Sex pheromones of Phyllonorycter acerifoliella and Ph. heegerella and communication peculiarities in three species of leafmining moths

Females of the leaf miner moth Phyllonorycter acerifoliella (Z.) [=Ph. sylvella (Hw.)] and Ph. heegerella (Z.) (Lepidoptera: Gracillariidae: Lithocolletinae) release their sex pheromone at the beginning of photophase. The periodicity of the `calling' behaviour of Ph. acerifoliella females was established. Three compounds from calling virgin Ph. heegerella females were collected by the Solid Phase Micro Extraction (SPME) technique and identified as (Z)-8-tetradecenyl acetate (Z8-14:OAc), tetradecyl acetate (14:OAc) and (Z)-8-tetradecenol (Z8-14:OH) in the ratio (88±3):(2±0.6):(10±5) by capillary gas chromatography and mass spectrometry. Field trapping experiments demonstrated that the first two compounds are important for the attraction of conspecific males. Z8-14:OAc was found to be attractive when tested separately, while 14:OAc acted as synergist. The attractivity of the three component blend was reduced by 10% admixture of either (E)-10-dodecenyl acetate (E10-12:OAc) or (Z)-10-tetradecenyl acetate (Z10-14:OAc).Field tests of Z10-, Z8- and E10-14:OAc, identified from Ph. acerifoliella females, demonstrated that the first two compounds were essential for the attraction of conspecific males; so both are sex pheromone components. The attractivity of the three component blend of Z10- Z8- and E10-14:OAc was reduced by 10% admixture of (E)-10-dodecenol (E10-12:OH). The following four semiochemical compounds, Z8-14:OAc, Z8-14:OH, E10-14:OAc and 14:OAc, identified from phyllonoryctid females, as well as two sex attraction antagonists for Ph. acerifoliella and Ph. heegerella males, E10-12:OAc and Z10-14:OAc, are new for the family Gracillariidae. The results of field trapping experiments revealed mechanisms ensuring the specificity of the chemocommunication systems in Ph. acerifoliella, Ph. heegerella and Ph. ulmifoliella (Hb.) moths.     

NOR Sites Detected by Ag-DAPI Staining of an Unusual Autosome Chromosome of Bradysia Hygida (Diptera:Sciaridae) Colocalize with C-banded Heterochromatic Region

The study of chromosomes in insects is a good tool in mitotic process analysis, zoographic localization and evolution investigation. Among them, the Sciaridae offers a karyotype with a small number of chromosomes, where the heterochromatin and nucleolar organizer region, NOR, are easily analyzed in metaphase chromosomes obtained from cerebral ganglia squashes. In this work, the heterochromatic regions on Bradysia hygida mitotic chromosomes, revealed by C-banding, were identified as centromeric blocks on A and C chromosomes and as dark interstitial region in B and X chromosomes. By Ag-DAPI staining, active nucleolus organizer region, NOR, was revealed associated to the constitutive heterochromatin in the end of the C autosome chromosome. The C-band regions and the unusual ribosomal site localization are discussed.     

Unequal X Chromosomes in Bradysia Hygida (Diptera:Sciaridae) Females: Karyotype Assembly and Morphometric Analysis

The mitotic chromosomes of Bradysia hygida(Diptera:Sciaridae) neuroblast cells are described together with their morphometric data. Giemsa-stained neuroblast chromosomes from female and male larvae confirm the chromosome number of this species, 2n=8 (XX) and 2n=7 (XO), respectively. The karyotype assembly reveals two metacentric autosomic pairs, the A and B chromosome; a subtelocentric, the C chromosome, the smallest one; and a sexual unequal metacentric pair, X chromosome, in female karyotype and a one sexual metacentric X chromosome in male. The implications of the unequal X chromosome pair are discussed.     

云南食用菌眼蕈蚊分类及优势种分析

本文报道了云南省各食用菌栽培地眼蕈蚊科（Sciaridae）害虫共6属11种, 其中异迟眼蕈蚊Bradysia difformis Frey为中国新纪录种, 独刺普眼蕈蚊Cosmosciara perniciosa Edwards为中国新记录属和种, 并提供了云南省食用菌眼蕈蚊种类检索表。研究标本均保存于云南农业大学昆虫系标本室。根据各地标本采集数据对不同食用菌上眼蕈蚊种类优势度进行了分析, 结果表明平菇厉眼蕈蚊Lycoriella pleuroti Yang et Zhang和异迟眼蕈蚊B. difformis Frey为云南省食用菌上眼蕈蚊的主要优势种类。     

