01675nas a2200217   4500008004100000020001400041245011500055210006900170260001200239300001000251490000700261520089400268653001301162100002401175700002601199700002901225700002501254700002401279700002001303856013401323       2007                        eng d  a0028-104200aAntennally mediated negative feedback regulation of pheromone production in the pine engraver beetle, Ips pini0 aAntennally mediated negative feedback regulation of pheromone pr  cJanuary  a61-640 v943 aBark beetles use monoterpenoid aggregation pheromones to coordinate host colonization and mating. These chemical signals are produced de novo in midgut cells via the mevalonate pathway, and pheromone production may be regulated by a negative feedback system mediated through the antennae. In this study, we explored the effect of antennectomy on pheromone production and transcript levels of key mevalonate pathway genes in juvenile hormone III-treated male pine engraver beetles, Ips pini (Say). Antennectomized males produced significantly greater amounts of pheromone than podectomized males and those with intact antennae. Likewise, mRNA levels of three mevalonate pathway genes important in pheromone biosynthesis were measured by quantitative real-time PCR and found to be induced to a greater extent with antennectomy, suggesting a transcriptional regulation of pheromone production10aIps pini1 aGinzel, Matthew, D.1 aBearfield, Jeremy, C.1 aKeeling, Christopher, I.1 aMcCormack, Colin, C.1 aBlomquist, Gary, J.1 aTittiger, Claus  uhttps://weevil.myspecies.info/antennally-mediated-negative-feedback-regulation-pheromone-production-pine-engraver-beetle-ips-pini02067nas a2200181   4500008004100000020001400041245007100055210006900126260000900195300001200204490000700216520149000223653001301713100001901726700002001745700002401765856009601789       2009                        eng d  a0028-104200aUnique animal prenyltransferase with monoterpene synthase activity0 aUnique animal prenyltransferase with monoterpene synthase activi  cJune  a731-7350 v963 aMonoterpenes are structurally diverse natural compounds that play an essential role in the chemical ecology of a wide array of organisms. A key enzyme in monoterpene biosynthesis is geranyl diphosphate synthase (GPPS). GPPS is an isoprenyl diphosphate synthase that catalyzes a single electrophilic condensation reaction between dimethylallyl diphosphate (C5) and isopentenyl diphosphate (C5) to produce geranyl diphosphate (GDP; C10). GDP is the universal precursor to all monoterpenes. Subsequently, monoterpene synthases are responsible for the transformation of GDP to a variety of acyclic, monocyclic, and bicyclic monoterpene products. In pheromone-producing male Ips pini bark beetles (Coleoptera: Scolytidae), the acyclic monoterpene myrcene is required for the production of the major aggregation pheromone component, ipsdienol. Here, we report monoterpene synthase activity associated with GPPS of I. pini. Enzyme assays were performed on recombinant GPPS to determine the presence of monoterpene synthase activity, and the reaction products were analyzed by coupled gas chromatography-mass spectrometry. The functionally expressed recombinant enzyme produced both GDP and myrcene, making GPPS of I. pini a bifunctional enzyme. This unique insect isoprenyl diphosphate synthase possesses the functional plasticity that is characteristic of terpene biosynthetic enzymes of plants, contributing toward the current understanding of product specificity of the isoprenoid pathway.10aIps pini1 aGilg, Anna, B.1 aTittiger, Claus1 aBlomquist, Gary, J.  uhttps://weevil.myspecies.info/unique-animal-prenyltransferase-monoterpene-synthase-activity02118nas a2200181   4500008004100000020001400041245010500055210006900160260000800229300001000237490000700247520146400254100002501718700002301743700002401766700002001790856012601810       2012                        eng d  a0965-174800aIpsdienol dehydrogenase (IDOLDH): A novel oxidoreductase important for Ips pini pheromone production0 aIpsdienol dehydrogenase IDOLDH A novel oxidoreductase important   cFeb  a81-900 v423 aIpsdienone (2-methyl-6-methylene-2,7-octadien-4-one) is an important intermediate in the biosynthesis of pheromonal ipsdienol (2-methyl-6-methylene-2,7-octadien-4-ol) and ipsenol (2-methyl-6methylene-7-octen-4-ol) in male pine engraver beetles. Ips pini (Say). A novel ipsdienol dehydrogenase (IDOLDH) with a pheromone-biosynthetic gene expression pattern was cloned, expressed, functionally characterized, and its cellular localization analyzed. The cDNA has a 762 nt ORF encoding a 253 amino acid predicted translation product of 28 kDa and pI 5.8. The protein has conserved motifs of the Cp2 subfamily of "classical" short-chain dehydrogenases. Transcript levels were highest in pheromone producing tissue: the anterior midgut of fed males. The protein was