(Z)-11-Hexadecenal and (3Z,6Z,9Z)-Tricosatriene: Sex Pheromone Components of the Red Banded Mango Caterpillar Deanolis sublimbalis

The sex pheromone of the red banded mango caterpillar, Deanolis sublimbalis (Lepidoptera: Crambidae), a serious pest of the mango Mangifera indica (Anacardiaceae) in India and Southeast Asia and a recent invader into northern Australia, has been identified. Three candidate compounds were identified from pheromone gland extracts of female moths, using gas chromatography (GC), GC-electroantennographic detection and GC-mass spectrometric analyses, in conjunction with dimethyldisulfide derivatization. Field bioassays established that both (Z)-11-hexadecenal (Z11-16:Ald) and (3Z,6Z,9Z)-tricosatriene (3Z,6Z,9Z-23:Hy) were required for attraction of male D. sublimbalis moths, and 1,000 μg of a 1:1 mix of Z11-16:Ald and 3Z,6Z,9Z-23:Hy was more attractive to male moths than caged virgin females. However, the binary blend was only attractive when the isomeric purity of the monounsaturated aldehyde was >99%, suggesting that the (E)-isomer was inhibitory. Although (Z)-11-hexadecen-1-ol (Z11-16:OH) was tentatively identified in gland extracts, the addition of this compound to the binary blend did not increase the numbers of moths captured. The pheromone can now be used in integrated pest management strategies.

Heterologous signals allow efficient targeting of a nuclear-encoded fusion protein to plastids and endoplasmic reticulum in diverse plant species

Approximately 30% of plant nuclear genes appear to encode proteins targeted to the plastids or endoplasmic reticulum (ER). The signals that direct proteins into these compartments are diverse in sequence, but, on the basis of a limited number of tests in heterologous systems, they appear to be functionally conserved across species. To further test the generality of this conclusion, we tested the ability of two plastid transit peptides and an ER signal peptide to target green fluorescent protein (GFP) in 12 crops, including three monocots (barley, sugarcane, wheat) and nine dicots (Arabidopsis, broccoli, cabbage, carrot, cauliflower, lettuce, radish, tobacco, turnip). In all species, transient assays following microprojectile bombardment or vacuum infiltration using Agrobacterium showed that the plastid transit peptides from tomato DCL (defective chloroplast and leaves) and tobacco RbcS [ribulose bisphosphate carboxylase (Rubisco) small subunit] genes were effective in targeting GFP to the leaf plastids. GFP engineered as a fusion to the N-terminal ER signal peptide from Arabidopsis basic chitinase and a C-terminal HDEL signal for protein retention in the ER was accumulated in the ER of all species. The results in tobacco were confirmed in stably transformed cells. These signal sequences should be useful to direct proteins to the plastid stroma or ER lumen in diverse plant species of biotechnological interest for the accumulation of particular recombinant proteins or for the modification of particular metabolic streams.

Decoration of Pasteurella multocida lipopolysaccharide with phosphocholine is important for virulence

Phosphocholine (PCho) is an important substituent of surface structures expressed by a number of bacterial pathogens. Its role in virulence has been investigated in several species, in which it has been shown to play a role in bacterial adhesion to mucosal surfaces, in resistance to antimicrobial peptides, or in sensitivity to complement-mediated killing. The lipopolysaccharide (LPS) structure of Pasteurella multocida strain Pm70, whose genome sequence is known, has recently been determined and does not contain PCho. However, LPS structures from the closely related, virulent P. multocida strains VP161 and X-73 were shown to contain PCho on their terminal galactose sugar residues. To determine if PCho was involved in the virulence of P. multocida, we used subtractive hybridization of the VP161 genome against the Pm70 genome to identify a four-gene locus (designated pcgDABC) which we show is required for the addition of the PCho residues to LPS. The proteins predicted to be encoded by pcgABC showed identity to proteins involved in choline uptake, phosphorylation, and nucleotide sugar activation of PCho. We constructed a P. multocida VP161 pcgC mutant and demonstrated that this strain produces LPS that lacks PCho on the terminal galactose residues. This pcgC mutant displayed reduced in vivo growth in a chicken infection model and was more sensitive to the chicken antimicrobial peptide fowlicidin-1 than the wild-type P. multocida strain

Factors affecting localized abundance and distribution of lesser mealworm in earth-floor broiler houses in subtropical Australia.

Factors that influence the localized abundance and distribution of lesser mealworm, Alphitobius diaperinus (Panzer), in litter of two compacted earth-floor broiler houses in subtropical Australia were studied using various experimental manipulations. Numbers of lesser mealworms substantially increased inside caged areas and under uncaged empty feed pans placed in open areas of the houses. These populations were found to be localized and independent of chicken-feed, manure, and high beetle populations that normally occur under existing feed pans. Substantial horizontal movement of larvae to under feed pans was recorded. Placing metal barriers around these pans significantly restricted this movement. In almost all treatments, lesser mealworms typically peaked in numbers during the middle of the flock time. This temporal pattern of abundance also was observed under pans within barriers, where relatively low insect numbers occurred, but it was not observed in uncaged open areas (where chickens had complete access). It is likely that larvae do not establish in open areas, but fluctuate in numbers as they either move to refuges away from chickens or suffer high rates of mortality. In these refuges, larvae peak in numbers and then leave the litter environment to pupate in the earth floor before the end of the flock time. This behavior might be exploited for management of lesser mealworm by targeting applications of control agents.

