(1<Emphasis Type="Italic">S</Emphasis>,3<Emphasis Type="Italic">R</Emphasis>)-<Emphasis Type="Italic">cis</Emphasis>-Chrysanthemyl Tiglate: Sex Pheromone of the Striped Mealybug, <Emphasis Type="Italic">Ferrisia virgata</Emphasis>

Derivatives of 2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropanecarboxylic acid (chrysanthemic acid) are classic natural pyrethroids discovered in pyrethrum plants and show insecticidal activity. Chrysanthemic acid, with two asymmetric carbons, has four possible stereoisomers, and most natural pyrethroids have the (1R,3R)-trans configuration. Interestingly, chrysanthemic acid–related structures are also found in insect sex pheromones; carboxylic esters of (1R,3R)-trans-(2,2-dimethyl-3-(2-methylprop-1-enyl)cyclopropyl)methanol (chrysanthemyl alcohol) have been reported from two mealybug species. In the present study, another ester of chrysanthemyl alcohol was discovered from the striped mealybug, Ferrisia virgata (Cockerell), as its pheromone. By means of gas chromatography–mass spectrometry, nuclear magnetic resonance spectrometry, and high-performance liquid chromatography analyses using a chiral stationary phase column and authentic standards, the pheromone was identified as (1S,3R)-(?)-cis-chrysanthemyl tiglate. The (1S,3R)-enantiomer strongly attracted adult males in a greenhouse trapping bioassay, whereas the other enantiomers showed only weak activity. The cis configuration of the chrysanthemic acid–related structure appears to be relatively scarce in nature, and this is the first example reported from arthropods.     

Efficient production of <Emphasis Type="SmallCaps">d</Emphasis>-1,2,4-butanetriol from <Emphasis Type="SmallCaps">d</Emphasis>-xylose by engineered <Emphasis Type="Italic">Escherichia coli</Emphasis> whole-cell biocatalysts

We have developed a whole-cell bioconversion system for the production of d-1,2,4-butanetriol (BT) from renewable biomass. A plasmid pETduet-xylB-yjhG-T7-adhP-T7-mdlC was constructed and transformed to Escherichia coli BL21(DE3) to obtain the whole cells of E. coli BL21-XYMA capable of bioconversion d-xylose to BT. Then, the factors including carbon sources, nitrogen sources, metal ions, and culture conditions (pH, temperature, IPTG) were identified, and their effects on the whole-cell activity for BT production were investigated. To obtain the highest whole-cell activity, the optimal cultivation parameters are: 15 g·L^–1 yeast extract, 5 g·L^–1 sucrose, 3 g·L^–1 KH₂PO₄, 5 g·L^–1 NaCl, 3 g·L^–1 NH₄Cl, 0.25 g·L^–1 MgSO₄·7H₂O and 1 mL·L^–1 the mixture of trace elements. With the optimized whole cells of E. coli BL21-XYMA, 60 g·L^–1 of xylose was converted to 28 g·L^–1 BT with a molar yield of 66 %, which is higher than those reported in the biotechnological system.

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Ultralow ppm <Emphasis Type="Italic">se</Emphasis>ATRP synthesis of PEO-<Emphasis Type="Italic">b</Emphasis>-PBA copolymers

Poly(ethylene oxide)-block-poly(butyl acrylate) copolymers were prepared via simplified electrochemically mediated atom transfer radical polymerization (seATRP) utilizing only 1 ppm of Cu^II complex, which is the limit of a successful well-controlled polymerization. The presented seATRP system works under potentiostatic and galvanostatic conditions. The polymerization results showed similar molecular weight evolution while maintaining a narrow molecular weight distribution throughout the reactions. ¹H NMR results confirm chemical structure of synthesized diblock copolymers. This ultralow ppm technique is promising candidate for polymerization from nanoparticles, flat surfaces, proteins, and DNA.     

