Engineered nickel bioaccumulation in Escherichia coli by NikABCDE transporter and metallothionein overexpression

Mine wastewater often contains dissolved metals at concentrations too low to be economically extracted by existing technologies, yet too high for environmental discharge. The most common treatment is chemical precipitation of the dissolved metals using limestone and subsequent disposal of the sludge in tailing impoundments. While it is a cost-effective solution to meet regulatory standards, it represents a lost opportunity. In this study, we engineered Escherichia coli to overexpress its native NikABCDE transporter and a heterologous metallothionein to capture nickel at concentrations in local effluent streams. We found the engineered strain had a 7-fold improvement in the bioaccumulation performance for nickel compared to controls, but also observed a drastic decrease in cell viability due to metabolic burden or inducer (IPTG) toxicity. Growth kinetic analysis revealed the IPTG concentrations used based on past studies lead to growth inhibition, thus delineating future avenues for optimization of the engineered strain and its growth conditions to perform in more complex environments.     

Comparative effects of Steinernema feltiae (Nematoda: Steinernematidae) and insecticides on yield and cropping of the mushroom Agaricus bisporus

In a mushroom crop (Agaricus bisporus) affected by a very low level of sciarid fly (Lycoriella auripila) infestation, the effects of an indigenous isolate of insect-parasitic nematode (Steinernema feltiae) and of two commonly used insecticides (diazinon and diflubenzuron) were studied. When compared with untreated plots, nematodes applied to the casing had no adverse effects on mushroom yields whereas insecticides decreased yields. At a rate of 3 × 10⁶ infective juveniles per tray (surface area = 0.56 m²), S. feltiae elicited increases of 28.5% and 19% in the mean total numbers and weights of mushrooms respectively. Treatment only with diflubenzuron resulted in 14.6% and 6% reductions in mean total numbers and weights of mushrooms, respectively; treatment with both diazinon and diflubenzuron caused 18.5% and 9.4% losses. Application of nematodes generally reduced the mean weight per mushroom whereas insecticides increased it; nematodes delayed the onset of mushroom production (first flush) whereas diflubenzuron delayed the third and fourth flushes. Nematode contamination of sporophores was minimal when S. feltiae was applied at casing. Although their numbers declined with time, the nematodes persisted, in the casing layer, throughout the cropping period of seven weeks. It is concluded that yield benefits associated with nematode application can result mainly from nematode effects on A. bisporus and not solely from suppression of a damaging pest population.     

Effects of Caenorhabditis elegans (Nematoda: Rhabditidae) on the spread of the bacterium Pseudomonas tolaasii in mushrooms (Agaricus bisporus)

The effects of mass-produced saprobic rhabditid nematodes, Caenorhabditis elegans on the spread of the bacterial blotch pathogen, Pseudomonas tolaasii , were studied in mushroom growth chambers. C. elegans significantly reduced the intensity of blotch on sporophores. Repeated isolations of the bacterial flora from the gut of C. elegans recovered from mushroom sporophores during cropping, revealed the presence of Pseudomonas fluorescens biovar reactans . All the isolates of P. fluorescens biovar reactans isolated from nematodes were antagonists of P. tolaasii .
C. elegans produced much larger populations in monoxenic cultures with P. fluorescens biovar reactans than with P. tolaasii . It is suggested that as C. elegans selects P. fluorescens biovar reactans rather than P. tolaasii as a food substrate it probably spreads the antagonist in the mushroom crop and may contribute to the control of bacterial blotch.     

Plant preference in relation to life history traits in the zoophytophagous predator Dicyphus hesperus

Dicyphus hesperus Knight (Heteroptera: Miridae) is an omnivorous predator used to control pests of greenhouse vegetables. Plant preferences and life history traits were studied using nine plant species: Lycopersicon esculentum Mill. (Solanaceae), Capsicum annuum L. (Solanaceae), Verbascum thapsus L. (Scrophulariaceae), Nepeta cataria L. (Lamiaceae), Stachys albotomentosa (Lamiaceae), Nicotiana tabacum L. (Solanaceae), Vicia sativa L. (Fabaceae), Zea mays L. (Gramineae), and Chrysanthemum coronarium L. (Asteraceae). Plants were selected from among potential target crops, natural hosts, plants used for mass rearing, and plants on which D. hesperus has not been reported. Plant preference was measured by multi‐choice host plant selection and oviposition assays. Development and reproduction were measured on each of the plant species on both a plant diet alone and on a plant diet supplemented with Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs. Dicyphus hesperus females and nymphs expressed a preference for some plants over others. Plant preference ranged from low preference plants, such as Z. mays, V. sativa, C. coronarium, and C. annuum, to high preference plants such as V. thapsus, N. tabacum, and S. albotomentosa. When E. kuehniella eggs were supplied, there were few differences in the development time and fecundity of D. hesperus among plants, with the exception of corn and broad bean, where fecundity was lower. On a plant diet alone, nymphs were able to complete their development on V. thapsus, C. annuum, and N. cataria. However, mortality and development time were much lower on V. thapsus than on C. annuum and N. cataria. On most of the plant species D. hesperus did not lay any eggs when fed on a plant diet alone. On V. thapsus, females laid a few eggs and lived longer than when fed on prey. Dicyphus hesperus females tended to prefer host plants on which nymph survival without prey was greatest.     

