Key biosynthetic gene subfamily recruited for pheromone production prior to the extensive radiation of Lepidoptera

<p>Abstract</p> <p>Background</p> <p>Moths have evolved highly successful mating systems, relying on species-specific mixtures of sex pheromone components for long-distance mate communication. Acyl-CoA desaturases are key enzymes in the biosynthesis of these compounds and to a large extent they account for the great diversity of pheromone structures in Lepidoptera. A novel desaturase gene subfamily that displays Δ11 catalytic activities has been highlighted to account for most of the unique pheromone signatures of the taxonomically advanced ditrysian species. To assess the mechanisms driving pheromone evolution, information is needed about the signalling machinery of primitive moths. The currant shoot borer, <it>Lampronia capitella</it>, is the sole reported primitive non-ditrysian moth known to use unsaturated fatty-acid derivatives as sex-pheromone. By combining biochemical and molecular approaches we elucidated the biosynthesis paths of its main pheromone component, the (<it>Z,Z</it>)-9,11-tetradecadien-1-ol and bring new insights into the time point of the recruitment of the key Δ11-desaturase gene subfamily in moth pheromone biosynthesis.</p> <p>Results</p> <p>The reconstructed evolutionary tree of desaturases evidenced two ditrysian-specific lineages (the Δ11 and Δ9 (18C>16C)) to have orthologs in the primitive moth <it>L. capitella </it>despite being absent in Diptera and other insect genomes. Four acyl-CoA desaturase cDNAs were isolated from the pheromone gland, three of which are related to Δ9-desaturases whereas the fourth cDNA clusters with Δ11-desaturases. We demonstrated that this transcript (<it>Lca</it>-KPVQ) exclusively accounts for both steps of desaturation involved in pheromone biosynthesis. This enzyme possesses a Z11-desaturase activity that allows transforming the palmitate precursor (C16:0) into (<it>Z</it>)-11-hexadecenoic acid and the (<it>Z</it>)-9-tetradecenoic acid into the conjugated intermediate (<it>Z,Z</it>)-9,11-tetradecadienoic acid.</p> <p>Conclusion</p> <p>The involvement of a single Z11-desaturase in pheromone biosynthesis of a non-ditrysian moth species, supports that the duplication event leading to the origin of the Lepidoptera-specific Δ11-desaturase gene subfamily took place before radiation of ditrysian moths and their divergence from other heteroneuran lineages. Our findings uncover that this novel class of enzymes affords complex combinations of unique unsaturated fatty acyl-moieties of variable chain-lengths, regio- and stereo-specificities since early in moth history and contributes a notable innovation in the early evolution of moth-pheromones.</p

Identification, synthesis and activity of sex pheromonegland components of the autumn gum moth (Lepidoptera:Geometridae), a defoliator of Eucalyptus

Summary.: The autumn gum moth, Mnesampela privata (Guenée) (Lepidoptera: Geometridae), is native to Australia and can be a pest of plantation eucalypts. Field-collected and laboratory-reared female autumn gum moths were dissected to remove glands likely to contain components of the sex pheromone. Using gas chromatography (GC) and combined gas chromatography-mass spectrometry (GC-MS), three compounds were identified from female extracts, namely (3Z,6 Z,9 Z)-3,6,9-nonadecatriene, 1-hexadecanol and 1-octadecanol (confirmed by comparison with synthetic samples). Nonadecatriene elicited an antennal response in male autumn gum moth during gas chromatographic analyses combined with electroantennographic detection (GC-EAD). In electroantennogram (EAG) recording male M. privata antennae responded to the nonadecatriene. Nonadecatriene was synthesised via Kolbe electrolysis, starting with (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid (linolenic acid) and propanoic acid or via an alternative four-step method also starting from linolenic acid. In field trials (3Z,6Z,9Z)-3,6,9-nonadecatriene proved attractive to male moths. Thus, we conclude that (3Z,6Z,9Z)-3,6,9- nonadecatriene is a sex pheromone component of autumn gum moth. This component has been identified in extracts from other geometrids in the same subfamily, Ennominae. However, to our knowledge this is the first example where (3Z,6Z,9Z)-3,6,9-nonadecatriene has been found in females and also proved attractive to male moths when presented on its own. Our results are discussed in relation to other geometrid pheromone