Ammonia Released by <Emphasis Type="Italic">Streptomyces aburaviensis</Emphasis> Induces Droplet Formation in <Emphasis Type="Italic">Streptomyces violaceoruber</Emphasis>

Streptomyces violaceoruber grown in co-culture with Streptomyces aburaviensis produces an about 17-fold higher volume of droplets on its aerial mycelium than in single-culture. Physical separation of the Streptomyces strains by either a plastic barrier or by a dialysis membrane, which allowed communication only by the exchange of volatile compounds or diffusible compounds in the medium, respectively, still resulted in enhanced droplet formation. The application of molecular sieves to bioassays resulted in the attenuation of the droplet-inducing effect of S. aburaviensis indicating the absorption of the compound. ¹H-NMR analysis of molecular-sieve extracts and the selective indophenol-blue reaction revealed that the volatile droplet-inducing compound is ammonia. The external supply of ammonia in biologically relevant concentrations of ≥8 mM enhanced droplet formation in S. violaceoruber in a similar way to S. aburaviensis. Ammonia appears to trigger droplet production in many Streptomyces strains because four out of six Streptomyces strains exposed to ammonia exhibited induced droplet production.     

Bioengineering functional copolymers: synthesis and characterization of poly(<Emphasis Type="Italic">N</Emphasis>-isopropyl acrylamide-<Emphasis Type="Italic">co</Emphasis>-3,4-2H-dihydropyran)s

Novel bioengineering copolymers were synthesized by radical copolymerization of N-isopropylacrylamide (NIPA) and 3,4-2H-dihydropyran (DHP) with 2,2′-azobisisobutyronitrile as an initiator in acetone solution at 70 °C under nitrogen atmosphere. Structure, tacticity and compositons of the copolymers prepared in a wide range of monomer feed were confirmed by FTIR, ¹H{¹³C} NMR-DEPT and elemental analyses. The monomer reactivity ratios (r ₁ and r ₂) were detected using known two methods: r ₁ (NIPA)?=?1.25 and r ₂?=?0.035 (DHP), and r ₁ (NIPA) ?=?0.97 and r ₂?=?0.022 (DHP) by Kelen-Tüdös and Jaacks methods, respectively. It was demonstrated that the studied monomer pair has a tendency to form H-bonding beween amide/ether groups through ?NH...O< complexation which played an important role in the stereoselective chain growth, and significant decrease of allyl degradative chain transfer reactions. This phenomenon is also confirmed by the observed relatively high molecular weights of copolymers (M _v ). The synthesized water-soluble stimuli-responsive poly(NIPA-co-DHP)s exhibit thermal stability, higher glass-transition temperature, polyelectrolyte, pH- and temperature-sensitive behavior and can be attributed to the class of bioengineering functional copolymers useful for various bio- and gene-engineering, and drug delivery applications.     

Sexual Deception in the <Emphasis Type="Italic">Eucera</Emphasis>-Pollinated <Emphasis Type="Italic">Ophrys leochroma</Emphasis>: A Chemical Intermediate between Wasp- and <Emphasis Type="Italic">Andrena-</Emphasis>Pollinated Species

Ophrys flowers mimic sex pheromones of attractive females of their pollinators and attract males, which attempt to copulate with the flower and thereby pollinate it. Virgin females and orchid flowers are known to use the same chemical compounds in order to attract males. The composition of the sex pheromone and its floral analogue, however, vary between pollinator genera. Wasp-pollinated Ophrys species attract their pollinators by using polar hydroxy acids, whereas Andrena-pollinated species use a mixture of non-polar hydrocarbons. The phylogeny of Ophrys shows that its evolution was marked by episodes of rapid diversification coinciding with shifts to different pollinator groups: from wasps to Eucera and consequently to Andrena and other bees. To gain further insights, we studied pollinator attraction in O. leochroma in the context of intra- and inter-generic pollinator shifts, radiation, and diversification in the genus Ophrys. Our model species, O. leochroma, is pollinated by Eucera kullenbergi males and lies in the phylogeny between the wasp and Andrena-pollinated species; therefore, it is a remarkable point to understand pollinator shifts. We collected surface extracts of attractive E. kullenbergi females and labellum extracts of O. leochroma and analyzed them by using gas chromatography with electroantennographic detection (GC-EAD) and gas chromatography coupled with mass spectrometry (GC-MS). We also performed field bioassays. Our results show that O. leochroma mimics the sex pheromone of its pollinator’s female by using aldehydes, alcohols, fatty acids, and non-polar compounds (hydrocarbons). Therefore, in terms of the chemistry of pollinator attraction, Eucera-pollinated Ophrys species might represent an intermediate stage between wasp- and Andrena-pollinated orchid species.     

