trypsins hydrolyze peptides with Arg at P1 at least 3 times more efficiently than peptides with Lys at P1, whereas Lepidoptera trypsins have no preference between Arg and Lys at that position. The hydrophobicities of each subsite were calculated from the efficiency of hydrolysis of the different amino acid replacements at that subsite. The results suggested that insect trypsin subsites become progressively more hydrophobic along evolution. Apparently, this is an adaptation to resist plant protein inhibitors, which usually have polar residues at their reactive sites. Results also suggested that, at least in lepidopteran trypsins, S3, S2, S1', and S2' significantly bind the substrate ground state, whereas in the transition state only S1' and S2' do that, supporting aspects of the presently accepted mechanism of trypsin catalysis. Homology modeling showed differences among those trypsins that may account for the varied kinetic properties.     

Phylogeny of the holometabolous insect orders: molecular evidence

Phylogenetic relationships among the holometabolous insect orders were reconstructed using 18S ribosomal DNA data drawn from a sample of 182 taxa representing all holometabolous insect orders and multiple outgroups. Parsimony analysis supports the monophyly of all holometabolous insect orders except for Coleoptera and Mecoptera. Mecoptera is paraphyletic with respect to Siphonaptera, which is nested within Mecoptera. Coleoptera is scattered as a paraphyletic assemblage across the tree topology. These data support a monophyletic Halteria (Strepsiptera + Diptera), Amphiesmenoptera (Trichoptera + Lepidoptera), Neuropterida (Neuroptera + (Megaloptera + Raphidioptera)), but Antliophora (Halteria + Mecoptera + Siphonaptera) and Mecopterida (Antliophora + Amphiesmenoptera) are paraphyletic. The limitations of using 18S ribosomal DNA as the sole phylogenetic marker for reconstructing insect ordinal relationships are discussed.     

Fruit sizes and the structure of frugivorous communities in a New Guinea lowland rainforest

A community of frugivorous insects was studied by rearing of 25 565 individual insects representing three orders (Coleoptera, Lepidoptera and Diptera except Drosophilidae) from 326 woody plant species in a lowland rainforest in Papua New Guinea. Fruits from 19.3% of plant species were not attacked by any insect order, 33.4% of plant species were attacked by a single order, 30% by two orders and 17.2% by all three orders. The likelihood of attack by individual orders was positively correlated so that a higher proportion of plant species than expected suffered either no attack at all or was attacked by all three insect orders. Fruits from most of the plant species exhibited low rates of attack and low densities of insects. One kilogram of fruit was attacked on average by 11 insects, including three to four Coleoptera, six Diptera and one Lepidoptera. Thus, we reared on average one insect from 10 fruits, including one Diptera from 14 fruits, one Coleoptera from 22 fruits and one Lepidoptera from 100 fruits. Only 72 out of the 326 plant species hosted more than one insect per 10 fruits, and only seven species supported a density of greater than one insect per fruit. Our results suggest that specialized insect seed predators are probably too rare to maintain the diversity of vegetation by density‐dependent mortality of seeds as suggested by the Janzen–Connell hypothesis. Fruit weight, fruit volume, mesocarp volume, seed volume and fleshiness had no significant effect on the probability that a fruit would be attacked by an insect frugivore. However, fruits attacked by Diptera were significantly larger and had larger volume of both mesocarp and seeds than fruits attacked by Coleoptera and Lepidoptera.     

Identification and characterization of digestive serine proteases from inhibitor-resistant Helicoverpa zea larval midgut

Protease inhibitors mediate a natural form of plant defence against insects, by interfering with the digestive system of the insect. In this paper, affinity chromatography was used to isolate trypsins and chymotrypsins from Helicoverpa zea larvae, which had been raised on inhibitor-containing diet. Sensitivity of the fractions to inhibition by plant proteinase inhibitors was tested, and compared to the sensitivity of proteinases found in insects raised on diet to which no inhibitor had been added. The isolated chymotrypsin activity was found to be less sensitive to plant protease inhibitors. The sensitivity of the isolated trypsin activity was found to be intermediate between completely sensitive trypsins and completely insensitive forms that have been previously described. Mass spectrometry was used to identify one trypsin and two chymotrypsins in the partially purified protease fraction. The sequence features of these proteases are discussed in relation to their sensitivity to inhibitors. The results provide insight in the enzymes deployed by Helicoverpa larvae to overcome plant defence.     

