Rapid assessment of the sex of codling moth Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae) eggs and larvae

Two different methods were tested to identify the sex of the early developmental stages of the codling moth Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae) with a WZ/ZZ (female/male) sex chromosome system. First, it was shown that the sex of all larval stages can be easily determined by the presence or absence of sex chromatin, which is formed by the female‐specific W chromosome in interphase nuclei. This trait can also be used to identify the sex of newly hatched larvae but it does require care and accuracy. Secondly, a new sexing technique was developed based on a molecular marker of the codling moth W chromosome. Flanking regions of an earlier described W‐specific sequence (CpW2) were isolated and sequenced and a 2.74 kb sequence (CpW2‐EcoRI), specific for the W chromosome, was obtained. Several PCR tests were conducted, which confirmed that the CpW2‐EcoRI sequence is a reliable marker for the sex identification in codling moth samples of different geographical origin. In addition, a fragment of a codling moth gene, period (Cpper) was isolated and sequenced. Results of southern hybridization of the Cpper probe with female and male genomic DNA suggested that the Cpper gene is located on the Z chromosome. Then a multiplex PCR assay was developed, which co‐amplified the CpW2‐EcoRI sequence to identify the W chromosome and the Z‐linked Cpper sequence, which served as a positive control of accurate processing of tested samples. The multiplex PCR provides an easy and rapid identification of the sex of embryos and early larval instars of the codling moth.     

Effect of Aedes aegypti (Diptera:Culicidae) Age on Sex Ratios in Romanomermis culicivorax (Nematoda:Mermithidae)

First, second, third, early and late fourth-instar larvae, and pupae of Aedes aegypti were infected with Romanomerrais culicivorax and reared at 20, 25, and 30 C. An increase in the ratio of male to female nematodes was observed with increase in host age at the time of infection at each temperature. The number of pupal and late fourth-instar infections was low, but R. culicivorax continued to develop in adult A. aegypti. Since male nematodes were recovered from both male and female hosts infected as late fourth instars or pupae, the sex of the host did not influence the sex of the nematode.     

Environmental determination of sex in Apistogrammai (Cichlidae) and two other freshwater fishes (Teleostei)

Environmental sex determination by temperature could be revealed significantly in 33 Apistogramma-species and in Poecilia melanogaster . In some, but not all, Apistogramma-species pH also influences the sex ratio, whereas neither temperature nor pH affect the sex ratio of Pseudocrenilabrus multicolor victoriae . The sex in offspring of A. trifasciata is determined within a sensitive period of about 30 to at least 40 days after spawning.     

Sex ratio variation as a function of host size in Pseudacteon flies (Diptera: Phoridae), parasitoids of Solenopsis fire ants (Hymenoptera: Formicidae)

Some Pseudacteon (Diptera: Phoridae) flies are parasitoids of Solenopsis (Hymenoptera: Formicidae) ant workers in North and South America. Laboratory studies of sex allocation revealed a pattern of sex ratio variation as a function of host size, with more females arising from larger hosts. Environmental sex determination is a possible mechanism for the observed pattern, and examination of Pseudacteon life history reveals several traits assumed to be important in models predicting conditions under which environmental sex determination is favoured. Sex allocation patterns of Pseudacteon are compared with theoretical predictions and empirical data from better-studied hymenoptcran parasitoids that have haplodiploid sex determination. The pattern of sex ratio variation observed has important implications for biocontrol efforts of imported Solenopsis fire ants by the introduction of Pseudacteon parasitoids.     

