Sex differentiation studies relating to releaser aggregation pheromones of the desert locust,Schistocerca gregaria

Behavioural responses of nymphs and adults in the gregarious phase of the desert locust,Schistocerca gregaria (Forskal) (Orthoptera: Acrididae) were investigated in a single-chamber bioassay system to a choice of two columns of air, one permeated with airborne volatiles emanating from either sex of nymphs or adults and the other untreated. There was no sexual differentiation in the production of or response to nymphal volatiles. Young adults of either sex did not produce a stimulus with significant activity. Of the older adults, only the males produced the aggregation stimulus to which both sexes were equally responsive. Charcoal-trapped volatiles from the two sexes of nymphs and adults evoked similar aggregation responses. Antennae of the older adults showed significantly higher EAG responses than those of fifth instar nymphs to all four volatile collections, of which volatiles from older adult males were the most stimulatory and evoked the highest EAG amplitudes.     

Comparative analysis of complete plastid genome reveals powerful barcode regions for identifying wood of Dalbergia odorifera and D. tonkinensis (Leguminosae)

Dalbergia odorifera T. C. Chen (Leguminosae), a rare and endangered tree species endemic to Hainan Island of China, produces the most expensive and rarest wood in China. The wood characteristics of D. odorifera are remarkably similar to those of D. tonkinensis (a much less sought-after species from Vietnam), and the DNA from wood is often highly degraded, making it very difficult to identify the two species using anatomical features or DNA barcoding based on regular DNA markers. To solve the confusion of identifying wood reliably from the two species, we built and analyzed the plastome library of 26 samples from 18 Dalbergia species, of which 12 samples from eight closely related species of D. odorifera are newly sequenced in this study. Phylogenomic analysis suggested that the relationships among the 26 samples are mostly well resolved, and conspecific individuals from different populations of D. odorifera and D. tonkinensis clustered together. Between the plastid genomes of the two species, we identified 129 indels and 114 single nucleotide polymorphisms. By assessing a subset of 20 nucleotide polymorphisms and 10 indels using 37 population-level samples (20 samples of D. odorifera and 17 samples of D. tonkinensis), we recovered eight species-specific barcode regions that could be suitable for identifying the wood D. odorifera and D. tonkinensis. To examine their utility in wood identification, we amplified the eight DNA barcodes using six wood samples and recovered an amplification success rate of 83.3%, demonstrating a reliable method for precise wood identification of the two species.     

Comparative responses of ovipositing Anopheles gambiae and Culex quinquefasciatus females to the presence of Culex egg rafts and larvae

Field observations have demonstrated that gravid Anopheles gambiae Giles s.s. (Diptera: Culicidae) are selective in their choice of oviposition sites. For example, immature stages of An. gambiae s.s. are rarely found in water that contains Culex quinquefasciatus Say immatures. The possibility that this may, in part at least, reflect a response by ovipositing An. gambiae s.s. females to volatile signals associated with Culex juveniles was evaluated by testing the response of An. gambiae s.s. females to varying densities of Cx. quinquefasciatus egg rafts and/or larvae in oviposition choice assays. For comparison, the oviposition choices of Cx. quinquefasciatus to conspecific egg rafts and/or larvae were similarly assayed. At a low density of Cx. quinquefasciatus egg rafts (1-15 egg rafts/100 mL water), An. gambiae s.s. females laid more eggs in the treatment water than in the control, with a maximum of twice as many in the treatment water at 5 egg rafts/100 mL water. At higher egg raft densities and in all treatments that included Cx. quinquefasciatus larvae, oviposition decreased significantly in the treatment dishes in a density-dependent manner. As previous studies have indicated, ovipositing Cx. quinquefasciatus females were attracted to and laid egg rafts in dishes containing conspecific egg rafts and, interestingly, also in dishes containing larvae.     

Isoschaftoside,a C-glycosylflavonoid from Desmodium uncinatum root exudate,is an allelochemical against the development of Striga

In East African small-holder farming of maize, the cattle forage legume, Desmodium uncinatum is used as an intercrop due to its allelopathic inhibition of parasitism by Striga hermonthica, an obligate parasitic weed that can devastate the maize crop. Bioassay-guided fractionation of the root extract of D. uncinatum revealed isoschaftoside to be the main compound in the most potent fraction inhibiting growth of germinated S. hermonthica radicles. Bioassays repeated with isoschaftoside isolated from a different plant source, Passiflora incarnata, proved it to be a biologically active component. Analysis of the root exudates produced by hydroponically grown D. uncinatum showed isoschaftoside to be present in the hydroponic media at biologically active concentrations of 10–100 nM.     