Semi-selective fatty acyl reductases from four heliothine moths influence the specific pheromone composition.

peer reviewed[en] BACKGROUND: Sex pheromones are essential in moth mate communication. Information on pheromone biosynthetic genes and enzymes is needed to comprehend the mechanisms that contribute to specificity of pheromone signals. Most heliothine moths use sex pheromones with (Z)-11-hexadecenal as the major component in combination with minor fatty aldehydes and alcohols. In this study we focus on four closely related species, Heliothis virescens, Heliothis subflexa, Helicoverpa armigera and Helicoverpa assulta, which use (Z)-11-hexadecenal, (Z)-9-tetradecanal, and (Z)-9-hexadecenal in different ratios in their pheromone blend. The components are produced from saturated fatty acid precursors by desaturation, β-oxidation, reduction and oxidation. RESULTS: We analyzed the composition of fatty acyl pheromone precursors and correlated it to the pheromone composition. Next, we investigated whether the downstream fatty-acyl reduction step modulates the ratio of alcohol intermediates before the final oxidation step. By isolating and functionally characterizing the Fatty Acyl Reductase (pgFAR) from each species we found that the pgFARs were active on a broad set of C8 to C16 fatty acyl substrates including the key pheromone precursors, Z9-14, Z9-16 and Z11-16:acyls. When presenting the three precursors in equal ratios to yeast cultures expressing any of the four pgFARs, all reduced (Z)-9-tetradecenoate preferentially over (Z)-11-hexadecenoate, and the latter over (Z)-9-hexadecenoate. Finally, when manipulating the precursor ratios in vitro, we found that the pgFARs display small differences in the biochemical activity on various substrates. CONCLUSIONS: We conclude that a pgFAR with broad specificity is involved in heliothine moth pheromone biosynthesis, functioning as a semi-selective funnel that produces species-specific alcohol product ratios depending on the fatty-acyl precursor ratio in the pheromone gland. This study further supports the key role of these in pheromone biosynthesis and emphasizes the interplay between the pheromone fatty acyl precursors and the Lepidoptera specific pgFARs in shaping the pheromone composition

Alarm Pheromones and Chemical Communication in Nymphs of the Tropical Bed Bug Cimex hemipterus (Hemiptera: Cimicidae)

The recent resurge of bed bug infestations (Cimex spp.; Cimicidae) and their resistance to commonly used pesticides calls for alternative methods of control. Pheromones play an important role in environmentally sustainable methods for the management of many pest insects and may therefore be applicable for the control of bed bugs. The tropical bed bug, Cimex hemipterus, is a temporary ectoparasite on humans and causes severe discomfort. Compared to the common bed bug, Cimex lectularius, little is known about the chemical signalling and pheromone-based behaviour of the tropical species. Here, we show that the antennal morphology and volatile emission of C. hemipterus closely resembles those of C. lectularius and we test their behavioural responses to conspecific odour emissions. Two major volatiles are emitted by male, female and nymph C. hemipterus under stress, (E)-2-hexenal and (E)-2-octenal. Notably, nymph emissions show contrasting ratios of these compounds to adults and are further characterized by the addition of 4-oxo-(E)-2-hexenal and 4-oxo-(E)-2-octenal. The discovery of this nymph pheromone in C. hemipterus is potentially the cause of a repellent effect observed in the bio-tests, where nymph odours induce a significantly stronger repellent reaction in conspecifics than adult odours. Our results suggest that pheromone-based pest control methods developed for C. lectularius could be applicable to C. hemipterus, with the unique nymph blend showing promising practical properties

Bland hårdhandskar och silkesvantar. – en etnografisk studie om väntrumsmiljöer på socialkontor i Göteborg