Odorant binding protein diversity and distribution among the insect orders, as indicated by LAP, an OBP-related protein of the true bug Lygus lineolaris (Hemiptera, Heteroptera)

Insect odorant binding proteins (OBPs) are thought to deliver odors to olfactory receptors, and thus may be the first biochemical step in odor reception capable of some level of odor discrimination. OBPs have been identified from numerous species of several insect orders, including Lepidoptera, Diptera, Coleoptera and Hymenoptera; all are holometabolous insects belonging to the monophyletic division of insects known as the Endopterygota. Recently, an antennal protein with OBP-like properties was identified from Lygus lineolaris, a hemipteran insect representing the Hemipteroid Assemblage, a sister division to the Endopterygota. The full length sequence of Lygus antennal protein (LAP) is presented in this report. In situ hybridization analysis revealed LAP expression in cell clusters associating with olfactory sensilla; expression was adult-specific, initiating in developing adult tissue during the transitional period that precedes the actual adult molt. Sequence analysis confirmed that LAP is homologous with the OBP-related protein family, and most similar to the OS-E and OS-F proteins of Drosophila, the ABPX proteins of Lepidoptera and the OBPRP proteins of the Coleoptera. Assuming that the OBP-related proteins represent one homologous family, the identification of LAP significantly expands the phylogenetic depth of that family and its underlying role in odor detection to encompass all members of the Endopterygota and Hemipteroid Assemblage, which comprise >90% of all insect species.     

Diet and trophic groups of an aquatic insect community in a tropical stream.

The diet and trophic groups of an assemblage of aquatic insects were studied in a tropical stream. Genera of the orders Ephemeroptera, Odonata, Plecoptera, Lepidoptera, and Hemiptera showed feeding specialization. Others, such as Trichoptera, Coleoptera, and Diptera, showed great diet variation with genera of different trophic groups. Seasonal variation of insect diet, evident only for some genera of the orders Trichoptera, Lepidoptera, Coleoptera, and Diptera, was due to the differences observed in community composition and to generalist habits of these genera. However, the seasonal comparison of trophic groups showed no significant statistical differences. The great importance of organic matter, a non-limited resource, in the diet of Ribeir?o do Atalho aquatic insects may be the explanation for the trophic stability in this community organization.     

Coevolution of insect trypsins and inhibitors

Many plant proteinase inhibitors have lysine at the P1 position, presumably to avoid hydrolysis by insect trypsins. Lepidopteran trypsins appear to have adapted to resist proteinase inhibitors through increased inhibitor hydrolysis and decreased binding to inhibitor hydrophilic reactive sites. Lepidopteran digestive trypsins prefer lysine at the P1 position and have substrate binding subsites more hydrophobic than trypsins from insects in other orders. All available sequences of sensitive and inhibitor-insensitive insect trypsins were aligned with porcine trypsin, for which interactions with Kunitz and Bowman-Birk inhibitor are known. After discounting conserved positions and positions not typical of sensitive or insensitive trypsins, the following residues were considered important to insect trypsin-PI interactions (chymotrypsin numbering): 60, 94, 97, 98, 99, 188, 190, 213, 215, 217, 219, 228. These residues support the Neighbor Joining analysis tree branches separating sensitive and insensitive trypsin sequences. Primary sequences interacting with PIs are around the active site, with some forming part of the S1 (188, 217, 219 and 228) or S4 (99, 215) pockets.     

Immunohistochemical evidence for the presence of tryptophan hydroxylase in the brains of insects as revealed by sheep anti-tryptophan hydroxylase polyclonal antibody

Immediately following the discovery of tryptophan hydroxylase in Drosophila, we demonstrated the presence of tryptophan hydroxylase in the brain of the beetle Harmonia axyridis (Coleoptera: Coccinellidae). However, whether tryptophan hydroxylase is present in the brains of other insects is still a matter of discussion. In the current study, sheep anti-tryptophan hydroxylase polyclonal antibody has been applied to test for tryptophan hydroxylase immunoreactivity in a broader taxonomic range of insect brains, including holometabolous and hemimetabolous insects: one species each of Coleoptera, Hymenoptera, Diptera, and Blattaria, and two species of Lepidoptera. All species show consistent tryptophan hydroxylase immunoreactivity with distribution patterns matching that of serotonin. The immuno-positive results of such an antibody in brains from diverse orders of insects suggest that specific tryptophan hydroxylase responsible for central serotonin synthesis is probably present in the brains of all insects. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. This work was supported by grants from the National Natural Science Foundation of China (grant no. 30470546) and the Natural Science Foundation of Jilin Province (grant no. 20030550–7).     