detected only in male midguts and localized in the cytosolic fraction of midgut cells. Recombinant IDOLDH was produced in Sf9 cells using a baculovirus expression system. Enzyme assays of protein preparations showed IDOLDH used both NAD+ and NADP+ as coenzymes with specific activities in the nanomole range. Enzyme assays and GC/MS analysis showed that IDOLDH catalyzed the oxidation of racemic ipsdienol and (4R)-(-)-ipsdienol to form ipsdienone, while (4S)-(+)-ipsdienol was not a substrate. These data strongly implicate IDOLDH as an enzyme involved in terminal pheromone-biosynthetic steps, likely functioning to "tune" ipsdienol enantiomeric ratios. (C) 2011 Elsevier Ltd. All rights reserved.1 aFigueroa-Teran, Rubi1 aWelch, William, H.1 aBlomquist, Gary, J.1 aTirtiger, Claus  uhttps://weevil.myspecies.info/ipsdienol-dehydrogenase-idoldh-novel-oxidoreductase-important-ips-pini-pheromone-production03689nas a2200241   4500008004100000020001400041245004100055210004100096260000800137300001200145490000700157520304700164653000803211100002403219700002503243700001303268700001703281700002203298700002303320700001603343700002003359856006803379       2010                        eng d  a0965-174800aPheromone production in bark beetles0 aPheromone production in bark beetles  cOct  a699-7120 v403 aThe first aggregation Pheromone components from bark beetles were identified in 1966 as a mixture of ipsdienol, ipsenol and verbenol. Since then, a number of additional components have been identified as both aggregation and anti-aggregation pheromones, with many of them being monoterpenoids or derived from monoterpenoids. The structural similarity between the major pheromone components of bark beetles and the monoterpenes found in the host trees, along with the association of monoterpenoid production with plant tissue, led to the paradigm that most if not all bark beetle pheromone components were derived from host tree precursors, often with a simple hydroxylation producing the pheromone. In the 1990s there was a paradigm shift as evidence for de novo biosynthesis of pheromone components began to accumulate, and it is now recognized that most bark beetle monoterpenoid aggregation pheromone components are biosynthesized de novo. The bark beetle aggregation pheromones are released from the frass, which is consistent with the isoprenoid aggregation pheromones, including ipsdienol, ipsenol and frontalin, being produced in midgut tissue. It appears that exo-brevocomin is produced de novo in fat body tissue, and that verbenol, verbenone and verbenene are produced from dietary cc-pinene in fat body tissue. Combined biochemical, molecular and functional genomics studies in Ips pini yielded the discovery and characterization of the enzymes that convert mevalonate pathway intermediates to pheromone components, including a novel bifunctional geranyl diphosphate synthase/myrcene synthase, a cytochrome P450 that hydroxylates myrcene to ipsdienol, and an oxidoreductase that interconverts ipsdienol and ipsdienone to achieve the appropriate stereochemistry of ipsdienol for pheromonal activity. Furthermore, the regulation of these genes and their corresponding enzymes proved complex and diverse in different species. Mevalonate pathway genes in pheromone producing male I. pini have much higher basal levels than in females, and feeding induces their expression. In I. dupIicatus and I. pini, juvenile hormone III (JH III) induces pheromone production in the absence of feeding, whereas in I. paraconfusus and I. confusus, topically applied JH III does not induce pheromone production. In all four species, feeding induces pheromone production. While many of the details of pheromone production, including the site of synthesis, pathways and knowledge of the enzymes involved are known for Ips, less is known about pheromone production in Dendroctonus. Functional genomics studies are under way in D. ponderosae, which should rapidly increase our understanding of pheromone production in this genus. This chapter presents a historical development of what is known about pheromone production in bark beetles, emphasizes the genomic and post-genomic work in I. pini and points out areas where research is needed to obtain a more complete understanding of pheromone production. [copyright]2010 Elsevier Ltd. All rights reserved.10aIps1 aBlomquist, Gary, J.1 aFigueroa-Teran, Rubi1 aAw, Mory1 aSong, Minmin1 aGorzalski, Andrew1 aAbbott, Nicole, L.1 aChang, Eric1 aTittiger, Claus  uhttps://weevil.myspecies.info/pheromone-production-bark-beetles02657nas a2200217   4500008004100000020001400041245009600055210006900151260000900220300001200229490000700241520193500248653000802183653001302191100002602204700002502230700002002255700002402275700002402299856011602323       2009                        eng d  a0098-033100aTwo regulatory mechanisms of