A semiochemical view of the ecology of the seed beetle Acanthoscelides obtectus Say (Coleoptera: Chrysomelidae, Bruchinae).

The dried bean beetle, Acanthoscelides obtectus, is an economically important pest of stored legumes worldwide. Tracking the human-aided dispersion of its primary hosts, the Phaseolus vulgaris beans, it is now widespread in most bean-growing areas of the tropics and subtropics. In temperate regions where it can only occasionally overwinter in the field, A. obtectus proliferates in granaries, having multiple generations a year. Despite its negative impact on food production, no sensitive detection or monitoring tools exist, and the reduction of local populations still relies primarily on inorganic insecticides as fumigating agents. However, in the quest to produce more nutritious food more sustainably and healthily, the development of environmentally benign crop protection methods is vital against A. obtectus. For this, knowledge of the biology and chemistry of both the host plant and its herbivore will underpin the development of, among others, chemical ecology-based approaches to form an essential part of the toolkit of integrated bruchid management. We review the semiochemistry of the mate- and host-finding behaviour of A. obtectus and provide new information about the effect of seed chemistry on the sensory and behavioural ecology of host acceptance and larval development.

Progress on reducing acrylamide levels in potato crisps in Europe, 2002 to 2019.

European Snacks Association (ESA) data on acrylamide in potato crisps from 2002 to 2019 (99704 observations) were analysed. Acrylamide levels have plateaued since 2011, although the lowest mean so far was attained in 2018 at 353 ± 2.7 ng g-1: a 54% reduction since 2002. The 85th, 90th and 95th quantiles did show evidence of continued downward progress, the 90th quantile being lower than the 750 ng g-1 European Benchmark Level from 2017 to 2019. A smaller dataset from the European Food Safety Authority (2124 observations) for 2011-2018 was also analysed. The yearly means were higher than those of the ESA data but showed a fall in average acrylamide from 715 ± 40.5 ng g-1 in 2015 to 505 ± 28.5 ng g-1 in 2018, as well as steep falls in the 85th, 90th and 95th quantiles. Nevertheless, even the 85th quantile remained above the 750 ng g-1 Benchmark Level. The ESA data showed a reduction in the proportion of samples with acrylamide exceeding 750 ng g-1, from over 40% in 2002 to 7.75% in 2019. Seasonality was evident, with highest acrylamide levels from November to May. Crisp type had little effect except that thicker types had a higher proportion of samples containing >750 ng g-1 acrylamide. Analysis of the region of origin in Europe of the final product revealed improvements in the east and north. Geographical factors combined with seasonality continued to be problematic but was also an aspect in which progress was most evident. The findings show that improvements have been made in reducing the number of samples with very high levels of acrylamide, but do not suggest that mean acrylamide levels could be reduced substantially below where they have been since 2011, or that levels could be kept consistently below the current Benchmark Level.

Wheat amino acid transporters highly expressed in grain cells regulate amino acid accumulation in grain.

Amino acids are delivered into developing wheat grains to support the accumulation of storage proteins in the starchy endosperm, and transporters play important roles in regulating this process. RNA-seq, RT-qPCR, and promoter-GUS assays showed that three amino acid transporters are differentially expressed in the endosperm transfer cells (TaAAP2), starchy endosperm cells (TaAAP13), and aleurone cells and embryo of the developing grain (TaAAP21), respectively. Yeast complementation revealed that all three transporters can transport a broad spectrum of amino acids. RNAi-mediated suppression of TaAAP13 expression in the starchy endosperm did not reduce the total nitrogen content of the whole grain, but significantly altered the composition and distribution of metabolites in the starchy endosperm, with increasing concentrations of some amino acids (notably glutamine and glycine) from the outer to inner starchy endosperm cells compared with wild type. Overexpression of TaAAP13 under the endosperm-specific HMW-GS (high molecular weight glutenin subunit) promoter significantly increased grain size, grain nitrogen concentration, and thousand grain weight, indicating that the sink strength for nitrogen transport was increased by manipulation of amino acid transporters. However, the total grain number was reduced, suggesting that source nitrogen remobilized from leaves is a limiting factor for productivity. Therefore, simultaneously increasing loading of amino acids into the phloem and delivery to the spike would be required to increase protein content while maintaining grain yield.