Accelerated evolution of mitochondrial but not nuclear genomes of Hymenoptera: new evidence from crabronid wasps

Mitochondrial genes in animals are especially useful as molecular markers for the reconstruction of phylogenies among closely related taxa, due to the generally high substitution rates. Several insect orders, notably Hymenoptera and Phthiraptera, show exceptionally high rates of mitochondrial molecular evolution, which has been attributed to the parasitic lifestyle of current or ancestral members of these taxa. Parasitism has been hypothesized to entail frequent population bottlenecks that increase rates of molecular evolution by reducing the efficiency of purifying selection. This effect should result in elevated substitution rates of both nuclear and mitochondrial genes, but to date no extensive comparative study has tested this hypothesis in insects. Here we report the mitochondrial genome of a crabronid wasp, the European beewolf (Philanthus triangulum, Hymenoptera, Crabronidae), and we use it to compare evolutionary rates among the four largest holometabolous insect orders (Coleoptera, Diptera, Hymenoptera, Lepidoptera) based on phylogenies reconstructed with whole mitochondrial genomes as well as four single-copy nuclear genes (18S rRNA, arginine kinase, wingless, phosphoenolpyruvate carboxykinase). The mt-genome of P. triangulum is 16,029 bp in size with a mean A+T content of 83.6%, and it encodes the 37 genes typically found in arthropod mt genomes (13 protein-coding, 22 tRNA, and two rRNA genes). Five translocations of tRNA genes were discovered relative to the putative ancestral genome arrangement in insects, and the unusual start codon TTG was predicted for cox2. Phylogenetic analyses revealed significantly longer branches leading to the apocritan Hymenoptera as well as the Orussoidea, to a lesser extent the Cephoidea, and, possibly, the Tenthredinoidea than any of the other holometabolous insect orders for all mitochondrial but none of the four nuclear genes tested. Thus, our results suggest that the ancestral parasitic lifestyle of Apocrita is unlikely to be the major cause for the elevated substitution rates observed in hymenopteran mitochondrial genomes.     

太阳能灭虫器对吐鲁番葡萄产区害虫的诱捕效果及对天敌安全性评价

本文研究了FWS-DBL-1新型太阳能灭虫器在320～580、360、400、460和520 nm 5种不同波长下对吐鲁番葡萄产区害虫的诱捕效果及对天敌安全性。结果表明:在5种波长下,灭虫器诱捕昆虫的种类相似,以鞘翅目、鳞翅目、半翅目、脉翅目、双翅目、直翅目和膜翅目等为主。诱捕的主要昆虫是蜉金龟Aphodius sp.、白云斑鳃金龟Polyphylla alba vicaria Semenov、额喙丽金龟Adoretus nigriforns Steven、毛喙丽金龟Adoretus hirsutus Ohaus、绣罗夜蛾Leucanitis picta Christoph和淘赏夜蛾Catocala puerpera Giorna等。其中:灭虫器在400 nm波长下的诱捕量最多,其次是320～580 nm和360 nm波长。综合分析比较:400 nm波长对鞘翅目害虫的诱捕效果最好,360 nm波长对鳞翅目和双翅目害虫的诱捕能力最强,520 nm波长对半翅目害虫的诱捕效果最好。对天敌的安全性研究表明:FWS-DBL-1太阳能灭虫器对天敌有一定的诱捕作用,益害比为1∶4～1∶5。诱捕量较多的是步甲科和草蛉科的天敌,占天敌总诱捕量的85.82%。在这5个波长中,360 nm波长下,灭虫器对天敌的诱捕作用最小,对天敌的安全性相对较高。     