Female resistance behaviour and progeny sex ratio in two Bradysia species (Diptera: Sciaridae) with paternal genome elimination

The relationship between female mating preferences and sex allocation has received considerable theoretical and empirical support. Typically, choosier females adjust their progeny sex ratio towards sons, who inherit the attractive traits of their father. However, in species with paternal genome elimination, where male sperm do not contain the paternal genome, predictions for the direction of progeny sex ratio biases and their relationship with female choosiness are atypical. Paternal genome elimination also creates a potential for male–female conflict over sex allocation, and any influence of female mate choice on sex ratio outcomes have interesting implications for sexually antagonistic coevolution. Within the Sciaridae (Diptera) are species that produce single‐sex progeny (monogenic species) and others in which progeny comprise both sexes (digenic species). Paternal genome elimination occurs in both species. We explore female mate resistance behaviour in a monogenic and digenic species of mushroom gnat from the genus Bradysia. Our experiments confirmed our theoretical predictions, revealing that in the monogenic and digenic species, females producing female‐biased progeny were more likely to have resisted at least one mating attempt.     

Pest species of a fungus gnat genus Bradysia Winnertz (Diptera: Sciaridae) injuring agricultural and forestry products in Japan,with a review on taxonomy of allied species

Agricultural and forestry pest species of the Bradysia tilicola group from Japan were revised taxonomically after examination of the type specimens. A sciarid pest of Welsh onion occurred from northern Saitama, Japan since 2014 and later from south‐eastern Gunma, Japan, was identified as B. odoriphaga Yang & Zhang, 1985, sp. rev. It was removed from synonymy of B. cellarum Frey, 1948, being the first record of the species from Japan. Bradysia agrestis Sasakawa, 1978, was assigned to be a junior synonym of B. impatiens (Johannsen, 1912). Bradysia paupera Tuomikoski, 1960, sp. rev. , was removed from synonymy of B. impatiens and was recognized as a distinct species. We also compiled their host plants and fungi hitherto reported from Japan. Bradysia impatiens was here recorded from an indoor facility of cloud ear fungus, Auricularia polytricha (Montagne) Sacc., for the first time. At least 19 cultural crops belonging to 12 plant and fungal families were reported as actual and potential hosts of B. impatiens. We suggested that B. impatiens and B. paupera might have been confused in studies of mushroom production in Japan. Further survey of distribution of B. paupera in Japan needs to be done.     

A female sex pheromone in Bruchidius atrolineatus (Pic) (Coleoptera, Bruchidae): analysis of the factors affecting production or emission of this pheromone

ABSTRACT. . The females of the tropical Bruchid Bruchidius atrolineatus (Pic) (Coleoptera) produce a sex pheromone that influences male behaviour. This pheromone, tested on males in an olfactometer, triggers characteristic antennal movements followed by a distinct chemo-anemotaxis. Sex pheromone production depends on the age of the females and is partly correlated with their ovarian activity. Females 0–24 h old are not attractive, but they begin to attract males on the second day of adult life. Vitellogenesis seems to correlate with the emission or the production of the pheromone but there is no precise synchrony between the two phenomena which are probably induced by the same endocrine factors. Within 1 h after mating there is a decrease in the females' attractiveness, which lasts at least 48 h. Under our experimental conditions no particular rhythm of production or emission of the sex pheromone in B. atrolineatus could be observed.     

山东不同地区韭菜迟眼蕈蚊共生菌Wolbachia 的检测及鉴定

【目的】为了揭示山东省韭菜迟眼蕈蚊Bradysia odoriphaga Yang et Zhang种群共生菌 Wolbachia 的感染率及其分类地位,探讨该共生菌对韭菜迟眼蕈蚊的潜在影响。【方法】利用线粒体细胞色素氧化酶I（mtCOI）基因引物（LCO1490/HCO2198）,通过扩增测序和序列比对对采自山东省12个地区的根蛆种群进行了分类鉴定。在上述基础上,利用 Wolbachia 的16S rDNA和 wsp 基因特异引物（分别为16S-F/16S-R和81F/691R）对鉴别出的11个韭菜迟眼蕈蚊种群体内Wolbachia 感染情况进行了PCR检测;对感染个体体内 Wolbachia 依据16S rDNA基因片段序列进行分类鉴定。【结果】山东省12个根蛆种群中,11个种群为韭菜迟眼蕈蚊种群。基于 Wolbachia 的16S rDNA基因特异引物检测结果发现,这些韭菜迟眼蕈蚊种群广泛感染 Wolbachia （感染率为6.67%~93.33%）,而利用wsp 基因特异引物检测的感染率（0.00%~40.00%）相对较低些。基于 Wolbachia 的16S rDNA基因构建系统发育树表明,这些韭菜迟眼蕈蚊种群感染的Wolbachia 全部属于A组。【结论】确定了 Wolbachia 在山东省韭菜迟眼蕈蚊体内的感染率及其分类地位,为研究 Wolbachia 对韭菜迟眼蕈蚊生物学及生态学的影响奠定了基础。     