Novel amphiphilic temperature responsive graft copolymers PCL-<Emphasis Type="Italic">g</Emphasis>-P(MEO<Subscript>2</Subscript>MA-<Emphasis Type="Italic">co</Emphasis>-OEGMA) via a combination of ROP and ATRP: synthesis,characterization, and sol-gel transition

A series of well-defined novel amphiphilic temperature-responsive graft copolymers containing PCL analogues P(αClεCL-co-εCL) as the hydrophobic backbone, and the hydrophilic side-chain PEG analogues P(MEO₂MA-co-OEGMA), designated as P(αClεCL-co-εCL)-g-P(MEO₂MA-co-OEGMA) have been prepared via a combination of ring-opening polymerization (ROP) and atom transfer radical polymerization (ATRP). The composition and structure of these copolymers were characterized by ¹H NMR and GPC analyses. The self-assembly behaviors of these amphiphilic graft copolymers were investigated by UV transmittance, a fluorescence probe method, dynamic light scattering (DLS) and transmission electron microscopy (TEM) analyses. The results showed that the graft copolymers exhibited the good solubility in water, and was given the low critical temperature (LCST) at 35(±1) °C, which closed to human physiological temperature. The critical micelle concentrations (CMC) of P(αClεCL-co-εCL)-g-P(MEO₂MA-co-OEGMA) in aqueous solution were investigated to be 2.0 × 10^?3, 9.1 × 10^?4 and 1.5 × 10^?3 mg·mL^?1, respectively. The copolymer could self-assemble into sphere-like aggregates in aqueous solution with diverse sizes when changing the environmental temperature. The vial inversion test demonstrated that the graft copolymers could trigger the sol-gel transition which also depended on the temperature.     

Identification of Crepenynic Acid in the Seed Oil of <Emphasis Type="Italic">Atractylodes lancea</Emphasis> and <Emphasis Type="Italic">A. macrocephala</Emphasis>

Atractylodes rhizome is widely used in traditional Chinese herbal medicine. Although the chemical composition of the root has been studied in detail, the oil content and fatty acid composition of the seeds of Atractylodes species have not been reported. Fatty acyl composition of seeds from Atractylodes lancea and A. macrocephala was determined by gas chromatography and mass spectrometry of fatty acid methyl esters and 3-pyridylcarbinol esters. The predominant fatty acid in the seeds of both species was linolenic acid, but the unusual acetylenic fatty acid, crepenynic acid (cis-9-octadecen-12-ynoic acid), was also observed at levels of 18% in A. lancea and 13–15% in A. macrocephala. Fatty acid content was 24% for the samples of A. lancea and 16–17% for samples from A. macrocephala. sn-1,3 regioselective lipase digestion of seed lipids revealed that crepenynic acid was absent from the sn-2 position of the seed triacylglycerol. Crepenynic acid was also found in the seed oil of Jurinea mollis at 24% and was not present in the sn-2 position of the TAG. A contrasting distribution of crepenynic acid was found in the oil of Crepis rubra, suggesting differences in crepenynic acid synthesis or TAG assembly between these species.     