Seasonal pattern of insect abundance in the Brazilian cerrado

Abstract In Brazil, a severe dry season lasting for approximately 5 months and frequent fires make life difficult for cerrado insects. In certain aspects, the cerrado can be considered to be an understudied ecosystem; even basic information such as knowledge about the annual peak in abundance of different insect orders is unknown. Insect abundance patterns have only been investigated for a few groups in the cerrado region. Thus, our study concerns the temporal distribution of insect abundance in the savanna‐like vegetation of the central Brazilian cerrado (sensu stricto) in Distrito Federal. The region has a well‐defined, long dry season between May and September. The insects were sampled by window, malaise tent and pitfall traps within 1 year. We used a multiple linear regression to analyse the relationship between abundance of insects of each order and climate variables. A total of 50 127 individuals from 15 orders was collected. The orders were Coleoptera (26%), Hymenoptera (23%), Diptera (20.5%), Isoptera (20%), Homoptera (4%), Lepidoptera (4%), Orthoptera (1.5%) and Hemiptera (1%). The abundance of Diptera, Homoptera, Lepidoptera and Orthoptera was randomly distributed over time, Isoptera peaked in the first half of the wet season, Coleoptera and Hemiptera in the second half of the wet season and Hymenoptera in each season. A significant correlation was found only between Coleoptera and delayed climatic variables. There were no obvious trends that might help explain the abundance patterns observed. The study provides baseline information about phenological patterns of insect abundance and permits evaluation of this group as a resource for various food chains and different trophic levels.     

Prospecting for cellulolytic activity in insect digestive fluids

Efficient cellulolytic enzymes are needed to degrade recalcitrant plant biomass during ethanol purification and make lignocellulosic biofuels a cost-effective alternative to fossil fuels. Despite the large number of insect species that feed on lignocellulosic material, limited availability of quantitative studies comparing cellulase activity among insect taxa constrains identification of candidate species for more targeted identification of effective cellulolytic systems. We describe quantitative determinations of the cellulolytic activity in gut or head-derived fluids from 68 phytophagous or xylophagous insect species belonging to eight different taxonomic orders. Enzymatic activity was determined for two different substrates, carboxymethyl cellulose (CMC) and microcrystalline cellulose (MCC), approximating endo-β-1,4-glucanase and complete cellulolytic activity, respectively. Highest CMC gut fluid activities were found in Dictyoptera, Coleoptera, Isoptera, and Orthoptera, while highest MCC gut fluid activities were found in Coleoptera, Hymenoptera, Lepidoptera, and Orthoptera. In most cases, gut fluid activities were greater with CMC compared to MCC substrate, except in Diptera, Hymenoptera, and Lepidoptera. In contrast, cellulolytic activity levels in most head fluids were greater on the MCC substrate. Our data suggests that a phylogenetic relationship may exist for the origin of cellulolytic enzymes in insects, and that cellulase activity levels correlate with taxonomic classification, probably reflecting differences in plant host or feeding strategies.     

中国半翅目等29目昆虫新分类单元2021年年度报告

本文以2021年度中国半翅目等29目昆虫(除昆虫纲鞘翅目、双翅目、鳞翅目、膜翅目之外的其他昆虫类群)发表的新分类单元为研究对象, 系统编汇了新分类单元信息、模式标本名录与文献目录, 并分析了描述新种的热点地区、模式标本存放情况以及发表期刊的特点等。2021年度中国半翅目等29目昆虫描述了新属35个, 新物种386个以及1个新种下分类单元, 共计422个新分类单元; 基于对新物种在省级行政区分布的分析, 新物种发现的热点地区分布于中国西南(云南、贵州以及四川等)及其邻近地区(广西和西藏等), 而以200 km × 200 km栅格为操作单位, 新物种发现的热点地区主要集中在山地地区。395位研究者参与了本年度新分类单元命名和描述工作, 相关结果发表在28种期刊上, 共205篇文章。本研究及时更新了2021年度中国半翅目等29目昆虫的生物多样性基础数据, 对于加快建设数据共享平台、支撑生物多样性保护和管理, 以及保障国门生物安全等具有重要的作用。     