The paternal‐sex‐ratio (PSR) chromosome in natural populations of Nasonia (Hymenoptera: Chalcidoidea)

Selfish genetic elements may be important in promoting evolutionary change. Paternal sex ratio (PSR) is a selfish B chromosome that causes all‐male families in the haplodiploid parasitic wasp Nasonia vitripennis, by inducing paternal genome loss in fertilized eggs. The natural distribution and frequency of this chromosome in North American populations of N. vitripennis was investigated using a combination of phenotypic and molecular assays. Sampling throughout North America failed to recover PSR except from populations in the Great Basin area of western North America. Extensive sampling of Great Basin populations revealed PSR in frequencies ranging from 0 to 6% at different collection sites, and extended its distribution to Idaho and Wyoming. Intensive sampling in upstate New York did not detect the chromosome. Frequencies of the maternal‐sex ratio distorter (MSR), son killer (SK) and virgin females ranged from 0 to 12%. Paternal sex ratio may be restricted to the Great Basin because its spread is hampered by geographical barriers, or because populations in other areas are not conducive to PSR maintenance. However, it cannot be ruled out that PSR occurs in other regions at very low frequencies. The apparent limited distribution and low frequency of PSR suggest that it will have relatively little impact on genome evolution in Nasonia.     

The importance of cytoplasmic male killing elements in natural populations of the two spot ladybird, Adalia bipunctata (Linnaeus) (Goleoptera: Goccinellidae)

Adalia bipunctata , the two spot ladybird, is polymorphic for a cytoplasmically inherited element which produces female-biased sex ratios by effecting the death of male offspring during embryogenesis. The levels of this element were assessed in Cambridge populations. Six out of 82 females tested showed both a female-biased sex ratio and low egg hatch rates consistent with the presence of this element. Population sex ratios were assessed by collecting pupae from the Cambridge area. The population sex ratio was found to be 1.15: 1, female biased, significantly different from 1:1, and consistent with predictions based on a model incorporating the observed level of the sex ratio element in the population.     

Control of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) in greenhouse tomato crops using the mating disruption technique

The effectiveness of mating disruption to control the tomato leafminer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), in greenhouse tomato crops was evaluated in four trials carried out in winter–spring and summer–winter growing seasons in Southwestern Sardinia (Italy). Pheromone dispensers loaded with 60 mg of the natural blend of the major and minor sex pheromone component (rate 90 : 10) were applied in disrupted greenhouses at a rate of 1000/ha (60 g of active ingredient/ha). Male captures in monitoring pheromone traps, percentage of tomato plants infested by T. absoluta and damage on leaves and fruits were monitored weekly and compared in disrupted and untreated (control) greenhouses. In greenhouses disrupted with 1000 dispensers/ha, a reduction of 93–97% in male trap captures was observed, compared with control. Leaf damage was significantly lower in greenhouses disrupted with 1000 dispensers/ha than in control ones, with a reduction of infestation throughout the growing season ranging from 57% to 85%. Pheromone dispensers applied at the density of 1000/ha significantly reduced the percentage of damaged fruits by 62–89%. In control greenhouses, the highest damage on leaves and fruits was generally observed in edge plants, while leaf and fruit infestation was uniformly distributed in pheromone‐treated greenhouses, indicating an even distribution of the pheromone cloud inside the greenhouse. Mating disruption showed to be an efficient strategy to control in greenhouse the tomato leafminer and can be included in the overall tomato integrated pest management programs.     

Parasitism of Leptasterias spp. (Echinodermata: Asteroidea) by the ciliated protozoan Orchitophrya stellarum (Scuticociliata)

Abstract. The parasitic ciliate Orchitophrya stellarum was found in the testes of brooding, winter-spawning Leptasterias spp. from San Juan Island, Washington, but not in the testes of Leptasterias spp. from the Lynn Canal, Alaska. Dense populations of the ciliate were localized within the fuller areas of testes, where sperm counts were significantly reduced. The ciliates were loaded with phagosomes, some of which contained sperms in various stages of digestion. Leptasterias spp. are not as severely impacted by this ciliated protozoan parasite as Pisaster ochraceus. Leptasterias spp. may serve as seasonal hosts for O. stellarum ; this ciliate lives in seawater and in ripe males of winter-brooding and spring-summer broadcasting sea stars. The wet weight of parasitized and non-parasitized males did not differ. The sex ratio did not deviate from the expected (1:1) in any population except at Point Louisa, Alaska, where males outnumbered females. Testis indexes of parasitized males from 2 of the 4 locations on San Juan Island, Washington were significantly reduced relative to non-parasitized males, indicating a loss of sperm output during spawning.     