Mapping sites of gibberellin biosynthesis in the Arabidopsis root tip.

Root elongation depends on the action of the gibberellin (GA) growth hormones, which promote cell production in the root meristem and cell expansion in the elongation zone. Sites of GA biosynthesis in the roots of 7-d-old Arabidopsis thaliana seedlings were investigated using tissue-specific GA inactivation in wild-type (Col-0) or rescue of GA-deficient dwarf mutants. Tissue-specific GA depletion was achieved by ectopic expression of the GA-inactivating enzyme AtGA2ox2, which is specific for C19 -GAs, and AtGA2ox7, which acts on C20 -GA precursors. In addition, tissue-specific rescue of ga20ox triple and ga3ox double mutants was shown. Furthermore, GUS reporter lines for major GA20ox, GA3ox and GA2ox genes were used to observe their expression domains in the root. The effects of expressing these constructs on the lengths of the root apical meristem and cortical cells in the elongation zone confirmed that roots are autonomous for GA biosynthesis, which occurs in multiple tissues, with the endodermis a major site of synthesis. The results are consistent with the early stages of GA biosynthesis within the root occurring in the meristematic region and indicate that the penultimate step of GA biosynthesis, GA 20-oxidation, is required in both the meristem and elongation zone.

Determination of the prebiotic activity of wheat arabinogalactan peptide (AGP) using batch culture fermentation.

PURPOSE: To test the prebiotic activity of wheat arabinogalactan-peptide (AGP), which is a soluble dietary fibre composed of arabinogalactan polysaccharide linked to a 15-residue peptide, which accounts for up to 0.4% of the dry weight of wheat flour. METHODS: The prebiotic activity of AGP prepared from white wheat flour was tested using in vitro fermentation by colonic bacteria in automated pH-controlled anaerobic stirred batch cultures and compared to fructooligosaccharide (FOS) and wheat flour arabinoxylan (AX). Bacterial populations were measured using fluorescence in situ hybridisation (flow-FISH) and short chain fatty acid (SCFA) concentrations were measured using HPLC. RESULTS: Fermentation of AGP resulted in a significant bifidogenic activity and increased concentrations of SCFAs, mainly acetate after 24 h of fermentation. CONCLUSIONS: These results were comparable to those obtained with AX and confirm the prebiotic potential of AGP. Furthermore, fermentation of a mixture of AGP and AX was faster compared to the single substrates and more similar to FOS, indicating that combinations of fermentable carbohydrates with different structures are potentially more effective as prebiotics than single substrates.

Foliar N Application at Anthesis Stimulates Gene Expression of Grain Protein Fractions and Alters Protein Body Distribution in Winter Wheat (Triticum aestivum L.).

The major components of wheat storage proteins are gliadins and glutenins, and as they contribute differently to baking quality, a balanced mixture of these components is essential. The application of foliar nitrogen (N) at anthesis is a common practice to improve protein concentration and composition. The aim of this study was to investigate the effects of a foliar N application at anthesis on storage protein gene expression during grain development and on the distribution of protein concentration and protein body size within the grain. In this experiment, an additional N application at anthesis stimulated the expression of genes of the majority of storage proteins when the N supply was low. Furthermore, it led to higher protein concentrations in the subaleurone layers, while in the center of the lobes, the protein concentrations were decreased. These changes will affect the protein recovery in white flours, as proportionally more protein might be lost during milling processes.

An Evidence Synthesis of Pharmaceuticals and Personal Care Products (PPCPs) in the Environment: Imbalances among Compounds, Sewage Treatment Techniques, and Ecosystem Types.