Mechanisms of programmed cell death in the midgut and salivary glands from Bradysia hygida (Diptera: Sciaridae) during pupal–adult metamorphosis

Programmed cell death is involved with the degeneration/remodeling of larval tissues and organs during holometabolous development. The midgut is a model to study the types of programmed cell death associated with metamorphosis because its structure while degenerating is a substrate for the formation of the adult organ. Another model is the salivary glands from dipteran because their elimination involves different cell death modes. This study aimed to investigate the models of programmed cell death operating during midgut replacement and salivary gland histolysis in Bradysia hygida. We carried out experiments of real‐time observations, morphological analysis, glycogen detection, filamentous‐actin localization, and nuclear acridine orange staining. Our findings allow us to establish that an intact actin cytoskeleton is required for midgut replacement in B. hygida and nuclear condensation and acridine orange staining precede the death of the larval cells. Salivary glands in histolysis present cytoplasmic blebbing, nuclear retraction, and acridine orange staining. This process can be partially reproduced in vitro. We propose that the larval midgut death involves autophagic and apoptotic features and apoptosis is a mechanism involved with salivary gland histolysis.     

黄杨绢野螟雌蛾性信息素的鉴定及诱蛾活性

【目的】研究旨在深入探讨中国黄杨绢野Diaphania perspectalis 的雌蛾性信息素组成及诱蛾活性。【方法】利用气质联用仪（GC-MS）对黄杨绢野螟正常型性成熟雌蛾的性腺体提取物和合成标样比较分析,并用反-11-十六碳烯醛(E11-16:Ald) 、顺-11-十六碳烯醛（Z-11-16:Ald）、顺-9-十六碳烯醛（Z-9-16:Ald)、顺-11-十六碳烯醇（Z-11-16:OH)等物质进行触角的电生理测定,最后开展田间诱集比较试验以筛选出最佳性信息素混合物。【结果】Z-11-16:Ald为中国黄杨绢野螟正常型性信息素主要组分,E-11-16:Ald的含量极低,Z-11-16:OH未检测到。正常型雄性黄杨绢野螟触角对Z-11-16:Ald, E-11-16:Ald, Z-9-16:Ald和Z-11-16:OH产生强烈的EAG反应,并随着浓度的提高而显著增加;而对Z-11-16:Ac和E-11-16:Ac的嗅觉反应较弱,低于对植物绿叶气味顺3-己烯乙酸酯（Z-3-6:Ac）的反应。单一Z-1-16:Ald对正常型雄性黄杨绢野螟具有强烈的诱集效果,加入E-11-16:Ald有一定的增效作用,但在统计上则不显著。单一Z-11-16:Ald组分对黑化型雄性黄杨绢野螟无引诱活性,必需加入一定比例的E-11-16:Ald才显示诱蛾活性。Z-11-16:Ald:E-11-16:Ald的比例为250 μg:250 μg时诱集到的黑化型雄性黄杨绢野螟数量最多,而Z-11-16:Ald:E-11-16:Ald的比例为429 μg:71 μg时则诱集到的正常型雄性黄杨绢野螟数量最多。同时,单一Z-11-16:Ald也可引诱大量雄性粘虫Mythimna separata,但E-11-16:Ald抑制其活性。【结论】中国黄杨绢野螟的性信息素主成分是Z-11-16:Ald,单一组分即可在田间强烈引诱雄蛾,E-11-16:Ald的功能只起到微弱的增效作用,但也可能起种的专一性的作用。正常型与黑化型黄杨绢野螟对性信息素的嗅觉反应存在差异,黑化型黄杨绢野螟的性信息素接近日本种,即性信息素组成为Z-11-16:Ald和E-11-16:Ald 的混合物,其比例为1:1,且E-11-16:Ald为必需。     

Olfactory Sex Attraction and Mating Behaviour in the Pine Sawyer Monochamus galloprovincialis (Coleoptera: Cerambycidae)