Variation in allocation to sexual and asexual reproduction among clones of cyclically parthenogenetic Daphnia pulex (Crustacea: Gladocera)

Organisms reproducing by cyclical parthenogenesis combine the benefits of both sexual and asexual reproduction within the same life cycle. Few studies have examined the evolution of variation in the pattern of investment in parthenogenetic compared to sexual reproduction. Seven clones of Daphnia pulex (Crustacea: Cladocera) varying in allocation to sexual reproduction, as measured by the production of males, were raised in isolation and together in a microcosm to study the pattern of sexual reproduction and the effect of this variation on clone fitness. Sex allocation for clones raised together a microcosm was similar to their allocation when raised in isolation, suggesting a genetic basis to the variation. Three clones showed a cost of producing males that lead to their extinction after about 30 days due to the lack of females required for the clones to persist by parthenogenetic reproduction. The remaining four clones persisted until the end of the 72-day experiment. Clones with little or no allocation to males showed no increased allocation to sexual females. The seven clones showed a greater variation in estimated fitness through male and female function than in total estimated fitness. The clone with the greatest total fitness gained most of its fitness through male function but also had a relatively high fitness through female function. Although one clone produced only females it had the next highest fitness. The three clones that went extinct because of a high investment in males had estimated fitness as high as some clones that persisted in the microcosm because of a higher investment in parthenogenetic reproduction. The similarity in total fitness among clones suggests that Daphnia pulex populations in temporary habitats maintain a sex polymorphism where different genotypes vary-in functional gender ranging from female to primarily male.     

The mating speed in Oligonychus gossypii (Zacher) (Tetranychidae: Acarina)

The number of mated pairs in laboratory populations of the spider mite, O. gossypii increases with time, but levels off by 30–35th min of a 60 min observation period. This levelling off results from a progressive decline in the numbers of sexually active adults available in the population. Non-maters constitute about one-fourth of the laboratory populations at all density levels. Irrespective of the density and reversal of sex ratio, the shape of the mating curve remains the same.     

Typhlocybinae of New Zealand (Homoptera: Cicadellidae)

Two tribes of the subfamily Typhlocybinae are represented in New Zealand, as follows. Erythroneurini: the cosmopolitan genus Zygina is represented by 4 endemic species—toetoe, ramsayi n.sp., dumbletoni, and agni n.sp.—plus zealandica, which occurs in Australia also. Typhlocybini: Typhlocyba froggatti, T. lethierryi, Ribautiana tenerrima, Eupteryx melissae, Kybos smaragdula, and K. betulicola are introduced Northern Hemisphere elements; Matatua montivaga n.gen.&sp. is endemic; M. maorica (formerly Dikraneura maorica), also endemic, is of uncertain identity in the absence of male specimens. Keys are given for the separation of all taxa, the genera and species are described, and all species except Matatua maorica are figured.     

Complete enzymatic deglycosylation of native sex steroid-binding protein (SBP or SHBG) of human and rabbit plasma: effect on the steroid-binding activity.