Pharmaceuticals and personal care products (PPCPs) garner increasing attention globally for both their usefulness as indicators of human waste and their potency as emerging organic toxicants. Three decades of rapid increase in PPCP study combined with an increasing number of PPCPs on the global market have created an opportunity (1) to review trends in diversity of compounds, sewage treatment techniques (STTs), and ecosystems investigated as well as (2) to identify knowledge gaps in the literature. We conducted a quantitative evidence synthesis of 6517 abstracts from primary articles in the environmental PPCP literature by examining relative abundance of specific PPCP classes, STTs, and ecosystem types. Our results demonstrate that non-prescription drugs and antibiotics dominated PPCP abstracts, appearing in 51% and 39% of reviewed abstracts, respectively, in comparison to hormones (18%), prescription drugs (18%), fragrances (0.3%), and antioxidants (0.0%), which can all elicit physiological and ecological responses even at low concentrations. References to centralized STTs (e.g., activated sludge, 37%) were more frequent than decentralized STTs (e.g., septic, 2%), despite decentralized STTs being common and frequently high impact sources of sewage pollution worldwide. Freshwater lotic systems (63%) were more prevalent than freshwater lentic (24%) and terrestrial (20%) systems. This discrepancy is notable because the longer residence times of lentic and terrestrial systems may enable PPCPs to concentrate and thus increase risk of biological consequences. These results highlight distinct opportunities to address knowledge gaps in the environmental PPCP literature, including underrepresented compounds (e.g., fragrances), sewage treatment techniques (e.g., septic systems), and ecosystem types (e.g., lakes).

Demand-Driven Model for Global Phosphate Rock Suggests Paths for Phosphorus Sustainability.

Phosphorus is required as a fertilizer for producing food, and there is no substitute. Losses between mine production and diet result in significant environmental harm. We used a demand-driven substance flow model to explore the sensitivity of global phosphorus production to interventions of the food system including: reduction in animal fraction in the diet (AFD); manure use efficiency (MUE); animal food yield (YA); phosphorus use efficiency (PUE); the fraction of food that is wasted (FWF); the fraction of food waste recycled (FRE); and the fraction of human waste recycled (WRE). The model indicated that AFD and YA are the most influential interventions, and they interact with PUE and MUE. Furthermore, there is a minimum in AFD and YA below which it becomes necessary to increase mining. Another result is that reducing food waste is about 80 times more effective than recycling food waste in reducing P demand. Finally, the model was used to explore the global carrying capacity for humans on the basis of P sources other than mining. These sources may satisfy P requirements for as many as 2.5 billion people. If significant improvements were made in all of the considered interventions simultaneously, as many as 14.7 billion people could be sustained.

Stream Transport and Retention of Environmental DNA Pulse Releases in Relation to Hydrogeomorphic Scaling Factors.

The DNA of aquatic organisms can be identified in water sampled from freshwater ecosystems to detect species presence. Because these DNA-based methods (termed environmental DNA, eDNA) confirm species presence by proxy of DNA in water, the processes influencing eDNA transport and removal from water are critical to the method's efficacy and interpretation of results. Previous studies of aquatic eDNA transport and fate have employed uncontrolled field experiments, controlled studies in experimental streams, and laboratory column tests. As a step toward understanding the processes controlling eDNA transport and retention, we released and tracked experimental pulses of white sturgeon eDNA (novel to the system) in five fourth-order stream reaches with varied hydrology and geomorphology. We found strong support that stream water transient storage controls eDNA areal uptake rate (or spiraling length). We calculated the median spiraling length to be ∼260 m. Down channel slope correlated with transient storage, suggesting that this slope could be used as a proximate measure of eDNA removal into the benthic zone. Our results suggest that sampling effort should be increased in reaches with longer transient storage (or lower slopes) to compensate for the increase in eDNA retention.

Molecular quantification of Plasmodium parasite density from the blood retained in used RDTs.