The architecture of gene regulatory variation across multiple human tissues: the MuTHER study

While there have been studies exploring regulatory variation in one or more tissues, the complexity of tissue-specificity in multiple primary tissues is not yet well understood. We explore in depth the role of cis-regulatory variation in three human tissues: lymphoblastoid cell lines (LCL), skin, and fat. The samples (156 LCL, 160 skin, 166 fat) were derived simultaneously from a subset of well-phenotyped healthy female twins of the MuTHER resource. We discover an abundance of cis-eQTLs in each tissue similar to previous estimates (858 or 4.7% of genes). In addition, we apply factor analysis (FA) to remove effects of latent variables, thus more than doubling the number of our discoveries (1,822 eQTL genes). The unique study design (Matched Co-Twin Analysis--MCTA) permits immediate replication of eQTLs using co-twins (93%-98%) and validation of the considerable gain in eQTL discovery after FA correction. We highlight the challenges of comparing eQTLs between tissues. After verifying previous significance threshold-based estimates of tissue-specificity, we show their limitations given their dependency on statistical power. We propose that continuous estimates of the proportion of tissue-shared signals and direct comparison of the magnitude of effect on the fold change in expression are essential properties that jointly provide a biologically realistic view of tissue-specificity. Under this framework we demonstrate that 30% of eQTLs are shared among the three tissues studied, while another 29% appear exclusively tissue-specific. However, even among the shared eQTLs, a substantial proportion (10%-20%) have significant differences in the magnitude of fold change between genotypic classes across tissues. Our results underline the need to account for the complexity of eQTL tissue-specificity in an effort to assess consequences of such variants for complex traits.     

Three-dimensional culture of leech and snail ganglia for studies of neural repair

Three-dimensional (3D) collagen gels provide a stable matrix in which isolated regenerating ganglia from leech and snail can be maintained for studies of the molecular and cellular mechanisms underlying the regenerative process. Segmental ganglia from leech, or supraoesophageal, suboesophageal or buccal ganglia from snail were maintained for up to 3 weeks in 3D matrices of mammalian Type I collagen. The collagen matrix supports the regenerative outgrowth of axon tracts as well as the migration of microglial cells, important elements in the repair process. Proteins or soluble factors or target tissue may be added to the basic collagen matrix to manipulate the environment of the regenerating tissue. We describe techniques for immunostaining of regenerating axons and microglial cells within the gel matrix in combination with staining of cell nuclei, and the use of intracellular labelling to distinguish axons of identified neurons within the regenerative outgrowth.E.J. Babington and J. Vatanparast contributed equally     

Characterisation of cattle anal odour constituents associated with the repellency of <Emphasis Type="Italic">Rhipicephalus appendiculatus</Emphasis>