Denna studie syftar till att undersöka mottagningsmiljön i några av socialkontorens väntrum inom Göteborgs Stad och hur utformningen av miljön påverkar mötet mellan klient och det myndighetssystem som socialtjänsten utgör. Studien granskar väntrummens utformning och interiör samt interaktioner mellan besökare och myndighetsutövare genom deltagande observation. Analysen sker med hjälp av ett reflexivt förhållningssätt till kunskapsframställning där empiri och slutsatser tolkas genom en abduktiv ansats. Fältstudierna baseras på en mikroetnografisk metod som genomfördes under åtta dagar, varav sex dagar ägnades åt djupstudier på tre utvalda socialkontor. Analys och resultat tolkas genom teorier om makt, skam och stigma. Resultaten visar på stora skillnader i väntrumsmiljöerna och mottagningen för klienter där interiören inte visar många tecken på samverkan mellan socialkontoren när det kommer till tankar om utformning. Vissa socialkontor har en vakt som central aktör medan andra har receptionister och socialsekreterare som tar emot klienter. Bemötandena varierar mellan socialkontoren från medvetet vänskapliga, med småprat och tecken på etablerade relationer som betonar trivsel, till snabba och korta hänvisningar som betonar funktionalitet. Samtliga socialkontor har i varierande grad anslag på väggar, utformning av rummet och bemötande som kan ha stigmatiserande effekt. Makten ger sig till uttryck genom olika former beroende på socialkontor och varierar från osynlig till tydlig och auktoritär

Genetically similar strains of Escherichia coli O157:H7 isolated from sheep, cattle and human patients

Conclusions: The same pathogenic subtypes of VTEC O157:H7, including the highly virulent clade 8, appear to be present in both sheep and cattle in Sweden, suggesting strains can circulate freely between ruminant reservoirs

Using Jupiter’s Moon Io as a Plasma Probe

The structure of the plasma in Jupiter’s vast magnetosphereis complicated and not fully understood. One way to study the plasma is to look at auroral emissions from the moonIo as it moves through different regions of the plasma torus that surrounds Jupiter. In this paper, the correlation between aurorabrightness on Io and the plasma density at the position of the moon is investigated. If a correlation exists, auroral emissonson Io could be used as a diagnostic for the current state ofJupiter’s plasma environment. For this purpose, a model of the Io plasma torus is developed, combining ideas from different existing models. The model is compared with observations of aurorae on Io made by the Hubble Space Telescope. Io’s position at the time of the observations is obtained with SPICE, a software developed by NASA. A moderate correlation is found when using the whole data set of observations. However, a strong correlation is found for observations on the dusk side of Jupiter. Strong correlations are also found when studying individual years and epochs.Strukturen på plasman i Jupiters vidsträckta magnetosfär är komplicerad och inte fullständigt känd. Ett sätt att studera plasman är att undersöka ljuset från polarsken på månen Io då den passerar genom olika regioner av det torusformade plasmamolnet som omsluter Jupiter. I denna artikel undersöks korrelationen mellan polarskenets ljusstyrka och plasmans densitiet kring månens position. Om ett sådant samband finns skulle ljusstyrkan hos månens polarsken kunna användas som diagnostik för plasmans aktuella tillstånd. För detta ändamål utvecklas en modell av plasmatorusen genom att kombinera ide´er från flera tidigare modeller. Modellen jämförs sedan med observationer av polarskenet på Io genomförda med rymdteleskopet Hubble. Månens position vid de olika tidpunkterna bestäms med hjälp av SPICE, en mjukvara utvecklad av NASA. En måttligt stark korrelation uppnås när hela datamängden används. När däremot endast data från Jupiters gryningssida används uppnås en korrelation. Det hittas även starka samband när enskilda år studeras.Kandidatexjobb i elektroteknik 2022, KTH, Stockhol

Pheromone mating disruption of the pine sawfly Neodiprion sertifer : is the size of the treated area important?