Oxidizable Phenolic Concentrations Do Not Affect Development and Survival of <Emphasis Type="Italic">Paropsis Atomaria</Emphasis> Larvae Eating <Emphasis Type="Italic">Eucalyptus</Emphasis> Foliage

Insect folivores can cause extensive damage to plants. However, different plant species, and even individuals within species, can differ in their susceptibility to insect attack. Polyphenols that readily oxidize have recently gained attention as potential defenses against insect folivores. We tested the hypothesis that variation in oxidizable phenolic concentrations in Eucalyptus foliage influences feeding and survival of Paropsis atomaria (Eucalyptus leaf beetle) larvae. First we demonstrated that oxidizable phenolic concentrations vary both within and between Eucalyptus species, ranging from 0 to 61 mg.g^?1 DM (0 to 81% of total phenolics), in 175 samples representing 13 Eucalyptus species. Foliage from six individuals from each of ten species of Eucalyptus were then offered to batches of newly hatched P. atomaria larvae, and feeding, instar progression and mortality of the first and second instar larvae were recorded. Although feeding and survival parameters differed dramatically between individual plants, they were not influenced by the oxidizable phenolic concentration of leaves, suggesting that P. atomaria larvae may have effective mechanisms to deal with oxidizable phenolics. Larvae feeding on plants with higher nitrogen (N) concentrations had higher survival rates and reached third instar earlier, but N concentrations did not explain most of the variation in feeding and survival. The cause of variation in eucalypt herbivory by P. atomaria larvae is therefore still unknown, although oxidizable phenolics could potentially defend eucalypt foliage against other insect herbivores.     

Sexual Isolation and Cuticular Hydrocarbon Differences between <Emphasis Type="Italic">Drosophila santomea</Emphasis> and <Emphasis Type="Italic">Drosophila yakuba</Emphasis>

Drosophila santomea and Drosophila yakuba are two sister species inhabiting Saõ Tomé island. Previous studies showed that both species display strong reproductive isolation, although they can produce a few viable hybrids. Our study tried to understand the mechanism of this ethological isolation between two allopatric strains. A strong sexual isolation was confirmed, with a marked asymmetry. Comparisons of latency times to either courtship or copulation suggest that males do not discriminate females, whereas D. yakuba females, but not D. santomea females, accept their homospecifics more quickly. Cuticular hydrocarbon compositions of both species and sexes were also established with gas chromatography (GC) and GC/mass spectrometry analysis. All have (Z)-7-tricosene as their major compound. There are several quantitative differences between species for few minor compounds. The largest difference concerns n-heneicosane, which is more abundant in D. santomea than in D. yakuba flies (up to seven times more between males). A similar quantitative difference was also found in a pair of sympatric strains. Furthermore, D. yakuba males artificially perfumed with n-heneicosane were discriminated negatively by D. yakuba females, suggesting a role for this compound in the sexual isolation between these two species.     

The Role of the Glucosinolate-Myrosinase System in Mediating Greater Resistance of <Emphasis Type="Italic">Barbarea verna</Emphasis> than <Emphasis Type="Italic">B. vulgaris</Emphasis> to <Emphasis Type="Italic">Mamestra brassicae</Emphasis> Larvae