南京地区外来植物一年蓬上访花昆虫的多样性及其访花选择性的影响因素分析

为揭示外来植物一年蓬Erigeron annuus上的本土访花昆虫多样性和影响访花行为的因素, 本研究在南京郊区进行了连续2年的野外调查, 采用跨栏模型分析了环境因素如何影响昆虫的访花选择性, 即接受概率（测度是否接受一年蓬花）和访问频数（测度接受一年蓬花的程度）。调查发现, 访问一年蓬花的昆虫共计9目54科145种, 其中, 科丰富度占优势的是膜翅目、 鳞翅目和鞘翅目（均占总科数的20.75%）, 其次是双翅目（18.87%）和半翅目（13.21%）; 物种丰富度占优势的是双翅目（26.39%）, 其次是膜翅目（18.75%）、 半翅目（18.75%）、 鞘翅目（17.36%）和鳞翅目（15.38%）。多数目的物种丰富度在6-7月最高, 9月最低, 仅双翅目（食蚜蝇为主）在5月最高。运用跨栏模型对物种优势度最大的半翅目、 膜翅目和双翅目等的访花个体数量及其影响因素的分析结果表明： 影响半翅目和膜翅目对一年蓬花访问倾向（接受概率）的因素多于影响其访问频数的因素, 由此预测这些访花昆虫可能参考较多因素做出是否接受一年蓬花, 而依据较少线索做出访问程度的访花行为决策; 一年蓬植株密度影响半翅目和膜翅目昆虫的接受概率, 而花密度影响半翅目和双翅目昆虫的接受概率和访问频数, 说明靶标植物花的特性可能对访花昆虫的访花行为决策起主要作用。     

Replication origin of mitochondrial DNA in insects

The precise position of the replication origin (O(R)) of mtDNA was determined for insect species belonging to four different orders (four species of Drosophila, Bombyx mori, Triborium castaneum, and Locusta migratoria, which belong to Diptera, Lepidoptera, Coleoptera, and Orthoptera, respectively). Since the free 5' ends of the DNA strands of mtDNA are interpreted as the O(R), their positions were mapped at 1-nucleotide resolution within the A + T-rich region by using the ligation-mediated PCR method. In all species examined, the free 5' ends were found within a very narrow range of several nucleotides in the A + T-rich region. For four species of Drosophila, B. mori, and T. castaneum, which belong to holometabolous insects, although the O(R)'s were located at different positions, they were located immediately downstream of a series of thymine nucleotides, the so-called T-stretch. These results strongly indicate that the T-stretch is involved in the recognition of the O(R) of mtDNA at least among holometabolous insects. For L. migratoria (hemimetabolous insect), on the other hand, none of the long stretches of T's was found in the upstream portion of the O(R), suggesting that the regulatory sequences involved in the replication initiation process have changed through insect evolution.     

Elongation factor-1 alpha occurs as two copies in bees: implications for phylogenetic analysis of EF-1 alpha sequences in insects

We report the complete sequence of a paralogous copy of elongation factor-1 alpha (EF-1 alpha) in the honeybee, Apis mellifera (Hymenoptera: Apidae). This copy differs from a previously described copy in the positions of five introns and in 25% of the nucleotide sites in the coding regions. The existence of two paralogous copies of EF-1 alpha in Drosophila and Apis suggests that two copies of EF-1 alpha may be widespread in the holometabolous insect orders. To distinguish between a single, ancient gene duplication and parallel, independent fly and bee gene duplications, we performed a phylogenetic analysis of hexapod EF-1 alpha sequences. Unweighted parsimony analysis of nucleotide sequences suggests an ancient gene duplication event, whereas weighted parsimony analysis of nucleotides and unweighted parsimony analysis of amino acids suggests the contrary: that EF-1 alpha underwent parallel gene duplications in the Diptera and the Hymenoptera. The hypothesis of parallel gene duplication is supported both by congruence among nucleotide and amino acid data sets and by topology-dependent permutation tail probability (T-PTP) tests. The resulting tree topologies are also congruent with current views on the relationships among the holometabolous orders included in this study (Diptera, Hymenoptera, and Lepidoptera). More sequences, from diverse orders of holometabolous insects, will be needed to more accurately assess the historical patterns of gene duplication in EF-1 alpha.      