Survival of graded scallops amusium balloti in Queensland's (Australia) trawl fishery.

In the Queensland, Australia, scallop fishery, the scallop catch is graded at sea using a specially designed grading machine called a "tumbler." Experiments were conducted to determine the effect of repeated trawl capture, grading, and discarding on the survival of sublegal saucer scallops Amusium balloti. Scallops were caught within an area closed to commercial fishing and known to contain dense scallop beds. The trawled scallops were randomly divided into 2 groups, tumbled and control, and subjected to up to 4 tumbles and/or trawls before being caged for 2.5 days adjacent to the trawl grounds. Increased levels of both trawling and tumbling were found to decrease significantly the survival of sublegal scallops. Although 83% of scallops survived repeated intensive trawling (4 consecutive tows), survival fell to 64% when scallops were also graded using a commercial tumbler. Survival was high for both tumbled and control sublegal scallops after 1 trawl (97% and 98%, respectively).

Effect of physiological and experiential state of Bactrocera tryoni flies on intra-tree foraging behavior for food (bacteria) and host fruit

Using caged guava trees in Queensland, Australia, provided with food and oviposition sites, the foraging behaviour of females of the tephritid Bactrocera tryoni was investigated in relation to hunger for protein, the presence or absence of bacteria as a source of protein, the degree of prior experience with host fruit and quality of host fruit for oviposition. One aim was to evaluate whether it is immature or mature B. tryoni females that are responsible for initially inoculating host fruit surfaces with "fruit-fly-type" bacteria, the odour of which is known to attract B. tryoni females. Three-week-old immature females provided with sucrose but deprived of protein from eclosion had a much greater propensity than 3-week-old protein-fed mature females to visit vials containing fruit-fly-type bacteria, irrespective of whether vials were associated with adjacent host fruit or not. In the absence of associated bacteria in vials, immature females had a much lower propensity than mature females to visit host fruit. In the presence of bacteria in vials, however, propensity of immature and mature females to visit fruit was about equal. Mature (but not immature) females were more inclined to visit fruit that ranked higher for oviposition (nectarines) than fruit that ranked lower (sweet oranges). Mature females that attempted oviposition during a single 3-min exposure period to a nectarine prior to release were much more likely to find a nectarine than were mature females naive to fruit or immature females with or without prior contact with fruit. Exposure to a nectarine before release did not affect the propensity of either mature or immature females to alight on an odourless visual model of a nectarine, however. As judged by numbers of leaves visited, protein-deprived immature females were more active than protein-fed mature females, irrespective of the sorts of resources on a tree. It was concluded that: the 1st B. tryoni females to arrive on the fruit of a host tree and therefore inoculate the fruit with fruit-fly-type bacteria were unlikely to be sexually immature, but to be mature as a result of having earlier acquired protein elsewhere; the odour of colonies of fruit-fly-type bacteria when associated with host fruit attracted protein-hungry but not protein-fed females; and the odour of the fruit itself attracted mature females (especially experienced ones) but not immature females.

The insecticidal spider toxin SFI1 is a knottin peptide that blocks the pore of insect voltage-gated sodium channels via a large β-hairpin loop

Spider venoms contain a plethora of insecticidal peptides that act on neuronal ion channels and receptors. Because of their high specificity, potency and stability, these peptides have attracted much attention as potential environmentally friendly insecticides. Although many insecticidal spider venom peptides have been isolated, the molecular target, mode of action and structure of only a small minority have been explored. Sf1a, a 46-residue peptide isolated from the venom of the tube-web spider Segesteria florentina, is insecticidal to a wide range of insects, but nontoxic to vertebrates. In order to investigate its structure and mode of action, we developed an efficient bacterial expression system for the production of Sf1a. We determined a high-resolution solution structure of Sf1a using multidimensional 3D/4D NMR spectroscopy. This revealed that Sf1a is a knottin peptide with an unusually large β-hairpin loop that accounts for a third of the peptide length. This loop is delimited by a fourth disulfide bond that is not commonly found in knottin peptides. We showed, through mutagenesis, that this large loop is functionally critical for insecticidal activity. Sf1a was further shown to be a selective inhibitor of insect voltage-gated sodium channels, consistent with its 'depressant' paralytic phenotype in insects. However, in contrast to the majority of spider-derived sodium channel toxins that function as gating modifiers via interaction with one or more of the voltage-sensor domains, Sf1a appears to act as a pore blocker.