Most malaria-endemic countries are heavily reliant upon rapid diagnostic tests (RDT) for malaria case identification and treatment. RDT previously used for malaria diagnosis can subsequently be used for molecular assays, including qualitative assessment of parasite species present or the carriage of resistance markers, because parasite DNA can be extracted from the blood inside the RDT which remains preserved on the internal components. However, the quantification of parasite density has not previously been possible from used RDT. In this study, blood samples were collected from school-age children in Western Kenya, in the form of both dried blood spots on Whatman filter paper, and the blood spot that is dropped into rapid diagnostic tests during use. Having first validated a robotic DNA extraction method, the parasite density was determined from both types of sample by duplex qPCR, and across a range of densities. The methods showed good agreement. The preservation of both parasite and human DNA on the nitrocellulose membrane inside the RDT was stable even after more than one year's storage. This presents a useful opportunity for researchers or clinicians wishing to gain greater information about the parasite populations that are being studied, without significant investment of resources.

Identification of a non-host semiochemical from tick-resistant donkeys (Equus asinus) against Amblyomma sculptum ticks.

Amblyomma sculptum is a tick affecting animal and human health across Argentina, Bolivia, Paraguay and Brazil. Donkeys, Equus asinus, are known to be resistant to A. sculptum, suggesting that they can produce non-host tick semiochemicals (allomones), as already demonstrated for some other vertebrate host/pest interactions, whereas horses, Equus caballus, are considered as susceptible hosts. In this study, we tested the hypothesis that donkeys produce natural repellents against A. sculptum, by collecting sebum from donkeys and horses, collecting the odour from sebum extracts, and identifying donkey-specific volatile compounds by gas chromatography (GC) and coupled GC-mass spectrometry (GC-MS). From the complex collected blends, five main compounds were identified in both species. Hexanal, heptanal and (E)-2-decenal were found predominantly in donkey extracts, whilst ethyl octanoate and ethyl decanoate were found predominantly in horse extracts. One compound, (E)-2-octenal, was detected exclusively in donkey extracts. In Y-tube olfactometer bioassays 36 different A. sculptum nymphs were tested for each extract, compound and concentration. The dry sebum extracts and the compounds identified in both species induced neither attraction nor repellency. Only (E)-2-octenal, the donkey-specific compound, displayed repellency, with more nymphs preferring the arm containing the solvent control when the compound was presented in the test arm across four concentrations tested (p < 0.05, Chi-square test). A combination of a tick attractant (ammonia) and (E)-2-octenal at 0.25 M also resulted in preference for the control arm (p < 0.05, Chi-square test). The use of semiochemicals (allomones) identified from less-preferred hosts in tick management has been successful for repelling brown dog ticks, Rhipicephalus sanguineus sensu lato from dog hosts. These results indicate that (E)-2-octenal could be used similarly to interfere in tick host location and be developed for use in reducing A. sculptum numbers on animal and human hosts.

Open science, reproducibility, and transparency in ecology.

Reproducibility is a key tenet of the scientific process that dictates the reliability and generality of results and methods. The complexities of ecological observations and data present novel challenges in satisfying needs for reproducibility and also transparency. Ecological systems are dynamic and heterogeneous, interacting with numerous factors that sculpt natural history and that investigators cannot completely control. Observations may be highly dependent on spatial and temporal context, making them very difficult to reproduce, but computational reproducibility can still be achieved. Computational reproducibility often refers to the ability to produce equivalent analytical outcomes from the same data set using the same code and software as the original study. When coded workflows are shared, authors and editors provide transparency for readers and allow other researchers to build directly and efficiently on primary work. These qualities may be especially important in ecological applications that have important or controversial implications for science, management, and policy. Expectations for computational reproducibility and transparency are shifting rapidly in the sciences. In this work, we highlight many of the unique challenges for ecology along with practical guidelines for reproducibility and transparency, as ecologists continue to participate in the stewardship of critical environmental information and ensure that research methods demonstrate integrity.

Host shift induces changes in mate choice of the seed predator Acanthoscelides obtectus via altered chemical signalling.