We investigated the influences of two structurally similar glucosinolates, phenethylglucosinolate (gluconasturtiin, NAS) and its (S)-2-hydroxyl derivative glucobarbarin (BAR), as well as their hydrolysis products on larvae of the generalist Mamestra brassicae (Lepidoptera: Noctuidae). Previous results suggested a higher defensive activity of BAR than NAS based on resistance toward M. brassicae larvae of natural plant genotypes of Barbarea vulgaris R. Br. (Brassicaceae) dominated by BAR. In the present study, the hypothesis of a higher defensive activity of BAR than NAS was tested by comparing two Barbarea species similarly dominated either by BAR or by NAS and by testing effects of isolated BAR and NAS on larval survival and feeding preferences. Larvae reared on leaf disks of B. verna (Mill.) Asch. had a lower survival than those reared on B. vulgaris P- and G-chemotypes. Leaves of B. verna were dominated by NAS, whereas B. vulgaris chemotypes were dominated by BAR or its epimer. In addition, B. verna leaves showed a threefold higher activity of the glucosinolate-activating myrosinase enzymes. The main product of NAS from breakdown by endogenous enzymes including myrosinases (“autolysis”) in B. verna leaves was phenethyl isothiocyanate, while the main products of BAR in autolyzed B. vulgaris leaves were a cyclized isothiocyanate product, namely an oxazolidine-2-thione, and a downstream metabolite, an oxazolidin-2-one. The glucosinolates BAR and NAS were isolated and offered to larvae on disks of cabbage. Both glucosinolates exerted similar negative effects on larval survival but effects of NAS tended to be more detrimental. Low concentrations of BAR, but not of NAS, stimulated larval feeding, whereas high BAR concentrations acted deterrent. NAS only tended to be deterrent at the highest concentration, but the difference was not significant. Recoveries of NAS and BAR on cabbage leaf disks were similar, and when hydrolyzed by mechanical leaf damage, the same isothiocyanate-type products as in Barbarea plants were formed with further conversion of BAR to cyclic products, (R)-5-phenyloxazolidine-2-thione [(R)-barbarin] and (R)-5-phenyloxazolidin-2-one [(R)-resedine]. We conclude that a previously proposed generally higher defensive activity of BAR than NAS to M. brassicae larvae could not be confirmed. Indeed, the higher resistance of NAS-containing B. verna plants may be due to a combined effect of rather high concentrations of NAS and a relatively high myrosinase activity or other plant traits not investigated yet.     

Modifying the Alkylglucosinolate Profile in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> Alters the Tritrophic Interaction with the Herbivore <Emphasis Type="Italic">Brevicoryne brassicae</Emphasis> and Parasitoid <Emphasis Type="Italic">Diaeretiella rapae</Emphasis>

Arabidopsis thaliana was used as an experimental model plant to investigate a tritrophic interaction between the plant, a specialist aphid herbivore, Brevicoryne brassicae, and its natural enemy, the parasitoid Diaeretiella rapae. The A. thaliana ecotype Col-5 was transformed with a functional 2-oxoglutarate dependent dioxygenase (BniGSL-ALK) that converts 3-methylsulfinylpropylglucosinolate and 4-methylsulfinylbutylglucosinolate to 2-propenylglucosinolate and 3-butenylglucosinolate, respectively. This transformation results in a change in the glucosinolate hydrolysis profile where 3-butenylisothiocyanate, 2-propenylisothiocyanate and 5-vinyloxazolidine-2-thione are produced in contrast to the wild-type plant where 4-methylsulfinylbutylisothiocyanate is the main product. Performance of B. brassicae was affected negatively by transforming Col-5 with BniGSL-ALK in terms of mean relative growth rates. In a series of behavioral bioassays, naïve D. rapae females were able to discriminate between B. brassicae infested and uninfested Col-5 plants transformed with BniGSL-ALK, with parasitoids showing a preference for B. brassicae infested plants. By contrast, naïve D. rapae females were unable to discriminate between aphid infested and uninfested Col-5 plants. Subsequent air entrainments of B. brassicae infested Col-5 plants transformed with BniGSL-ALK further confirmed the presence of 3-butenylisothiocyanate in the headspace. By contrast, no glucosinolate hydrolysis products were recorded from similarly infested Col-5 plants.     