Few attempts to control forest insects by means of pheromone mating disruption have been reported. The first such experiments with the European pine sawfly, Neodiprion sertifer (Hymenoptera: Diprionidae), resulted in nearly complete trap catch reduction, but no effects on population density or sex ratio were noted. Unmated females lay eggs, which develop into males only. Therefore, if mating disruption would be successful, a more male-biased sex ratio is expected the next generation. One possible explanation for the early results is that mated females disperse into the treated area, and thus obscure the effects of the treatment. In order to reduce the effect of such immigrating females, the treated area was increased in the experiment described here from the earlier used 0.5 – 4 ha to 25 ha. The acetate of erythro-3,7-dimethyl-2- pentadecanol was used for disruption and released from dispensers every 10 m. The influence on male orientation was monitored by pheromone traps, baited with the acetate of (2S,3S,7S)-3,7- dimethyl-2-pentadeanol and placed at 100 m interval along two perpendicular, 1500 m lines intersecting the treated area. The trap catch reduction was near 100% during the first month, but then declined to around 90% during the second month. Mating frequencies were checked by comparing the sex ratio of the next generation from within and outside the treatment area. No effect of the treatment on the sex ratio was detected, and the frequency of mated females could be assumed to be independent of treatment. Alternative hypotheses to explain the failure of pheromone mating disruption in N. sertifer are discussed

SPME collection and GC-MS analysis of volatiles emitted during the attack of male Polygraphus poligraphus (Coleoptera, Curcolionidae) on Norway spruce

Tree mortality caused by bark beetles has increased in recent decades in both Europe and North America. In a large recent outbreak in central Sweden the bark beetle Polygraphus poligraphus was often found together with the spruce bark beetle Ips typographus in killed trees. To increase the understanding of the aggregation behavior of P. poligraphus we used solid phase microextraction (SPME) to collect volatile organic compounds (VOCs) released from single P. poligraphus males, with and without added females, colonizing Norway spruce stem sections and analyzed the sampled compounds by combined gas chromatography and mass spectrometry (GC-MS). High amounts of terpinen-4-ol, a substance found in the hindguts of P. poligraphus males in earlier studies, were released by colonizing males. The emission of both enantiomers of terpinen-4-ol was monitored by GC-MS over time as the males aged in the absence and presence of females. Single males emitted (R)-(-)-terpinen-4-ol for up to 60 days in high enantiomeric purity but the enantiomeric excess (ee) varied between males, and also for the same individual, over time from 96.3% to 99.3% ee. In the presence of females, males also emitted terpinen-4-ol for up to 50 days but now in lower amounts and with lower enantiomeric purity varying from 67.7% ee to 99.3% ee. Small quantities of other volatile compounds were emitted from the colonizing beetles including cis-and trans-4-thujanol, both of which were previously shown to be present in the hindguts of males. In earlier studies frontalin was found to attract P. poligraphus, but in our study it was not identified among emitted compounds from colonizing beetles

Stereoisomeric separation of derivatised 2-alkanols using gas chromatography – mass spectrometry: sex pheromone precursors found in pine sawfly species

Abstract in UndeterminedStereoisomers of long-chain secondary alcohols are used as sex pheromone precursors among pine sawflies and some species can be severe pine forest pests. To use their pheromone in environmentally friendly pest management, methods are needed that can determine the stereochemistry of the precursor alcohols. Combinations of 11 acid chloride derivatives and 10 GC columns were evaluated for separation of stereoisomers of 3,7-dimethylundecan-2-ol, 3,7-dimethyldodecan-2-ol, 3,7-dimethyltridecan-2-ol, 3,7-dimethyltetradecan-2-ol, and 3,7-dimethylpentadecan-2-ol. Derivatization with (S)-2-acetoxypropionyl chloride in combination with a Chiraldex B-PA column separated all eight stereoisomers of 3,7dimethylundecan-2-ol, 3,7-dimethyldodecan-2-ol, and 3,7-dimethyltridecan-2-ol. A combination of two different derivatization methods was needed to separate all stereoisomers of the longer chained alcohols 3,7-dimethyltetradecan-2-ol and 3,7-dimethylpentadecan-2-ol. A female extract of the pine sawfly Neodiprion lecontei was also analyzed and the stereochemistry of the sex pheromone alcohol precursor was determined to be (2S,3S,7S)-3,7-dimethylpentadecan-2-ol at an amount of about 7 ng/female. This paper presents the first GC-MS separation of all eight stereoisomers of 3,7-dimethylundecan-2-ol, 3,7dimethyldodecan-2-ol, and 3,7-dimethyltridecan-2-ol in a single analytical run and also the first GC-MS determination of the stereochemistry of the sex pheromone precursor found in females of N. lecontei