The pine sawyer Monochamus galloprovincialis Olivier (Coleoptera: Cerambycidae) is a vector of the pine wood nematode Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle. Male and females of this species have a kairomonal attraction to host volatiles and Ips semiochemicals. Once on the host tree, males and females copulate and oviposition occurs. Bioassays using Y-tube olfactometry revealed that females were attracted to volatile compounds produced by males, but not to volatiles produced by females. However, immature males did not seem to release attractants for mature females and immature females did not show any attraction for mature males. A experiment designed to know about mating behaviour in this specie revealed that most of the males encountered females while they were walking. In all pairs both sexes contacted with antennae before copulation, however, in some occasions copulation did not proceed just after antennal contact. On the contrary, in all pairs observed, the male licked the elytra of the female with his mouth palpi just before copulation (licking); immediately afterwards, the male grabbed the female with his forelegs and mounted her suggesting a chemically mediated mate recognition by males.     

Molecular characterization and phylogenetic analysis of three odorant binding protein gene transcripts in Dendrolimus species (Lepidoptera: Lasiocampidae)

Pine caterpillar moths, Dendrolimus spp. (Lepidoptera: Lasiocampidae), are serious economic pest of pines. Previously, phylogenetic analyses of Dendrolimus using different methods yielded inconsistent results. The chemosensory systems of insects may play fundamental roles in promoting speciation. Odorant‐binding proteins (OBPs) participate in the first step of odor detection. Studying the evolution of OBPs in closely related species may help us to identify their role in speciation. We identified three OBPs – one pheromone‐binding protein and two general odorant‐binding proteins – from male antennae of four Dendrolimus species, D. superans (Butler), D. punctatus (Walker), D. kikuchii Matsumura, and D. houi Lajonquiere, the olfactory recognition systems of which had not been previously investigated. We analyzed their molecular characteristics and compared their sequences to those of OBPs in D. tabulaeformis Tsai et Liu. Ka/Ks ratio analyses among the five Dendrolimus species indicate that PBP1 genes experienced more evolutionary pressure than the GOBPs. Phylogenetic relationships of PBP1 and GOBP1 both indicated that D. houi was the basal species, then branched D. kikuchii, while D. tabulaeformis, D. punctatus, and D. superans evolved more recently. These relationships are consistent with the changes in sex pheromone components of these five species. Dendrolimus tabulaeformis and D. punctatus are closely related sister species. However, the distances among GOBP2 sequences in the five Dendrolimus were very short, and the relationships of D. houi and D. kikuchii could not be resolved. Integrating our results with those of previous studies, we hypothesized that D. kikuchii, D. punctatus and D. superans evolved from the basal ancestor because of sex pheromone mutations and environmental pressure.     

Specificity of accessory gland extracts. in three Delia fly species (Diptera: Anthomyiidae)

Abstract. Extracts of testes and male accessory (paragonial) glands made from three species of Delia (onion fly {D.antiqua) , seedcorn fly (D.platura) , and cabbage fly (D.radicum)) were injected into conspecific virgin females. Extracts of paragonial glands, but not testes, from onion, seedcorn and cabbage fly males stimulated oviposition and suppressed mating when injected into conspecific virgin females. When extracts of paragonial glands from males of these species were injected into heterospecific virgin females, the extracts of D.antiqua and D.platura were fully cross-reactive with respect to oviposition; interspecific injection stimulated oviposition at the level of the conspecific mated controls. Injection of D.radicum extract fully activated the D.antiqua and D.platura ovipositional response. D.antiqua extract caused mating inhibition and partial oviposition in D.radicum; that of D.platura had no effect on either oviposition or mating inhibition in D.radicum. These results suggest that D.antiqua and D.platura are more closely related to one another than either is to D.radicum , and agree with published anatomically-based phylogenies and a genetic distance calculation based on eight enzyme loci. The occurrence of sex peptide cross-reactivity, though asymmetrical, between D.radicum versus D.antiqua and D.platura indicates that, functionally, sex peptides have changed little during the evolution of this genus. An emerging pattern of broad cross-reactivity within genera suggests that sex peptides are not an initiator of reproductive isolation.     