Microstructure and <Emphasis Type="Italic">in vitro</Emphasis> Bioactivity of Silicon-Substituted Hydroxyapatite

Silicon-substituted hydroxyapatite has shown superior biological performance compared to its stoichiometric counterpart both in vitro and in vivo. In the present study, single-phase silicon-substituted hydroxyapatite was successfully synthesized by the precipitation method. Chemical composition, crystalline phase, microstructure, and morphology of the materials were characterized by XRF, XRD, FT-IR, solid-state NMR and SEM. The results showed that hydroxyapatite kept its original structure with silicon up to a level of 0.9 wt%. The precipitation method was proved to be an efficient way to synthesize single-phase silicon-substituted hydroxyapatite. Solid-state NMR combined with other techniques gave direct evidence for the isomorphous substitution of PO₄ ^3- by SiO₄ ^4- in the hydroxyapatite structure. Silicon-substituted hydroxyapatite showed better bioactivity than stoichiometric hydroxyapatite in the in vitro bioactivity experiment. The higher the silicon content in the hydroxyapatite structure, the better the in vitro bioactivity. The enhanced bioactivity of silicon-substituted hydroxyapatite over pure hydroxyapatite has been attributed to the effect of silicate ions in accelerating dissolution.     

Identification of Floral Volatiles and Pollinator Responses in Kiwifruit Cultivars, <Emphasis Type="Italic">Actinidia chinensis</Emphasis> var. <Emphasis Type="Italic">chinensis</Emphasis>

Volatiles emitted from unpollinated in situ flowers were collected from two male cultivars, ‘M33’, ‘M91’, and one female cultivar ‘Zesy002’ (Gold3) of kiwifruit (Actinidia chinensis var. chinensis). The samples were found to contain 48 compounds across the three cultivars with terpenes and straight chain alkenes dominating the headspace. Electrophysiological responses of honey bees (Apis mellifera) and bumble bees (Bombus terrestris) to the headspace of the kiwifruit flowers were recorded. Honey bees consistently responded to 11 floral volatiles from Gold3 pistillate flowers while bumble bees consistently responded to only five compounds from the pistillate flowers. Nonanal, 2-phenylethanol, 4-oxoisophorone and (3E,6E)-α-farnesene from pistillate flowers elicited responses from both bee species. Overall, honey bees were more sensitive to the straight chain hydrocarbons of the kiwifruit flowers than the bumble bees, which represented one of the main differences between the responses of the two bee species. The floral volatiles from staminate flowers of the male cultivars ‘M33’ and ‘M91’ varied greatly from those of the pistillate flowers of the female cultivar Gold3, with most of the bee active compounds significantly different from those in the Gold3 flower headspace. The total floral emissions of ‘M33’ flowers were significantly less than those of the Gold3 flowers, while the total floral emissions of the ‘M91’ flowers were significantly greater than those of the Gold3 flowers.     

Interspecific Cross-Attraction between the South American Cerambycid Beetles <Emphasis Type="Italic">Cotyclytus curvatus</Emphasis> and <Emphasis Type="Italic">Megacyllene acuta</Emphasis> is Averted by Minor Pheromone Components

During field screening trials conducted in Brazil in 2015, adults of both sexes of the cerambycid beetles Cotyclytus curvatus (Germar) and Megacyllene acuta (Germar) (subfamily Cerambycinae, tribe Clytini) were significantly attracted to racemic 3-hydroxyhexan-2-one and racemic 2-methylbutan-1-ol, chemicals which previously have been identified as male-produced aggregation-sex pheromones of a number of cerambycid species endemic to other continents. Subsequent analyses of samples of beetle-produced volatiles revealed that males of C. curvatus sex-specifically produce only (R)-3-hydroxyhexan-2-one, whereas males of M. acuta produce the same compound along with lesser amounts of (2S,3S)-2,3-hexanediol and (S)-2-methylbutan-1-ol. Follow-up field trials showed that both sexes of both species were attracted to synthetic reconstructions of their respective pheromones, confirming that males produce aggregation-sex pheromones. The minor pheromone components of M. acuta, (S)-2-methylbutan-1-ol and (2S,3S)-2,3-hexanediol, synergized attraction of that species, but antagonized attraction of C. curvatus to (R)-3-hydroxyhexan-2-one. Beetles of other cerambycine species also were attracted in significant numbers, including Chrysoprasis linearis Bates, Cotyclytus dorsalis (Laporte & Gory), and Megacyllene falsa (Chevrolat). Our results provide further evidence that 3-hydroxyhexan-2-one is a major component of attractant pheromones of numerous cerambycine species world-wide. Our results also highlight our increasing understanding of the crucial role of minor pheromone components in imparting species specificity to cerambycid pheromone blends, as is known to occur in numerous species in other insect families.     