The applicability of the plant vigor and resource regulation hypotheses in explaining Epiblema gall moth–Parthenium weed interactions

A basic question in insect–plant interactions is whether the insects respond to, or regulate plant traits, or a complex mixture of the two. The relative importance of the directions of the influence in insect–plant interactions has therefore been articulated through both the plant vigor hypothesis (PVH) and the resource regulation hypothesis (RRH). This study tested the applicability of these hypotheses in explaining the interactions between Parthenium hysterophorus L. (Asteraceae) and its stem‐galling moth, Epiblema strenuana Walker (Lepidoptera: Tortricidae). Parthenium plants exposed to galling were sampled at three sites in north Queensland, Australia, over a 2‐year period, and the relationship between gall abundance and plant vigor (plant height, biomass, flowers per plant, and branches per plant) was studied. To test the predictions of PVH and RRH, the vigor of parthenium plants protected from galling using insecticides was compared to galled plants and plants that escaped from galling. The vigor of ungalled plants was less than the vigor of galled plants. The higher plant vigor in galled plants was not due to galling, as was evident from insecticide exclusion trials. The insect seemed to preferentially gall the more vigorous plants. These findings support the predictions of the PVH and are contrary to those of RRH. Since gall abundance is linked to plant vigor, galling may have only a limited impact on the vigor of parthenium. This has implications for weed biological control. If the objective of biological control is to regulate the population of a plant by a galling insect, a preference for more vigorous plants by the insect is likely to limit its ability to regulate plant populations. This may explain the paucity of successes against biocontrol of annual weeds using gall insects.     

Non‐target habitat exploitation by Trichogramma brassicae (Hym. Trichogrammatidae): what are the risks for endemic butterflies?

Abstract 1 Trichogramma brassicae Bezdenko is inundatively released against the European corn borer in Switzerland. Because parasitoids dispersing from the release fields might pose a threat to native butterflies, the searching efficiency of T. brassicae was investigated in nontarget habitats. 2 In field studies, T. brassicae was released at rates of 120 000 females/ha. Parasitism of sentinel Ephestia kuehniella egg clusters was 1.6–3.6% in meadows and 2.0–4.0% in flower strips. The respective figures were 57.6–66.7% and 19.2–46.9% in maize, significantly higher than the parasitism rates in the nontarget habitats. Experiments carried out in small field cages confirmed these results: Again, significantly higher parasitism rates were found in maize compared to meadows and flower strips, and also compared to hedgerows (in sleeve cages). 3 To elucidate potential factors underlying the low searching efficiency in nontarget habitats, the behaviour of individual T. brassicae females was investigated on four meadow plants comparatively to maize and a filter paper control. Mean (±SE) walking speed on maize was 2.2 ± 0.2 mm/s, similar to three of the plants tested and filter paper but significantly higher than on Trifolium pratense (0.85 mm/s). A higher turning rate was found on T. pratense, Viola wittrockiana and Plantago lanceolata, in contrast to the longer leaved maize and Alopecurus pratensis. The number of wasps leaving the plant within the observation period differed significantly between plant species, and was twice as high for T. pratense (and the filter paper control) compared to the other plant species. 4 In a choice experiment carried out in a climate cabinet with all five host plant species in cages, we obtained the highest parasitism rates on maize and the lowest parasitism on T. pratense, thus confirming the behavioural observations. 5 In conclusion, there is evidence for a decreased searching efficiency on plants in nontarget habitats compared to maize. However, the data explain only part of the differences found between parasitism in maize compared to nontarget habitats. Other factors, such as the structural complexity of a habitat, may also play a role. We conclude that the risk for butterfly populations in the tested nontarget habitat due to mass released T. brassicae is low.     

COMPLEMENTARY SEX DETERMINATION IN HYMENOPTERAN PARASITOIDS AND ITS IMPLICATIONS FOR BIOLOGICAL CONTROL

Abstract In haplodiploid Hymenoptera, unfertilized eggs produce haploid males while fertilized eggs lead to diploid females under most circumstances. Diploid males can also be produced from fertilization under a system of sex determination known as complementary sex determination (CSD). Under single-locus CSD, sex is determined by multiple alleles at a single sex locus. Individuals heterozygous at the sex locus are female while hemizygous and homozygous individuals develop as haploid and diploid males, respectively. In multiple-locus CSD, two or more loci, each with two or more alleles, determine sex. Diploid individuals are female if one or more sex loci are heterozygous, while a diploid is male only if homozygous at all sex loci. Diploid males are known to occur in 43 hymenopteran species and single-locus CSD has been demonstrated in 22 of these species. Diploid males are either developmentally inviable or sterile, so their production constitutes a genetic load. Because diploid male production is more likely under inbreeding, CSD is a form of inbreeding depression. It is crucial to preserve the diversity of sex alleles and reduce the loss of genetic variation in biological control. In the parasitoid species with single-locus CSD, certain precautionary procedures can prevent negative effects of single-locus CSD on biological control.     