Sex pheromones and olfactory proteins in Antheraea moths: A. pernyi and A. polyphemus (Lepidoptera: Saturniidae)

Olfaction is essential for regulating the physiological and behavioral actions of insects with specific recognition of various odors. Antheraea moths (Lepidoptera: Saturniidae) possess relatively large bodies and antennae so that they are good subjects for exploring molecular aspects of insect olfaction. Current knowledge of the molecular aspects of Antheraea olfaction is focused on the Chinese tussah silkmoth A. pernyi Guérin-Méneville and another species A. polyphemus (Cramer) in their pheromones, odorant-binding proteins (OBPs), odorant receptors (ORs), odorant receptor coreceptors (ORCOs), sensory neuron membrane proteins (SNMPs), and odorant-degrading enzymes (ODEs). The first insect OBP, SNMP, and ODE were identified from A. polyphemus. This review summarizes the principal findings associated with the olfactory physiology and its molecular components in the two Antheraea species. Three types of olfactory neurons may have specific ORs for three respective sex-pheromone components, with the functional sensitivity and specificity mediated by three respective OBPs. SNMPs and ODEs are likely to play important roles in sex-pheromone detection, inactivation, and degradation. Identification and functional analysis of the olfactory molecules remain to be further performed in A. pernyi, A. polyphemus, and other Antheraea species.     

Parasitism and decreased response to sex pheromones in malePeriplaneta americana (Dictyoptera: Blattidae)

Parasites are known to alter the behavior of their hosts, but little is known about their effects on responses to sexual stimuli. Periplaneta americanainfected with the acanthocephalan Moniliformis moniliformiswere compared to uninfected animals in their behavioral and electroantennogram responses to a synthetic P. americanapheromone component, periplanone-B, and a pheromone mimic, bornyl acetate. In a t-maze there was no significant difference between infected animals' responses to periplanone-B and a random binomial distribution; the responses of uninfected animals were significantly nonrandom. The electroan-tennogram responses of infected and uninfected animals to bornyl acetate or periplanone-B did not differ significantly, however, indicating that the alteration probably does not occur at the peripheral level but at a central nervous system level.     

Sublethal effects of chlordimeform on chemical communication and other reproductive behaviors in the female cabbage looper moth (Lepidoptera: Noctuidae)

Insecticides can affect the complex coordination of activities associated with reproduction through their sublethal impact on the nervous system. Our objective was to document the effects of a sublethal dose (1% mortality) of chlordimeform on reproductive events in the female cabbage looper moth, Trichoplusia ni. A significantly higher percentage of females treated with chlordimeform initiated calling at some time during the scotophase. Chlordimeform stimulated pheromone emission early in the scotophase. However, late in the scotophase pheromone emission was significantly lower in chlordimeform-treated females when compared with control females. Overall, a lower amount of pheromone was detected in glands of chlordimeform-treated females. This effect was significant only during the second half of the scotophase. We suggest that chlordimeform stimulates pheromone transport to the gland surface and calling behavior resulting in depletion of pheromone reserves over the course of the scotophase. Chlordimeform also decreased the mating success of males. Oviposition and egg hatch were also affected by chlordimeform. Mated females treated with chlordimeform laid significantly fewer eggs than acetone-treated females. In addition, hatchability of eggs laid by mated female T. ni treated with chlordimeform was significantly lower than for eggs laid by control females.     

苹果蠹蛾性信息素的研究和应用进展

苹果蠹蛾Cydia pomonella (L.)是我国重要的检疫害虫, 在我国仅分布于新疆和甘肃以西局部地区, 但一直保持向我国东部扩张的趋势。在国际上, 利用性信息素监测和迷向防治苹果蠹蛾已经成为一种切实可行并广泛应用的害虫管理技术。本文综述了苹果蠹蛾性信息素的成分鉴定、人工合成和应用情况的研究进展, 指出了目前存在的问题和应用前景, 以期为我国苹果蠹蛾的防控策略的研究和应用提供参考。     

Sex pheromone levels in pheromone glands and identification of the pheromone and hydrocarbons in the hemolymph of the moth Scoliopteryx libatrix L. (Lepidoptera: Noctuidae)

The hydrocarbon sex pheromone (13-methyl-Z6-heneicosene) of Scoliopteryx libatrix L. (Lepidoptera: Noctuidae) was found to reach its highest levels on pheromone glands of 3-day-old females. Pheromone levels were not different between the time of maximum calling (end of scotophase) and at the middle of photophase. Overwintering females collected in October had sex pheromone present. Decapitation did not lower the amount of pheromone present, indicating that a head factor is not involved in maintaining pheromone titers. Hemolymph also contained the pheromone, indicating that it is made by oenocytes and transported to the sex pheromone gland. Longer chain length hydrocarbons were also identified from the hemolymph and on the cuticular surface. Quantitative differences in hydrocarbon profiles were found with more methyl-branched hydrocarbons found in the hemolymph than on the cuticular surface. Arch.