长白山苔原带昆虫群落组成与时间动态

【目的】苔原是一种极地景观类型,本研究调查了长白山苔原带昆虫的物种组成与时间动态,以期为苔原带昆虫保护以及昆虫与苔原植物协同进化研究提供基础依据。【方法】2005-2007年每年6-9月,在长白山北坡、西坡苔原带利用网捕、灯诱、巴氏罐法采集昆虫标本。【结果】2005-2007年从长白山苔原带共获得昆虫标本4 634头,隶属于11目105科550种,其中鳞翅目、鞘翅目、膜翅目、双翅目的种类和个体数量较为丰富,这4个类群的物种数与个体数的最高峰都出现在7月份。对应分析显示,鳞翅目昆虫对7月份,鞘翅目昆虫对8和9月,膜翅目和双翅目昆虫对6月的适应性较强。苔原带7月份昆虫的物种数(382种)、个体数(2 571个个体)和多样性指数(4.673)都最高,物种数和个体数在9月份最低,仅22种265个个体。不同月份昆虫的物种数与个体数之间呈显著正相关(R=0.992)。不同月份间昆虫的相似性低(相似性系数＜0.20)。【结论】长白山苔原带昆虫多样性较低,7月为昆虫的活跃高峰期,9月昆虫的个体数和物种数均较少。鳞翅目昆虫的物种最丰富,对7月份气侯的适应性相对较强,鞘翅目对季节变化的适应能力强于其他类群,在维持苔原带生态平衡中起着重要作用。     

Long-Term Storage of Infective Microsporidian Spores in Liquid Nitrogen

Thirty-one species of microsporidia, isolated from insects and stored in liquid nitrogen for up to 25 yr, were infectious when removed from liquid nitrogen. The natural hosts of all of these microsporidia were terrestrial insects, representing six different insect orders: Coleoptera, Diptera, Hemiptera, Hymenoptera, Lepidoptera, and Orthoptera. All microsporidia from terrestrial insects that were tested survived storage in liquid nitrogen, while Nosema algerae , a microsporidium from aquatic mosquito hosts did not survive freezing in liquid nitrogen. A Nosema species from the alfalfa weevil, Hypera postica , lost some infectivity in a water storage medium after 25 yr in liquid nitrogen. Liquid nitrogen storage of microsporidian spores in 50% and 100% glycerol media reduced loss of infectivity and is recommended for extended storage of microsporidia from terrestrial insect hosts.     

The complete nucleotide sequence and gene organization of the mitochondrial genome of the oriental mole cricket, Gryllotalpa orientalis (Orthoptera: Gryllotalpidae)

The complete nucleotide sequences of the mitochondrial genome (mitogenome) of the oriental mole cricket, Gryllotalpa orientalis (Orthoptera: Gryllotalpidae), were determined. The 15,521-bp-long G. orientalis mitogenome contains typical gene complement, base composition, and codon usage found in metazoan mitogenomes. The G. orientalis mitogenome contains the third lowest A+T content (70.5%) among the complete insects mt genome sequences. The initiation codon for the G. orientalis COI gene appears to be ATG, instead of the tetranucleotides, which have been postulated to act as initiation codon for Locusta migratoria and some lepidopteran COI genes. The initiation codon for ND2 appears to be GTG, which is rare, but has been designated as an initiator of Tricholepidion gertschi ND2. All anticodons of G. orientalis tRNAs were identical to Drosophila yakuba and L. migratoria. The tRNA(Ser)(AGN) could not form a stable stem loop structure in the DHU arm as shown in many other insect tRNA(Ser)(AGN). Phylogenetic analysis of nucleotide sequence information from all mt genes supported a monophyletic Diptera, a monophyletic Lepidoptera, a monophyletic Coleoptera, a monophyletic Mecopterida (Diptera+Lepidoptera), and a monophyletic Endopterygota (Diptera+Lepidoptera+Coleoptera), suggesting that the complete insect mitogenome sequence has a resolving power to the diversification events within Endopterygota. However, the relationships of ancient insect orders were unstable, indicating the limited use of mitogenome information at deeper phylogenetic depth.     

Occurrence of proctolin in six orders of insects

Nine representatives of six orders of insects (Orthoptera, Diptera, Coleoptera, Hemiptera, Hymenoptera, Lepidoptera) were extracted and partially processed by means used in the recent isolation of proctolin, a pentapeptide transmitter candidate in insects. Each insect yielded a substance with pharmacological activity on cockroach proctodeal muscle similar to that of proctolin. Like the responses evoked by proctolin and nerve stimulation, responses to the purified extracts were inhibited by tyramine. All of the active substances behaved as proctolin when subjected to paper chromatography or high voltage paper electrophoresis at pH 6.4 and 3.5. Proctolin appears to be widely and perhaps universally present in the Insecta occurring in most at levels of 2 to 9 μg/kg body weight.