An enzymatic procedure for the complete removal of the N-linked and O-linked oligosaccharide side chains of the sex steroid-binding proteins (SBP or SHBG) of human and rabbit plasma under native conditions is described. Deglycosylation was catalyzed by N-glycanase, neuraminidase, and O-glycanase and was monitored by SDS-PAGE, lectin blotting, and molecular weight analyses by electrospray mass spectrometry. Digestion of rabbit SBP with N-glycanase generated a major 39,777-Da protein and two minor ones of 39,389 and 39,545 Da. The molecular weight of the major protein agrees with the molecular weight calculated from the sequence of the sugar-free polypeptide monomer (39,769 Da: Griffin, P.R., Kumar, S., Shabanowitz, J., Charbonneau, H., Namkung, P.C., Walsh, K.A., Hunt, D.F., & Petra, P.H., 1989, J. Biol. Chem. 264, 19066-19075), whereas the other two are deglycosylated proteolytic cleavage products lacking the TQR and TQ sequences at the amino-terminus. The N- and O-linked side chains of human SBP were removed by sequential digestion with N-glycanase and neuraminidase/O-glycanase. A 38,771-Da protein was generated, which agrees well with the molecular weight of the sugar-free polypeptide monomer (Walsh, K.A., Titani, K., Kumar, S., Hayes, R., & Petra, P.H., 1986, Biochemistry 25, 7584-7590). N-deglycosylation of human and rabbit SBP has no effect on the steroid-binding activity, but removal of the O-linked side chains of N-deglycosylated human SBP results in an apparent 50% loss of steroid-binding activity and an increase in the Kd for the binding of 5 alpha-dihydrotestosterone from 0.3 mM to 0.9 nM.(ABSTRACT TRUNCATED AT 250 WORDS)     

烟夜蛾性肽受体基因的克隆与表达模式分析

【目的】克隆烟夜蛾Helicoverpa assulta (Guenée)性肽受体基因并分析其表达模式, 为深入研究性肽与交配后反应的关系奠定基础。【方法】采用RT-PCR方法, 从烟夜蛾雌蛾性信息素腺体中得到性肽受体基因cDNA全序列。利用荧光定量PCR方法, 分析该基因的表达模式。【结果】序列分析结果显示, 烟夜蛾性肽受体基因cDNA全长2 048 bp, 命名为HassSPR（GenBank登录号: AFH53182.1）。该基因的开放阅读框长1 275 bp, 编码424个氨基酸残基, 序列中含有7个跨膜域结构, 预测分子量和等电点分别为48.6 kDa和9.25。序列比对分析表明, HassSPR与近缘种棉铃虫H. armigera和其他蛾类性肽受体的氨基酸序列一致性分别达98.35%和超过84%, 与已经报道的其他昆虫的性肽受体的氨基酸序列一致性也在64%以上。不同组织表达分析表明, HassSPR在测定的1日龄雌蛾不同组织中均有表达, 以在脑中的表达量最高。时序表达分析表明, 在羽化前1 天至羽化后6日龄雌蛾的信息素腺体中均有表达, 以3日龄表达量最高。雌蛾交配后, HassSPR在性信息素腺体和脑中的表达量显著上调, 而在交配囊和卵巢中的表达量显著下调。【结论】从烟夜蛾雌蛾性信息素腺体中克隆得到性肽受体基因HassSPR, 其表达模式提示该基因的表达水平与雌蛾的生殖生理和生殖行为有关。     

Isolation of polymorphic microsatellite markers from the malaria vector Anopheles marajoara (Diptera: Culicidae)

Microsatellite‐containing regions were isolated and characterized in Anopheles (Nyssorhynchus) marajoara, a primary vector of malaria parasites in northeastern Amazonia, Brazil. An enrichment protocol yielded 500 positive clones. We designed primers to amplify 40 unique microsatellites, 11 of which amplified cleanly and were polymorphic. A survey of 323 individuals showed that these loci are highly variable with the number of alleles ranging from 11 to 52, and expected heterozygosity ranging from 0.64 to 0.95. These markers will be useful for studies of population structure and intraspecific variation in A. marajoara.     