Improvement of bacterial cellulose production in <Emphasis Type="Italic">Acetobacter xylinum</Emphasis> using byproduct produced by <Emphasis Type="Italic">Gluconacetobacter hansenii</Emphasis>

A single sugar α-linked glucuronic acid based oligosaccharide (SSGO) is water soluble oligosaccharides (WSOS) obtained by Gluconacetobacter hansenii PJK (KCTC 10505BP) as a byproduct during bacterial cellulose (BC) production. In this study, SSGO was used for the improvement of BC production by the vinegar bacterium, Acetobacter xylinum, which produces heteropolysaccharides as a byproduct. The addition of 1.0% SSGO to the chemically defined medium (CDM) resulted in an 89.3% increase in BC production by A. xylinum after 15 days of cultivation under static condition, and a 52.3% increase in BC production by G. hansenii. Both the dry cell weight and live cell density of A. xylinum increased 50% with the addition of 1.0% SSGO. SSGO successfully improved BC production by A. xylinum.     

Microbial Conversion of Arachidonic Acid to Arachidonyl Alcohol by a New <Emphasis Type="Italic">Acinetobacter</Emphasis> Species

Arachidonyl alcohol rarely occurs in natural oils. It can be used as a substrate for production of several ether lipids possessing beneficial functions. Although arachidonyl alcohol has been produced on a laboratory scale by the chemical reduction of arachidonic acid, it will be difficult to scale up this process for industrial application. The aim of this study was to develop a new bioprocess for converting arachidonic acid to arachidonyl alcohol. Screening was conducted using 11 wax ester- (esters of fatty acids and fatty alcohols) producing strains reported in our previous study, and a single-cell oil containing arachidonic acid. A new strain, Acinetobacter species N-476-2, most effectively converted arachidonic acid to arachidonyl alcohol, which accumulated inside the cells as a wax ester. GC–MS, FT–IR, and NMR analyses showed that this strain reduced the carboxyl group of 5-cis,8-cis,11-cis,14-cis-arachidonic acid to a hydroxyl group without altering the position or configuration of the double bonds; the product was identified as 5-cis,8-cis,11-cis,14-cis-arachidonyl alcohol. A time-course study of cultivation showed that the amount of arachidonyl alcohol produced by the strain after 4 days was 2.2 mg/mL culture. The bioprocess using Acinetobacter sp. N-476-2 can be applied to the large-scale production of arachidonyl alcohol.     

Naphthoquinone Metabolites Produced by <Emphasis Type="Italic">Monacrosporium ambrosium</Emphasis>, the Ectosymbiotic Fungus of Tea Shot-Hole Borer, <Emphasis Type="Italic">Euwallacea fornicatus,</Emphasis> in Stems of Tea, <Emphasis Type="Italic">Camellia sinensis</Emphasis>