Effect of temperature on life table parameters of Podisus nigrispinus (Het., Pentatomidae) fed with Alabama argillacea (Lep., Noctuidae) larvae

Abstract: The influence of temperature on life table parameters of Podisus nigrispinus (Dallas) (Het., Pentatomidae) fed with Alabama argillacea (Hübner) (Lep., Noctuidae) larvae was studied. This predator was kept at constant temperatures of 20, 23, 25, 28, 30 and 33±0.2°C, at relative humidity of 60±10% and photoperiod of L : D 14 : 10. Gross (GRR) and net ( R ₀) reproductive rates of P. nigrispinus ranged from 1.6 to 366.6 and from 0.02 to 189.5 females/female at temperatures of 33 and 28°C, respectively; generation time ( T  ) ranged from 33.3 (33°C) to 85.5 (20°C) days; doubling time ( D ) from 0.82 (33°C) to 17.8 (20°C) days; intrinsic rate of increase ( r _m ) from −0.13 (33°C) to 0.12 (28°C) per day; and the finite rate of increase ( λ ) from 0.88 (33°C) to 1.12 (28°C) females/female added to the population per day. The ideal age to release P. nigrispinus should be when this predator presents higher reproductive values (VR _x ); that is, its adults are about 7 days old, independent of prevailing temperature. Population growth of P. nigrispinus was affected by temperature with maximum numerical response between 28 and 30°C. The negative population growth shown at 33°C may not occur in natural conditions due to milder microclimate in the cotton agroecosystem and due to oscillations of temperature in the course of the day.     

Profiling a besieged flora: endemic and threatened plants of the Cape Peninsula, South Africa

The Cape Peninsula (area: 471 km²), situated at the south-western extremity of the Cape Floristic Region, has exceptionally high plant species richness (2285 species and infraspecific taxa) and numbers of endemic (90; 88 species and two infraspecific) and threatened (141; 138 species and three infraspecific) taxa (termed species from here on). This biodiversity is threatened by urban development and the spread of invasive alien plants. Peninsula endemics are concentrated in a few, predominantly species-rich families and these correspond well with endemic-rich families in other areas of the Cape Floristic Region. A high level of similarity exists between families with threatened and families with endemic species. A frequency analysis of the biological traits of both endemic and threatened species shows that low growing, ant-dispersed shrubs are over-represented in both groups. Endemics are most likely to be non-sprouters, but threatened plants do not have a specific post-fire regeneration strategy. Threatened species have higher frequencies of geophytes, sprouters and wind-dispersed species compared to endemic species. Numbers of endemic and threatened species are not randomly distributed with regard to occurrence in vegetation types and patterns are similar for both groups. The habitat and biological profiles of both endemic and threatened species suggest that they are highly vulnerable to extinction as a result of increasing rates of alien plant infestation, urbanization and inappropriate fire regimes.     

Conformational analysis and clustering of short and medium size loops connecting regular secondary structures: a database for modeling and prediction.

Loops are regions of nonrepetitive conformation connecting regular secondary structures. We identified 2,024 loops of one to eight residues in length, with acceptable main-chain bond lengths and peptide bond angles, from a database of 223 protein and protein-domain structures. Each loop is characterized by its sequence, main-chain conformation, and relative disposition of its bounding secondary structures as described by the separation between the tips of their axes and the angle between them. Loops, grouped according to their length and type of their bounding secondary structures, were superposed and clustered into 161 conformational classes, corresponding to 63% of all loops. Of these, 109 (51% of the loops) were populated by at least four nonhomologous loops or four loops sharing a low sequence identity. Another 52 classes, including 12% of the loops, were populated by at least three loops of low sequence similarity from three or fewer nonhomologous groups. Loop class suprafamilies resulting from variations in the termini of secondary structures are discussed in this article. Most previously described loop conformations were found among the classes. New classes included a 2:4 type IV hairpin, a helix-capping loop, and a loop that mediates dinucleotide-binding. The relative disposition of bounding secondary structures varies among loop classes, with some classes such as beta-hairpins being very restrictive. For each class, sequence preferences as key residues were identified; those most frequently at these conserved positions than in proteins were Gly, Asp, Pro, Phe, and Cys. Most of these residues are involved in stabilizing loop conformation, often through a positive phi conformation or secondary structure capping. Identification of helix-capping residues and beta-breakers among the highly conserved positions supported our decision to group loops according to their bounding secondary structures. Several of the identified loop classes were associated with specific functions, and all of the member loops had the same function; key residues were conserved for this purpose, as is the case for the parvalbumin-like calcium-binding loops. A significant number, but not all, of the member loops of other loop classes had the same function, as is the case for the helix-turn-helix DNA-binding loops. This article provides a systematic and coherent conformational classification of loops, covering a broad range of lengths and all four combinations of bounding secondary structure types, and supplies a useful basis for modelling of loop conformations where the bounding secondary structures are known or reliably predicted.