甜菜夜蛾交配行为和能力

在(27±1)℃,光周期L14∶D10的条件下对甜菜夜蛾I>Spodoptera exigua的交配行为及能力进行了研究。结果表明：成虫在羽化当晚即可进行交配,交配率以羽化后头三个晚上的较高(>82%),但从第4天起则显著下降。成虫一天中的交配时间出现于23:30～05:30之间,交配高峰出现在01:30～02:30和03:00～04:00 之间, 其中以第1高峰的发生频率较高。成虫交配持续时间从22～191 min不等,但以30～60 min的为多(40.8%, n=97), 60～90 min的次之(19.4%),超过180 min的较少(10.2 %)。另外,交配持续时间与蛾龄紧密相关。蛾龄越大,交配持续的时间越长,且差异显著。雄蛾一生的交配能力由1～11次不等,但受性比的影响显著：在性比为1∶1的条件下,雄蛾平均交配次数仅为3.0 次,而在2♀∶1♂至5♀∶1♂时,则增加到5.1～6.0 次。雌蛾交配比例及次数受性比的影响也很大：没有交配的雌蛾比例从1∶1时的8.3%增加到5♀∶1♂时的32%,仅交配一次的比例从16.7%增加到38.7%,而交配≥5 次的比例则从 25%下降到0。最后,对这些结果在甜菜夜蛾防治中应用的可能性进行了讨论。     

Cytological study of Rubenstrema exasperatum (Trematoda: Omphalometridae)

Cytological studies on Rubenstrensa exasperatum were carried out. The number of chromosomes in diploid cells (2n=16), and the morphological characteristics of the karyotype were determined. All the chromosomes (from Nos 2 to 8) had a well-defined short arm and, in accordance with the I^c values, were determined to be meta-submetacentric. Polymorphism was established in the length of the 2 bomologues of the 1st chromosome. This pair is probably linked with sex determination.     

Offspring production and development in the parasitoid wasp Melittobia clavicornis (Cameron) (Hymenoptera: Eulophidae) from Japan

The development time, sex ratio and offspring production of Melittobia clavicornis reared with wild, facultative and factitious hosts are presented. Known hosts and different biological aspects presented in previously published reports are summarized and clarified. Immature development time was equivalent or slightly longer than that of other Melittobia species, and the sex ratio was approximately 97% female. Total offspring numbers were considerably lower than that of other Melittobia species using the same hosts. We report female dimorphism in this species for the first time. The number and relative proportion of brachypterous morph females produced was higher than that in other species of the genus.     

Population structure and morphology of Canarium (Canarium) incisum and Canarium (Canarium) esculentum (Mollusca: Neostromboidae: Strombidae) from the Philippines with preliminary notes on aperture colouration based on DArTseq data

Canarium (Canarium) incisum and Canarium (Canarium) esculentum are small members of the molluscan Strombidae family. Little is known of their population structure. Therefore, we explored this using samples from a population of each. The first sample from Corong Corong Beach, El Nido, Philippines, consisted of 81 adult C. incisum, of which 33 were female and 48 were male. The second sample from Olango Island, Philippines consisted of 73 adult C. esculentum, of which 40 were female and 33 were male. Bias in sex ratio between species was not significant. However, there was bias in sex ratio within species, where males from both species were smaller in axial length than females. We found no evidence of pseudohermaphroditism. The black colouration of the aperture is a phenotype shared by many stromboidians, and 7.4% of C. incisum population exhibited this trait, while the C. esculentum population contained 50.1% black apertures specimens. Preliminary DArTseq analysis indicates that organisms with the black aperture colouration are nested within the populations. Our study fills a knowledge gap on C. incisum and C. esculentum population structure, and gives greater insights to size dynamics of stromboidian taxa in general.     

Facultative sex allocation in snow skink lizards (Niveoscincus microlepidotus)

Mathematical models suggest that reproducing females may benefit by facultatively adjusting their relative investment into sons vs. daughters, in response to population‐wide shifts in operational sex ratio (OSR). Our field studies on viviparous alpine skinks (Niveoscincus microlepidotus) document such a case, whereby among‐ and within‐year shifts in OSR were followed by shifts in sex allocation. When adult males were relatively scarce, females produced male‐biased litters and larger sons than daughters. The reverse was true when adult males were relatively more common. That is, females that were courted and mated by few males produced mainly sons (and these were larger than daughters), whereas females that were courted and mated by many males produced mainly daughters (and these were larger than sons). Maternal body size and condition also covaried with sex allocation, and the shifting pattern of sexual size dimorphism at birth may reflect these correlated effects rather than a discrete component of an evolved sex‐allocation strategy.