Adults of the Brown Ear Tick (Rhipicephalus appendiculatus) have a predilection for feeding inside and around the ears of cattle and other hosts. A previous study has shown that the tick locates the host ears by ‘push–pull’ deployment of a repellent blend emitted at the anal region and an attractant blend emitted at the ears. Interestingly, the two odours play reverse roles with Rhipicephalus evertsi, which prefer to feed around the anal region. The present study was undertaken to characterize the major constituents of the cattle anal odour and to evaluate their repellence to R. appendiculatus. The anal odour was trapped with reverse-phase C₁₈-bonded silica, Porapak Q and Super Q placed in an oven bag attached at the anal region of the cattle for 6 h. The adsorbents were then removed and extracted with dichloromethane, and the extracted compounds analyzed by linked gas chromatography-mass spectrometry (GC–MS). The major constituents of the odour were o-xylene, 4-hydroxy-4-methyl-2-pentanone, 4-methyl-2-methoxyphenol, ethylbenzene, 2,6,6-trimethyl-[1S(1α,β,5α)]bicycloheptanes, 5-ethoxydihydro-2(3H)-furanone, 3-methylene-2-pentanone, 5-methyl-2-phenyl-1H-indole, and 3-pentanone. The repellency of the available compounds (o-xylene, 4-hydroxy-4-methyl-2-pentanone, 4-methyl-2-methoxyphenol, ethyl benzene, 3-methylene-2-pentanone, and 3-pentanone) and blends was evaluated using a dual choice tick climbing assay at different doses. The anal odour showed repellence with RD₇₅ of 0.39. Of the compounds tested, 4-methyl-2-methoxyphenol was found to be most repellent (RD₇₅?=?0.56) and 3-pentanone least repellent (RD₇₅?=?622.7). The blend of the six constituents showed RD₇₅ of 0.34, comparable to that of the crude anal odour blend. A series of subtractive bioassays with one constituent of the 6-component blend missing was also carried out. Subtraction of 3-methylpentanone gave the most repellent blend (RD₇₅?=?0.097), whereas subtraction of 4-methylguaiacol gave the least repellent blend (RD₇₅?=?160.7) consistent with the high individual activity of this phenol. The study lays down useful groundwork for on-host deployment of controlled-release of a selected repellent or blend to disrupt the tick’s ability to locate its preferred feeding site.     

Larvicidal activities of limonoids from Turraea abyssinica (Meliaceae) on Tuta absoluta (Meyrick)

Preliminary screening of extracts of the leaves and stems of four Turraea plant species (Meliaceae) on second instar larvae of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) showed that the methanolic extract of Turraea abyssinica leaves possess good toxic potential with LD₅₀ of 270.7 ppm. Fractionation of this extract led to isolation of a new limonoid derivative, 12α‐diacetoxywalsuranolide (1) and three other known limonoids [1α,7α,12α‐triacetoxy‐4α‐carbomethoxy‐11β‐hydroxy‐14β,15β‐epoxyhavanensin (2), 11‐epi‐21‐hydroxytoonacilide (3), 11β,12α‐diacetoxycedrelone (4)]. The structures of the compounds were established by IR, UV, mass spectrometry and 1D/2D NMR analyses, and in the case of the known compounds, also by comparison with reported data. All the isolates were tested for their larvicidal activities at different dose range against second instar larvae of T. absoluta. They were found to be more active (with LD₅₀ < 7.0 ppm) compared to the reference compound azadirachtin (LD₅₀ value of 7.8 ppm).     

Cleome monophylla essential oil and its constituents as tick (Rhipicephalus appendiculatus) and maize weevil (Sitophilus zeamais) repellents

The repellency of the essential oil of the shrubCleome monophylla (Family: Capparidaceae) and identified constituents of the oil were evaluated against the livestock tick,Rhipicephalus appendiculatus and the maize weevil,Sitophillus zeamais. In a tick climbing repellency bioassay, the oil ofC. monophylla exhibited repellency which, at the highest dose, was comparable to that of the commercial arthropod repellent N,N-diethyl toluamide (DEET). In a Y-tube olfactometer bioassay,C. monophylla oil showed higher or comparable repellency againstS. zeamais relative to DEET at all the doses tested. 14 Compounds were identified in theC. monophylla oil by GC, GC-MS and coinjection with authentic samples. Terpenolene was found to occur in largest quantity (14%) followed by 1-α-terpeneol (10%), pentacosane (9%), (α+β)-humulene (8%), phytol (5%) and 2-dodecanone (4%). The most repellent components againstR. appendiculatus andS. zeamais were 1-α-terpeneol and 2-dodecanone. The overall pattern of repellency activity of theC. monophylla constituents with respect to the two arthropods was, however, different. The potential ofC. monophylla in tick and maize weevil control is discussed.     

Repellent and acaricidal properties of Ocimum suave against Rhipicephalus appendiculatus ticks

An oil extracted from the leaves of a tropical shrub Ocimum suave was found to repel as well as kill all stages of the tick Rhipicephalus appendiculatus. In an in vitro bioassay for the larvae, the LC₅₀ of the oil in liquid paraffin was 0.024%. A 10% solution was found to kill all immatures and more than 70% of adults feeding on rabbits. Rabbits were protected for 5 days against attaching larvae using a 10% solution. Preliminary experiments undertaken with cattle kept in the field suggest that the oil may have potential in tick control, and a role in integrated tick management.