The mechanisms of host shift in phytophagous insects are poorly understood. Among the many proposed processes involved, sexual selection via semiochemicals has recently been suggested. This hypothesizes that sexual communication using pheromones is modified as a result of development on a new host, and such plant-induced phenotypic divergence in mate recognition cues can lead to reproductive isolation between host lines. We tested this hypothesis on Acanthoscelides obtectus, an oligophagous bruchid of Phaseolus vulgaris beans worldwide, which also develops in acceptable non-hosts, such as chickpea (Cicer arietinum L.). Male sex pheromone blends of the bean, chickpea and chickpea/bean host lines during artificially induced host shifts showed different composition. Bean-reared females did not distinguish between blends, whereas chickpea and chickpea/bean females preferred the chickpea male pheromone. However, electrophysiological (EAG) responses to male odour of antennae of the three female host lines were similar, all preferring bean-reared males. Egg-laying choice tests revealed a uniform preference for bean seeds across female host lines, even after multiple generations, whereas larvae did not distinguish between bean and chickpea seeds. We conclude that the development of divergent chemical signalling systems during host shifts does not facilitate the evolution of host races in A. obtectus, because oviposition preferences remain unaffected.

Managing Diffuse Phosphorus at the Source versus at the Sink.

Judicious phosphorus (P) management is a global grand challenge and critical to achieving and maintaining water quality objectives while maintaining food production. The management of point sources has been successful in lowering P inputs to aquatic environments, but more difficult is reducing P discharges associated with diffuse sources, such as nonpoint runoff from agriculture and urban landscapes, as well as P accumulated in soils and sediments. Strategies for effective diffuse-P management are imperative. Many options are currently available, and the most cost-effective and practical choice depends on the local situation. This critical review describes how the metrics of P quantity in kg ha-1 yr-1 and P form can influence decision-making and implementation of diffuse-P management strategies. Quantifying the total available pool of P, and its form, in a system is necessary to inform effective decision-making. The review draws upon a number of " current practice" case studies that span agriculture, cities, and aquatic sectors. These diverse examples from around the world highlight different diffuse-P management approaches, delivered at the source in the catchment watershed or at the aquatic sink. They underscore workable options for achieving water quality improvement and wider P sustainability. The diffuse-P management options discussed in this critical review are transferable to other jurisdictions at the global scale. We demonstrate that P quantity is typically highest and most concentrated at the source, particularly at farm scale. The most cost-effective and practically implementable diffuse-P management options are, therefore, to reduce P use, conserve P, and mitigate P loss at the source. Sequestering and removing P from aquatic sinks involves increasing cost, but is sometimes the most effective choice. Recovery of diffuse-P, while expensive, offers opportunity for the circular economy.

Final grain weight is not limited by the activity of key starch-synthesising enzymes during grain filling in wheat.

Since starch is by far the major component of the mature wheat grain, it has been assumed that variation in the capacity for starch synthesis during grain filling can influence final grain weight. We investigated this assumption by studying a total of 54 wheat genotypes including elite varieties and landraces that were grown in two successive years in fields in the east of England. The weight, water content, sugars, starch, and maximum catalytic activities of two enzymes of starch biosynthesis, ADP-glucose pyrophosphorylase and soluble starch synthase, were measured during grain filling. The relationships between these variables and the weights and starch contents of mature grains were analysed. Final grain weight showed few or no significant correlations with enzyme activities, sugar levels, or starch content during grain filling, or with starch content at maturity. We conclude that neither sugar availability nor enzymatic capacity for starch synthesis during grain filling significantly influenced final grain weight in our field conditions. We suggest that final grain weight may be largely determined by developmental processes prior to grain filling. Starch accumulation then fills the grain to a physical limit set by developmental processes. This conclusion is in accord with those from previous studies in which source or sink strength has been artificially manipulated.

Dry matter losses and quality changes during short rotation coppice willow storage in chip or rod form.

This study compares dry matter losses and quality changes during the storage of SRC willow as chips and as rods. A wood chip stack consisting of approximately 74 tonnes of fresh biomass, or 31 tonnes dry matter (DM) was built after harvesting in the spring. Three weeks later, four smaller stacks of rods with an average weight of 0.8 tonnes, or 0.4 tonnes DM were built. During the course of the experiment temperature recorders placed in the stacks found that the wood chip pile reached 60 °C within 10 days of construction, but the piles of rods remained mostly at ambient temperatures. Dry matter losses were calculated by using pre-weighed independent samples within the stacks and by weighing the whole stack before and after storage. After 6 months the wood chip stack showed a DM loss of between 19.8 and 22.6%, and mean losses of 23.1% were measured from the 17 independent samples. In comparison, the rod stacks showed an average stack DM loss of between 0 and 9%, and between 1.4% and 10.6% loss from the independent samples. Analysis of the stored material suggests that storing willow in small piles of rods produces a higher quality fuel in terms of lower moisture and ash content; however, it has a higher fine content compared to storage in chip form. Therefore, according to the two storage methods tested here, there may be a compromise between maximising the net dry matter yield from SRC willow and the final fine content of the fuel.