monoterpenoid pheromone production in Ips spp. of bark beetles0 aTwo regulatory mechanisms of monoterpenoid pheromone production   cJune  a689-6970 v353 aBark beetles use aggregation pheromones to coordinate host colonization and mating. These monoterpenoid chemical signals are produced de novo in midgut cells via the mevalonate pathway, and pheromone production is induced when an adult beetle feeds on phloem of a host tree. In Ips pini, juvenile hormone (JH) III influences key regulatory enzymes along the mevalonate pathway that leads to pheromone production. In fact, topically applied JH III is sufficient to stimulate pheromone production in unfed males. In this study, we explore the influence of feeding and JH III treatment on pheromone production in male Ips confusus, the pinyon Ips. We also characterize the influence of feeding and JH III treatment on transcript levels and activity of three key enzymes involved in pheromone biosynthesis: 3-hydroxy-3-methylglutaryl-CoA (HMG) synthase (HMGS), HMG-CoA reductase (HMGR) and geranyl diphosphate synthase (GPPS). We also extend the current understanding of the regulation of pheromone biosynthesis in I. pini, by measuring the influence of feeding and JHIII treatment on enzymatic activity of HMGS and GPPS. Feeding on host phloem alone strongly induces pheromone production in male I. confusus, while JH III treatment has no effect. However, feeding and JH III both significantly up-regulate mRNA levels of key mevalonate pathway genes. Feeding up-regulates these genes to a maximum at 32 h, whereas with JH III-treatment, they are up-regulated at 4, 8, and 16 h, but return near to non-treatment levels at 32 h. Feeding, but not JH III treatment, also increases the activity of all three enzymes in I. confusus, while both feeding or treatment with JH III increase HMGS and GPPS activity in I. pini. Our data suggest that pheromone production in Ips is not uniformly controlled by JH III and feeding may stimulate the release of some other regulatory factor, perhaps a brain hormone, required for pheromone production.10aIps10aIps pini1 aBearfield, Jeremy, C.1 aHenry, Anastasia, G.1 aTittiger, Claus1 aBlomquist, Gary, J.1 aGinzel, Matthew, D.  uhttps://weevil.myspecies.info/two-regulatory-mechanisms-monoterpenoid-pheromone-production-ips-spp-bark-beetles02579nas a2200229   4500008004100000020001400041245009700055210006900152260001300221300001000234490000700244520185100251653002802102100001602130700002002146700002902166700001802195700002602213700002402239700002002263856006602283       2010                        eng d  a1471-216400aFunctional genomics of mountain pine beetle (Dendroctonus ponderosae) midguts and fat bodies0 aFunctional genomics of mountain pine beetle Dendroctonus pondero  c30 March  a12pp.0 v113 aBackground: The mountain pine beetle (Dendroctonus ponderosae) is a significant coniferous forest pest in western North America. It relies on aggregation pheromones to colonize hosts. Its three major pheromone components, trans-verbenol, exo-brevicomin, and frontalin, are thought to arise via different metabolic pathways, but the enzymes involved have not been identified or characterized. We produced ESTs from male and female midguts and associated fat bodies and used custom oligonucleotide microarrays to study gene expression patterns and thereby made preliminary identification of pheromone-biosynthetic genes. Results: Clones from two un-normalized cDNA libraries were directionally sequenced from the 5' end to yield 11,775 ESTs following sequence cleansing. The average read length was 550 nt. The ESTs clustered into 1,201 contigs and 2,833 singlets (4,034 tentative unique genes). The ESTs are broadly distributed among GO functional groups, suggesting they reflect a broad spectrum of the transcriptome. Among the most represented genes are representatives of sugar-digesting enzymes and members of an apparently Scolytid-specific gene family of unknown function. Custom NimbleGen 4-plex arrays representing the 4,034 tentative unique genes were queried with RNA from eleven different biological states representing larvae, pupae, and midguts and associated fat bodies of unfed or fed adults. Quantitative (Real-Time) RT-PCR (qRT-PCR) experiments confirmed that the microarray data accurately reflect expression levels in the different samples. Candidate genes encoding enzymes involved in terminal steps of biosynthetic pathways for exo-brevicomin and frontalin were tentatively identified. Conclusions: These EST and microarray data are the first publicly-available functional genomics resources for this devastating forestry pest10aDendroctonus ponderosae1 aAw, Tidiane1 aSchlauch, Karen1 aKeeling, Christopher, I.1 aYoung, Sharon1 aBearfield, Jeremy, C.1 aBlomquist, Gary, J.1 aTittiger, Claus  uhttp://www.biomedcentral.com/content/pdf/1471-2164-11-215.pdf