The tea shot-hole borer beetle (TSHB, Euwallacea fornicatus) causes serious damage in plantations of tea, Camellia sinensis var. assamica, in Sri Lanka and South India. TSHB is found in symbiotic association with the ambrosia fungus, Monacrosporium ambrosium (syn. Fusarium ambrosium), in galleries located within stems of tea bushes. M. ambrosium is known to be the sole food source of TSHB. Six naphthoquinones produced during spore germination in a laboratory culture broth of M. ambrosium were isolated and identified as dihydroanhydrojavanicin, anhydrojavanicin, javanicin, 5,8-dihydroxy-2-methyl-3-(2-oxopropyl)naphthalene-1,4-dione, anhydrofusarubin and solaniol. Chloroform extracts of tea stems with red-colored galleries occupied by TSHB contained UV active compounds similar to the above naphthoquinones. Laboratory assays demonstrated that the combined ethyl acetate extracts of the fungal culture broth and mycelium inhibited the growth of endophytic fungi Pestalotiopsis camelliae and Phoma multirostrata, which were also isolated from tea stems. Thus, pigmented naphthoquinones secreted by M. ambrosium during spore germination may prevent other fungi from invading TSHB galleries in tea stems. The antifungal nature of the naphthoquinone extract suggests that it protects the habitat of TSHB. We propose that the TSHB fungal ectosymbiont M. ambrosium provides not only the food and sterol skeleton necessary for the development of the beetle during its larval stages, but also serves as a producer of fungal inhibitors that help to preserve the purity of the fungal garden of TSHB.     

Olfactory Preferences of the Parasitic Nematode <Emphasis Type="Italic">Howardula aoronymphium</Emphasis> and its Insect Host <Emphasis Type="Italic">Drosophila falleni</Emphasis>

Many parasitic nematodes have an environmental infective stage that searches for hosts. Olfaction plays an important role in this process, with nematodes navigating their environment using host-emitted and environmental olfactory cues. The interactions between parasitic nematodes and their hosts are also influenced by the olfactory behaviors of the host, since host olfactory preferences drive behaviors that may facilitate or impede parasitic infection. However, how olfaction shapes parasite-host interactions is poorly understood. Here we investigated this question using the insect-parasitic nematode Howardula aoronymphium and its host, the mushroom fly Drosophila falleni. We found that both H. aoronymphium and D. falleni are attracted to mushroom odor and a subset of mushroom-derived odorants, but they have divergent olfactory preferences that are tuned to different mushroom odorants despite their shared mushroom environment. H. aoronymphium and D. falleni respond more narrowly to odorants than Caenorhabditis elegans and Drosophila melanogaster, consistent with their more specialized niches. Infection of D. falleni with H. aoronymphium alters its olfactory preferences, rendering it more narrowly tuned to mushroom odor. Our results establish H. aoronymphium-D. falleni as a model system for studying olfaction in the context of parasite-host interactions.     

Aqueous radical polymerization of <Emphasis Type="Italic">N</Emphasis>,<Emphasis Type="Italic">N</Emphasis>-dimethylacrylamide redox-initiated by aerobically catalytic oxidation of water-soluble tertiary amines

Catalytic oxidation of water-soluble tertiary amines by complexes of Cu^II, Fe^III and Co^II was utilized to initiate radical polymerization of N,N-dimethylacrylamide (DMAAm) in aqueous solution at 70–80 °C. The oxidation of tertiary amines by Cu^II was studied by proton nuclear magnetic resonance spectroscopy and online ultraviolet–visible spectrophotometry. The polymerization kinetics was monitored by gas chromatography, and molecular weight of the PDMAAm was measured by gel-permeation chromatography coupled with multi-angle laser light scattering. Oxidation of tertiary amines occurs predominantly via formation of C_alpha·radicals to initiate polymerization of electron-deficient monomers and N-dealkylation, and redox equilibrium between Cu^I/L and Cu^II/L is established at a faster rate in aqueous media. Fe^III and Cu^II complexes are efficient catalysts as each catalyst molecule could generate above 10 propagating radicals in 5 h, while Co^II complex might involve in oxidation of tertiary amines in non-radical pathway, leading to a low catalytic efficiency. Water-soluble tertiary amines such as N,N-dialkylethanolamine (alkyl = methyl, ethyl etc.) are reducing agents of a higher activity in aqueous media than those primary or secondary analogues. Our strategy renders it possible to prepare polymer of alpha-amino functionality via one-pot process from commercially available commodity reagents under practical conditions with negligible catalyst residue.