Plasmodium-associated changes in human odor attract mosquitoes.

Malaria parasites (Plasmodium) can change the attractiveness of their vertebrate hosts to Anopheles vectors, leading to a greater number of vector-host contacts and increased transmission. Indeed, naturally Plasmodium-infected children have been shown to attract more mosquitoes than parasite-free children. Here, we demonstrate Plasmodium-induced increases in the attractiveness of skin odor in Kenyan children and reveal quantitative differences in the production of specific odor components in infected vs. parasite-free individuals. We found the aldehydes heptanal, octanal, and nonanal to be produced in greater amounts by infected individuals and detected by mosquito antennae. In behavioral experiments, we demonstrated that these, and other, Plasmodium-induced aldehydes enhanced the attractiveness of a synthetic odor blend mimicking "healthy" human odor. Heptanal alone increased the attractiveness of "parasite-free" natural human odor. Should the increased production of these aldehydes by Plasmodium-infected humans lead to increased mosquito biting in a natural setting, this would likely affect the transmission of malaria.

Temperature and nitrogen supply interact to determine protein distribution gradients in the wheat grain endosperm.

Gradients exist in the distribution of storage proteins in the wheat (Triticum aestivum) endosperm and determine the milling properties and protein recovery rate of the grain. A novel image analysis technique was developed to quantify both the gradients in protein concentration, and the size distribution of protein bodies within the endosperm of wheat plants grown under two different (20 or 28 °C) post-anthesis temperatures, and supplied with a nutrient solution with either high or low nitrogen content. Under all treatment combinations, protein concentration was greater in the endosperm cells closest to the aleurone layer and decreased towards the centre of the two lobes of the grain, i.e. a negative gradient. This was accompanied by a decrease in size of protein bodies from the outer to the inner endosperm layers in all but one of the treatments. Elevated post-anthesis temperature had the effect of increasing the magnitude of the negative gradients in both protein concentration and protein body size, whilst limiting nitrogen supply decreased the gradients.

Isolation and identification of floral attractants from a nectar plant for the dried bean beetle, Acanthoscelides obtectus (Coleoptera: Chrysomelidae, Bruchinae).

BACKGROUND: The response of virgin females of the legume pest Acanthoscelides obtectus (Coleoptera: Bruchidae) to headspace extracts of volatiles collected from flowers of a nectar plant, Daucus carota, was investigated using behaviour (four-arm olfactometry) and coupled gas chromatography-electroantennography (GC-EAG). RESULTS: Odours from inflorescences were significantly more attractive to virgin female beetles than clean air. Similarly, a sample of volatile organic compounds (VOCs) collected by air entrainment (dynamic headspace collection) was more attractive to beetles than a solvent control. In coupled GC-EAG experiments with beetle antennae and the VOC extract, six components showed EAG activity. Using coupled GC-mass spectrometry (GC-MS) and GC peak enhancement with authentic standards, the components were identified as α-pinene (S:R 16:1), sabinene, myrcene, limonene (S:R 1:3), terpinolene and (S)-bornyl acetate. Females preferred the synthetic blend of D. carota EAG-active volatiles to the solvent control in bioassays. When compared directly, odours of D. carota inflorescences elicited stronger positive behaviour than the synthetic blend. CONCLUSION: This is the first report of behaviourally active volatiles linked to pollen location for A. obtectus, and development of the six-component blend is being pursued, which could underpin the design of semiochemical-based field management approaches against this major pest of stored products. © 2018 Society of Chemical Industry.